ISSN Vol. 8, No. 1, January-June, Appl. Hort. Journal of THE SOCIETY FOR ADVANCEMENT OF HORTICULTURE

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1 ISSN Vol. 8, No. 1, Jnury-June, 2006 Appl Hort Journl of THE SOCIETY FOR ADVANCEMENT OF HORTICULTURE

2 JOURNAL OF APPLIED HORTICULTURE A Journl of THE SOCIETY FOR THE ADVANCEMENT OF HORTICULTURE (Registered under Act XXI of 1860) Registrtion No ISSN or The Journl of Applied Horticulture publishes rticles in English on ll spects of horticulturl crops. Send rticles for publiction strictly typed s per formt. Ppers tht report novel dt with scientific nd/or prcticl importnce nd from which the science is sound, will be ccepted for publiction. Ppers of restricted prcticl or scientific importnce will not be ccepted. CO-ORDINATING EDITORS AND REFEREES A. Ptks (Greece) Fbio Pinto Gomes (Brzil) Penn Suprsnn (Indi) Akos Mthe (Hungry) Freddy Lel (Venezuel) Petel Gilles (Frnce) Alberto C.Q. Pinto (Brzil) Fure-Chyi Chen (Tiwn) Piet Stssen (South Afric) Alberto Prdossi (Itly) G. Sivkumr (South Kore) Po-Yung Li (Tiwn) Alex-Aln Furtdo de Almeid (Brzil) Hugo S. Grci (Mexico) R.P. Awsthi (Indi) Amnollh Jvnshh (Irn) Inuw S. Usmn (Nigeri) Rm Kishun (Indi) Amos Mizrch (Isrel) Jn Verschoor (News Zelnd) Rnvir Singh (Indi) Antonio de Almeid Hunter (Portugl) Kee-Yoeup Pek (Kore) Reginldo Bez Snudo (Mexico) Atill Eris (Turkey) Koutsik-Sotiriou Metxi (Greece) Riccrdo d Andri (Itly) B. Ssikumr (Indi) Leon A. Terry (UK) Richrd E. Litz (USA) Bingru Hung (USA) Ling Chen (Chin) Robert Prnge (Cnd) Bletsos Fotios (Greece) Luis Romero (Spin) R.T. Gunjte (Indi) C.P.A. Iyer (Indi) Mjeed Mohmd (West Indies) S. Adniy (Jpn) Cengiz Ky (Turkey) Mshru Msud (Jpn) Slih Kfks (Turkey) Chng-Hung Chou (Tiwn) Mohmed Ibrhim El Tmzini (Egypt) Smuel Kwme Offei (Ghn) Dniel Vlero Grrido (Spin) Muchit Th Ozky (Turkey) Sisir K. Mitr (Indi) Dvid W.M. Leung (New Zelnd) Nryn R. Bht (Kuwit) Teres Terrzs Slgdo (Mexico) Der-Ming Yeh (Tiwn) Nzim Grud (Germny) Tzong-Shyn Lin (Tiwn) Duong Tn Nhut (Vietnm) N.A. Nikolou (Greece) Uri Lvi (Isrel) E. Lhv (Isrel) Norbert Keutgen (Germny) V.S. Motil (Indi) Ekterini Trk-Mvron (Greece) P.K. Chttophdhyy (Indi) Vsileios Noitskis (Greece) Elhdi M. Yhi (Mexico) P.L. Tndon (Indi) Yueming Jing (Chin) Enzo Mgliulo (Itly) Polo Inglese (Itly) Zeev Wiesmn (Isrel) ES du Toit (South Afric) Pedro Correi (Portugl) Zen-hong Shu (Tiwn) Eun Joo Hhn (South Kore) Pedro Mrtinez Gomez (Spin) Zor Singh (Austrli) Chief Editor: R.P. Srivstv Associte Editor: S. Rjn Mnging Editor: V. Rjn Editoril nd Business Office A-859, Indirngr, Lucknow , INDIA Phones: , Fx: / e-mil: jh@hortreserch.net or jh@horticutlurereserch.net Currently Journl of Applied Horticulture is included in the following Abstrcting Services Horticulturl Abstrcts, Agroforestry Abstrcts, Agriculturl Engineering Abstrcts, Crop Physiology Abstrcts, Forestry Abstrcts, Irrigtion nd Dringe Abstrcts, Indin Science Abstrcts, Ornmentl Horticulture, Plnt Breeding Abstrcts, Plnt Genetic Resources Abstrcts, Posthrvest News nd Informtion, Review of Plnt Pthology, Seed Abstrcts, Soils nd Fertilizers, Review of Aromtic nd Medicinl Plnts, World Agriculturl Economics nd Rurl Sociology Abstrcts, Rurl Development Abstrcts, Review of Agriculturl Entomology nd Review of Medicl nd Veterinry Entomology, Indin Science Abstrcts. Online bstrcts re lso vilble t nd See Guideline for uthors on inner lst cover of the journl. For more detils plese visit or Vist for online submission. Author Pge Chrges Becuse of the high cost of publishing rticles in Journl of Applied Horticulture, pyment of pge chrges is mndtory nd chrges re subject to chnge without notice. Current chrges re $10 (Rs.150 in Indi) per printed pge(a4 Size, pprox. 800 words).

3 JOURNAL OF APPLIED HORTICULTURE Vol. 8, No. 1, Jnury-June, 2006 CONTENTS Effect of grfting on growth nd yield of tomto (Lycopersicon esculentum Mill.) in greenhouse nd open-field -E.M. Khh, E. Kkv, A. Mvromtis, D. Chchlis nd C. Gouls (Greece) 3 Use of chlorophyll meter nd plnt visul spects for nitrogen mngent in tomto fertigtion -Pulo Cezr Rezende Fontes nd Chrles de Arujo (Brzil) 8 Biodegrdble pper/polymerized vegetble oil mulches for tomto nd pepper production - Rndl. L. Shogren nd Monic Dvid (USA) 12 Compct 3U s novel lighting source for the propgtion of some horticulturl plnts - D.T. Nhut, M.T. Ngoc Huong, D.V. Khiem nd J.A. Teixeir d Silv (Vietnm) 15 Effect of slow relese fertiliser on the growth of continerised flnnel flower (Actinotus helinthi Lbill.) - Lotte von Richter nd Ctherine A. Offord (Austrli) 21 Growth nd flowering response of snpdrgons fter relese from picl dominnce - Muhmmd Munir nd Frht Nz (United Kingdom & Pkistn) 25 Posthrvest control of soft-rot fungi on grpe berries by fungicidl tretment nd Trichoderm -Y. A. Btt (Isrel) 29 Nitrogen metbolism of Aloe ver under long-term diluted sewter irrigtion - Liu Zho-Pu, Zho Geng-Mo, Liu Ling, Zheng Qing-Song (Chin) 33 Reltionship between soil nd lef minerl nutrient concentrtion nd yield of selected citrus species - L. Andrews nd R.A.I. Brthwite (West Indies) 37 Studies with thidizuron on the vse life of cut rose flowers -Esmeil Chmni, Donld E. Irving, Dryl C. Joyce, Mos Arshd (Austrli) 42 Growth behviour of pple cctus (Cereus species) in hyper-rid environment - Ahmed A. ElObeidy (Egypt) 45 Assessment of genetic diversity nd reltionships mong some grpe vrieties using ISSR mrkers -Mnjush Dhne, S.A.Tmhnkr, S.G. Ptil, G.S. Kribspp nd V.S. Ro (Indi) 50 Impct of polyethylene glycol-induced wter stress on growth nd development of shoot tip cultures from different bnn (Mus spp.) cultivrs -K.H. Mohsen, Ebrhim, Ibrhim A. Ibrhim, Hmdy A. Emr nd Ewld Komor (Egypt nd Germny) 53 Effect of gibberellin tretment on prthenocrpic bility nd promotion of fruit swelling in ppy - Fredh. K. Rimberi, S. Adniy, M. Kwjiri, N. Urski, S. Kwno, T. Etoh nd Y. Ishimine (Jpn) 58 Effect of minerl concentrtion on in vitro explnt growth of lmond (Prunus mygdlus vr. Binzir) -Mohmmd E. Amiri (Irn) 62 Chemicl effect of reclimed wter on soil nd rose plnt grown in soil nd tuff medi -M.I..Sfi, A. Frdous. M. Muddber. S. El-Zuriqi L. Al-Hdidi. I. Bshbsheh (Jordn) 65 Prtil ringing nd liquid nitrogen effects on shoot growth nd fruit qulity of pech -J. M. Onguso, F. Mizutni, A.B.M. Shrif Hossin nd A. R. El-Shereif (Jpn) 70 Performnce of three sweet ornge vrieties grfted on four rootstocks under Jordn Vlley conditions -J. Muhtseb, H. Ghnim nd A. Sheikh (Jordn) 75 Comprison of bnns ripened by two methods for texturl sle-grdes -Sunit Singh nd S.D. Kulkrni (Indi) 78 Constrints in production nd mrketing of pistchio in Irn nd the policies concerned -Rez Sedght nd S. Suryprksh (Irn nd Indi) 82

4 Forthcoming Ppers Trnsgenic tomto (Lycopersicon esculentum) overexpressing capx exhibits enhnced tolernce to UV-B nd het stress-yueju Wng, Richrd Meiln, Michel Wisniewski, Minggng Cui, nd Leslie Fuchigmi (USA). Effects of busive tempertures on the posthrvest qulity of lettuce leves: scorbic cid loss nd microbil growth- Mrí del R. Moreir, Alejndr G. Ponce, del Vlle, E. Crlos, R. Ansoren nd S.I. Rour (Argentin). Posthrvest ppliction of exogenous sucrose in broccoli: Some physiologicl responses nd chnges in the ctivities of glutmine synthetse- Dewoowoogen P. Bclyon, Toshiyuki Mtsui, Hruo Suzuki nd Yusuke Kosugi (Jpn). Developmentl influence of in vitro light qulity nd crbon dioxide on photochemicl efficiency of PS II of strwberry leves (Frgri x nnss)- John H. Mirnd nd Richrd Willims (Austrli). Effect of forcing t different times on bud burst, flowering, nd fruit development of low-chill pech cultivr Premier -Sutsinee Mneethon, Kenji Beppu, Noko Kozi, Ryosuke Mochiok nd Ikuo Ktok (Jpn). Ectopic expression of Mn-SOD in Lycopersicon esculentum leds to enhnced tolernce to slt nd oxidtive stress-yueju Wng, Richrd Meiln, Michel Wisniewski, Sndr L. Urtsu, Minggng Cui, Abhy Dndekr nd Leslie Fuchigmi (USA). Induction of phenolic compounds biosynthesis with light irrdition in the flesh of red nd yellow pples-d. Bkhshi nd O. Arkw (Jpn). Thidizuron-induced in vitro plnt regenertion from immture seed cotyledon explnts of mcdmi (Mcdmi tetrphyll L. Johnson)-Richrd M.S. Mulw nd Prem L. Bhll-(Austrli). Supplement of 2,4-D nd NAA mitigtes utotoxicity of strwberry in hydroponics- H. Kitzw, T. Aso, T. Bn, Y. Hshimoto, T. Hosoki (Jpn). Sucrose synthse nd cid invertse ctivities in reltion to the florl structures bortion in pepper (Cpsicum nnuum L.) grown under low night temperture-néji Trchoun, Slh Rezgui nd Abdelziz Mougou (Tunisi). Effects of genotype nd grft type on the hydrulic chrcteristics nd wter reltions of sweet melon-smuel Agele nd Shbti Cohen (Isrel nd Nigeri). Tolernce of lilyturf (Liriope muscri) nd four perennil ornmentl grsses to preemergent herbicides-jmes T. Cole nd Jnet C. Cole (USA). Effect of exogenous ppliction of nti-stress substnces nd elementl sulfur on growth nd stress tolernce of tissue culture derived plntlets of dte plm (Phoenix dctylifer L.) c.v. Khls during cclimtiztion-mohmed. A. Awd, A. A. Soud nd S. M. El-Konissi (UAE). Effect of onion cultivrs on storge losses under hot conditions-nser Alemzdehnsri Ansri (Irn) Shelf-life nd qulity of pple fruits in response to posthrvest ppliction of UV-C rdition-syvsh Hemmty, Noorollh Mollemi nd Lotfli Nseri (Irn). vlues to follow lycopene concentrtion during ripening nd storge of tomto (cv. Cruso)-Axelle Schuwers, Ad M.C.N Roch nd Alcin M.M.B Moris (Portugl). N-NO 3 from cellulr extrct s indictor of nutritionl sttus of cntloupe muskmelon in fertigtion-mrí Remedios Cigles Rivero nd Octvio Pérez Zmor(Mexico). Economic rtionle of commercil orgnic fertilizer technology in vegetble production in Osun Stte of Nigeri- T. Alimi; O.C. Ajewole, O.O. Olubode-Awosol nd E.O. Idowu (Niger). Thinning response of Abbé Fetel per to lime sulphur-p.i. Grriz, H.L. Alvrez, G.M. Colvit nd M.S. Gjdos (Argentin). Growth, fruit yield nd qulity of Golden Delicious pple trees under fixed prtil root zone drying-g. Tlluto, V. Frin nd R. Lo Binco (Itly). A one step in vitro cloning procedure for grpevine: The influence of bsl medi nd plnt growth regultors- M.S. Brreto, A. Nookrju, N.V.M. Hrini nd D.C. Agrwl (Indi) Soil, plnt nd cnopy resistnce to wter flow in bell pepper (Cpsicum nnuum L.) s ffected by fertigtion regimes- S.O. Agele (Nigeri). Chlorine disinfection: Effects on hydroponics lettuce- Zdenk Premuzic, Hemilse E. Plmucci, Jun Tmborene, Mrtin Nkm (Argintin). Identity nd efficiency of pollintors of wtermelon (Citrullus lntus (Thunb.) Mnsf.) t Ytt, Keny-G.N. Njoroge, B. Gemmill, R. Bussmnn, L.E.Newton nd V.W Ngumi (Keny) Effects of orgnic mnure on okr (Abelmoschus esculentus L. moench) production-ofosu-anim, E. T. J. Bly nd M.E. Frempong (Ghn).

5 Journl of Applied Horticulture, 8(1): 3-7, Jnury-June, 2006 Effect of grfting on growth nd yield of tomto (Lycopersicon esculentum Mill.) in greenhouse nd open-field Appl Journl E.M. Khh*, E. Kkv*, A. Mvromtis*, D. Chchlis** nd C. Gouls* *University of Thessly, School of Agriculturl Sciences, Deprtment of Agriculture, Crop Production nd Agriculturl Environment, Fytoko Street, 38446, N. Ioni, Mgnesis, Volos, Greece. **Ntionl Agriculturl Reserch Foundtion (N.AG.RE.F.), PlntProtection Institute of Volos, P.O. Box 1303, Fitoko, Volos 38001, Greece. E-mil: Abstrct Seedlings of tomto (Lycopersicon esculentum Mill.) cv. Big Red were used s scion nd rootstock (self-grfted) nd non-grfted control, while two hybrid tomtoes Hemn nd Primver were used s rootstocks. Grfted nd non-grfted plnts were grown in the greenhouse nd in the open-field. Grfted plnts (BH nd BP) were more vigorous thn the non-grfted ones in the greenhouse s well s in the open-field. Plnts grfted onto Hemn nd Primver produced 32.5, 12.8% nd 11.0 nd 11.1% more fruit thn the control (B) in the greenhouse nd the open-field, respectively, wheres self-grfted plnts BB hd lower yield in both cultivtion conditions. However, the self-rooted plnts B presented erliness in their performnce, probbly due to the lck of stress tht followed the grfting opertion. Qulity nd qulittive fruit chrcteristics were not ffected by grfting. Key words: Lycopersicon esculentum, Lycopersicon hirsutum, grfting, rootstock, scion, tomto, yield. Introduction Tomto (Lycopersicon esculentum Mill.) is crop of high importnce in mny countries; ccording to FAO (1998), in Greece, 1.8 millions MT were produced. In the Mediterrnen re, where lnd use is very intensive nd continuous cropping is in common prctice, vegetble grfting is considered n innovtive technique with n incresing demnd by frmers. Viewing recent dt concerning the Mediterrnen re by Leonrdi nd Romno (2004) it ws reported tht Spin is the most importnt country for the spreding of vegetble grfting with minly tomto nd wtermelon, with 40 nd 52% of the totl of 154 million plnts in 2004, respectively. They lso indicted tht in Itly n incresing dissemintion of the grfting technique incresed the number of the vegetble grfted plnts from 4 million in 1997 to 14 million in In Greece, grfting is becoming highly populr, especilly in southern res, where the rtio of the production re using grfted plnts to the totl production re, mounts to lmost % for erly cropping wtermelons, 40-50% for melons under low tunnels, 5-10% for cucumbers nd 2-3% for tomto nd eggplnt. In contrst, in northern Greece, the cultivtion of grfted fruit-bering vegetbles is rre (Trk-Mvron et l., 2000). Although in the beginning, tomto grfting ws dopted to limit the effects of Fusrium wilt (Lee, 1994; Scheffer, 1957), the resons for grfting hve incresed drmticlly over the yers. For exmple, grfts hve been used to induce resistnce ginst low (Bulder et l., 1990) nd high (Rivero et l., 2003) tempertures; to enhnce nutrient uptke (Ruiz et l., 1997); to improve yield when plnts re cultivted in infected soils (Bersi, 2002; Kcjn-Mrsic nd Osvld, 2004); to increse the synthesis of endogenous hormones (Proebsting et l. 1992); to improve wter use (Cohen nd Nor, 2002); to increse flower nd seed production (Lrdizbl nd Thompson, 1990); to enhnce vegetble tolernce to drought, slinity nd flooding (AVRDC, 2000; Estn et l., 2005). Moreover, mny reserchers reported tht n interction between rootstocks nd scions exists resulting in high vigor of the root system nd greter wter nd minerl uptke leding to incresed yield nd fruit enhncement (Lee, 1994; Od, 1995; Bersi, 2002; White, 1963; Leoni et l., 1990; Ionnou, et l., 2002; Kcjn-Mrsic nd Osvld, 2004). On the contrry, Romno nd Prtore (2001) stted tht vegetble grfting does not improve the yield when the selection of the rootstock is not suitble, for exmple the self-grfted plnt Rit x Rit hd lower yield thn the non-grfted plnts. Also there re some contrdictory results bout the fruit qulity trits nd how grfting ffects them. For exmple Trk-Mvron et l. (2000) report tht the solutes ssocited with fruit qulity re trnslocted in the scion through the xylem, wheres Lee (1994) sttes tht qulity trits e.g. fruit shpe, skin colour, skin or rind smoothness, flesh texture nd colour, soluble solids concentrtion etc. re influenced by the rootstock. However, other reserchers showed tht grfting did not ffect fruit qulity (Leoni et l., 1990; Romno nd Prtore, 2001). The im of this study ws to evlute populr Greek commercil hybrid tomto, self-grfted nd grfted on two new improved tomto rootstocks, for gronomic performnce, yield nd fruit qulity ttributes. Mterils nd methods Plnt mteril: The commercil tomto (L. esculentum Mill.) hybrid cv. Big Red ws used s self-grfted nd non-grfted control, while two hybrid tomtoes Hemn (L. hirsutum) nd Primver (L. esculentum Mill.) were used s rootstocks.

6 4 Effect of grfting on growth nd yield of tomto Hemn possesses resistnce to Pyrenochet lycopersici nd nemtodes, wheres Primver is resistnt to Verticillium nd nemtodes. Grfting combintions were s follows: BB (scion nd rootstock Big Red ), BP (scion Big Red nd rootstock Primver ), BH (scion Big Red nd rootstock Hemn ) nd B (non-grfted, control). The seeds of the scion cultivrs were sown 5 dys erlier thn the seeds of the 2 rootstocks to ensure similr stem dimeters t the grfting time becuse of the differences in growth vigour. Seedlings were grfted by hnd, pplying the splice grfting method when the scion hd 2 rel leves nd the rootstock rel leves. Then the grfted plnts were kept for 7 dys under controlled conditions (90-95% RH, o C nd 45% shding). Plnts were trnsplnted to the soil in greenhouse on 4/3/2004 nd to the open-field on 13/5/2004 t the Velestino Frm (Mgnesi, Greece) of the University of Thessly, t density of plnts h -1. Norml culturl prctices were followed for irrigtion, fertilizer nd pesticide ppliction. A rndomised complete block design ws dopted with 4 replictions, ech consisting of 8 plnts. Plnts were cultivted in 4 replicted plots ech of which contined 8 plnts spced t 0.6x1.0m. Four plnts from ech replicte were evluted for height, flowering nd yield, one ws used for dry nd wet weight mesurements, while the others remined s gurd plnts nd were not included in the evlutions. Mesurements: Men mximum nd minimum ir temperture, reltive humidity nd the mount of rinfll were recorded dily throughout the two cultivtions. Plnt height ws recorded between 8-96 DAT (Dys After Trnsplnttion) in the greenhouse cultivtion nd between DAT in the open-field cultivtion. In order to obtin flowering dt, flowers of 5 clusters ws considered. The fresh weight ws determined for plnts tht were hrvested t ground level nd seprted into leves, stem, flowers nd fruits. For the dry weight determintion the plnt tissues were dried in ventilted oven t 90 o C for 48h. Due to the different environmentl condition in field nd greenhouse, plnts from both conditions were hrvested lmost in the sme opticl size nd ssessment ws mde t 107 DAT nd 121 DAT for greenhouse nd open-field, respectively. Totl lef re ws mesured by Portble Are Meter (model LI3000A, LI-COR). Yield mesurements were recorded on ripe fruits, which were hnd-hrvested, counted nd weighed. For the greenhouse cultivtion, 16 hrvests were crried out between DAT, while for the open-field cultivtion 8 hrvests were crried out between DAT. Finlly 6 fruits were rndomly hrvested from ech repliction nd were used for qulittive mesurements i.e., firmness (penetrometer FT327-8mm), soluble solids (refrctometer), ph, titrtble cidity, lycopene concentrtion (spectrophotometer t 600 nm) nd concentrtion of Zn, Cu, Mn, Fe nd C (tomic bsorption spectrophotometer). Dt nlysis: Sttisticl nlysis ws performed using SPSS 11.0 for Windows nd the differences between the mens were compred using the criterion of the Duncn s multiple rnge test nd LSD (P=0.05). Results nd discussion Plnt height ws not significntly ffected by grfting under greenhouse conditions, wheres in the open-field cultivtion t 130 DAT the height of BH ws significntly greter thn the control nd BP (Tble 1). This result grees with the results of Lee (1994) nd Ionnou et l. (2002) who found tht grfted plnts were tller nd more vigorous thn self-rooted ones nd hd lrger centrl stem dimeter. Tble 1. Plnt height of non-grfted (B) nd 3 grfted tomto plnts (BH, BP, BB) over different growth periods in greenhouse nd openfield conditions DAT Plnt height (cm) BH BP BB B Greenhouse b 48.44c b Open-field bc b 56.81c b b b b Mens followed by the sme letter re sttisticlly not significnt ccording Duncn s multiple rnge test (P=0.05). DAT: Dys After Trnsplnting, BH: Big Red x Hemn, BP: Big Red x Primver, BB: Big Red x Big Red, B: Big Red. It ws observed tht in both greenhouse nd open field cultivtions flowering begn erlier in the self-rooted plnt, probbly due to the fct tht grfting cused stress nd delyed flower formtion. However, by the 5 th cluster, grfted plnts generlly ppered to hve lrger number of flowers but no significnt differences between ll the tretments with respect to the totl number of flowers per plnt were found. Also, it is worth mentioning tht the number of flowers in the open field were lmost 50 % less thn in the greenhouse in ll the tretments (Tble 2). Tble 2. The men number of flowers per cluster nd totl number of flowers per plnt of non-grfted (B) nd 3 grfted tomto plnts (BH, BP, BB) t different growth periods under greenhouse nd open-field conditions Cluster number DAT Number of flowers/cluster BH BP BB B Greenhouse 1 st nd b b 3 rd th b 4.88b b 5 th Totl 5 th Open-field 1 st nd rd th th Totl lowers 5 th Mens followed by the sme letter re sttisticlly not significnt ccording Duncn s multiple rnge test (P=0.05) From the dt presented in Tble 3, it is seen tht there were no significnt differences between the fresh nd dry weights of stems, leves nd fruits both in the greenhouse nd in the openfield fter 107 nd 121 DAT respectively, with the exception of the BH plnts, which hd significntly lower fresh nd

7 Effect of grfting on growth nd yield of tomto 5 Tble 3. Fresh nd dry weight, plnt height nd totl lef re of non-grfted (B) nd 3 grfted tomto plnts (BH, BP, BB) t 107 DAT nd 121 DAT under greenhouse nd open-field conditions, respectively Chrcterstics/ prt Greenhouse dry weight of flowers thn BP in the greenhouse cultivtion. However, the rtio of totl dry weight to totl fresh weight ws not significntly different between grfted plnts nd the control in both cultivtions (Tble 3). Moreover, in the greenhouse, grfted plnts of BH nd BP hd hevier fresh nd dry weight thn the open field cultivtion. Tble 3 shows tht lthough the distribution of dry mtter in the vrious prts of the plnt ws even in greenhouse cultivtion, grfted plnts hd higher ccumultion of dry mtter. It is worth mentioning tht Romno nd Prtore (2001) lso reported tht the dry weight of the eril orgns of grfted tomto plnts ( Rit x Beufort ) ws greter thn tht of the self-rooted plnts. Lef re mesurements t 107 DAT nd 121 DAT in the greenhouse nd in the open-field, respectively (Tble 3) reveled tht the plnts of BH grfting hd lrger lef re thn the other tretments. However, there ws no significnt difference. Also Pulgr et l. (1998) observed incresed production of leves in grfted plnts s result of n incresed uptke of wter nd nutrients. In the greenhouse s well s in the open-field during the hrvest period 0-84 DAT, the self-rooted plnts B hd greter yield thn the grfted plnts. This could be due to the fct tht grfted plnts were initilly subjected to stress following the grfting opertion. This erly negtive effect of grfting hs lso been reported by other uthors (Ginoux, 1974; Tsouvltzis et l., 2004). However, during the 2 nd hrvest period the grfted plnts BH nd BP hd greter yield thn the self-rooted B, while during the 3 rd hrvest period the three types of grfted plnts hd greter yield thn the self-rooted control (Tble 4). It seems tht the 4 tretments produced higher quntity of fruits per plnt t the 2 nd hrvest period when the plnts hd more fvourble environmentl conditions for growth. Men dily tempertures for the first, second nd third hrvesting periods were 22.3, 27.8, 3 nd 33.1 o C for the greenhouse nd 20.3, 26.8 nd 23.5 o C for the open field cultivtions respectively. Finlly, these increses in the totl fruit yield of the BH nd BP plnts of the greenhouse cultivtion, t Open-field BH BP BB B BH BP BB B Stem FW DW Leves FW , DW Flowers FW b 20.40b 14.93b DW b 3.73b 3.03b Fruits* FW DW Totl DW/FW % Totl lef re (cm 2 ) Plnt height (cm) *Ripe nd Unripe. Mens followed by the sme letter re sttisticlly not significnt (Duncn s multiple rnge test, P=0.05) 192 DAT resulted into 32.5% nd 10% more fruit weight per plnt thn the control B, respectively, wheres self-grfted plnts gve lmost the sme yield s the control. Similr results were found for the open-field cultivtion where higher totl fruit weight of BH nd BP t 130 DAT were obtined (12.8 nd 11.1% higher thn in the control, respectively) (Tble 4). Regrding fruit qulittive chrcteristics (Tble 5) there were no significnt differences between the 4 tretments in ph, Brix (%), concentrtion of lycopene or firmness. However, fruit cidity in grfted plnts of BH cultivted in the open field ws higher thn in BB nd B plnts. The bove results in generl gree with other reserchers who found tht fruit descriptive nd qulittive chrcteristics were not ffected by grfting. (Leoni et l., 1990; Romno nd Prtore, 2001). The fruit Cu, Mn nd Fe contents were not significntly different Tble 4. Yield t different hrvest periods nd totl of non-grfted (B) nd 3 grfted tomto plnt types (BH, BP, BB) under greenhouse nd open-field conditions DAT Fruit weight (g) plnt -1 BH BP BB B Greenhouse 1 st b b b 2 nd rd Totl b b b Open-field 1 st nd rd b b b Totl Mens followed by the sme letter re sttisticlly not significnt ccording Duncn s multiple rnge test (P=0.05)

8 6 Effect of grfting on growth nd yield of tomto Tble 5. Qulittive fruit prmeters of non-grfted (B) nd 3 grfted tomto plnts (BH, BP, BB) under greenhouse nd open-field conditions Cultivrs ph BRIX (%) Greenhouse Acidity (% citric cid) Lycopene (mg/ 100gDW) between the grfted plnts nd the control plnts, either in the greenhouse or in the open-field. However, nlyses showed tht the fruit concentrtion of C in grfted plnts BH ws greter thn in the fruits of the grfted plnts BB nd B in the greenhouse cultivtion. The bsorption of C could be ssocited strongly with the higher rte of bsorption of wter nd minerls from the soil by roots of the rootstock Hemn nd therefore this could improve the bsorption of C. Tsouvltzis et l. (2004) recorded similr results, when tomto cv. Scos F1 ws grfted on Primver rootstock nd fruit yield nd minerl concentrtion incresed. Also Lee (1994) found n increse in yield which ws ttributed to the vigour of the rootstock nd the higher uptke of wter nd nutrients. Pssm et l. (2005) found tht eggplnts grfted on to two tomto rootstocks gve higher yield nd bigger fruit size thn those grfted on to two eggplnt rootstocks, but the minerl composition of fruits from grfted plnts did not differ from tht of non grfted plnts. This study showed tht in both the greenhouse nd the open-field, tomto cv. Big Red grfted on tomto rootstock Hemn gve higher totl yield without hving significnt effects on the qulity of the fruits produced. The results showed tht tomto grfting on suitble rootstocks hs positive effects on the cultivtion performnce, especilly in the greenhouse conditions. The use of improved genotypes for rootstocks is required so s to improve yields under vriety of climtic nd soil conditions. It is well known tht the root system of the plnts ffects vegettive growth nd yield. So, the effects of grfting recorded in most reserch ppers re obviously relted to the differences in the root system between grfted nd nongrfted plnts, i.e. to the efficiency of wter nd nutrient uptke by the roots, or even to the distribution of growth regultors. In Greece, where the vegetble cultivtion is still crried out mostly by trditionl methods nd modern cultivted techniques re dopted slowly, the grfting technique could help in the solution of mny problems. Therefore, we consider the dvntges of grfted plnts, which offer incresed yield nd Firmness (kg) Zn (ppm) consequently higher profit, to be of vlue for frmers. Finlly, the use of grfting is simple step for more developed cultivtion forms, like hydroponics. References Cu (ppm) Mn (ppm) Fe (ppm) C (ppm) BH b BP b BB B Open-field BH b b BP b b BB B b Mens followed by the sme letter re sttisticlly not significnt (Duncn s multiple rnge test, P=0.05) AVRDC, Grfting tkes root in Tiwn. Center point, the qurterly Newsletter of the Asin Vegetble Reserch nd Development Centre. September 2000: 1-3. Bersi, M Tomto grfting s n lterntive to methyl bromide in Mrocco. Institut Agronomieque et Veterinire Hsn II. Mrocco. Bulder, H.A.M., P.R. vn Hsselt., P.J.C. Kuiper., E.J. Speek nd A.P.M. Den Nijs, The effect of low root temperture in growth nd lipid composition of low temperture tolernt rootstock genotypes for cucumber. Journl of Plnt Physiology, 138: Cohen, S. nd A. Nor, The effect of three rootstocks on wter use, cnopy conductnce nd hydrulic prmeters of pple trees nd predicting cnopy from hydrulic conductnce. Plnt, Cell nd Environment, 25: Estn, M.T., M.M. Mrtinez-Rodrigues, F. Perez-Alfoce, T.J. Flowers nd M.C. Bolrin, Grfting rises the slt tolernce of tomto through limiting the trnsport of sodium nd chloride to the shoot. J. Experimentl Botny, 56(412) : FAO, Production yerbook, Agriculturl Sttistics Series. FAO, Rome. Vol. 52. Ginoux, G Biln de qutre nnée de expérimenttion sur le greffge de solncées dns le Sud-Est. Pépiniéristes Horticultures Mrîchers, 152: Ionnou, N., M. Ionnou nd K. Hdjiprskevs, Evlution of wtermelon rootstocks for off-seson production in heted greenhouses. Act Horticulture, 579: Kcjn-Mrsic, N. nd J. Osvld, The influence of grfting on yield of two tomto cultivrs (Lycopersicon esculentum Mill.) grown in plstic house. Act Agriculture Slovenic, 83(2): Lrdizbl, R.D. nd P.G. Thompson, Growth regultors combined with grfting increse flower number nd seed production in sweet potto. HortScience, 25: Lee, J.M Cultivtion of grfted vegetbles I, current sttus, grfting methods nd benefits. HortScience, 29: Leonrdi, C. nd D. Romno, Recent issues on vegetble grfting. Act Horticulture, 631:

9 Effect of grfting on growth nd yield of tomto 7 Leoni, S., R. Grudin, M. Cdinu, B. Mdeddu nd M.C. Grletti, The influence of four rootstocks on some melon hybrids nd cultivr in greenhouse. Act Horticulture, 287: Od, M New grfting method for fruit-bering vegetbles in Jpn. Jpn Agriculturl Reserch Qurterly, 29: Pssm, H.C., M. Stylinoy nd A. Kotsirs, Performnce of Eggplnt Grfted on Tomto nd Eggplnt Rootstocks. Europen Journl of Horticulturl Science, 70(30): Proebsting, W.M.P., M.J. Hedden, S.J. Lewis nd L.N. Croker- Proebsting, Gibberellin concentrtion nd trnsport in genetic lines of pe. Plnt Physiology, 100: Pulgr, G., R.M. Rivero, D.A. Moreno, L.R. Lopez-Lefebre, G. Villor, M. Bghour nd L. Romero, Micronutrientes en hojs de sndí injertds. In: VII Simposio ncionl-iii Ibérico sobre Nutrición Minerl de ls Plnts. Gárte A. (Ed.), Universidd Autónom de Mdrid, Mdrid., Rivero, R.M., J.M. Ruiz nd L. Romero, Role of grfting in horticulturl plnts under stress conditions. Food, Agriculture & Environment, 1(1): Romno, D. nd A. Prtore, Effects of grfting on tomto nd eggplnt. Act Horticulture, 559: Ruiz, J.M., L. Belkbir., J.M. Rgl nd L. Romero, Response of plnt yield nd lef pigments to sline conditions: effectiveness of different rootstocks in melon plnts (Cucumis melo L.). Soil Science Plnt Nutrition, 43: Scheffer, R.P Grfting experiments with Fusrium wilt resistnt nd susceptible tomto plnts. Phytopthology, 47: 30. Trk-Mvron, E., M. Koutsik-Sotiriou nd T. Prits, Response of sqush (Cucurbit Spp.) s rootstock for melon (Cucumis melo L.). Scienti Horticulture, 83: Tsouvltzis, P.I., A.S. Siomos nd K.C. Dogrs, The effect of the two tomtoes grfting on the performnce, erliness nd fruit qulity. Proc. 21 st Pn-Hellenic Congress of the Greek Society for Horticulturl Science. Ionnin, Greece, 8-10 October Vol. 11: White, R.A.J Grfted greenhouse tomtoes give hevier crops. New Zelnd Journl Agriculture, 106:

10 Journl of Applied Horticulture, 8(1): 8-11, Jnury-June, 2006 Use of chlorophyll meter nd plnt visul spect for nitrogen mngement in tomto fertigtion Appl Journl Pulo Cezr Rezende Fontes nd Chrles de Arujo Deprtmento de Fitotecni, Universidde Federl, , Viços-MG. Brzil. Bolsists do CNPq. Apoio FAPEMIG, e-mil: Abstrct This study evluted the fesibility of using SPAD-502 chlorophyll meter nd plnt visul spect for N mngement in drip fertirrigted tomto plnts (Lycopersicon esculentum Mill.) under unheted greenhouse. Two seprte experiments were crried out t Universidde Federl de Viços - MG Brzil in leched nd non-leched soils under greenhouse. Six tretments were evluted in rndomised complete-block design with four replictes. In tretment 1, N ws pplied t the time SPAD reding in lef dropped below criticl vlue previously estblished for the specific plnt physiologicl stge (SPAD-1). In tretments 2 nd 3, SPAD criticl vlues were incresed 20 % (SPAD-2) nd decresed 10% (SPAD-3), respectively. In tretment 4, the visul spect of tomto plnt (PVA) ws utilized s criterion of N mngement. In tretments 5 nd 6 (check), N rtes were 280 nd 0 kg N h -1, respectively. Totl pplied N rtes rnged from 0 to 594 kg N h -1. In both the experiments, totl nd mrketble fruit yields were highest in SPAD-1 tretment which only differed from the check plot. All five criteri llowed high totl tomto fruit yields but, s experiments verge, N use efficiency ws highest with the PVA tretment. The highest net income ws obtined with SPAD-1 tretment nd ws ssocited with the highest yield. The results indicte tht SPAD meter cn provide quntittive mesure of the N requirement of the tomto plnts s long s pproprite SPAD criticl vlues re estblished. Visul rtings of plnt cnopy needs to be more evluted nd improved. Key words: Lycopersicon esculentum Mill, unheted greenhouse, drip irrigtion, SPAD, plnt nutrition Introduction Usully, nitrogen (N) fertilizer recommendtion to tomto crop re derived from nlysis of yield response to different N rtes from group of experiments (Fontes nd Guimrães, 1999). In intensive vegetble cropping systems, s greenhouse tomto production (Fyd et l., 2000), growers tend to dd excessive N fertilizer. However, economic, environmentl nd sfety considertions demnd tht N fertilizer should be pplied only in quntities which re strictly justified. Mtching greement between crop demnd nd supply is one of the prerequisites for efficient N use. Approches bsed on N contents in leves hve been used to increse N fertilizer use efficiency. N mngement progrm in tomto production cn be ttined by suitble evlution of plnt N sttus (Coltmn, 1988; Smith nd Lonergn, 1997) which is usully ccomplished by quntittive nlysis of the N concentrtion in the plnt dry mtter. Alterntively, quick procedures hd been proposed s the tomto lef greenness determintion by hnd-held device Minolt SPAD-502 meter (Sndovl-Vill et l., 1999; Guimrães et l., 1999) The chlorophyll meter SPAD-502 is for simple, rpid, nd non destructive estimtion of chlorophyll contents in tomto leves (Guimrães et l., 1999). As severl uthors hve shown reltionship between chlorophyll nd N contents in plnt leves (Scheepers et l., 1992; Sexton nd Crol, 2002; Wng et. l, 2004), chlorophyll contents cn be used s n lterntive mesure of plnt N sttus (Fontes, 2001). Timely nd nondestructive lef N sttus detection could llow rel time decision nd improvement in N mngement. Chlorophyll meter utiliztion to evlute plnt N sttus t rel time is suitble for precision griculture nd cnopy greenness might serve s useful dignostic tool to ssess plnt N demnd (Wiesler et l., 2002). This is lso vlid for plnt visul spect s long s evlution criterion could be estblished. Very few ppers del with the theme (Ronchi et l., 2001). The objective of this study ws to evlute the fesibility of using SPAD-502 chlorophyll meter nd plnt visul spect for N mngement in drip fertirrigted tomto plnt under unheted greenhouse conditions. Mterils nd methods Two experiments were crried out in unheted greenhouse t the Federl University of Viços MG Brzil. One experiment ws set in previously leched re (experiment 1) nd the other one ws set in non-leched re (experiment 2), in the sme greenhouse conditions. Leching ws ccomplished by pplying excessive wter in the soil during 15 dys immeditely before tomto plnt trnsplnttion. Six tretments were evluted in rndomised complete-block design with four replictes. In three tretments, Minolt SPAD-502 meter ws utilized for mesurements on five leflets of the lef closest to ech specific cluster, t the sme dy time, from 7:00 to 9:00.m., immeditely fter drip irrigtion. A men SPAD vlue ws clculted for ech plot t 28, 42, 56, 70 nd 98 dys fter trnsplnttion (DAT) coinciding to the flowering time of the first, second, third, fourth, fifth, nd sixth cluster, respectively. Ech SPAD vlue ws the men of the mesurement in 10 leflets. In tretment 1, (SPAD- 1), N ws pplied t the time SPAD reding dropped below

11 Use of chlorophyll meter nd plnt visul spect for nitrogen mngent in tomto fertigtion 9 Tble 1. Previously estblished SPAD criticl vlues (CV) nd SPAD redings t selected tomto plnt physiologicl stges 1 (dys fter trnsplnttion-dat) in experiments (Experiment 1 & 2) DAT 1 Tretments criticl vlue previously estblished for the specific physiologicl stge of the plnt. In tretments 2 nd 3, SPAD criticl vlues were incresed 20% (SPAD-2) nd decresed 10% (SPAD-3), respectively (Tble 1). SPAD criticl vlues (Y) utilized in the experiment were previously estblished from the eqution Ŷ = x, derived from Guimrães (1998), where x vlues were 28, 42, 56, 70, 84, nd 98 DAT (Tble 1). Plnts in ll three SPAD tretments received 50 kg N h -1 t trnsplnting nd the remining N ws pplied s necessry set by SPAD criticl vlues (Tble 1) t the rtes clculted by equtions given in Tble 2. Tble 2. Equtions utilized to clculte nitrogen fertilizer rte in SPAD tretments 1 Tretment Eqution SPAD-1 F={[50.7-(d x 0.17)]-C}x 70 SPAD-2 F={[60.8-(d x 0.20)]-C}x 70 SPAD-3 F={[45.7-(d x 0.15)]-C}x 70 SPAD-1 SPAD-2 SPAD-3 CV Exp. 1 Exp. 2 CV Exp. 1 Exp. 2 CV Exp. 1 Exp From the first to the sixth cluster. 1 F = N rte (kg N h -1 ); d = plnt ge (dys fter trnsplnttion) t the moment of SPAD reding; C = SPAD criticl vlues t selected physiologicl stge; 0.17, 0.20, nd 0.15 = dily decreses in the SPAD criticl vlue with tomto plnt ging; 70 = N rte (kg N h -1 ) to increse 1 SPAD unit. In tretment 4, tomto plnt visul spect (PVA) ws utilized s criterion for N mngement. The severity of lef chlorosis ws chrcterized using visul rting index (Tble 3). Every 14 dys, depending on the plnt visul rting index it ws decided on N sidedress ppliction. Nitrogen rte of 30, 22.5, 15 or 7.5 kg N h -1 ws dded whenever PVA where bd, regulr, good or very good, respectively. A pre-plnting 50 kg N h -1, t the trnsplnting time, ws pplied. In tretment 5 (REFE), N ws 280 kg N h -1 following recommendtion supported by locl experimentl results (Fontes nd Guimrães, 1999). In the tretment 6 (Check), plnts were not fertilized with N. At the trnsplnting time, N fertilizer (mmonium sulphte) ws plced in open furrows, under the tomto plnt. In sidedress, N fertilizer ws pplied by drip irrigtion. N rtes pplied during the experiment re given in Tble 4. The experiments were conducted using recommended culturl prctices (Fontes nd Silv, 2002) which includes 25 dys old seedlings (hybrid Crmen), plnt stems verticlly trined with plstic twine, stnd of 1.66 plnts m -2, drip irrigtion, stem tip pruned t 9 cluster, 10 hrvests (during 65 dys) nd 143 dys fter trnsplnttion cycle, from 10 September to 30 Jnury. Hrvested fruits were seprted s mrketble nd nonmrketble; the mrketble ones were grded ccording to Brzilin grde stndrds for big, medium, nd smll fruit. Bsed on different mrket prices for these three tomto fruit clsses, yield ws lso expressed s weighted yield tking into ccount the big, medium nd smll fruits being 1, 0.658, nd 0.396, respectively. Dt were sttisticlly evluted by nlysis of vrince nd tretment verges were compred with Tukey test (P=0.05). Results nd discussion In both experiments, tretments led to different N sidedress rtes nd ppliction dtes (Tble 4). Totl N rtes rnged from 0 to 594 kg N h -1. N requirement for high-yielding tomto fruit (> 80 t h -1 ), t field conditions, rnged from 125 to 351 kg N h -1 (Scholberg et l., 2000). In both experiments, incresing (SPAD- 2) or decresing (SPAD-3) SPAD criticl vlues in reltion to SPAD-1, led to higher or lower N fertilizer pplictions rtes, respectively (Tble 4). In experiments 1 nd 2 (Tbles 5 nd 6), totl nd mrketble fruit yields were highest t SPAD-1 tretment which only differed significntly from the check plot. Totl, mrketble, nd weighted yield vlues in this tretment were higher thn 97, 75, nd 45 previously obtined in the sme plce (Guimrães et l., 1999). Weighted yield indictes the production csh vlue s it tkes into ccount the price reltionships between ech fruit size grde (Fontes, 1997). All five criteri llowed high totl tomto fruit yields but with the PVA tretment, s experiments verge, due to lower N ddition, the nitrogen use efficiency (NUE) ws highest (Tble 7). NUE ws expressed s: (totl fruit yield t ech tretment - totl fruit yield t check plot)/(n rte in the tretment). Adjusting N rte in ssocition with visul spect nd eliminting evlutor bis my turn the PVA pproch useful. The highest net income ws obtined with SPAD-1 tretment (Exp. 2) nd ws ssocited with both the highest yield nd the highest NUE (Tble 7). SPAD-1 tretment led to pply N t 70 dys fter trnsplnttion (DAT), t lmost mid tomto plnt cycle, t the beginning of fruit hrvest which strted t 77 DAT. This ws probbility due to high N demnd by the tomto fruit enlrgement. At this time, N demnd increses (Tpi nd Gutierrez, 1997; Fyd et l., 2000) nd soil N contents plus 50 kg N h -1 dded t trnsplnttion time were not sufficient to mintin SPAD reding bove the criticl vlue. N rte pplied in function of SPAD tretment ws clculted bsed upon the criterion to pply 70 kg N h -1 to increse 1 SPAD unit. To increse 1 SPAD

12 10 Use of chlorophyll meter nd plnt visul spect for nitrogen mngent in tomto fertigtion Tble 3. Tomto plnt visul spect (PVA) utilized s criterion for N mngement in the tretment number 4 nd ssocited chrcteristics determined during plnt cycle PVA Chrcteristic Dys fter trnsplntting Bd Regulr Good Very good Cnopy greenness YE YE YE YE YE YE Lef number Plnt height (cm) Cnopy greenness YG YG YG YG YG YG Lef number Plnt height (cm) Cnopy greenness LG LG LG LG LG LG Lef number Plnt height (cm) Cnopy greenness DG DG DG DG DG DG Lef number Plnt height (cm) YE = yellow; YG = yellow green; LG = light green; DG = drk green. Tble 4. Sidedress N rtes (kg N h -1 ) pplied during the tomto plnt growth cycle in experiments 1 nd 2 Tretment Experiment Dys fter trnsplnting Totl SPAD SPAD SPAD PVA REFE CHECK Tble 5. Totl, mrketble nd weighted yields of tomto s function of tretments in experiment 1 Tretment Yield (t h -1 ) Totl Mrketble Weighted SPAD SPAD b 77.3 b 45.4 SPAD b 78.7 b 43.4 PVA 84.5 b 81.0 b 44.5 REFE 93.7 b 91.5 b 53.7 CHECK 68.3 b 64.5 b 37.5 Tble 6. Totl, mrketble nd weighted yields of tomto s function of tretments in experiment 2 Tretment Yield (t h -1 ) Totl Mrketble Weighted SPAD SPAD b 82.3b 49.2b SPAD b 74.9b 40.9b PVA 93.1b 88.5b 50.3b REFE 94.3b 89.8b 55.5b CHECK 71.7b 68.2b 40.3b In ech column, mens followed by the sme letter were not different by Tukey test (P=0.05) unit in cotton nd potto plnts it ws necessry 25 or 61 kg N h -1, respectively (Feibo et l., 1998; Gil et l., 2002). Vrvel et l. (1997) utilized 30 kg N h -1 when SPAD reding ws below the criticl level to obtin the highest corn yield. In SPAD-1 tretment, commercil verge yield ws 688 kg h -1 dy -1. Usully, tomto plnt cycle in the field is dys. But, it cn be grown for in the field for longer time nd in such cses the fruit productivity will be higher. So, expressing fruit productivity per dy plnt sty in the field, llow pproprite comprison mong reserch results (Fontes, 1997). Vlues rnging from 700 (Vooren et l., 1986) to kg h -1 dy -1 (Fontes et l., 1997, Ppodopoulos nd Ho, 1997) hve been reported. Finlly, the result suggests SPAD meter cn provide quntittive mesure of the requirement of tomto plnts s long s pproprite SPAD criticl vlue re estblished. To estblish precise nd universl criticl SPAD index is complex process due to the nrrow vlues seprting N deficiency from surplus nd gret number of vribles ffecting the index, s chnges in lef irrdince nd wter sttus (Mrtinez nd Guimet, 2004), environmentl conditions nd sttisticl procedures (Fontes nd Ronchi, 2002). Cution is needed regrding the universlity of SPAD nd N clibrtions cross geogrphicl

13 loctions nd sesons. To counter these potentil problems, users should estblish the SPAD criticl vlues for specific environmentl condition. Visul rtings of plnt cnopy needs to be more evluted. This my fcilitte more precise N fertilizer recommendtions nd thereby help to minimize nitrte contents in the soil. References Use of chlorophyll meter nd plnt visul spect for nitrogen mngent in tomto fertigtion 11 Tble 7. Totl nitrogen fertilizer rte nd cost, net income, nitrogen use efficiency (NUE), gronomic nitrogen efficiency (ANE) for ech tretment in experiments 1 nd 2 1 Tretment Experiment Totl N N cost NPI 2 NUE 3 ANE 4 (kg N h -1 ) (US$ h -1 ) (US$ h -1 ) (kg kg -1 ) (kg kg -1 ) SPAD , , SPAD , , SPAD , , PVA , , REFE , , CHECK , , N price: US$ 1.25 kg -1 ; selling price of high grded fruit (weighted yield): US$ 0.30 kg -1 2 Net prtil income: (weighted yield x 0.30) (N fertilizer cost). 3 NUE: (totl fruit yield t ech tretment - totl fruit yield t check plot)/(n rte t tretment). 4 NE: (totl fruit yield t ech tretment)/(n rte t ech tretment). Coltmn, R.R Yield of greenhouse tomtoes mnged to mintin specific petiole sp nitrte. HortScience, 23(1): Fyd, J.A., P.C.R Fontes, A.A. Crdoso, F.L. Finger nd F.A. Ferreir, Absorção de nutrientes pelo tomteiro cultivdo em condições de cmpo e de estuf. Horticultur Brsileir, 20(1): Feibo, W., W. Linghun nd X. Fuhu, Chlorophyll meter to predict nitrogen sidedress requirements for short-seson cotton (Gossypium hirsutum L.). Field Crops Res., 56: Fontes, P.C.R Produtividde do tomteiro: kg/h ou kg/h/di?. Horticultur Brsileir, 15(2): Fontes, P.C.R Dignóstico do estdo nutricionl ds plnts. Viços: UFV, 122p. Fontes, P.C.R., E.N. Dis, S.R. Znin nd F.L. Finger, Produção de cultivres de tomte em estuf cobert com plástico. Revist Ceres, 44(252): Fontes, P.C.R. nd T.G. Guimrães, Mnejo dos fertilizntes ns culturs de hortliçs cultivds em solo, em mbiente protegido. Informe Agropecuário, 20(200&201): Fontes, P.C.R. nd C.P. Ronchi, Criticl vlues of nitrogen indices in tomto plnts grown in soil nd nutrient solution determined by different sttisticl procedures. Pesquis Agropecuári Brsileir, 37(10): Fontes, P.C.R. nd D.J.H. Silv, Produção de tomte de mes. Viços, MG: Aprend Fácil, 195p. Gil, P.T., P.C.R. Fontes, P.R. Cecon nd F.A. Ferreir, Índice SPAD pr o dignóstico do estdo de nitrogênio e pr o prognóstico d produtividde de btt. Horticultur Brsileir, 20(4): Guimrães, T.G Nitrogênio no solo e n plnt, teor de clorofil e produção do tomteiro, no cmpo e n estuf, influencidos por doses de nitrogênio. Viços-MG: UFV, 184p. (DS Thesis). Guimrães, T.G., P.C.R. Fontes, P.R.G. Pereir, V.H.V. Alvrez nd P.H. Monnert, Teores de clorofil determindos por medidor portátil e su relção com forms de nitrogênio em folhs de tomteiro cultivdos em dois tipos de solo. Brgnti, 58(1): Mrtinez, D.E. nd J.J. Guimet, Distortion of the SPAD 502 chlorophyll meter redings by chnges in irrdince nd lef wter sttus. Agronomie, 24(1): Ppdopoulos, A.P. nd X. Ho, Effects of three greenhouse cover mterils on tomto growth, productivity, nd energy use. Scienti Hortic., 69: Ronchi, C.P., P.C.R. Fontes, P.R.G. Pereir, J.C.S. Nunes nd H.E.P. Mrtinez, Índices de nitrogênio e de crescimento do tomteiro em solo e em solução nutritiv. Revist Ceres, 48: Sndovl-Vill, M., C.W. Wood nd E.A. Guertl, Ammonium concentrtion in solution ffects chlorophyll meter redings in tomto leves. J. Plnt Nut., 22(11): Scheepers, J.S., D.D. Frncis, M. Vigil nd F.E. Below, Comprison of corn lef-nitrogen concentrtion nd chlorophyll meter redings. Commun. Soil Sci. Plnt Anl., 23(17&20): Scholberg, J., B.L. McNel, K.J. Boote, J.W. Jones, S.J. Locscio nd S.M. Olson, Nitrogen stress effects on growth nd nitrogen ccumultion by field-grown tomto. Agronomy Journl, 92: Sexton, P. nd J. Crrol, Comprison of SPAD chlorophyll meter redings vs. petiole nitrte concentrtion in sugrbeet. J. Plnt Nut., 25(9): Smith, F.W. nd J.E. Lonergn, Interprettion of plnt nlysis: concepts nd principles. In: Plnt nlysis - n interprettion mnul. Reuter, D.J., Robinson, J.B. (Eds). Collingwood: CSIRO Publishing, p Tpi, M.L. nd V. Gutierrez, Distribution pttern of dry weight, nitrogen, phosphorus, nd potssium thought tomto ontogenesis. J. Plnt Nut., 20(6): Vrvel, G.E., J.S. Schepers nd D.D. Frncis, Ability for in-seson correction of nitrogen deficiency in corn using chlorophyll meters. Soil Sci. Soc. Am. J., 61: Vooren, J., G.W.H. Welles nd G. Hymn, Glsshouse crop production. In: The tomto crop scientific bsis for improvement. Atherton, J.G., Rudich, J. (Eds.). London: Chpmn nd Hll, p Wng, Q., J. Chen nd Y. Li, Nondestructive nd rpid estimtion of lef chlorophyll nd nitrogen sttus of pece lily using chlorophyll meter. J. Plnt. Nut., 27: Wiesler, F., M. Buer, M. Kmh, T. Engels nd S. Reusch, The crop s indictor for sidedress nitrogen demnd in sugr beet production limittions nd perspectives. J. Plnt Nutr. Soil Sci., 165:

14 Journl of Applied Horticulture, 8(1): 12-14, Jnury-June, 2006 Biodegrdble pper/polymerized vegetble oil mulches for tomto nd pepper production Appl Journl Rndl L. Shogren 1* nd Monic Dvid 2 1 Plnt Polymer Reserch Unit, Ntionl Center for Agriculturl Utiliztion Reserch, USDA/ARS, 1815 N. University St., Peori, IL 61604, 2 University of Illinois Extension, 1201 South Dorner Drive, Urbn, IL 61801, *Corresponding uthor. e-mil: shogrerl@ncur.usd.gov Abstrct This project ws undertken to compre the efficcy of biodegrdble pper/cured vegetble oil mulch with newspper/strw nd bre soil for reducing weed growth nd promoting vegetble yields. There were no significnt differences in totl tomto (Lycopersicon esculentum) or pepper (Cpsicum nnum) yields between the different mulch types. The coted pper nd newspper/strw mulches were effective in preventing weed growth round the plnts while hnd weeding ws required for the bre soil plots. After 3 months, there ws slight degrdtion ( few crcks, holes) of the coted pper mulches but not enough to llow noticeble weed penetrtion or detchment of the buried edge. Pper/cured oil mulch rolls pper to be convenient nd effective lterntive to lborious hnd weeding or spreding of newspper nd strw for vegetble grdening. Key words: Degrdble mulch, soyben oil, sustinble griculture, vegetble production Introduction There hs been growing interest recently in the use of biodegrdble mulch films for suppression of weeds, incresing soil tempertures nd yields of vegetbles nd fruits (Greer nd Dole, 2003; Hlley et l., 2001; Weber, 2003). A biodegrdble mulch llows growers to till the mulch into the field t the end of the growing seson rther thn hving to remove nd dispose of non-degrdble polyethylene mulches, often t considerble cost (Anderson et l., 1995). Vrious types of biodegrdble mulches hve been considered including strch-bsed films (Hlley et l., 2001), polyester films (Dever et l., 1998), fiber slurries (Olsen nd Gounder, 2001) nd coted pper. Pper lone begins to ter nd blow wy within 2-3 weeks fter field ppliction due to rpid biodegrdtion nd loss of strength when wet (Anderson et l., 1995; Shogren, 2000). Therefore, number of cotings or lmintes on pper hve been exmined to increse wet strength nd slow biodegrdtion. These include tr (Rivise, 1929), wx, polyethylene (Vndenberg nd Tiessen, 1972), ltexes (Brult et l., 2002), polyesters (Rngrjn, 2000) nd vegetble oils (Anderson et l., 1995). Vegetble oil cotings re ttrctive becuse they re inexpensive, renewble, produced in lrge quntity in the U.S. nd cn be polymerized (cured) into wter resistnt, biodegrdble films (Shogren, 1999). Previous studies hve shown tht krft pper coted with soyben or linseed oils then llowed to cure vi sun nd ir in the field were effective s polyethylene mulches for growing wtermelon (Shogren nd Hochmuth, 2004) nd cottonwood trees (Shogren nd Rousseu, 2005). One disdvntge of these oil sturted pper mulches ws the messiness ssocited with hndling oily pper in the field (Shogren nd Hochmuth, 2004). In order to void this problem, krft pper ws coted with resin mde from epoxidized soyben oil (ESO) nd citric cid nd then thermlly cured (Shogren, 1999; Shogren, 2000). Previous work hs shown these compositions to be fully biodegrdble in soil but over longer time spn thn uncoted pper (severl months) (Shogren et l., 2004). The objective of this study ws to ssess whether mulches mde from pper coted with ESO/citric cid resins would serve s effective weed brriers, promote the growth of tomto nd pepper plnts nd sustin good yields when compred to other weed control methods commonly used in grden or smll frm setting. Mterils nd methods Mterils: Brown krft pper, 30 nd 40 lb weights (3000 ft -2 ), mde from 100% recycled fiber ws obtined from Crter Pper nd Pckging, Peori, IL. Epoxidized soyben oil (ESO, Prplex G-62) ws from C. P. Hll, Bedford Prk, IL. Citric cid (99%) ws from Aldrich, Milwukee, WI. Strw ws mixture of whet (Triticum estivum) nd rye (Secle cerele) stems from locl frm. Tomto (Lycopersicon esculentum) nd sweet green pepper (Cpsicum nnum) plnts were donted by Greenview Nursery, Peori, IL. Preprtion of coted pper: ESO (2950 g) ws heted to 100 o C in 11.5 l stinless steel beker nd 110 g of crbon blck (colornt) ws dded with motorized propeller stirrer. Crbon blck ws dded to reduce light trnsmission through the pper mulches. A hot (102 o C) solution of 850 g citric cid in 280 g deionized wter ws next dded to the ESO with stirring. After the mixture reched 105 o C, the beker ws removed from het nd plced in n ice bth to cool to 40 o C. * Product nmes re necessry to report fctully on vilble dt; however the USDA neither gurntees nor wrrnts the stndrd of the product, nd the use of the nme. USDA implies no pprovl of the product to the exclusion of others tht my lso be suitble.

15 Biodegrdble pper/polymerized vegetble oil mulches for tomto nd pepper production 13 Pper coting nd het curing ws ccomplished using simple in-house design. This consisted of stinless steel tble with supply nd motorized tke-up rolls on the lower shelf, flt luminum sheet on the top shelf for coting, nd n ttched flow-through sheet oven for het-curing. The oven consisted of 2 x 4.5 ft. luminum sheets seprted by 1 in. heted by electricl resistnce mts nd insulted with 2 in. of fiberglss top nd bottom. ESO/citric cotings were dripped onto one side of the pper nd were spred into thin lyer by neoprene rubber blde clmped between 2 luminum brs. Tempertures nd residence times in the oven were o C nd ~1 min, respectively t coting speeds of 2-3 ft./min. Coting weights were 37 nd 42% of pper weight for the 30 nd 40 lb ppers, respectively. Field studies: There were four different mulch tretments (30 lb coted pper, 40 lb coted pper, newspper/strw, bre soil). The field site ws locted in Peori, IL nd run by the University of Illinois Mster Grdeners of Peori County s prt of the Plnt--Row-for-the-Hungry progrm. Tomto nd bell pepper plnts were plnted in djcent 75 ft. long rows with spcings of pproximtely 1.5 ft. between plnts nd 4 ft. between row centers. Plnting took plce on 26 My nd mulches were lid on 3 June Coted pper mulch smples were cut into 6 x 2 ft. lengths then cut from the side with scissors to llow the mulch to be plced round the plnts (coted side up). Edges of the mulches were then weighted down with soil (pepper) or held in plce by steel cges (tomto). For the newspper/strw tretment, 3 lyers of newspper were plced round the plnts followed by pproximtely 2 inches of strw on top on the newspper to help hold the newspper in plce. There were 3 replictions for ech tretment nd were rrnged in rndomised order. The bre soil plots were hnd weeded once per week over the summer. Vegetbles were hrvested weekly beginning 21 July until 15 September. Totl number s well s weight of vegetbles deemed mrketble were mesured weekly. Soil tempertures were mesured in duplicte on 18 June, 24 June, 7 July nd 26 August t 8 AM nd 2 PM using digitl temperture probe model 4045 (Control Co., Friendswood, TX). Mesurements were tken t depth of pproximtely 4 in. (10.2 cm) below the surfce. Soil smples for moisture determintion were collected on 18 June, 24 June, 7 July nd 26 Tble 1. Totl men yields of tomto nd pepper August. Mulches were crefully lifted from the edge nd the top 3-4 in. of soil (~100 g) ws scooped into plstic zip lock bgs. Edges of the mulches were reburied fter smple collection. Soil smples were then trnsported to the lb where wter content ws mesured grvimetriclly fter heting 10 g soil to 105 o C for 20 min. using n Ohus moisture nlyzer, model MB200 (Ohus Co., Florhm Prk, NJ). Counts of weeds growing through the mulches were mde from detiled photogrphs of the plots tken on 24 June, 24 July, 4 August nd 18 September. Sttisticl nlyses: A Levene's homogeneity of vrince test ws crried out to determine if trnsformtion of the vegetble number nd weight dt were necessry. Four, single-fctor Anlyses of Vrince (ANOVA) were performed compring the four mulch tretments for number nd weight of hrvested tomtoes nd green peppers t the end of ech week nd for the seson. In cse significnt F-test ws obtined in ANOVA, Duncn's multiple rnge test ws used for multiple comprison procedure, t the P=0.05 level, for determining pirwise differences between the mulch tretments. Results nd discussion As shown in Tble 1, there were no significnt differences in the totl number or weight of tomtoes or peppers between plnts grown on the different mulch types. Soil tempertures for the different mulch tretments re shown in Tble 2. Soil tempertures underneth the newspper/strw were less thn the coted pper or bre soil, especilly during the hottest prt of the dy (fternoon). This is probbly due to the thick, insultive properties of the strw s well s its light colour which tends to reflect solr rdition. There ws noticeble lightening in colour (bleching) of the coted ppers over the summer nd this would likely tend to lessen the soil wrming lter on. This lightening effect hs been noted previously for coted pper mulches (Brult et l., 2002). Soil moisture, s shown in Tble 2, ws generlly higher under the newspper/strw mulch thn under the bre soil or coted pper. This is likely due to the lower soil temperture under the newspper/strw nd hence lower evportion rtes. Wter permebility of the coted pper mulches hs not been mesured but simple test of drop of wter plced on the mulch shows it will pss through within n Mulch Totl men tomto yield Totl men pepper yield Number (number/plot) Weight (kg/plot) Number (number/plot) Weight (kg/plot) 30 lb pper/oil 59 z lb pper/oil Newspper/strw Bre soil z Mens with the sme letter within column re not significntly different t P=0.05. Tble 2. Men soil tempertures nd moistures on 7 July Mulch 8 AM 2 PM Temperture ( o C) Moisture (%) Temperture ( o C) Moisture (%) 30 lb pper/oil 25.4 z lb pper/oil Newspper/strw 24.0 b b 17.9 b Bre soil z Mens with the sme letter within column re not significntly different t P=0.05. Ambient ir tempertures were 30 nd 34.8 o C t 8 AM nd 2 PM, respectively

16 14 Biodegrdble pper/polymerized vegetble oil mulches for for tomto nd pepper production Tble 3. Men y number of weeds from six 1.8 x 0.6 m plots Mulch Dys post-plnting lb pper/oil 0 z lb pper/oil Newspper/strw Bre soil 4 b 6 b 14 b 11 b y Men of six replictions (dt from tomto nd pepper replictes combined) z Mens with the sme letter within column re not significntly different t P=0.05. hour or so, indicting some permebility. Men number of weeds penetrting different mulch tretments re given in Tble 3. Even fter lmost 4 months, there ws only n verge of 1 weed per 6 ft plot penetrting the coted pper mulches. The exposed nd buried tuck res of the coted pper mulches were lrgely intct, with few holes. In contrst, there were >10 weeds per plot in the bre soil tretments, even with weekly hnd pulling of weeds. There ws no significnt difference between the 30 nd 40 lb coted pper so the thinner pper could be used for better economics. Mnul weed removl required bout 1 hour per week for the six 6-ft bre ground plots or 16 hours totl for the 16 week growing seson. The time required for the initil coted pper mulch lying ws bout 1 hour for six 6-ft plots. Thus, pper/eso/citric cid mulches were effective in preventing weed growth nd plnts grown on these gve yields of tomtoes nd green peppers similr to the newspper/strw mulch or hndweeded control plots. For mny home or community grdeners, hnd-weeding is tedious nd difficult tsk so use of weedblocking mulch would be desirble. Appliction of the pper/oil mulch would be little esier thn newspper/strw since the former is in roll form. It could be unrolled first then seedlings plnted in cut holes or plced round existing plnts s in this study. For lrger-scle vegetble growers, the pper/oil rolls would work with conventionl plstic mulch-lying equipment, s hs been shown previously (Shogren nd Hochmuth, 2004). The ESO/citric cid coted pper used here might be more redily ccepted commercilly since it hs been het cured to give hrd surfce rther thn the oily pper described previously (Shogren nd Hochmuth, 2004). Pper coted with other types of biodegrdble polymers, especilly polyesters, hve been tested recently (Rngrjn, 2000). These biodegrdble polyesters, such s polycprolctone, polylctic cid, poly(butylene succinte-dipte), re currently rther expensive ($1.5-3/lb.) Epoxidized soyben oil nd citric cid re less expensive ($ /lb.), mking them more economiclly ttrctive. Pper, on n re bsis, is however more expensive thn polyethylene mulch due to the greter thickness of the pper. Thus the coted pper mulches would be more suited to higher vlue pplictions such s home grdening or smll frmers or where biodegrdble, wter permeble mulch is required. Acknowledgements We thnk Elizbeth Krietemeyer, Lynn Webb, Helen Nixon nd Ptrise Swnson for help in field work. We lso thnk Debr Plmquist, USDA/ARS Midwest re sttisticin for sttisticl nlyses. References Anderson, D.F., M. Gristo, J. Bourrut, M.W. Schonbeck, R. Jye, A. Wurzberger nd R. DeGregorio, Evlution of pper mulch mde from recycled mterils s n lterntive to plstic film mulch for vegetbles. J. Sustinble Agriculture, 7: Brult, D., K.A. Stewrt nd S. Jenni, Opticl properties of pper nd polyethylene mulches used for weed control in lettuce. HortScience, 37: Dever, M., P. Lmbdin nd B. Ownley, Biodegrdble plstic mulch: An option in vegetble production? Tennessee Agriculturl Science, 186: 16. Greer, L. nd J.M. Dole, Aluminum foil, luminum-pinted, plstic, nd degrdble mulches increse yields nd decrese insectvectored diseses of vegetbles. HortTechnology, 13: Hlley, P., R. Rutgers, S. Coombs, J. Kettels, J. Grlton, G. Christie, M. Jenkins, H. Beh, K. Griffin, R. Jysekr nd G. Lonergn, Developing biodegrdble mulch films from strch-bsed polymers. Strch/Stärke, 53: Olsen, J.K. nd R.K. Gounder, Alterntives to polyethylene mulch film - field ssessment of trnsported mterils in cpsicum (Cpsicum nnuum L.). Austrlin J. Experimentl Agriculture, 41: Rngrjn, A Pper mulch: Cn it replce plstic? hort.cornell.edu-commercilvegetbles. Rivise, C.W Mulch pper. Pper Trde J., 89: Shogren, R.L Preprtion nd chrcteriztion of biodegrdble mulch: pper coted with polymerized vegetble oils. J. Appl. Polym. Sci., 73: Shogren, R.L Biodegrdble mulches from renewble resources. J. Sustinble Agriculture., 16: Shogren, R.L. nd R.C. Hochmuth, Field evlution of wtermelon grown on pper-polymerized vegetble oil mulches. HortScience, 39: Shogren, R.L. nd R.J. Rousseu, Field testing of pper/ polymerized vegetble oil mulches for enhncing growth of estern cottonwood trees for pulp. Forest Ecology nd Mngement, 208: Shogren, R.L., Z. Petrovic, Z. Liu nd S.Z. Erhn, Biodegrdtion behviour of some vegetble oil-bsed polymers. J. Polymers nd the Environment, 12: Vndenberg, J. nd H. Tiessen, Influence of wx-coted nd polyethylene-coted pper mulch on growth nd flowering of tomto. HortScience, 7: Weber, C.A Biodegrdble mulch films for weed suppression in the estblishment yer of mtted-row strwberries. HortTechnology, 13:

17 Journl of Applied Horticulture, 8(1): 15-20, Jnury-June, 2006 Compct 3U s novel lighting source for the propgtion of some horticulturl plnts Appl Journl D.T. Nhut 1*, M.T. Ngoc Huong 1, D.V. Khiem 1 nd J.A. Teixeir d Silv 2 1 Deprtment of Plnt Biotechnology, Dlt Institute of Biology, 116 Xo Viet Nghe Tinh, Dlt, Lm Dong, Vietnm. 2 Fculty of Agriculture, Kgw University, Miki-cho, Kgw, , Jpn. *Corresponding uthor; e-mil: duongtnnhut@yhoo.com Abstrct A novel lighting system (Compct 3U) ws successfully pplied to the micropropgtion of some horticulturl plnts. Cymbidium Tim Hot, Lilium longiflorum nd Frgri vesc cv. My D shoots were used for this study. To compre in vitro growth of plntlets plced under Neon nd Compct 3U lighting systems, Frgri vesc cv. My D shoots were cultured on ½ MS medium supplemented with 1 gl -1 ctivted chrcol, 30 gl -1 sucrose nd 8 gl -1 gr under two lighting sources t 45 µmolm -2 s -1. After three weeks of culture, the shoot nd root length, lef re nd fresh weight of strwberry plntlets under Compct 3U system were significntly higher thn those grown under Neon system. To clrify the effect of irrdince of Compct 3U system on the development of plntlets, Cymbidium Tim Hot shoots were cultured on MS medium supplemented with 0.5 mgl -1 NAA, 1 gl -1 ctivted chrcol, 100% coconut wter, 25 gl -1 sucrose nd 8 gl -1 gr, Lilium longiflorum nd Frgri vesc cv. My D shoots were cultured on ½ MS medium supplemented with 1 gl -1 ctivted chrcol, 30 gl -1 sucrose nd 8 gl -1 gr t different irrdinces: (1) Neon t 45 µmolm -2 s -1 (control), nd Compct 3U t: (2) 45 µmolm -2 s -1, (3) 60 µmolm -2 s -1, nd (4) 75 µmolm -2 s -1. The results showed tht plntlets of the three gener dpted differently to irrdinces nd lighting sources, but in ll, the growth of plntlets were better under the Compct 3U system. Futhermore, ex vitro plntlets derived from Compct 3U system lso developed better thn those from Neon system. Key words: Compct 3U, Neon, Cymbidium Tim Hot, Lilium longiflorum, Frgri vesc cv. My D Introduction Now--dys, in vitro multipliction is primry method to rpidly mss-produce horticulturl plnts. The demnd for high qulity plnting mteril hs been incresing quickly worldwide for reforesttion, foods/forge production, urbn/indoor horticulture nd globl environment protection (Kozi et l., 1992). In mny cses, since micropropgtion gve some superior trnsplnt qulities to seedling production nd conventionl vegettive production, billions of micropropgted plntlets were produced nnully world-wide (Debergh nd Zimmermn, 1990). Tissue culture hs been crried out in more thn 600 compnies ll over the world. However, the widespred use of micropropgtion for mjor crops in griculture nd horticulture ws restricted becuse of its reltively high production costs cused by high lbour cost (Kozi et l., 1992), especilly electricl energy consumption. Control of plntlet growth nd morphology is importnt in micropropgtion to obtin high plntlet qulity t different growth nd developmentl stges, nd to sve lbour by utomtion or robotics (Miyshit, 1995). Mny of the growth nd morphologicl chrcteristics of plnts in nd ex vitro re influenced by environmentl fctors, such s light (qulity, intensity, durtion nd direction), temperture, gseous composition (CO 2, O 2, H 2 O nd C 2 H 4 ), nd medium composition (Schwbe, 1963; Kozi et l., 1992). Light qulity hd significnt influence on the growth nd morphology of plnts in nd ex vitro (Wrrington nd Michell, 1976; Morgn nd Smith, 1981; Smith, 1982; Tibbitts et l., 1983; Mortensen nd Stromme, 1987; Economou nd Red, 1987; Agrwl, 1992). The totl quntity of light tht plnt received during illumintion directly ffected photosynthesis s well s plnt growth nd yield (Kim nd Kozi, 2000). Hence, mny lighting systems tht effectively used electricl energy in the multipliction of horticulturl plnts hve been studied intensively such s fluorescent, incndescent, luminescent (Sodium high pressure) lighting systems, nd recently, light-emitting diode (LED) lighting source. However, tissue cultured plnts re lmost invribly grown under fluorescent illumintion (Collin et l., 1988), especilly under cool white fluorescent lmps with high proportion of its output in the blue nd red regions (Hrt, 1988). Previous studies hd been done on the effect of light qulity nd intensity of different lighting sources on the growth nd morphology of in nd ex vitro plntlets (Sebrook, 1987; Hyshi et l., 1992; Iwnmi et l., 1992; Kozi et l., 1992; Kirdmnee et l., 1993; Gbrkiewicz et l., 1997; Wulster nd Jnes, 1997; Ms nd Bkx, 1997; Kunnemn nd Ruesink, 1997; Moe, 1997; Fust nd Heins, 1997; Murkmi et l., 1997; Gbryszewsk nd Rudnicki, 1997; Wlz nd Horn, 1997; Miyshit et l., 1997; Nhut, 2002). In our study, Compct 3U lmps were used s promising lighting source for propgting some horticulturl plnts such s Cymbidium, Lilium nd strwberry. These plnts re highly vluble economic crops in Vietnm s well s ll over the world. In this report, we focused on the effects of two different lighting sources (Neon nd Compct 3U) s well s some different intensities of Compct 3U lmps (45, 60, nd 75 µmolm -2 s -1, respectively) on the growth nd morphology of these in vitro plntlets, nd Neon lmp (with cool white emission) s control system. Compct 3U lmp (Fig. 1), which sves 80% electricl energy

18 16 Compct 3U s novel lighting source for the propgtion of some horticulturl plnts s compred to incndescent lmps, hs compct size, long life (>6.000 h), nd reches one-fifth the brightness of conventionl incndescent lmps. Hence, plnt production cost could be decresed. Mterils nd methods Plnt mterils nd culture medi: Cymbidium Tim Hot shoots (4 cm length), derived from protocorm-like bodies (PLBs) cultured on MS (Murshige nd Skoog, 1962) medium contining 0.5 mgl -1 α-nphthlenecetic cid (NAA), 2 mgl -1 6-benzyldenine (BA), 1 gl -1 ctivted chrcol (AC), 20% coconut wter (CW), 30 gl -1 sucrose nd 8 gl -1 gr (Hiphong Co., Vietnm), were cultured on MS medium contining 0.5 mgl -1 NAA, 1 gl -1 AC, 10% CW, 25 gl -1 sucrose nd 8 gl -1 gr. Lilium longiflorum bulb scles, derived from in vitro bulblets cultured on MS medium contining mgl -1 BA, 30 gl -1 sucrose nd 8 gl -1 gr, were cultured on ½ MS medium supplemented with 1 gl -1 AC, 30 gl -1 sucrose nd 8 gl -1 gr. Frgri vesc cv. My D shoots (1.5 cm), derived from meristems cultured on MS medium contining vitmin B 5, 0.2 mgl -1 BA, 30 gl -1 sucrose nd 8 gl -1 gr, were cultured on ½ MS medium supplemented with 1 gl -1 AC, 30 gl -1 sucrose nd 8 gl -1 gr. For ll experiments, explnts were cultured in vessels (500 ml) contining 60 ml medium. ph of medi ws djusted to 5.7 before utoclving t 121 o C, 1 tm for 40 min. Lighting systems: Cool white fluorescent lmps (Neon tubes) (40 W ech; Rng Dong Light source nd Vcuum Flsk Co., Vietnm, FL-40W/T10) nd wrm white fluorescent lmps (Compct 3U lmps) (18 W ech; Rng Dong Light source nd Vcuum Flsk Co., Vietnm, CFH-3U18W) were used s lighting sources in ech experiment. Irrdinces were 45 µmolm -2 s -1 under Neon light or 45, 60, 75 µmolm -2 s -1 for Compct 3U system ccording to ech experiment. Photosynthetic photon flux density (PPFD) ws mesured with n illumintion meter (Tokyo photoelectric Co., LTD., Jpn, ANA-F11) on the empty culture shelf. Experimentl designs Fig. 1 Fig. 2 Effect of Compct 3U lighting source on the in vitro development of Frgri vesc cv. My D plntlets: Five strwberry shoots were cultured in ech culture vessel, nd ten vessels were plced on the shelf in one row under the Compct 3U lighting system with three lmps per shelf, rrnged in one row. Ten other vessels were plced in one row on nother shelf under the Neon lighting system t 45 µmol m -2 s -1 (the control lighting system). After three weeks of culture, some morphologicl prmeters (plnt height nd fresh weight, root length, lef re) were recorded nd the in vitro plntlets were trnsplnted to the greenhouse. Effect of different irrdinces of Compct 3U on the in vitro development of Cymbidium, Lilium nd strwberry plntlets: Ech vessel contined five shoots of ech plnt (Cymbidium, Lilium nd strwberry). There were four shelves (ten vessels per shelf) with different irrdinces: three shelves with the Compct 3U lighting system t either 45, 60 or 75 µmolm -2 s -1, nd the remining shelf with the Neon lighting system t 45 µmolm - 2 s -1. Some morphologicl prmeters of strwberry plntlets (plnt height, lef re, root length, nd plnt fresh weight) were recorded fter three weeks of culture, of Cymbidium (plnt height, root length, lef re, number of newly formed roots, number of bulbs, bulb dimeter, bulb cluster fresh weight nd bulb fresh weight) nd of Lilium (plnt height, root length, lef width, number of roots nd plnt fresh weight) were recorded fter six weeks of culture. The process to set up the Compct 3U nd Neon lighting sources for studying in vitro development of some horticulturl plnts is depicted in Fig. 2. The in vitro plntlets were therefter trnsplnted to greenhouse. This subsequent stge of development of Lilium nd Cymbidium plntlets were plced under 6h/dy supplementl Compct 3U lighting source. After one nd hlf months of culture, plnt Explnts (shoot, bulb) After culture period, trnsplnting plntlets to greenhouse Glss vessel contining medium Neon light Compct 3U t 45 Compct 3U t 60 Compct 3U t 75 Fig. 1. Compct 3U lmp. In vitro explnts Fig. 2. Setting up the Compct 3U nd Neon lighting sources for studying in vitro growth nd development of some horticulturl plnts.

19 height, lef width, number of leves, number of roots, root length nd plnt fresh weight of Cymbidium nd plnt height, bulblet dimeter, lef width, number of leves, number of roots, root length, nd plnt fresh weight of Lilium were collected. Culture conditions (in nd ex vitro): In vitro cultures were incubted t 25 ± 2 o C with ten-hour photoperiod nd 75-80% reltive humidity under different lighting systems s tretment. After six weeks of culture, Cymbidium nd Lilium plntlets were trnsplnted to the greenhouse nd cultured on tree fern fiber substrte in spongy trys t 25 ± 2 o C, 80-85% reltive humidity nd under 6h/dy supplementl Compct 3U lighting source. Plntlets were spryed with n ntifungl solution contining 5 gl -1 Dithne M-45 (Dow AgroSciences Co., USA) twice week. In ddition, these plntlets were spryed with pesticide solution contining 150 gl -1 Sumi lph (Omo Chemicl Ltd., Co., Jpn) nd fertilizer solution contining 100 gl -1 NPK, 50 gl -1 Komix BFC 201 (Thien Sinh Biochemicl Agriculture nd Trde Co., Vietnm) nd 15 gl -1 Mircle Fort (Phu Hung Foundtion, Vietnm) once week. Moreover, plntlets were lso wtered twice dily. Sttisticl nlysis: Ech tretment ws repeted three times nd dt ws recorded t the 3 rd or 6 th week of culture. The explnts in experiments were rrnged in rndomized complete block design with five shoots per tretment nd three blocks. The dt were nlyzed for significnce using nlysis of vrince with the men seprtion by Duncn s multiple rnge test (Duncn, 1995). Results nd discussion Compct 3U s novel lighting source for the propgtion of some horticulturl plnts 17 Effect of Compct 3U lighting system on the in vitro development of Frgri vesc cv. My D plntlets: The effect of the Compct 3U nd Neon lighting source on the in vitro growth nd development of strwberry plntlets re described in Tble 1 nd Fig. 3. There ws significnt difference in plnt height, root length, lef re nd plnt fresh weight between plntlets plced under two lighting systems t 45 µmolm -2 s -1. The morphologicl prmeters of strwberry cultured under Compct 3U were higher thn those under Neon lighting system. Tble 1. Effect of the Compct 3U lighting source on the in vitro development of Frgri vesc cv. My D plntlets fter 3 weeks of culture Lighting Plnt height Lef re Root length Plnt fresh system (cm) (mm 2 ) (cm) weight (mg) Neon (control) 3.7b b 200b Compct 3U b Different letters within column indicte significnt differences (P = 0.05) by Duncn s multiple rnge test. Effect of different irrdinces of Compct 3U lighting source on the in vitro development of Cymbidium, Lilium nd strwberry plntlets Frgri vesc cv. `My D : The effect of different Compct 3U irrdinces on the in vitro development of strwberry is shown in Tble 2. In generl, in vitro strwberry shoots plced under the Neon lighting system hd slower growth thn those plced under the Compct 3U lighting system. Two lighting sources hd different effects on the morphology nd biomss of strwberry, wheres different Compct 3U irrdinces virtully did not ffect the morphologicl prmeters of strwberry (Fig. 3). Strwberry plntlets were best t 75 μmol m -2 s -1. becuse of the mximum growth nd development. However, for commercil purposes, we recommended the use of Compct 3U light t 45 μmol m -2 s -1 for the micropropgtion of strwberry becuse of sving electricl energy nd still remining the reltively good growth nd development. Tble 2. Effect of different irrdinces of Compct 3U on the in vitro development of Frgri vesc cv. My D plntlets fter 3 weeks of culture Irrdince (Compct 3U) (µmolm -2 s -1 ) Plnt height (cm) Lef re (mm 2 ) Root length (cm) Plnt fresh weight (mg) Neon (45) 3.7c x 40.5d 2.5d 200d b 45.4b 3.5b 250c b 41.4c 3.0c b x Different letters within column indicte significnt differences (P = 0.05) by Duncn s multiple rnge test. Cymbidium cv. Tim Hot : The effect of different irrdinces of Compct 3U on the in vitro growth nd development of Cymbidium plntlets is shown in Tble 3. Dt showed tht there ws no significnt difference in root length of plntlets plced under the different lighting systems. However, the plntlets were significntly tller t 60 µmolm -2 s -1, with considerbly more roots t 45 µmolm -2 s -1 s compred to Neon t 45 µmolm -2 s -1 (Fig. 3b). Besides, the dt indicted tht the plntlets plced under the Compct 3U lighting system hd better growth thn those under the Neon lighting system. The irrdince in this cse did not ffect the root growth but plntlet fresh weight, which ws highest t 60 µmolm -2 s -1. The most suitble irrdince of Compct 3U (60 µmolm -2 s -1 ) for growth nd morphology of Cymbidium, chrcterized by long shoots nd roots, wide leves nd high fresh weight (Fig. 3b), ws lso the most pproprite intensity Tble 4. Effect of different irrdinces of Compct 3U on the in vitro development of Lilium longiflorum plntlets fter 6 weeks of culture Irrdince (Compct 3U) (µmolm -2 s -1 ) Plnt height (cm) Root length (cm) Lef re (mm 2 ) Number of new formed roots Number of bulbs Bulb dimeter (mm) Bulb cluster fresh weight (mg) Bulb fresh weight (mg) Neon (45) 9.1c * 2.5c 26.1d 11.1b c 510c c 1.6c 5.5d 810b b 2.1d 30.7b 9.7d 1.8b 5.8c 830b 470b c 3.0b 30.0c b b * Different letters within column indicte significnt differences (P = 0.05) by Duncn s multiple rnge test

20 18 Compct 3U s novel lighting source for the propgtion of some horticulturl plnts for inducing vigorous growth of the plntlets trnsplnted to greenhouse. Tble 3. Effect of different irrdinces of Compct 3U on the in vitro development of Cymbidium cv. Tim Hot plntlets fter 6 weeks of culture Irrdince (Compct 3U) (µmolm -2 s -1 ) Plnt height (cm) Root length (cm) Lef width (mm) Number of roots Plnt fresh weight (mg) Neon (45) 10.1b w b 2.0c 620c b b b 740 3U lighting source t night is given in Tble 5. The results showed tht Cymbidium plntlets grown in the greenhouse under supplementl Compct 3U lighting source hd better development thn those derived from Neon lighting source (except for lef width) (Fig. 4 1, 4 2 ). Lilium longiflorum: Results in Tble 6 show tht the development of Lilium plntlets when trnsplnted to greenhouse ws ffected by different lighting sources nd intensities. Neon lighting sourcederived plntlets hd lower growth (plnt height nd lef width) but greter number of leves nd better root length thn those derived from the Compct 3U lighting source, which yielded vrible results under different irrdinces (Fig. 4b 1, 4b 2 ) b 2.6b 4.4c c w Different letters within column indicte significnt differences (P = 0.05) by Duncn s multiple rnge test. Lilium longiflorum:the effect of different irrdinces of Compct 3U on the in vitro growth nd development of L. longiflorum plntlets is indicted in Tble 4 nd Fig. 3c. The results suggest tht the lighting source s well s its PPFD significntly ffected the fresh weight of new bulblet clusters derived from initil bulb scles. These results show tht L. longiflorum bulb scles could be cultured t 75 µmolm -2 s -1 under Compct 3U for multiplying new high qulity bulblets nd under Neon t 45 µmolm -2 s -1 for incresing biomss nd producing vigorous bulblets before trnsplnting to greenhouse. In ddition, there ws considerble effect of Compct 3U t 45 µmolm -2 s -1 on the in vitro L. longiflorum morphology. This might be result of the increse in red light spectrum ssocited with low PPFD of Compct 3U lighting source which plyed certin role in incresing plnt height nd lef re. On the other hnd, the L. longiflorum root morphology ws not ffected by different lighting sources. Subsequent growth of Cymbidium nd Lilium plntlets Cymbidium cv. Tim Hot : The subsequent growth of Compct 3U-derived Cymbidium plntlets in the greenhouse fter one nd hlf months culture under 6h/dy supplementl Compct In summry, the irrdince of Compct 3U hd positively stimulted impct tht ffected significntly on the development of these three plnts. The Compct 3U lighting source hd positive effect on the plnt height, nd the plnt fresh weight of Lilium, Cymbidium, the number of strwberry shoots cultured in vitro s well s on the root length of strwberry nd the number of roots of Cymbidium cultured in vitro. The results obtined in this study showed tht the Compct 3U lighting source ffected the morphology of these plnts, incresed their biomss, nd enhnced plntlet growth before trnsplnting to the greenhouse. Furthermore, this dt lso showed tht different plnts dpted differently to different lighting sources, but different irrdinces from two lighting sources did not ffect root development including root fresh weight, root elongtion (except for strwberry) nd the number of roots (except for Cymbidium) of these plnts. In most cses, different Compct 3U irrdinces hd no obvious impct on plntlet development s compred to those of Neon light. Different irrdinces ffected Lilium nd Cymbidium plnt height nd Lilium plnt fresh weight. In these plnts, lower intensity gve higher plnt qulity. Except for the plnt fresh weight of Lilium tht ws enhnced when cultured under Compct 3U t 75 µmolm -2 s -1, the remining cses showed tht plntlets developed well under lower intensities (45 or 60 µmol m -2 s -1 ). Consequently, Compct 3U confirmed the positive effect Tble 5. Subsequent growth of Compct 3U-derived Cymbidium plntlets in the greenhouse fter one nd hlf months of culture under 6h/dy supplementl Compct 3U lighting source Lighting systems (µmolm -2 s -1 ) Plnt height (cm) Lef width (mm) Number of leves Number of roots Root length (cm) Plnt fresh weight (g) Neon b * c 3.7b 2.9b 1.7b Compct 3U b 5.5b 4.4c 3.6b b b c 4.6b b 1.6b * Different letters within column indicte significnt differences (P = 0.05) by Duncn s multiple rnge test. Tble 6. Subsequent growth of Compct 3U-derived Lilium plntlets in the greenhouse fter 2 months of culture under 6h/dy supplementl Compct 3U lighting source Lighting systems (µmol m -2 s -1 ) Plnt height (cm) Bulblet dimeter (cm) Lef width (mm) Number of leves Number of roots Root length (cm) Plnt fresh weight (mg) Neon d * 0.72c 4.5d c 0.7c 410c Compct 3U b c 4.0c b 0.72c 6.0b 3.7b 4.5b 1.0b c c 3.7b b 490b * Different letters within column indicte significnt differences (P = 0.05) with the men seprtion by Duncn s multiple rnge test.

21 Compct 3U s novel lighting source for the propgtion of some horticulturl plnts 19 Fig. 3. Strwberry, Cymbidium nd Lilium plntlets cultured under Compct 3U t different irrdinces. (): Strwberry cultured under Neon light t 45 µmolm -2 s -1 (top left), Compct 3U t 45 µmolm -2 s -1 (top right), 60 µmolm -2 s -1 (bottom left), or 75 µmolm -2 s -1 (bottom right) fter three weeks of culture. (b): Cymbidium plntlets cultured under Neon light t 45 µmolm -2 s -1 (top left), Compct 3U t 45 µmolm -2 s -1 (top right), 60 µmolm -2 s -1 (bottom left), or 75 µmolm -2 s -1 (bottom right) fter six weeks of culture. (c): Lilium plntlets cultured under Neon light t 45 µmolm -2 s -1 (top left), Compct 3U t 45 µmolm -2 s -1 (top right), 60 µmolm -2 s -1 (bottom left), or 75 µmolm -2 s -1 (bottom right) fter six weeks of culture. Fig. 4. Cymbidium, Lilium plnts fter trnsplnted in greenhouse. ( 1 ): Cymbidium plnts trnsplnted in greenhouse fter one nd hlf months. Left to right: Cymbidium plnts derived from Neon light t 45 µmolm -2 s -1, Compct 3U t 45, 60 or 75 µmolm -2 s -1. ( 2 ): Cymbidium plnts in spongy try fter one nd hlf months trnsplnted in greenhouse. Left to right: Cymbidium plnts derived from Neon light t 45 µmolm -2 s -1, Compct 3U t 45, 60 or 75 µmolm -2 s -1. (b 1 ): Lilium plnts derived from Neon light t 45 µmolm -2 s -1 (bottom left), Compct 3U t 45 µmolm -2 s -1 (bottom right), 60 µmolm -2 s -1 (top left), or 75 µmolm -2 s -1 (top right) fter two months in greenhouse. (b 2 ): Lilium plnts in spongy try fter two months trnsplnted in greenhouse. Left to right: Lilium plnts derived from Neon light t 45 µmolm -2 s -1, Compct 3U t 45, 60 or 75 µmolm -2 s -1. on the in vitro development of plntlets before trnsplnting to greenhouse. These bove results were similr to those of Wrrington nd Michell (1976), Morgn nd Smith (1981), Smith (1982), Tibbitts et l. (1983), Mortensen nd Stromme (1987), Economou nd Red (1987), nd Agrwl (1992), who ll confirmed the significnt effects of light qulity (relted to different lighting sources) on the growth nd morphology of in nd ex vitro plnts. The effect of irrdince of different lighting sources on the development of plnts were lso the concern of some studies of Gilslerod nd Mortensen (1997), Miyshit et l. (1997), nd Nhut (2002). In these studies, higher irrdinces gve the best plnt growth. But in this report, we suggest for use of lower intensities (45 or 60 µmolm -2 s -1 ) of Compct 3U for plnt propgtion owing to the reduction of electricl energy consumption s well s the increse in the development of some horticulturl plnts. The Compct 3U lighting source hd highly significnt effect on the development of Cymbidium Tim Hot, Lilium longiflorum nd Frgri vesc cv. My D s in this study. The results indicte tht these plnts dpted differently with different light sources nd intensities. Strwberry shoots hd better growth nd morphology when cultured under Compct 3U lighting system thn those under Neon light system. The preeminence of Compct 3U lighting sytem ws lso expressed when shoots of three plnts were cultured under different irrdinces of two lighting system. Though in most cses, different irrdinces of Compct 3U hd no obvious effect on the development of plntlets s compred to the Neon light, lower irrdince gve higher Cymbidium nd Lilium plntlets qulity. Moreover, plntlets derived from Compct 3U lighting system developed better thn those from Neon lighting system. Hence, the Compct 3U lmp, hving suitble light spectrum, resulting in good, high qulity plnts, sving 75-80% of electricl energy consumption s compred to incndescent lmps, being chep, nd subsequently retrieving initil investments quickly, ws expected to be novel lighting system used in the successful micropropgtion nd subsequent ex vitro growth of some horticulturl plnts. In fct, becuse of obvious dvntges, the Compct 3U lmp hs been used s the lighting source for propgtion of mny horticulture plnts in Dlt, Lm Dong, Vietnm. References Agrwl, S.B Effects of supplementl UV-B rdition on photosynthetic pigment, protein nd glutthione contents in green lge. Env. Exp. Bot., 32(2): Collin, R.N., E.N. Mrgret, H. Ted nd P. Dvid, Light qulity nd light pipes in the micropropgtion of woody ornmentl plnts. Act Hortic., 226: Debergh, P.C. nd R. Zimmermn, Micropropgtion: technology nd ppliction. Kluwer Acdemic Publishers, Dordrecht, pp. 484.

22 20 Compct 3U s novel lighting source for the propgtion of some horticulturl plnts Duncn, D.B Multiple rnge nd multiple F test. Biomet., 11: Economou, A.S. nd P.E. Red, Light tretments to improve efficiency of in vitro propgtion systems. HortSci., 22(5): Fust, J.E. nd R.D. Heins, Quntifying the influence of hightpressure dodium lighting on shoot-tip temperture. Act Hortic., 418: Gbrkiewicz, B., E. Gbryszewsk, R. Rudnicki nd D. Goszcynsk, Effects of light qulity on in vitro growing of Dieffenbchi cv. compct. Act Hortic., 418: Gbryszewsk, E. nd R.M. Rudnicki, The effects of light qulity on the growth nd development of shoots nd roots of Ficus benjmin in vitro. Act Hortic., 418: Gilslerod, H.R. nd L.M. Mortensen, Effect of light intensity on growth nd qulity of cut roses. Act Hortic., 418: Hrt, J.W Light nd Plnt Growth. In: Topics in Plnt Physiololgy, 1: Hyshi, M., N. Fujit, Y. Kity nd T. Kozi, Effect of sidewrd lighting on the growth of potto plntlets in vitro. Act Hortic., 319: Iwnmi, Y., T. Kozi, Y. Kity nd S. Kino, Effects of supplementl red nd fr-red lighting using light emitting diodes on stem elongtion nd growth of potto plntlets in vitro (Abstr.). Interntionl Symposium Trnsplnt production Systems, Yokohm, Jpn, pp Kim, H.H. nd T. Kozi, Production of vlue-dded trnsplnts in closed systems with rtificil lighting. In: Trnsplnt Production in the 21st Century. C. Kubot nd C. Chun (eds.), Kluwer Acdemic Publishers, Kirdmnee, C., Y. Kity nd T. Kozi, Effect of supplementl fr-red lighting nd photosynthetic photon flux density on stem elongtion nd dry weight increse of Euclyptus cmldulensis in vitro plntlets (Abstr.). XVth Interntionl Botnicl Congress Yokohm, Jpn, pp Kozi, T., K. Fujiwr, M. Hyshi nd J. Aitken-Christie, The in vitro environment nd its control in micropropgtion. Trnsplnt production systems. Kluwer Acdemic Publishers. Dordrech, pp Kunnemn, B.P.A.M. nd J.B. Ruesink, Interctions between light, temperture nd CO 2 in rooting of conifer cuttings. Act Hortic., 418: Ms, F.M. nd E.J. Bkx, Growth nd flower development in roses s ffected by light. Act Hortic., 418: Miyshit, Y Growth nd morphology of potto plntlets in vitro s ffected by light qulity, nd size nd nodl position of the explnt. Grdute School of Science nd Technology. Chib University, pp Miyshit, Y., T. Kimur, Y. Kity, C. Kubot nd T. Kozi, Effects of red light on the growth nd morphology of potto plntlets in vitro: Using light emitting diodes (LEDs) s lighting source for micropropgtion. Act Hortic., 418: Moe, R Physiologicl spects of supplementry lighting in horticulture. Act Hortic., 418: Morgn, D.C. nd H. Smith, Non-photosynthetic responses to light qulity. In: Encyclopedi of Plnt Physiology. Springer-Verlg, Berlin, pp Mortensen, L. M. nd E. Stromme, Effects of light qulity on some greenhouse crops. Sci. Hort., 33: Murkmi, K., I. Aig, K. Horguchi nd M. Morit, Red/frred photon flux rtio used s n index number for morphologicl control of plnt growth under rtificil lighting conditions. Act Hortic., 418: Murshige, T. nd F. Skoog, A revised medium for rpid growth nd biossys with tobcco tissue culture. Plnt Physiol., 15: Nhut, D.T In vitro growth nd physiologicl spects of some horticulturl plntlets cultured under red nd blue light emitting diodes (LEDs). PhD Thesis, Kgw University, pp Schwbe, W.W Morphogenetic responses to climte. In: Environmentl Control of Plnt Growth. Acdemic Press, New York, pp Sebrook, J.E.A Chnging the growth nd morphology of potto plntlets in vitro by vrying the illumintion source. Act Hortic., 212: Smith, H Light qulity, photoperception nd plnt strtegy. Annul Rev. Plnt Physiol., 33: Tibbitts, T.W., D.C. Morgn nd I.J. Wrrington, Growth of lettuce, spinch, mustrd, nd whet plnts under four combintions of high-pressure sodium, metl hlide, nd tungsten hlogen lmps t equl PPFD. J. Amer. Soc. Hort. Sci., 108(4): Wlz, F. nd W. Horn, The influence of light qulity on gs exchnge of dendrnthem. Act Hortic., 418: Wrrington, I.J. nd K.J. Michell, The influence of blue- nd red-bised light spectr on the growth nd development of plnts. Agr. Met., 16: Wulster, G. nd H.W. Jnes, Effects of supplementl light qulity, quntity, nd differentil temperture on growth nd development of Ester Lilium (Lilium longiflorum). Act Hortic., 418:

23 Journl of Applied Horticulture, 8(1): 21-24, Jnury-June, 2006 Effect of slow relese fertiliser on the growth of continerised flnnel flower (Actinotus helinthi Lbill.) Appl Journl Lotte von Richter nd Ctherine A. Offord Botnic Grdens Trust, Sydney, Mount Annn Botnic Grden, Mount Annn Drive, Mount Annn, NSW 2567, Austrli. E-mil: Abstrct Two controlled-relese fertiliser (CRF) formultions, Nutricote Totl 13N : 5.7P : 10.8K (N13) nd Nutricote Totl 18N : 2.6P : 6.6K (N18), were pplied t 0, 1.25, 2, 2.5, 5 nd 10 kg m - ³, to flnnel flower (Actinotus helinthi Lbill.) seedlings grown in soil-less potting mix in continers. After five months, during pek spring flowering, number of chrcters relting to the qulity of the cut flower product of this species were ssessed. As the rte of fertiliser ppliction incresed, the plnt height, totl number of stems, number of flowering stems nd number of flowers nd buds incresed. There were significntly more stems nd flowers overll, nd more flowering (sleble) stems, in the N18 tretments t ll ppliction rtes. Plnt height ws not ffected by fertiliser formultion. Bsl folir necrosis, which scored highly in the control tretment (0 fertiliser), ws reduced by fertiliser ppliction. Key words: Nutrition, controlled-relese fertiliser, nitrogen, Actinotus helinthi, flnnel flower, cut flower Introduction The flnnel flower (Actinotus helinthi Lbill.) is n erect nnul or perennil herb tht is covered with woolly indumentum (Powell, 1992) giving the plnt nd prticulrly the inflorescence its chrcteristic flnnel ppernce. It occurs nturlly in estern Austrli in New South Wles nd southern Queenslnd. This species is generlly found on sndy nd rocky soils long the cost nd lso in smll sndy ptches in the western extent of its distribution. Belonging to the Apicee fmily, with umbels subtended by lrge involucrl brcts giving n inflorescence reminiscent of disies, the long-stemmed selections re considered useful cut flower feture-filler product. Ten yers go, the species ws primrily bush hrvested for the cut flower mrket, but in recent yers it hs been cultivted nd export sles re stedily incresing (Worrll et l., 2004). Originlly there were severl significnt limittions to production of flnnel flower, including low seed germintion nd vegettive propgtion rtes. These problems hve lrgely been overcome (Offord nd Tyler, 1996; von Richter nd Offord, 1997, 2000) nd ttention hs recently turned to improving knowledge bout cultivtion of this species including nutrition, substrte requirements nd disese interctions. Little is known bout the nutritionl requirements of flnnel flowers or mny other Austrlin species tht occur on low nutrient soils (Brennn et l., 1998). This pper exmines the significnce of the effect of two controlled-relese fertiliser (CRF) formultions, t incresing ppliction rtes, on severl reported nd unreported growth chrcteristics of continerised flnnel flowers (von Richter nd Offord, 1997). Nutricote Totl ws used for this experiment s it ws redily vilble in our nursery t tht time; however, severl other CRF products commercilly vilble would hve been eqully suitble for this work. CRFs re commonly used source of nitrogen nd other mjor s well s minor nutrients becuse they relese the nutrients more evenly thn conventionl soluble fertilisers reducing problems ssocited with burning or leching, nd without the more lbour intensive liquid fertiliser pplictions tht lso deliver good plnt growth (Cresswell nd Weir, 1997; Oliet et l., 2004). Mterils nd methods Plnt mteril: Actinotus helinthi seeds were collected in spring (November) from Te Grdens on the NSW Centrl Cost (Ltitude, S Longitude 152.( E All work on seeds, seedlings nd plnts ws crried out t Mount Annn Botnic Grden, S 04 34º04 ).( E º46 Potting mix, fertiliser formultion nd rte: Seeds were sown soon fter hrvest onto seed rising mix (snd /perlite 1:1 v/v) nd the seedlings pricked out t the two lef stge nd plnted into 50 mm tubes contining snd/coir mix (4:1 v/v) nd 0.5 g L -1 FeSO 4 nd 0.5 g L -1 lime. When the seedlings were 80 mm high, 220 seedlings were plnted into the snd/coir mix in 140 mm slimline blck plstic pots, but with vrying ppliction rtes of either Nutricote Totl 13N : 5.7P : 10.8K (N13) or Nutricote Totl 18N : 2.6P : 6.6K (N18). The relese time for ech fertiliser is 270 dys t 25ºC. The following rtes were pplied: 0, 1.25, 2, 2.5, 5 nd 10 kg m - ³. The rte recommended by the mnufcturer is 2 kg m - ³. Experimentl design: There were 20 replictes of ech tretment. The plnts were rrnged in completely rndom design on rised benches in full sun. Wtering ws by hnd s nd when required. Assessment: At pek flowering time (mid-november), when the plnts hd been in the different fertiliser tretments for five months, the following mesurements were mde: plnt height,

24 22 Effect of slow relese fertiliser on the growth of continerised flnnel flower (Actinotus helinthi Lbill.) totl stem number, flowering stem number nd totl number of flowers nd buds. Folir lef necrosis ws scored (1 = bsl leves green; 2 = one bsl lef yellow; 3 = two bsl leves yellow or brown; 4 = ll bsl leves brown). Sttisticl nlysis: Min effects nd interctions were nlysed by ANOVA nd differences between the mens compred using LSD (P=0.05). Reponses to the two fertiliser formultions to ppliction rtes were nlysed by liner regression. All nlyses were performed using SYSTAT 11 (SPSS Inc. 2004). Results Plnt height: Plnt height ws lrgely unffected by the fertiliser formultion, but the rte effect ws significnt (Tble 1), probbly minly due to the much lower zero fertiliser control tretment (Fig. 1A). The regression slopes in Fig. 1A were highly significnt for N13 ( P = 0.001) nd N18 (P = 0.001), indicting tht the slopes were not equl to zero nd tht fertiliser ppliction rte hs some effect on plnt height. However, the proportions of the totl vrince explined by the ppliction rtes were very low (R 2 = 0.12 nd 0.13) (Fig. 1A). Stem number: Overll, the number of stems produced ws greter in the N18 tretment, when compred to the N13 tretments (Fig. 1B). The regression slopes (Fig. 1B) were highly significnt for N13(P <0.001) nd N18 (P < 0.001), which indictes tht the slopes were not equl to zero nd tht there is reltionship between fertiliser ppliction rte nd the stem number produced. Mximum stem numbers were found t N18 t 5 nd 10 kg m - ³ (verges of 6.5 nd 8.5 stems); the next highest stem number ws t N13 t 10 kg m - ³ (verge of 6 stems). Fertiliser types nd rtes of ppliction were significnt min effects for this vrible, nd there ws lso significnt interction effect detected (Tble 1). At the time of mesurement, fewer thn hlf of the stems hd produced flowers, with the lest flowering stems produced t zero fertiliser ppliction (Fig. 1C). The highest number of flowering stems ws produced t 10 kg m - ³ N18 which represented 35% of the totl number of stems (Fig. 1B nd C). There ws little difference between the formultions in the number of flowering stems produced t 2.5 nd 5 kg m - ³, but there were smll significnt differences between the formultions t 1.25 nd 2 kg m - ³ (Fig. 1C). This ccounts for the significnt nd highly significnt min effect responses detected for fertiliser formultion nd rte respectively (Tble 1). Agin, the regression slopes for this vrible were highly significnt for N13 (P <0.001) nd N18 (P < 0.001), Fig. 1C), which indictes tht the slopes were not equl to zero nd tht there is reltionship between fertiliser ppliction rte nd the flowering stem numbers, but the direction of the regression slopes were less steep (R 2 = 0.2%) (Fig. 1C), thn when compred to the totl number of stems (R 2 = 0.4 nd 0.6%) (Fig. 1B). Number of flowers nd buds: The number of flowers nd buds produced ws gretly ffected by both fertiliser type nd rte, lthough there ws no interction observed between the two (Tble 1). The regression slopes were highly significnt for N13 (P = 0.001) nd N18 (P = 0.001), which indictes tht the slopes were not equl to zero nd tht there is reltionship between fertiliser ppliction rte nd flower/bud numbers. Mximum flower numbers were observed t the 10 kg m - ³ level of both fertilisers (verge of 28 nd 22 flowers), with the slope of the response being similr (R 2 = 0.49 nd 0.5) for both formultions over the rnge of pplictions tested (Fig. 1D). Bsl folir necrosis: Bsl folir necrosis ws significntly ffected by fertiliser formultion nd rte (Tble 1), with significntly less necrosis observed for N18, when compred to N13 t the 2 nd 2.5 kg m - ³ ppliction rtes (Fig. 1 E), which my ccount for the significnt interction term for fertiliser x rte. The regression slopes for folir necrosis were highly significnt for N13 (P = 0.001) nd N18 (P = 0.001), indicting reltionship between fertiliser ppliction rte nd the degree of necrosis observed, despite the wek R 2 vlues. Overll, fertiliser ppliction, especilly N18 greter thn 2 kg m - ³ nd N13 t 5 nd 10 kg m - ³, significntly improved (decresed) the necrosis rting when compred with the control (zero fertiliser). Discussion In terms of the qulity of the flowering stems produced, flnnel flowers ppered to respond positively to ll the imposed fertiliser tretments in this study when compred with the control tretment (zero fertiliser ppliction), with no dverse ffects observed t ny pplied level. The recommended rte of 2 kg m - ³ ws exceeded up to five times (to 10 kg m - ³) nd the plnts produced more stems nd more flowers s the ppliction rtes incresed. This high tolernce of pplied nutrients is in contrst to some other Austrlin nd South Africn ntive species which require little fertiliser to chieve optiml results (Brennn et l., 1998; Clrk nd Burge, 1999),while others responded in similr fshion to A. helinthi (Lmont et l.,1990; Bennell nd Willims, 1992; Brennn et l., 2000). It hs commonly been observed tht flnnel flowers re found in bundnce fter bushfire, t which time there my be incresed nutrient levels re often vilble depending on the nutrient sttus of the plnt mteril tht hs been burnt (Ashton nd Mrtin, 1996; Enright et l., 1997). Further work is required to determine the ecologicl significnce of this response. Reltive differences between the two fertiliser tretments re of gretest horticulturl interest. The plnts responded to the higher nitrogen formultion by producing more sleble product i.e. more stems nd flowers. Nutricote N13 is recommended for potted flowering plnts becuse of its blnce of lower nitrogen when compred to phosphorus nd potssium, while the N18 formultion is recommended by the mnufcturer (Ytes Pty Ltd) Tble 1. Significnce (P vlue) of min effects (fertiliser type nd rte) nd interctions for chrcteristics of flnnel flower plnts five months fter fertiliser ppliction Height of plnt Number of stems Stems with flowers Flower number Bsl folir necrosis Fertiliser type Rte Fertiliser x Rte

25 Effect of slow relese fertiliser on the growth of continerised flnnel flower (Actinotus helinthi Lbill.) 23 Plnt height (mm) Number of flowering stems Necrosis rting R 2 = R 2 = Nutricote N18 Nutricote N Fertilizer ppliction rte (kg m ) R 2 = R 2 = Nutricote N18 Nutricote N Fertilizer ppliction rte (kg m ) R 2 = E C for folige plnts becuse of its higher nitrogen rtio. Certinly, greter flowering stem production is desired im in cut flower production in this species, nd it would pper tht the high nitrogen formultion chieved the best result in this respect. Although the overll number of flowering stems ws gretest t high nitrogen, the number of flowers produced per flowering stem (derived from Figs 1 C nd D) ws very similr for the two fertiliser tretments t ech rte. For exmple, t 2 kg m - ³ the N18 tretment hd n verge of 8.0 flowers per flowering stem nd N13 hd 7.1; t 10 kg m - ³ N18 hd 9.4 nd N13 hd A R 2 = Nutricote N18 Nutricote N Fertilizer ppliction rte (kg m ) Totl number of stems Number of flowers nd buds B R 2 = R 2 = Nutricote N18 Nutricote N Fertilizer ppliction rte (kg m ) D R 2 = R 2 = Nutricote N18 Nutricote N Fertilizer ppliction rte (kg m ) Fig. 1. Responses of continerised flnnel flower (Actinotus helinthi) to pplied slow relese fertilisers, Nutricote Totl 13N: 5.7P: 10.8K nd Nutricote Totl 18N: 2.6P: 6.6K, t different ppliction rtes (0, 1.25, 2, 2.5, 5 nd 10 kg m - ³). A. plnt height; B. totl number of stems; C. number of flowering stems; D. number of flowers nd buds per plnt; E. necrosis rting (1 = bsl leves green; 2 = one bsl lef yellow; 3 = two bsl leves yellow or brown; 4 = ll bsl leves brown). Regression slopes were ll highly significnt from zero t P < level. The br represents the LSD of the mens t P = Therefore, it ppers tht the higher nitrogen tretment did not suppress flower number per stem. Stem height ws not gretly ffected by the fertiliser formultion or ppliction rtes greter thn zero, which is likely to reflect tht plnt height in this species is primrily determined geneticlly, rther thn environmentlly. In wild stnds, we hve observed very tll plnts, with few stems, reflecting the low nutrient sttus of the soil in most of these res nd the dpttion of the species to vrible nutrient conditions. Plnt height is mjor selection criterion for Flnnel flower development, nd we hve documented the differences between popultions, which re mintined by the plnts in cultivtion (Offord nd Tyler, 1996). For exmple, short hedlnd-growing vrieties re being developed for the pot plnt mrket, while inlnd forest forms re useful for cut flower production becuse of their nturlly long stems. There re mny forms of Actinotus helinthi nd some of the tll growing cut flower types show bsl lef necrosis much sooner thn short pot plnt types. Bsed on evidence from tissue cultured

26 24 Effect of slow relese fertiliser on the growth of continerised flnnel flower (Actinotus helinthi Lbill.) plnts (von Richter, unpublished) there re differences in nutrient requirements for these different forms. Considering the xenic nture of tissue culture, the necrosis is likely to be cused by the movement of nutrients, prticulrly nitrogen wy from the bsl leves to be reutilised in the picl region rther thn the effects of pthogenic infection. The fster growing vrieties re likely to hve higher nutritionl requirement thn smller, slower growing ones. The optiml fertiliser regime could not be determined by this study, but it would be expected to not exceed the highest ppliction rte nd my be less, especilly in terms of the cost of ppliction (Obrez et l., 1999) or leching into the soil/wter environment, minly in the form of nitrte (Cox, 1993; Huett, 1997). The results of this study re bsed on seedlings from one popultion only. The recommendtions for fertiliser use found here is only n indictor of the nutrient usge of one flnnel flower type nd t one point in time. The temporl effects of fertiliser formultions nd rtes to enble growers to optimise their usge of CRFs lone, or in combintion with wter soluble fertilisers depends on the vrieties being produced nd the timing of production required in nursery situtions (Jcobs et l., 2005) or in the field (Brennn et l., 1998). Acknowledgements We grtefully cknowledge the finncil support of the Rurl Industries Reserch nd Development Corportion (RIRDC). References Ashton, D.H. nd D.G. Mrtin, Regenertion in pole-stge forest of Euclyptus regnns subjected to different fire intensities in Austrlin Journl of Botny, 44: Bennell, M.R. nd R.R. Willims, Cultivtion of the pink mull mull Ptilotus exlttus Nees. 2. Nutrition nd growth regultion. Scienti Horticulture, 51: Brennn, R.F., A.M. Crowhurst nd M.G. Webb, The growth of Yellow Bells (Geleznowi verrucos) seedlings in response to dditions of nitrogen, potssium nd phosphorus fertiliser. Austrlin Journl of Experimentl Agriculture, 38: Brennn, R.F., M.G. Webb nd A.M. Crowhurst, Yield responses of mull mull (Ptilotus exlttus Nees.) seedlings to dditions of nitrogen, potssium nd phosphorus fertiliser. Austrlin Journl of Experimentl Agriculture, 40: Clrk, G.E. nd G.K. Burge, Effects of nitrogen nutrition on Sndersoni cut flower nd tuber production in soil-less medium. New Zelnd Journl of Crop nd Horticulturl Science, 27: Cox, D.A Reducing nitrogen leching-losses from continerized plnts: the effectiveness of controlled-relese fertilizers. Journl of Plnt Nutrition, 16: Cresswell, G.C. nd R.G. Weir, Plnt nutrient disorders 5, Ornmentl Plnts nd Shrubs. Inkt Press, Melbourne. Enright, N.J., D. Goldblum, P. At nd D.H. Ashton, The independent effects of het, smoke nd sh on emergence of seedlings from the soil seed bnk of helthy Euclyptus woodlnd in Grmpins (Griwerd) Ntionl Prk, western Victori. Austrlin Journl of Ecology, 22: Huett, D.O Fertiliser use efficiency by continerised nursery plnts. 2. Nutrient leching. Austrlin Journl of Agriculturl Reserch, 48: Jcobs, D.F., D.L. Hsse nd R. Rose, Growth nd folir nutrition of Dougls-fir seedlings provided with supplementl polymer-coted fertilizer. Western Journl of Applied Forestry, 20(1): Lmont, G.P., G.C. Cresswell nd G.J. Griffith, Nutritionl studies of Christms Bell. HortScience, 25: Obrez, T.A., R.E. Rouse nd J.B. Sherrod, Economics of controlled-relese fertilizer use on young citrus trees. Journl of Production Agriculture, 12: Offord, C.A. nd J.L. Tyler, Flnnel Flowers. In: Horticulture of Austrlin Plnts. Burchett, M. nd Johnston K. (Eds), UNSW Press, Sydney, pp Oliet, J., R. Plnelles, M.L. Segur, F. Artero nd D.F. Jcobs, Minerl nutrition nd growth of continerized Pinus hlepensis seedlings under controlled-relese fertilizer. Scienti Horticulture, 103: Powell, J.M Apicee. In: Flor of NSW. G.J. Hrden (Ed). Vol. 3. New South Wles University Press, Sydney, pp von Richter, L. nd C. Offord, Flnnel flower (Actinotus helinthi) s cut flower crop. RBG 1A. Rurl Industries Reserch nd Development Corportion, Cnberr, 66 pp. von Richter, L. nd C.A. Offord, Development of Flnnel flower production system. Rurl Industries Reserch nd Development Corportion, Cnberr, Publiction NO. 00/106, 54pp. Worrll, R., C. Offord nd L. von Richter, Flnnel flower. In: The New Crop Industries Hndbook. Slvin, S., Bourke, M. nd Byrne, T. (Eds), Rurl Industries Reserch nd Development Corportion, Cnberr, pp

27 Journl of Applied Horticulture, 8(1): 25-28, Jnury-June, 2006 Growth nd flowering response of snpdrgons fter relese from picl dominnce Appl Journl Muhmmd Munir 1 nd Frht Nz 2 1 School of Plnt Sciences, The University of Reding, Reding, UK. 2 Fculty of Agriculture, Goml University, D.I. Khn. Pkistn. Abstrct Plnts of n erly flowering Antirrhinum cultivr Chimes White were pinched t 4, 5, 6, nd 7 lef-pir stge to observe the effects on flowering time nd plnt qulity. Though control plnts flowered erlier (81 dys) thn the pinched ones, they produced less number of flower buds. Flower time nd rte of progress to flowering in pinched plnts incresed linerly nd significntly. The qulity of pinched plnts regrding brnch numbers, lef re, plnt height, plnt fresh weight etc. ws significntly improved in ll tretments. Mny plnt growth prmeters were successfully fitted by the second degree polynomil model wheres liner model indicted good fit in reproductive development. Key words: Antirrhinum mjus, snpdrgon, picl dominnce, growth, flowering. Introduction Snpdrgons, prt from been grown s bedding plnts nd s cut flower crop, cn lso be grown s pot plnts, s they hve high flower bud to vegettive growth rtio, mture quickly nd rnge of brightly coloured cultivrs re vilble. Disdvntges when grown s pot plnts re tht they my grow too tll nd so re not self-supporting nd lrge centrl flowering inflorescence initilly domintes the plnt (Winwright nd Irwin, 1987). One possible method of overcoming these problems nd producing well-proportioned flowering pot plnt could be to prctice pinching technique. Pinching is the removl of the picl bud to relese the lower xillry buds from picl dominnce in order to increse brnching nd stimulte xillry bud development. Apicl dominnce hs been linked to uxins produced in the picl bud, which my indirectly inhibit lterl bud growth. Gocl et l. (1991) mesured the concentrtion of IAA in xillry buds fter pinching nd found tht IAA concentrtions, firstly, incresed nd then remined constnt. This ws ccompnied by n increse in the fresh weight of the buds. However, uxins re not the only hormones tht control xillry bud growth. Cytokinin content hs lso been shown to increse fter pinching (Mder et l., 2003; Bngreth, 1994; Turnbull et l., 1997). Exogenously pplied cytokinin ws found to hve the sme effect s pinching (Bngreth, 1994; Chen et l., 1997). Therefore, there my be n ntgonistic reltionship between uxin nd cytokinin in the control of picl dominnce. Relese of picl dominnce by pinching hs been shown to hve significnt effects on yield in mny crops. In rooted cuttings of chrysnthemum, pinching is used to produce mrketble spry shpe. The time pinch must be crried out t prticulr time in order to llow flower bud to initite in the centre of the spry. This will develop into n rrngement of inflorescences on peduncles of desired length (Kofrnek, 1981). Post (1949) suggested tht xillry meristems of chrysnthemum cn show period of insensitivity to photoperiod when relesed from picl dominnce, but contrdictory result ws reported by Adms et l. (1998), who found tht chrysnthemum plnts were cpble of responding to short dys immeditely following pinching. This method of removing picl dominnce is lso found to dely flowering but did not ffect plnt height or plnt width (Strmn nd Fust, 1999). In winter cultivrs of Antirrhinum (Coronette Yellow nd Coronette Scrlet), pinching produces plnts with significntly reduced plnt height, shorter but incresed number of flower spikes nd lengthened flowering time. The pinching stge does not seem to influence plnt height, but pinching crried out t lter stge of plnt development reduced the spike length nd prolonged flowering time. Pinching t the third pir of leves from the bse of plnt produced mximum flower spikes in the shortest time. These effects of pinching re result of more even distribution of ssimiltes between severl growing points rther thn just one. The dely in flowering is result pushing the plnt bck to the juvenile phse. In this wy, pinching ppers to mintin the juvenile phse for longer nd the precise cuse of the flowering dely, which my be ttributed to the xillry shoots being in less dvnced physiologicl phse thn picl shoot (Winwright nd Irwin, 1987). The stge t which the plnts re pinched, though, ws found to hve significnt effects on the time of flowering nd the length of spike, the following experiment ws however undertken to evlute the effect of pinching t different stges of plnt development (4, 5 nd 6 lef-pir stge) on the flowering nd plnt growth of Antirrhinum. Mterils nd methods Seeds of Antirrhinum mjus L. cultivr Chimes White were obtined from Colegrve Seeds Ltd., Bnbury, U.K., nd were sown on 2 nd Februry 2000 into module trys (P135, volume of ech cell, 20ml; Plntpk Ltd., Mldon, U.K.) contining pet-bsed modulr compost (SHL, Willim Sinclir

28 26 Growth nd flowering response of snpdrgons fter relese from picl dominnce Horticulture Ltd., Lincoln, U.K.). Seed trys were wtered nd held for germintion t 20 ± 1 C in growth room providing photosynthetic photon flux density of 72 µmol m -2 s -1 t pprox. one meter bove try height from mixture of white fluorescent nd tungsten bulbs (6.3% tungsten by nominl wttge), with 16 h d -1 photoperiod. After 70% seed germintion, plnts were trnsplnted into 9 cm pots (volume 370ml) contining mixture of pet-bsed compost (SHL) nd perlite (3:1 v/v) nd kept in glsshouse (7.3m x 11.3m). Ten plnts in ech tretment were pinched when they produced 4, 5, 6, nd 7 pir of leves. Sme number of plnts were left without pinched s control. In the glsshouse, ll plnts were eqully spced to void the light competition mong them nd were subjected to 20 C constnt temperture (19.8 C ctul temperture) until flowering. The set point temperture ws mintined with ventiltion nd wter pipe heting system bove 3 C. Temperture ws recorded inside the glsshouse comprtments using sensor situted in n spirted Dysto flowering No. of flower buds Lef number SED Con trol 4pir s SED Con trol 4pir s SED 5pir s 5pir s 6pir s 6pir s A R 2 = pi rs C R 2 = pir s Control 4pirs 5pirs 6pirs 7pirs Pinching tretments E Rteof progress to flowering Brnches per plnt Fig. 1. Effect of different pinching tretments on (A) dys to flowering, (B) rte of progress to flowering (1/f), (C) Number of flower buds, (D) No. of brnches per plnt, (E) lef numbers, nd (F) lef re (cm2). Verticl brs (where lrger thn the points on lines) represent the stndrd error (SE) of vribility within replictes, wheres the seprte ones represent the stndrd error of difference (SED) between mens Cont rol SED Cont rol 4pir s 4pir s SED 5pir s 5pir s screen ttched to dt-logger. In the temperture-controlled comprtment, PT100 4 wire pltinum resistnce sensor ws connected to dt-logger (Dttker 500, Dt Electronics, Letchworth Grden City, U.K.) recording temperture t every 15s. Tube solrimeters were used to mesure the verge light trnsmission into the glsshouse nd pproximtely 6.88MJ. m -2.d -1 light integrl from emergence to flowering were received by the plnts during this experiment. Plnts were irrigted by hnd to void Pythium ttck nd nutrient solution (Sngrl 111, Willim Sinclir Horticulture Ltd, Lincoln, U.K.) ws pplied twice week with the irrigtion t conductivity of 1500 µs cm 2 (182 ppm N; 78 ppm P; 150 ppm K), nd 5.8 ph. Plnts in ech tretment were dily observed until first flower opening (coroll fully opened). Flowering nd vegettive prmeters were recorded t hrvest. Dt were nlysed by using regression nd nlysis of vrince technique of GENSTAT-5, Relese 4.1 (Lwes Agriculturl Trust, Rothmsted Experimentl Sttion, U.K.). 6pir s 6pi rs B R 2 = pir s D R 2 = pir s R 2 = 0.98 Control 4pirs 5pirs 6pirs 7pirs Pinching tretments Lef re (cm 2 ) SED F Results Directly Mesured Prmeters Flowering prmeters: Fig. 1A show tht time to flowering incresed linerly when pinched lef-pir numbers were incresed (P<0.05). An erly flower nthesis (81 dys) ws recorded in control plnts followed by 91 dys in 4 lef-pir. However, plnts pinched t 7 lef-pir stge took mximum time to flower (126 dys). Similrly, the rte of progress to flowering (1/f) ws the inverse function to the pinching tretments i.e. when plnts were pinched lte, rte of progress to flowering decresed (Fig. 1B). Similr to time of flowering, liner reltionship between pinching tretments nd number of flower buds ws observed (P<0.05). Fig. 1C reveled tht minimum flower buds (26) were produced by the control plnts. Among pinching tretments, 33 flower buds were noted in 4 lef-pir tretment whilst 58 flower buds were produced in 7 lefpir pinching tretment. Plnt qulity prmeters: A liner increse in number of brnches ws observed between control nd 7 lef-pir pinching tretments. Minimum brnches per plnt (77) were produced by control plnts followed by 84 in 4 lef-pir tretment. Brnch numbers incresed significntly (P<0.05) in ll subsequent tretments nd up to 119 brnches were produced in 7 lef-pir tretment (Fig. 1D). Lef numbers below inflorescence did not chnge following pinching fter pinching the plnts mintined the sme lef number present t the time of pinching. However, mximum lef number (20) ws produced in control (Fig. 1E). Control plnts nd those pinched t 4 lef-pir stge produced minimum lef re (79 cm 2 ), which ws significntly (P<0.05) different thn the other tretments (Fig. 1F). However, it ws observed mximum in plnts

29 pinched t 7 lef-pir stge (90 cm 2 ). As control plnts were not pinched therefore they continued their picl growth nd ttined mximum height (21 cm) s compred to the pinched plnts (12-17 cm), which were forced to hve limited size (Fig. 2A). A curviliner response ws observed in plnt fresh (Fig. 2B) nd dry weight (Fig. 2C). Plnts pinched t 7 lef-pir stge produced mximum plnt fresh (34.12 g) nd dry (5.25 g) weights. This trend significntly (P<0.05) declined in ll preceding tretments including control plnts, which produced minimum fresh nd dry weights (15.12 nd 3.37 g, respectively). Derived Prmeters A grdul but significnt (P<0.05) increse in lef re rtio (LAR) ws seen between control nd 7 lef-pir pinched plnts (Fig. 2D). Un-pinched plnts (control) hve minimum LAR ( cm 2 g -1 ), which grdully incresed in ll subsequent tretments. However, mximum LAR ( cm 2 g -1 ) ws estimted in plnt pinched t 7 lef-pir stge. A firly similr trend (Fig. 2E) ws observed in reltive growth rte (RGR) where minimum RGR ws observed in control plnts (15.12 d -1 ) wheres it ws mximum (34.12 d ) in plnts received 7 lef-pir pinching tretment. 20 Net ssimiltion rte (NAR) showed n opposite 18 curviliner response to those in LAR nd RGR prmeters (Fig. 2F). As plnts in control 16 tretment took minimum time to flower, they hd mximum NAR (0.07 g cm -1 d -1 ) wheres 14 it ws estimted minimum (0.03 g cm -1 d -1 ) in 12 plnts pinched t 7 lef-pir stge nd they took more time to flower. 10 Discussion Growth nd flowering response of snpdrgons fter relese from picl dominnce 27 Flowering ws generlly delyed by pinching, s ll of the pinched tretments produced their first flower lter thn the control plnts (Fig. 1A). This dely in flowering seems to be result of pushing the plnts bck to the juvenile phse fter pinching. The lter the pinching occurred, the lter the plnts flowered. The precise cuse of flowering dely my be ttributed to the xillry shoots being in less dvnced physiologicl phse thn the picl shoot, s they begin to develop only fter pinching. Present results of summer cultivr Chimes White re coincided with winter cvs. Coronette Yellow nd Coronette Scrlet (Winwright nd Irwin, 1987). Another possible cuse could be the interception of developmentl signls from leves to the picl meristem, which normlly crries genes to chnge growing meristem from vegettive to reproductive phse. If plnts re pinched before the perception of development signl in the leves, induction will never occur nd plnts never become competent to relese the stimulus towrds pex (Munir, 2003; McDniel, 1996). Similrly, in roses, pinching of the picl meristem enhnced the suitbility of xillry meristem to determine florlly fter obtining specific nodes/lef number (Cockshull nd Plnt height (cm) Plnt dry weight (g) Reltive growth rte (g g -1 d -1 ) Horridge, 1977). The difference in flowering time cn be directly linked to the brnch numbers, lef re, plnt fresh nd dry weights. It cn be ssumed from the present study tht when plnts produced more brnches, lrger lef re, mximum fresh nd dry weights, they took more time to flower. This indicte tht fter pinching, plnts remined juvenile for longer time thn the un-pinched ones nd s consequence the relese of developmentl stimulus ws delyed. The co-ordintion of promotive root-sourced cytokinin (CK) nd inhibitory shoot pex-sourced uxin (IAA) is centrl to ll current models on lterl bud dormncy relese. For exmple, in chickpe it hs been observed tht three potentil lterl bud growth inhibitors, IAA, ABA, nd cis-zetin 9-riboside (ZR), declined shrply in the relesed buds nd xylem following pinching. This is in contrst to potentil dormncy breking CKs like trns-zr nd trns-zetin 9-riboside 5 phosphte (ZRMP), which represented strongest correltive chnges by incresing 3.5 fold in xylem sp nd 22-fold in buds. However, polymines hd SED Control 4pirs 5pirs 6pirs 7pirs SED A R 2 = 0.99 Control 4pirs 5pirs 6pirs 7pirs SED R 2 = 0.99 Control 4pirs 5pirs 6pirs 7pirs Pinching tretments E C Plnt fresh weight (g) Lef re rtio (cm 2 g -1 ) Net ssimilte rte (g cm -1 d -1 ) R 2 = 0.99 Fig. 2. Effect of different pinching tretments on (A) plnt height (cm), (B) plnt fresh weight (g), (C) plnt dry weight (g), (D) lef re rtio cm 2 g -1, (E) reltive growth rte (d -1 ), nd (F) net ssimilte rte (g cm -1 d -1 ). Verticl brs (where lrger thn the points on lines) represent the stndrd error (SE) of vribility within replictes, wheres the seprte ones represent the stndrd error of difference (SED) between mens SED Control 4pirs 5pirs 6pirs 7pirs SED Control 4pirs 5pirs 6pirs 7pirs SED B D R 2 = 0.99 R 2 = 0.99 Control 4pirs 5pirs 6pirs 7pirs Pinching tretments F

30 28 Growth nd flowering response of snpdrgons fter relese from picl dominnce not chnged significntly in buds or other tissues, so they were not directly involved in the breking of bud dormncy (Mder et l., 2003). This evidence could be relted to the present study on Antirrhinum tht wht would hppen t the pex nd in the xylem when plnt is relesed from picl dominnce. The mjor dvntge of pinching is tht the pinched plnts produced mximum flower buds t hrvest (Fig. 1C). This could be result of more even distribution of ssimiltes between severl growing points (in xillry brnches) rther thn just the picl one (s in control plnts). The lter the pinching the greter ws the number of flower buds, lthough they took longer to flower. Similr findings were obtined in Antirrhinum, chrysnthemum nd cowpe (Winwright nd Irwin, 1987; Strmn nd Fust, 1999; Argll nd Stewrt, 1984). Pinching is method tht cn be used to produce wellproportioned plnts, suitble for selling s high qulity pot-plnts nd lso s method of controlling the time of flowering, when plnts re required for selling them flowered t certin time nd occsion. Similrly, plnt qulity ws much improved in plnts pinched t 5-7 lef-pir stge t the expense of flowering time. Therefore, proper mnipultion of the bove-mentioned protocol cn be used commercilly to get mximum outcome. References Adms, S.R., S. Person nd P. Hdley, An pprisl of the use of reciprocl trnsfer experiments: ssessing the stges of photoperiod sensitivity in chrysnthemum cv. Snowdon (Chrysnthemum morifolium Rmt.). J. Exp. Bot., 49: Argll, J.F. nd K.A. Stewrt, Effects of decpittion nd benzyl denine on growth nd yield of cowpe. Ann. Bot., 54: Bngreth, F Response of cytokinin concentrtion in the xylem exudte of ben (Phseolus vulgris L.) plnts to decpittion nd uxin tretment nd reltionship to picl dominnce. Plnt, 194: Cockshull, K.E. nd J.S. Horridge, Apicl dominnce nd flower initition in the rose. J. Hort. Sci., 52: Chen, J.G., Y.H. Zho, X. Zhou, L.S. Mo nd X.X. Chen, Fluctution in levels of endogenous hormones fter decpittion nd 6-benzyl mino purine tretment in Azle nd their reltionship to picl dominnce. Scienti Hortic., 71: Gocl, F.W.G., P.R. Phris, C.E. Yeung nd D. Perce, Chnges fter decpittion in concentrtions of indole-3-cetic cid nd bscisic cid in the lrger xillry bud of Phseolus vulgris L. cv. Tender Green. Plnt Physiol., 95: Kofrnek, M.A Affecting the spry shpe of pompon chrysnthemum during nturl seson flowering. Act Hortic., 125: Mder, J.C., R.J.N. Emery nd C.G.N. Turnbull, Sptil nd temporl chnges in multiple hormone groups during lterl bud relese shortly following pex decpittion of chickpe (Cicer rietinum) seedlings. Physiologi Plntrum, 119: McDniel, C.N Developmentl physiology of florl initition in Nicotin tbcum L. J. Exp. Bot., 47: Munir, M A study on the effects of environmentl fctors ffecting the phses of flower development in Antirrhinum mjus L. Ph.D. Thesis. Deprtment of Horticulture nd Lndscpe, School of Plnt Sciences, The University of Reding, U.K. Ono, M., K.S. Ono, K. Ymd, M. Ysui nd M. Okzki, Axillry bud flowering fter picl decpittion in Phrbitis in reltion to photoinduction. Physiologi Plntrum, 87: 1-6. Post, K Pompon growth fter pinching. New York Stte Flower Growers Bulletin, 41: 3. Strmn, W.T. nd E.J. Fust, Field-grown grden chrysnthemum responses to pinching nd plnt dte. HortScience, 34: Turnbull, G.N.C., A.A.M. Rymond, C.I. Dodd nd E.S. Morris, Rpid increses in cytokinin concentrtion in lterl buds of chickpe (Cicer rietinum L.) during relese of picl dominnce. Plnt, 202: Winwright, H. nd H.L. Irwin, The effects of pclobutrzol nd pinching on Antirrhinum flowering pot plnts. J. Hortic. Sci., 62:

31 Journl of Applied Horticulture, 8(1): 29-32, Jnury-June, 2006 Appl Journl Posthrvest control of soft-rot fungi on grpe berries by fungicidl tretment nd Trichoderm Y.A. Btt Lbortory of Plnt Protection, Deprtment of Plnt Production nd Protection, Fculty of Agriculture, An-Njh Ntionl University, P.O. Box 425 (Tulkrm), West Bnk, Plestine, Vi Isrel. E-mil: Abstrct The present reserch dels with the effect of posthrvest tretment of grpe berries with four commonly used fungicides nd two forms of Trichoderm hrzinum on the infection with soft-rot fungi- Rhizopus stolonifer nd Mucor piriformis. This effect ws evluted by comprison of the externl dimeter of rot-lesion in treted nd untreted berries, in ddition to comprison of percent reduction in externl rot-lesion dimeter reltive to control. Results indicted tht the infection with R. stolonifer nd M. piriformis ws significntly reduced (P<0.05) in ll treted berries in comprison with untreted control berries. The highest reduction in men externl rot-lesion dimeter ws obtined for both R. stolonifer nd M. piriformis when inoculted berries were treted with Score (difenoconzole) pplied t 0.35%(v/v) or Switch (cyprodinil + flodioxonil) pplied t 0.20%(w/v) or formulted T. hrzinum conidi in invert emulsion pplied t 9.6x10 8 conidi/ml of formultion (13.5, 13.2, nd 19.3 mm, respectively for R. stolonifer; 7.2, 7.5, nd 19.2mm, respectively for M. piriformis). The gretest decrese in percent reduction in externl rot-lesion dimeter reltive to control ws lso obtined for both the fungl species when inoculted berries were treted with the sme type of fungicides (Score nd Switch ) nd Trichoderm (formulted T. hrzinum in invert emulsion) (60.9, 61.7, nd 44.1%, respectively for R. stolonifer; 74.5, 73.4, nd 31.9%, respectively for M. piriformis). Overll results indicte tht the most effective tretment obtined on grpe berries could be integrted with other control mesures being usully used in grpe berry-rot mngement plns by lternting fungicidl tretment (e.g. Score or Switch ) with ppliction of formulted T. hrzinum conidi in invert emulsion. Key words: Grpe, Rhizopus stolonifer, Mucor piriformis, Trichoderm hrzinum, difenoconzole, cptn, cyprodinil + flodioxonil, metlxyl + mncozeb, posthrvest. Introduction The rpid nd extensive deteriortion of the tble grpes is minly cused by the fungl decy. The mjor orgnisms tht re involved in this decy re: Botrytis cinere, Aspergillus niger, Rhizopus stolonifer nd Mucor piriformis (Nelson, 1979). Of these orgnisms, R. stolonifer nd M. piriformis re the two known fungl species which cuse severe losses to tble grpes during mrketing nd export in mny countries (Lisker et l., 1996). The reson for these losses is the incidence of soft-rot symptoms on the infected berries during storge t tempertures >8 o C. Mechnicl wounding drmticlly increses the susceptibility of berries to soft-rot fungi nd fcilittes their penetrtion into the ttcked berries (Lichter et l., 2002). The conventionl method used to void the fungl decy in tble grpes is by fumigting SO 2 or relesing it from genertor pds contining metbisulfite slt, nd pckging of the fruit in polyethylene liners (Lichter et l., 2002). Immersion of the detched berries in 70% ethnol lso elimintes most of the fungl nd bcteril popultion on the berry surfce. This immersion results in inhibition of berry decy tht is equivlent to or better thn tht chieved by SO 2 relesed from genertor pds (Lichter et l., 2002). High levels of SO 2 cn lso result in fruit dmge, unplesnt ftertste nd llergies (Lisker et l., 1996). Other methods such s cetic cid fumigtion (Sholberg et l., 1996), cetylldehyde vpors (Avissr nd Pesis, 1991), nd modified tmosphere pckging nd generting chlorine gs (Zoffoli et l., 1999) cn be used insted of SO 2 fumigtion. Benomyl ws one of the most widely used fungicides to control posthrvest fungl decy especilly Mucor rot on grpe berries nd other fruits in pckinghouse (Spotts nd Cervntes, 1986). Two other chemicls, orthophenyl phnte s fog nd clcium hypochlorite s chlorine vpor tht were pplied to the surrounding tmosphere, significntly decresed posthrvest decy in rtificilly inoculted berries (Lisker et l., 1996). Vinclozolin (Roniln 50 WP) cn be used effectively ginst R. stolonifer nd B. cinere on tble grpes, strwberries nd kiwifruit (Lim et l., 1997). Finlly, Ozone tretment cn be considered possible substitute for SO 2 fumigtion for the control of R. stolonifer on tble grpes (Srig et l., 1996). The subtmospheric pressures (0.25, 0.50, nd 0.75 tmos) for different times (1 to 24h) were used to reduce Rhizopus rot lesion on tble grpes nd other fruit types in comprison with the control tretment (1 tmos) (Romnzzi et l., 2001). Biologicl control s substitute to chemicl control or s prctice tht could be integrted with other control prctices my result in cceptble levels of fruit decy with reduced levels of pesticide use such s using the following ntgonistic yest species: Pichi membrnefciens to control posthrvest Rhizopus rot on nectrine fruit (Fn nd Tin, 2000), Kloecker picult nd Cndid guilliermondii to control posthrvest diseses of mny fruits including tble grpe (McLughline et l., 1992), Cryptococcus lurentii, C. flvus, C. lbicus to control Mucor rot on per fruit (Roberts, 1990), Aureobsidium pullulns nd Cndid oleophil to control Botrytis cinere nd Rhizopus

32 30 Posthrvest control of soft-rot fungi on grpe berries by fungicidl tretment nd Trichoderm stolonifer on strwberry, tble grpe nd kiwifruit (Lim et l., 1997). Also, using the following ntgonistic bcteril species: Pseudomons cepci to control Mucor rot on pple (Jnisiewicz nd Roitmn, 1987), Pntoe gglomerns to control Rhizopus stolonifer nd Monilini lx on pech, pricot nd nectrine (Bonterr et l., 2003), nd Enterobcter cloce to control Rhizopus rot on pech (Wilson et l., 1987). Although ntgonistic fungus, T. hrzinum is in widespred use ginst mny fungl plnt pthogens such s root rot fungi (e.g. Pythium, Sclerotini, Rhizoctoni nd Fusrium) (Frvel, 1998), nd posthrvest fungl pthogens on different types of fruit (e.g. Botrytis cinere, Alternri lternt, Penicillium expnsum) (Btt, 1999, 2000, 2001, 2004, nd 2004b), no previous study ws crried out to control soft-rot fungi of tble grpes using formulted or unformulted T. hrzinum. Therefore, the objectives of the present reserch were: i) to test the efficcy of posthrvest tretment of grpe berries with four types of commonly used fungicides nd two forms of Trichoderm (unformulted nd formulted T. hrzinum) on the infection with soft-rot fungi: R. stolonifer nd M. piriformis, ii) to compre the efficcy of this tretment with fungicides nd T. hrzinum forms. Mterils nd methods Grpe berries used in the experiments: Helthy mture grpe berries (tble grpe cultivr: Zini) were used for biossy of the tretment effect of fungicides nd T. hrzinum ginst Rhizopus nd Mucor soft-rot. Fungl strins used in the inocultion nd tretment: The following fungl strins were used in the experiments: strin RS1 of Rhizopus stolonifer, strin MP3 of Mucor piriformis, nd strin Th 2 of T. hrzinum. The first two strins were isolted from infected grpe berries (cultivr: Beiruti) then subcultured on pltes with potto dextrose gr (PDA) medium for production of sporngi nd sporngiospores to be used in the inocultion tests. The third strin which belongs to the ntgonistic fungus T. hrzinum ws obtined from Fculty of Agriculture in Gembloux (Belgium) nd subcultured on pltes with ot mel gr (OMA). Young cultures of 14-dy old of the bove-mentioned strins were used to crry out the vrious tests of biossy. The concentrtion of the conidi or sporngiospores in the suspensions prepred from these cultures ws 11.7x10 6 sporngiospores of strin RS1 per ml, 5.5x10 5 sporngiospores of strin MP3 per ml, nd 9.6x10 8 conidi of strin Th 2 per ml. Tretments with fungicides nd T. hrzinum used in the experiments: Four types of tretments with fungicides were pplied ginst Rhizopus nd Mucor soft-rot inocultions on grpe berries. They were: 0.30% (W/V) metlxyl + mncozeb (sold s Ridomil MZ 63.5 WP, concentrtion of.i=7.5% metlxyl + 56% mncozeb); 0.35% (V/V) difenoconzole (sold s Score 250 EC); 0.35% (W/V) cptn (sold s Merpn 50 WP) nd 0.20% (W/V) cyprodinil + flodioxonil (sold s Switch 62.5 WG,.i.=375g/kg cyprodinil g/kg flodioxonil). Two types of tretments with T. hrzinum (strin Th 2 ) were pplied ginst Rhizopus nd Mucor soft-rot inocultions on grpe berries viz., conidil suspension in sterile distilled wter; nd formulted conidi in invert emulsion. Concentrtion of the conidi in both the tretments ws 9.6x10 8 conidi/ml. Ingredients of the invert emulsion (wter-in-oil type) used in the experiments were identicl to those used by Btt (2004). The conidi of T. hrzinum (strin Th 2 ) hrvested from 14-dy old culture, were introduced into the invert emulsion ccording to the technique developed by Btt (2004). Two dditionl nontreted control tretments were included in the tests: one with blnk formultion of invert emulsion nd the other with sterile distilled wter only (wter control tretment). Rhizopus nd Mucor inocultion nd tretment effect ssessment: Inocultion of R. stolonifer nd M. piriformis ws ccomplished by depositing 25-µl droplet of spore suspension contining 292,500 sporngiospores of R. stolonifer nd 13,750 sporngiospores of M. piriformis (originl suspensions contined 11.7x10 6 nd 5.5x10 5 sporngiospores/ml, respectively) on the fruit surfce fter being superficilly wounded using sterile sclpel. All fruits used in the tests were disinfected with 0.025% sodium hypochlorite, nd then rinsed with sterile distilled wter 3 times before inocultion. For the ssessment of tretment effect with fungicides nd T. hrzinum on the rot-lesion development cused by the bovementioned fungi, the preventive effect (tretment ppliction t the sme time of the inocultion) ws studied. The sme droplet size (25µl) of fungicides solution (mentioned erlier) or T. hrzinum conidi formulted in invert emulsion or suspended in sterile distilled wter ws used in the tretments. The droplet ws deposited t the sme site of inocultion with sporngiospores of R. stolonifer or M. piriformis on the fruit surfce immeditely fter the inocultion. Inoculted fruits were incubted in closed plstic continers (9.5cm dimeter 6.5cm deep) with 6 fruits per continer for 3 dys t 20±1 o C under humid conditions. Evlution of the tretment effect with fungicides nd T. hrzinum on the soft-rot development: Such effect ws evluted by mesuring the bility of ech type of fungicides or T. hrzinum form to reduce the development of rot-lesion cused by R. stolonifer or M. piriformis on grpe berries. The externl rot-lesion dimeter ws mesured in ll replictes 3 dys fter the tretment t 20±1 o C. Men of externl rot-lesion dimeter in ech tretment type ws clculted to be used in the comprison of tretment effect. Men percent reduction in the externl rotlesion dimeter reltive to non-treted wter control ws lso clculted in ech tretment for comprison of their effect. Sttisticl nlyses: Dt were nlyzed using completely rndomized design (CRD) with 6 replictes representing 6 fruits per tretment. Anlysis of vrince (ANOVA) nd Duncn s multiple rnge test (DMRT) were used to test the significnt differences between tretments representing the mens of externl rot-lesion dimeter or the mens of percent reduction in externl rot-lesion dimeter reltive to non-treted wter control. Results nd discussion Effect of fungicides nd T. hrzinum on Rhizopus soft-rot development on grpe berries: Significnt differences (t P<0.05) were obtined between tretments with four types of fungicides or two forms of T. hrzinum when used ginst Rhizopus soft-rot on grpe berries in comprison with the nontreted control (Tble 1). Tretments with Switch (cyprodinil + flodioxonil) nd Score (difenoconzole) resulted in the

33 Posthrvest control of soft-rot fungi on grpe berries by fungicidl tretment nd Trichoderm 31 lest vlues of externl rot-lesion dimeter (13.2 nd 13.5mm, respectively), followed by the tretment with formulted conidi of T. hrzinum in invert emulsion (19.3mm). The other tretments used in the experiment resulted in significnt higher vlues of mens of externl rot-lesion dimeter rnging from 24.2 to 34.5mm. The tretment efficcy, in descending order, ws s follows: Switch or Score, formulted conidi of T. hrzinum in invert emulsion, conidil suspension of T. hrzinum in sterile distilled wter, Merpn or Ridomil, blnk formultion of invert emulsion or sterile distilled wter s non-treted control tretments. Similrly, significnt differences (t P<0.05) were obtined between mens of percent reduction in the externl rot-lesion dimeter reltive to control with sterile distilled wter (Tble 1). Therefore, the highest mens of the percent reduction (60.9 nd 61.7) were recorded with Score nd Switch, respectively, followed by formulted T. hrzinum conidi in invert emulsion (44.1). The rest of the tretments significntly cused lower mens of the percent reduction rnging between 29.8 nd 3.5. The tretment efficcy, in descending order, ws similr to the bove-mentioned trend. Effect of fungicides nd T. hrzinum on Mucor soft-rot development on grpe berries: Significnt differences (P<0.05) were obtined between tretments with four types of fungicides or two forms of T. hrzinum when used ginst Mucor soft-rot on grpe berries in comprison with the non-treted control (Tble 2). Tretment with Score (difenoconzole) nd Switch (cyprodinil + flodioxonil) gve the lest vlues of externl rotlesion dimeter (7.2 nd 7.5mm, respectively), followed by the tretment with Merpn (cptn) (15.7mm), formulted conidi of T. hrzinum in invert emulsion (19.2mm). The rest of the tretments used in the experiment gve significntly higher vlues of mens of rot-lesion dimeter rnging from 24.7 to 28.2mm. The tretment efficcy in descending order, ws s follows: Score or Switch, Merpn, formulted conidi of T. hrzinum in invert emulsion, conidil suspension of T. hrzinum in sterile distilled wter or Ridomil, blnk formultion of invert emulsion, nd sterile distilled wter s nontreted control tretment. Significnt differences (P<0.05) were lso obtined between mens of percent reduction in the externl rot-lesion dimeter reltive to control with sterile distilled wter (Tble 2). The highest mens of the percent reduction (74.5 nd 73.4) were thus cused by the tretment with Score nd Switch, respectively, followed by Merpn (44.3) nd formulted conidi of T. hrzinum in invert emulsion (31.9). The rest of the tretments significntly cused lower mens of the percent reduction rnging between 12.4 nd 1.8 nd similr to bove mentioned trend of the tretment efficcy. Overll results obtined in the present investigtion indicte tht the posthrvest tretment of grpe berries with Switch (cyprodinil + flodioxonil) or Score (difenoconzole) gve the highest percent reduction in the externl rot-lesion dimeter cused by R. stolonifer nd M. piriformis. This effectiveness of tretment is comprble with tht obtined by SO 2 fumigtion, conventionlly prcticed to prevent fungl decy in tble grpe berries (Lisker et l., 1996; Lichter et l., 2002). SO 2 tretment remins effective s long s SO 2 level is sufficiently high, but the high levels cn result in fruit dmge s fruit bleeching, unplesnt ftertste nd llergies (Lisker et l., 1996; Lichter et l., 2002). To void the undesirble side-effects of chemicl tretment ginst fungl decy of hrvested fruit especilly in tble grpe berries, tretment with biocontrol gents is recommended by mny investigtors (Eld, 1994; Hrmnn et l., 1996). In the present study, the posthrvest tretment of grpe berries with T. hrzinum formulted in invert emulsion lso gve high percent reduction in externl rot-lesion dimeter cused by R. stolonifer nd M. piriformis. However, this tretment is less Tble 1. Effect of tretment with four types of fungicides nd two forms of T. hrzinum (strin Th 2 ) on the soft-rot development cused by Rhizopus stolonifer (strin RS1) on grpe berries 3 dys fter inocultion nd tretment t 20±1 o C under humid conditions Fungicides nd T. hrzinum tretments Externl rot-lesion dimeter (mm) developed on berries (Men±SE) Ridomil (metlxyl + mncozeb) 30.2±2.8 d 1) 12.5 b 1) Score (difenoconzole) 13.5± e Merpn (cptn) 28.5±3.1 d 17.4 b Switch (cyprodinil + flodioxonil) 13.2± e Formulted conidi of T. hrzinum in invert emulsion 19.3±2.4 b 44.1 d Blnk formultion of invert emulsion s control tretment 2) 33.3±3.3 e 3.5 Reduction(%) in externl rot-lesion dimeter reltive to wter (control) Conidil suspension of T. hrzinum in sterile distilled wter 24.2±2.9 c 29.8 c Sterile distilled wter s control tretment 34.5±3.1 e Tble 2. Effect of tretment with four types of fungicides nd two forms of T. hrzinum (strin Th 2 ) on the soft-rot development cused by Mucor piriformis (strin MP3) on grpe berries 3 dys fter inocultion nd tretment t 20±1 o C under humid conditions Fungicides nd T. hrzinum tretments Externl rot-lesion dimeter (mm) developed on berries (Men±SE) Reduction(%) in externl rot-lesion dimeter reltive to wter (control) Ridomil (metlxyl + mncozeb) 24.8±2.5 d 1) 12.1 b 1) Score (difenoconzole) 7.2± e Merpn (cptn) 15.7±2.1 b 44.3 d Switch (cyprodinil + flodioxonil) 7.5± e Formulted conidi of T. hrzinum in invert emulsion 19.2±1.9 c 31.9 c Blnk formultion of invert emulsion s control tretment 2) 27.7±3.1 de 1.8 Conidil suspension of T. hrzinum in sterile distilled wter 24.7±2.8 d 12.4 b Sterile distilled wter s control tretment 28.2±3.7 e 1) Mens of externl rot-lesion dimeter within ech column followed by different letters re significntly different t P<0.05 ccording to ANOVA nd Duncn s multiple rnge test (DMRT). 2) Invert emulsion is composed of the following ingredients (W/W): sterile distilled wter (45.25%), glycerine (4.00%), wter-soluble wx or Dehymuls K (0.75%), Tween 20 (2.50%), nd mixture of 19.0% coconut oil % soyben oil.

34 32 Posthrvest control of soft-rot fungi on grpe berries by fungicidl tretment nd Trichoderm effective thn the fungicidl tretment with Switch or Score, it hs no undesirble side-effects e.g. leving toxic residues or bd flvors or smells in the treted berries. To overcome the problem of hrmful side-effects tht pper in ssocition with the chemicl tretment of grpe berries, mny investigtors hve recommended lterntion of chemicl nd biologicl tretment pplictions keeping menwhile the sme level of tretment efficcy. The pplictions of biologicl tretment could be lso integrted with other control mesures prcticed within the decy-mngement progrm of grpe berries. For the other types of grpe berry decy (e.g. Botrytis bunch rot cused by Botrytis cinere), the decy-mngement progrm ws performed successfully using both chemicl nd biologicl mens such s g L -1 of T. hrzinum dry conidi g L -1 of vinclozolin or iprodione or 0.25 g L -1 of diethofencrb plus crbendzim resulted in up to 78% disese reduction in grpe berries (Eld, 1994). Also, tretment with T. hrzinum dry conidi could replce some pplictions of iprodione or vinclozolin with little reduction in efficcy ginst B. cinere on grpe berries (Hrmnn et l., 1996). The effectiveness of posthrvest tretment with T. hrzinum formulted in invert emulsion ws proved by us in controlling other types of fruit decy on vrious types of fruits t posthrvest stge (e.g. gry mold on strwberry nd pple; blue mold on pple; blck fruit spot of persimmon) (Btt, 1999, 2001, 2004 nd 2004b). We hve lso proved in our previous reserch tht this type of posthrvest biocontrol tretment hs no hrmful side-effects since the ingredients used for the invert emulsion re especilly oils nd emulsifiers which re sfe nd hve no phytotoxic effect on treted fruits. These ingredients re lso likely to be non-toxic to humns s they re lso used s food dditives nd in the mnufcture of cosmetics. References Avissr, I. nd E. Pesis, The control of posthrvest decy in tble grpes using cetylldehyde vpor. Ann. Appl. Biol., 118: Btt, Y Biologicl effect of two strins of microorgnisms ntgonistic to Botrytis cinere: cusl orgnism of gry mold on strwberry. An-Njh Univ. J. Res. Nturl Sci., 13: Btt, Y Alternri lef spot disese on fig trees: vrietl susceptibility nd effect of some fungicides nd Trichoderm. The Islmic Univ. J. Gz, 8: Btt, Y Effect of fungicides nd ntgonistic microorgnisms on the blck fruit spot disese on persimmon. Dirst: Agric. Sci., 28: Btt, Y., Posthrvest biologicl control of pple gry mold by Trichoderm hrzinum Rifi formulted in invert emulsion. Crop Prot., 23: Btt, Y. 2004b. Effect of tretment with Trichoderm hrzinum Rifi formulted in invert emulsion on posthrvest decy of pple blue mold. Int. J. Food Microbiol., 96: Bonterr, A., M. Mri, L. Cslini nd E. Montesinos, Biologicl control of Monilini lx nd Rhizopus stolonifer in posthrvest of stone fruits by Pntoe gglomerns EPS 125 nd puttive mechnisms of ntgonism. Int. J. Food Microbiol., 84: Eld, Y Biologicl control of grpe gry mold by Trichoderm hrzinum. Crop Prot., 13: Fn, Q. nd S.P. Tin, Posthrvest biologicl control of Rhizopus rot of nectrine fruits by Pichi membrnefciens. Plnt Dis., 84: Frvel, D Commercil biocontrol products for use ginst soil borne diseses (dfrvel@srr.rsusd.gov). Hrmnn, G.E., B. Ltorre, E. Agosin, R. Sn Mrtin, D.G. Riegel, P.A. Nielson, A. Tronsomo ndr.c. Person, Biologicl nd Integrted control of Botrytis bunch rot of grpe using Trichoderm spp. Biologicl Control, 7: Jnisiewicz, W. J. nd J. Roitmn, Posthrvest mucor rot control on pple with Pseudomons cepci. Phytopth., 77: Lichter, A., Y. Zutkhy, L. Sonego, O. Dvir, T. Kplunov, P. Srig nd R. Ben-Arrie, Ethnol controls posthrvest decy of tble grpes. Posthrvest Biol. Technol., 24: Lim, G., A. Ippolito, F. Nigro nd M. Slerno, Effectiveness of Aureobsidium pullulns nd Cndid oleophil ginst posthrvest strwberry rots. Posthrvest Biol. Technol., 10: Lisker, N., Z. Keren-Shchmn, P. Srig, Y. Zutkhy nd R. Ben-Arie, The biology nd pthology of the fungus Rhizopus stolonifer, cuse of blck mold disese of tble grpes in Isrel. Plnt Pthol., 45: McLughline, R.J., C.L. Wilson, S. Droby, R. Ben-Arie nd E. Chlutz, Biologicl control of posthrvest diseses of grpe, pech nd pple with yests Kloecker picult nd Cndid guilliermondii. Plnt Dis., 76: Nelson, K.E Hrvesting nd hndling Cliforni tble grpe for mrket. Univ. Cliforni Bull. No Roberts, R.G Biologicl control of Mucor rot of per by Cryptococcus lurentii, C. flvus nd C. lbicus. Phytopthology, 80: Romnzzi, G., F. Nigro, A. Ippolito nd M. Slerno, Effect of short hypobric tretment on posthrvest rots of sweet cherries, strwberries nd tble grpes. Posthrvest Biol. Technol., 22: 1-6. Srig, P., Y. Zutkhy, T. Zhvi, S. Ynni, N. Lisker nd R. Ben-Arie, Ozone for control of posthrvest decy of tble grpes cused by Rhizopus stolonifer. Physoil. Mol. Plnt Pthol., 48: Sholberg, P.L., A.G. Reynolds nd A.P. Gunce, Fumigtion of tble grpes with cetic cid to prevent posthrvest decy. Plnt Dis., 80: Spotts, R.A. nd L.A. Cervntes, Popultion pthogenicity nd benomyl resistnce of Botrytis spp., Penicillium spp. ndmucor piriformis in pckinghouse. Plnt Dis., 70: Wilson, C.L., J.D. Frnklin nd P.L. Pusey, Biologicl Control of Rhizopus rot of pech with Enterobcter cloce. Phytopth., 77: Zoffoli, J.P., B.A. Ltorre, E.J. Rodrigues nd P. Aldnce, Modified tmosphere pckging using chlorine gs genertors to prevent Botrytis cinere on tble grpes. Posthrvest Biol. Technol., 15:

35 Journl of Applied Horticulture, 8(1): 33-36, Jnury-June, 2006 Nitrogen metbolism of Aloe ver under long-term diluted sewter irrigtion Appl Journl Liu Zho-Pu, Zho Geng-Mo, Liu Ling nd Zheng Qing-Song College of Resources nd Environmentl Sciences, Nnjing Agriculturl University, Nnjing, , P.R. Chin. Abstrct Diluted sewter such s 10% (10 volumes of sewter nd 90 volumes of freshwter), 25%, 50%, 75% nd 100% were used to irrigte Aloe ver L. during four successive yers in Ledong region, Hinn Province of Chin. The effect of sewter irrigtion on nitrogen metbolism of loe plnt ws studied. Totl nitrogen content of loe leves rnged from 1.48 to 1.56 % of dry mtter, nd no significnt differences were observed between control (freshwter irrigtion) nd sewter tretments. The totl nitrogen content of loe roots, in the rnge of 0.74 to 0.85 % of dry mtter, ws much lower thn tht in the leves. There ws no significnt difference in totl nitrogen content of roots between control nd sewter tretments. It is suggested tht sewter tretments do not ffect nitrogen uptke nd trnsport in loe plnt. The nitrte content in loe leves irrigted with sewter ws much lower thn tht with fresh wter irrigtion, nd continuous decline in nitrte content ws noted with incresing sewter concentrtion. The nitrte/totl nitrogen rtio lso tends to decrese in leves suggesting tht nitrte hs been ssimilted into osmoregulted substnces under sewter stress. The mino cid content of loe plnt ws not ffected, while the rtios of mino cid/totl nitrogen significntly incresed under sewter stress s compred with control. The protein content nd protein/totl nitrogen rtios were not ffected by sewter tretment except for 100%, suggesting tht there ws fvourble trnsformtion from mino cids to proteins under slt stress. It is concluded tht long term irrigtion by diluted sewter on lechble sndy soil with excessive nnul rin precipittion could effectively mintin yield nd improve the qulity of loe. Key words: Amino cid, loe qulities, nitrte, protein, totl nitrogen Introduction The shortge of freshwter resources for irrigtion is well known problem of griculture (Hmdy et l., 2005). The chllenge is to mintin crop production without impiring the blnce of good qulity wter: n obvious solution is consequently to explore the sustinble use of non-conventionl wter resources (Pereir, 2002). When freshwter supply is limited, there is n incresing demnd for the use of non-conventionl wters. Sewter, s n mple non-conventionl wter source, ws proposed to be used for crop production long the costl desert 40 yers go (Glenn, 1998). loe is commonly used to tret number of skin irrittions, such s dry skin nd irritnt contct dermtitis (West nd Zhu, 2003), heling of burns (Visuthikosol et l., 1995) nd cure of certin cncers. loe is widely cultivted in costl regions of southern Chin. Since severl qulities of loe plnt were relised recently, loe becme priority subject of mny studies (Zheng, 2004; Liu, 2003; Sun et l., 2003; Wu et l., 2003). However, studies on nitrogen metbolism of loe plnt under slt stress re rre. The effect of sewter irrigtion on nitrogen metbolism nd qulities of loe plnt hs not been studied. The objectives of the present study were (1) to study the nitrogen metbolism of loe plnt under long-term irrigtion with diluted sewter, nd (2) to ssess the qulities of loe plnt under sewter irrigtion vis vis increse/decrese of metbolic products. Mterils nd methods Study re nd experiment lyout: The experiments were conducted t the 863 Reserch Sttion, Ledong County, southest of Hinn Province, Chin (18º9 N, 108º56 E). The climte is tropicl monsoon with men nnul temperture of o C, men nnul precipittion mm, most of which occurs from lte My to October, nd men nnul evportion mm. A. ver used in this experiment hs slt stress tolernce since the tissue cultured plnts used for cultivtion were from plnts growing under sewter stress conditions. It ws grown in 20 m 2 plots (5 4 m). The soil contined 69, 30 nd 1% snd, silt nd cly, respectively. The totl soluble slts content nd bulk density were 0.002% nd 1.62 g cm -3, respectively. There were six tretments i.e. CK (freshwter irrigtion) nd 10% (EC=4.2 ds m -1 ), 25% (EC=10.9 ds m -1 ), 50% (EC=19.1 ds m -1 ), 75% (EC=28.4 ds m -1 ) nd 100% (EC= 39.2 ds m -1 ) sewter Tble 1 Some chemicl properties of the irrigtion wter ) Wrer used ph EC (ds m -1 ) TN (mg L -1 ) IN (mg L -1 ) TP (mg L -1 ) RP (mg L -1 ) Sewter Tp wter ) EC: Electricl conductivity; TN: Totl nitrogen; IN: Inorgnic nitrogen; TP: Totl phosphorus; RP: Rective phosphorus.

36 34 Nitrogen metbolism of Aloe ver under long-term diluted sewter irrigtion tretments, with rndomised complete block design, nd ech tretment hd three replictes. Sewter used for irrigtion hd slinity of bout 31.0 g L -1. Prior to irrigtion, the sewter nd tp wter were mixed in tnk, to which compound fertilizer (N:P 2 O 5 :K 2 O=15:15:15) ws dded to give the finl concentrtion of 0.2%. The prepred mixtures were delivered to the experimentl plots by plstic tubes. Bsic chemicl properties of irrigted wter re shown in Tble 1. On August, 2001, plots were seprted from ech other with deeply buried plstic film (40 cm depth) so s to prevent lterl permetion of sewter between plots. Before strt, the experimentl plots were pre-irrigted with sufficient freshwter to keep the soil field cpcity moisture. Bse fertilizers were pplied t the rtes of 600 nd kg h -1 for superphosphte nd pig mnure, respectively. Aloe ws grown in the plots with 60 cm between rows nd 50 cm between plnts in the row. In one of the plots within the replictes, tensionmeter ws instlled t the depth of 0~40 cm. On December 10, the first irrigtion ws given using freshwter, nd fter tht irrigtion with the sewter tretments ws done whenever soil wter suction t depth of 0~40 cm exceeded the vlue of 3.8 MP. In ll, 218 irrigtions were given during the growth stges of loe in 4 yers. The totl mount of wter supplied in the irrigtion reched bout 1 m 3 per plot. Smpling nd nlysis: Before irrigtion, sewter nd freshwter smples were nlysed for electricl conductivity, ph, bicrbonte nd levels of the ions: potssium, sodium, clcium, mgnesium, chlorine nd sulphte. Electricl conductivity nd ph were mesured in the field with EC 214 Conductivity Meter (HANNA instruments) nd ph Meter (Cyerscn 510 ), respectively. Certin wter prmeters nd initil soil smples which couldn t be mesured in the field were nlysed in the Resources nd Environmentl Lbortory, Nnjing Agriculturl University (NJAU). Aloe ws hrvested during lte April 2004, nd three individul plnts in ech plot were collected for nlyzing plnt biomss, totl nitrogen, nitrite, mino cid nd protein content. Anlysis of mino cid nd protein content ws s per Li et l. (1999). All other nlyses were ccording to Lu (1999). Sttisticl methods: The nlysis of vrince ws performed by the stndrd procedures using MS-EXCEL 2000 softwre. The mens of different tretments were compred by pplying Lest Significnt Difference Test (P=0.05) using SPSS 13.0 softwre. Results nd discussion Effect of sewter irrigtion on totl nitrogen content in leves nd roots: Totl nitrogen content in leves nd roots of loe under sewter irrigtion during four yers re shown in Fig.1. The nitrogen content of loe leves rnged from 1.48 to 1.56 % of dry mtter, nd no significnt differences were noted between CK- nd sewter-tretments. The nitrogen content of loe in roots, in the rnge of 0.74 to 0.85 % of dry mtter, ws much lower thn tht in leves. There ws no significnt difference in nitrogen content of roots between the CK- nd (%DW) Totl nitrogen b Lef b b Root CK 10% 25% 50% 75% 100% Tretment Fig. 1. The totl nitrogen content in leves nd roots of Aloe ver under diluted sewter stress. CK represents freshwter (tp wter) tretment, nd sewter tretment, for exmple, 10% mixture of 10% sewter nd 90% fresh wter on volume bsis. The sme letters on the brs re not significntly different (P=0.05) by Lest Significnt Difference Test. Aloe biomss (kg h -1. DW) Tretment Fig. 2. The loe biomss t hrvest in 2004 under diluted sewter stress. CK represents freshwter (tp wter) tretment, nd sewter tretment, for exmple, 10% mixture of 10% sewter nd 90% fresh wter on volume bsis. The sme letters on the brs re not significntly different (p=0.05) by Lest Significnt Difference Test. CK 10% 25% 50% 75% 100% the sewter- tretments. These results suggest tht sewter irrigtion didn t significntly influence the nitrogen uptke nd trnsport. This my be ttributed to high irrigtion frequency nd good wter dringe in the sndy soil. Totl nitrogen uptke by loe plnt decresed t 75 nd 100% sewter tretments due to the decresed biomss under severe sewter stress (Fig. 2). These results re in greement with erlier studies with different plnts species, like Alflf (Khn et l., 1994), pumpkin (Aroiee et l., 2005) nd ben (Rbie nd Almdini, 2005) under slt stress conditions. Effect of sewter irrigtion on nitrte content nd rtios of nitrte to totl nitrogen in leves of loe: Sewter irrigtion significntly decresed nitrte content in leves of loe when compred with the CK (Fig. 3A). Among the tretments, the highest vlue of nitrte content in leves ws obtined in 25%, followed by 10, 50, 75 nd 100%. The rtio of nitrte/nitrogen in leves re presented in Fig. 3B. Sewter irrigtion mrkedly lowered the rtio of nitrte/totl nitrogen in the leves compred to the CK. With 10, 25, 50, 75 nd 100% sewter irrigtion, nitrte/nitrogen in leves decresed by 20.8, 11.8, 22.9, 26.0 nd 32.3%, respectively s compred to CK. Decrese in the nitrte level of leves with incresing slinity could be ttributed to build-up of orgnic osmoregultory substnces. A decresed nitrte level in leves my lso be due to incorportion of b b b

37 Nitrogen metbolism of Aloe ver under long-term diluted sewter irrigtion 35 nitrte in sugrs nd mino cid (proline) (Boggess et l., 1976; Stewrt, 1981; Rhodes et l., 1986; Rhodes nd Hnd, 1989), nd this trnsformtion my be n dpting mechnism of plnt for mintining their norml growth under slt stress. It is lso likely tht n increse in Cl - concentrtion in soil fter sewter irrigtion hd reduced the nitrte uptke s reported for other plnts (Weigel et l., 1973; Kfkfi et l., 1982; Eli et l., 2004). Effect of sewter irrigtion on orgnic nitrogen nd rtio of orgnic nitrogen to totl nitrogen: Sewter irrigtion resulted in significnt increse in mino cid concentrtion in loe leves. The highest content of mino cid ws obtined in the 25% tretment. It ws 1.16, 1.15, 1.07 nd 1.04-folds respectively, when compred with the tretments 10, 50, 75 nd 100% (Fig. 4A). Sewter irrigtion hd incresed the rtio of mino cid to totl nitrogen s compred with the CK (Fig. 4B). Nitrte content (mg g -1.DW) c b CK 10% 25% 50% 75% 100% b Tretment d e Tretment Fig. 3. The nitrte content (A) nd the rtio of nitrte/totl nitrogen (B) in loe leves under diluted sewter irrigtion. CK represents freshwter tretment. Sewter tretment, for exmple, 10% is mixture of 10% sewter nd 90% fresh wter on volume bsis. The sme letters on the brs re not significntly different (P=0.05) by Lest Significnt Difference Test. Amino cid content (mg 100g -1.DW) Fig. 4. The mino cid content (A) nd the rtio of mino/totl nitrogen (B) in loe leves under diluted sewter stress. CK represents freshwter tretment, nd sewter tretment, for exmple, 10% is mixture of 10% sewter nd 90% fresh wter on volume bsis. The sme letters on the brs re not significntly different (P=0.05) by Lest Significnt Difference Test. Protein content (mg g -1.DW) c bc CK 10% 25% 50% 75% 100% A c Tretment Tretment b A A b CK 10% 25% 50% 75% 100% b Fig. 5. The protein content (A) nd the rtios of protein/totl nitrogen (B) in loe leves under diluted sewter stress. CK represents freshwter tretment, nd sewter tretment, for exmple, 10% is mixture of 10% sewter nd 90% fresh wter on volume bsis. The sme letters on the brs re not significntly different (P=0.05) by Lest Significnt Difference Test. Nirte / Totl nitrogen rtio Protein / Totl nitrogen rtio Amino cid / Totl nitrogen rtio bc b CK 10% 25% 50% 75% 100% B b b b CK 10% 25% 50% 75% 100% b b c Tretment b Tretment b c B B b CK 10% 25% 50% 75% 100% c

38 36 Nitrogen metbolism of Aloe ver under long-term diluted sewter irrigtion Protein content: A significnt increse in protein content of loe leves ws observed under sewter tretments (except for 100%) indicting tht the protein content hd not been influenced under moderte sewter stress (Fig. 5A). Sewter irrigtion enhnced the rtio of protein/totl nitrogen s compred with CK (except for 100%) (Fig. 5B). It my be due to the fct tht the presence of slt ions my ctivte the biosynthesis of glycinebetine which in turn my improve the protein synthesis, mechnism dopted by the plnts ginst slinity stress s ws suggested by Nizi et l. (2004). Totl nitrogen content of loe did not decline under diluted sewter irrigtion s compred to freshwter irrigtion. However, nitrte content s well s the rtio of nitrte to nitrogen in loe leves ws significntly reduced due to sewter stress, suggesting ssimiltion of nitrte into osmoregultory substnces like soluble sugrs nd orgnic cids. Subsequent results showed tht sewter irrigtion significntly incresed the mino cid content of leves suggesting tht nitrte in leves ws lrgely ssimilted into mino cid to resist severe slt stress. However, protein content of loe in leves ws not ffected by sewter except the pure sewter tretment, demonstrting tht there is trnsformtion of mino cids into proteins under slt stress. This study demonstrtes tht long term irrigtion by diluted sewter on lechble sndy soil with excessive nnul rin precipittion could effectively mintin yield nd improve the qulity of loe. Acknowledgement The reserch ws funded by the Ntionl High-Tech 863 Project (Nos. 2002AA2Z4061 nd 2003AA627040) of Chin. We cknowledge the ssistnce of LIU Lin, CHEN Jin-Mio, YUN Yn nd FU Yong-Sheng in tril mngement, s well s Sun X for creful review of this mnuscript. References Aroiee, H., M. Azizi nd R. Omibigi, Effect of slinity nd nitrogen nutrition on free proline nd oil content of medicinl pumpkin (Cucurbit pepo subsp. Pepo convr. Pepo vr. styric). Act Hortic., 676: Boggess, S.F., D. Aspinll nd L.G. Pleg, Stress metbolism. IX. The significnce of end-product inhibition of proline biosynthesis nd of comprtmenttion in reltion to stress-induced proline ccumultion. Aust. J. Plnt Physiol., 3: Eli, A., P. Sntmri nd F. Serio, Artichoke response to nitrogen form rtio under sline conditions. Act Hortic., 660: Glenn, E P., J.J. Brown nd J.W. Lery, Irrigting crops with sewter. Scientific Americn, 279(2): Hmdy, A., V. Srdob nd K.A. Frrg Ghnem, Sline wter in supplementl irrigtion of whet nd brley under rinfed griculture. Agriculturl Wter Mngement, 78: Kfkfi, U., N. Vlors nd J. Letey, Chloride interction with nitrte nd phosphte nutrition in tomto. J. Plnt Nutr., 5: 1369 Khn, M.G., M. Silberbush nd S.H. Lips, Physiologicl studies on slinity nd nitrogen interction in lflf. 1. Biomss production nd root development. J. Plnt Nut., 17: Li, H.S Principles nd Techniques of Plnt Physiologicl Biochemicl Experiment. Higher Eduction Press, Beijing. 123p. Liu, L., Liu, L., Liu, Z.P., Fu, Y.S., Yun, Y., Deng, L.Q. nd Chen, M.D., Study on Aloe ver irrigted by sewter in the se-bech of South Chin. Journl of Nturl Resources, 18(5): Lu, R.K Soil nd grochemistry nlyses. Beijing: Agriculturl Science nd Technology of Chin Press. Nizi, B.H., M. Athr nd J. Rozem, Slt tolernce in fodderbeet nd sebeet: nlysis of biochemicl reltions. Bulg. J. Plnt Physiol., 30(1-2): Pereir, L.S., T. Oweis nd A. Ziri, Irrigtion mngement under wter scrcity. Agriculturl wter mngement, 57: Rbie, G.H. nd A.M. Almdini, Role of bioinoculnts in development of slt-tolernce of Vici fb plnts under slinity stress. Africn Journl of Biotechnology, 4(3): Rhodes, D. nd S. Hnd, Amino cid metbolism in reltion to osmotic djustment in plnt cells. In: Environmentl Stress in Plnts: Biochemicl nd Physiologicl Mechnisms, NATO ASI Series, Vol. G19 (JH Cherry ed), Springer, Berlin, pp Rhodes, D., S. Hnd nd R.A. Bressn, Metbolic chnges ssocited with dpttion of plnt cells to wter stress. Plnt Physiol., 82: Stewrt, C.R Proline ccumultion: Biochemicl spects. In: "Physiology nd Biochemistry of Drought Resistnce in Plnts," (Pleg, L.G. nd Aspinll, D. eds), Acdemic Press, Sydney, pp Sun, S.B., Q.R. Shen, J.M. Wn nd Z.P. Liu, Induced expression of the gene for NADP-mlic enzyme in leves of Aloe ver L.under slt stress. Act Biomic et Biophysic Sinic., 35(5): Visuthikosol, V., B. Chowchuen, Y. Sukwnrt, S. Sriurirtn nd V. Boonpucknvig, Effect of Aloe ver gel to heling of burn wound: clinicl nd histologic study. J Med Assoc Thi., 78: Weigel, R.C., J.A. Schillinger, B.A. Mc Cw, H.G. Guch nd E. Hsio, Nutrient-nitrte levels nd ccumultion of chloride in leves of snpbens nd root of soybens. Crop Sci., 13: West, D.P. nd Y.F. Zhu, Evlution of Aloe ver gel gloves in the tretment of dry skin ssocited with occuptionl exposure. Am. J. Infect. Control, 31: Wu, J.M., Z.P. Liu nd G.M. Zho, Effects of tretment with different concentrtions of sewter on chlorophyll content nd ultrstructure of Aloe. Journl of Nnjing Agriculturl University. 26(3): Zheng, Q.S., Z.P. Liu,, Y. L. Liu nd L. Liu, Effects of iso-osmotic slt nd wter stresses on growth nd ionic distribution in Aloe seedlings. Act Phytoecologic Sinic, 28(6):

39 Journl of Applied Horticulture, 8(1): 37-41, Jnury-June, 2006 Reltionship between soil nd lef minerl nutrient concentrtion nd yield of selected citrus species Appl Journl L. Andrews nd R.A.I. Brthwite b Croni Reserch Sttion, Wterloo Rod, Crpichim, Republic of Trinidd nd Tobgo, e-mil: lpndrews@hotmil.com; b Fculty of Science nd Agriculture, The University of the West Indies, St. Augustine Cmpus, e-mil: rib@crib-link.net. Abstrct Low yields of citrus in Trinidd prompted n investigtion to determine whether infield yield vrition in citrus ws due to differences in plnt nutrition induced by field vribility. Selected trees of three cultivrs (Vlenci ornge (Citrus sinensis), Portugl mndrin (C. reticult) nd Ortnique tngor (C. sinensis x reticult) were monitored for one to two yers nd indictors of yield such s percent fruit set, fruit count nd fruit qulity mesured. Lef nutrient content of the trees nd nutritive fctors of the soil in the root zone were lso determined. Nutrient deficiencies were found in the fields of ll the three cultivrs. The most common deficiencies were of clcium, zinc nd mgnesium. There ws limited evidence of yield correltion with soil ph (P = 0.012), nd lef phosphorus content (P = 0.02), Zn (P = 0.005) nd N (P = 0.001). DRIS nlysis supported the notion tht infield yield vribility ws ssocited with nutrients tht were limiting. Percent fruit set ws ssocited with C/Mg rtio (r = 0.542, P = 0.045; r = 0.607, P = 0.016) nd folir concentrtion of micro elements Cu (r = 0.738, P = 0.003; r = 0.667, P = 0.007) nd Fe (r = 0.507, P = 0.064; r = 0.573, P = 0.026) in 1997 for one field ech of Vlenci ornge nd Portugl mndrin, respectively. The most commonly derived reltionship for fruit qulity ws negtive reltionship of lef nitrogen concentrtion with fruit weight. A positive reltionship between lef concentrtion of mngnese nd peel thickness occurred in Portugl mndrin for the two yers of the study. Key words: Citrus, Vlenci, Ortnique, mndrin, minerl nutrition, yield, fruit set, fruit qulity Introduction Men yield for ornges in Trinidd is low (Lucie-Smith, 1953; Cooper, 1956; Ali et l., 1973; Andrews et l., 2001), less thn 1.5 crtes per tree (41 kg per crte). It hs been observed s well tht there is gret vrition in yielding level between trees in the sme field (Andrews, 1994). Incresed yield cn be ccomplished by reduction of tree-to-tree vrition nd lso by incresing the yield of individul trees. It is hypothesized in this study tht vrition in yield, percent fruit set nd fruit qulity re mirrored by corresponding vrition in nutrient content in lef or soil. As in other crops, the routine method of ssessing nutritionl sttus nd needs hve been lef nd soil nlysis in citrus (Weir, 1965; Rodriguez et l., 1997). The use of sufficiency rnges nd criticl vlues hve been most common for folir nlyses (Br-Akiv et l., 1968; Jorgensen, 1978; Embleton nd Lbnusks, 1982; Dey nd Singh, 1998). An lterntive method of interpreting dt from the soil nd folir nlyses is the use of minerl nutrient rtios (Weir, 1969). The more recent development of this pproch is the use of the Dignosis nd Recommendtion Integrted System (DRIS) tht llows for determintion of reltive deficiency or excess (Beverly et l., 1984; Wlworth nd Sumner, 1987; Rodriguez et l., 1997; Vrlkshmi nd Bhrgv, 1998). DRIS indictes the reltive limiting order mong the nutrients nlyzed nd hs been pplied successfully to mny nnul nd perennil crops (Wlworth nd Sumner, 1987). A study ws strted in 1996 on citrus t the Todds Rod estte of Croni (1975) Limited with view to determine the mjor fctors influencing yield levels. This report dels with the nutrient levels in soil nd lef nd their reltionship with yield, fruit set nd fruit qulity. This informtion will be useful in developing strtegy for reduction of yield vrition nd the increse of crop yield. Mterils nd methods Tree selection: The cultivrs used were Vlenci ornge (Citrus sinensis), Portugl mndrin (C. reticult) nd Ortnique tngor (C. sinensis x reticult), the former two selected in two fields ech on different soil types (Tble 1). Five fields representing three cultivrs were used nd fifteen trees of similr size were selected from ech field for smpling. At the time of selection, the trees represented groups of high, medium nd low yielders of the 1996/1997 crop. These trees were locted t the Todds Rod estte in centrl Trinidd on Sevill/L Ebrnche (Dystric gleysol) nd Tlpro cly (Eutric vertisol) soils, respectively. Smpling: Soil smpling ws done in 1997 in ll plots t two depths, 0 to 23 cm nd 23 to 46 cm, within the drip circle of the tree. Lef smples were tken rndomly from non-fruiting terminls nd were 4-7 months old. Leves from plots were collected in Jnury - Februry 1997 nd dditionlly for Portugl mndrin in one plot in December 1997 (representtive of 1998). Soil nlyses: Anlysis ws done for ph, K nd P only. Smples were extrcted with mmonium cette nd red for K with flme photometer. Phosphorus ws extrcted with Troug's solution nd red t 660 nm using spectrophotometer. Folir nlysis: Folir nlysis ws conducted on oven dried nd ground mteril by digestion in sulphuric cid for N, P nd K. A smple ws treted with hydroquinone nd mmonium

40 38 Reltionship between soil nd lef minerl nutrient concentrtion nd yield of selected citrus species Tble 1. Field chrcteristics of study plots t Todds Rod Estte Cultivr/Field ID Soil type Are (h) Number of trees Yer of plnting Topogrphy Portugl L'Ebrnche cly Flt Portugl Tlpro cly Slope Vlenci Sevill/L'Ebrnche Flt Vlenci Tlpro cly Slope Ortnique Sevill/L'Ebrnche Flt molybdte nd P red on spectrophotometer t 660 nm. The smple for N ws treted with Nessler's regent nd red with spectrophotometer t 408 nm. The K smple ws diluted with wter nd red with Flme Photometer. Anlysis of trce elements ws done fter dry shing using the method of Richrd (1993). Levels of C, Mg, Fe, Zn, nd Cu were red using n Atomic Absorption Spectrophotometer. Microelement nlyses were done in 1997 nd 1998 on leves from the Portugl mndrin plot t field nd in 1997 only for Vlenci ornge, field Yield nd fruit set: Yield level ws initilly determined subjectively by visul inspection. All levels were ssocited with yield rnges nd were determined by technicins skilled in visul counts nd hving cceptble levels of consistency (Bekele nd Andrews, 1997). In 1998 nd 1999, on-tree fruit, counts were used in determining yield levels in Vlenci nd Ortnique but not Portugl. Percent fruit set ws determined by compring flower counts of lbeled brnches with fruit number fter three months. Fruit qulity: Fruit qulity tests were conducted in order to determine men volume, men fruit weight, percent juice, brix, percent titrtble cidity, number of seeds nd peel thickness. Fruit colour ws grded on scle of 1 to 4 bsed on colour photogrphs of fruit t different mturity nd colour development stges. Sugr:cid rtio ws clculted from the preceding dt. Fruit volume ws determined by wter displcement nd brix determined using n Atgo hnd held refrctometer. The percentge titrtble cidity ws done using method B (Wrdowski et l., 1991) with cidity expressed s percentge nhydrous citric cid. Juice percentge ws clculted s volume over weight of fruit. Dt nlysis: The reltionship between nutrients nd fruit qulity ws explored for the crop recently hrvested or on-tree t the time of lef nd soil smpling. However, the yield nd fruit set dt were tken for the crop following smpling, s fertilizer ppliction tht influenced folir nutrient levels ws pplied too lte in the yer to hve ffected either fruit set or fruit count of the crop used for fruit qulity nlyses. Sttisticl nlysis ws done using Minitb Relese nd DRIS nlysis ws ccomplished with the use of the Potsh nd Phosphte Institute softwre (Version Bet 1.1) vilble on the Brzil (POTAFOS) site ( DRIS mkes use of nutrient concentrtion rtios to interpret tissue nlysis in reltion to dtbse of nlyticl vlues of high yielding trees. Results nd discussion Effect of soil nd lef nutrient levels on yield Vlenci: A regression eqution ws derived for fruit count of Vlenci in field : Fruit count (1998) = % Mg 1451 % P ph Mn ppm %C 85.1 Zn ppm 26.4 P ppm 2. 2 refers to the cm soil depth. P < 0.01, Adjusted R 2 = 99.5 The regression indictes tht minerl nutrition ws the min cuse of yield vrition in Vlenci field 12069, however, yield vrition due to tristez infection hs lso been observed in tht field (Andrews et l., 2005). Positive yield response to correction of deficient lef levels of Mg, C nd Mn hve been reported (Chpmn, 1968; Embleton et l., 1973) nd support the regression formul presented for Vlenci. A previous nutrition survey in citrus producing res in Trinidd reveled widespred symptoms of mgnesium, zinc nd mngnese deficiency (Weir, 1965). According to sufficiency stndrds (Embleton et l., 1973; Jorgensen, 1978) zinc, mgnesium, clcium nd mngnese were deficient or considerbly low in ll three of the yield levels. When yield ws considered s high, medium nd low (3 levels), nlysis of vrince showed significnt differences for lef concentrtions of zinc (Tble 2). An pprisl of lef nutrient levels of Zn t the three yield levels suggests tht reducing levels of this nutrient re ssocited with incresed yield. One possible explntion reltes to the prctice of pplying fertilizer more liberlly to high yielding trees. The pplied N nd K my hve ggrvted the existing Zn deficiency (Reuther nd Smith, 1950). The lef content of Fe showed similr trend s Zn (Tble 2) nd this my hve been induced by high K ppliction which cn be ssocited with Fe vilbility (Chpmn, 1968). Anlysis of vrince for soil nd lef nutrient concentrtions showed no significnt differences between yield levels in field 12071; however, soil phosphorus levels were in the rnge of 4 6 ppm nd re considered low. DRIS nlysis of folir nutritionl levels for high yielding nd low yielding trees in Vlenci field show tht in both the high nd low yielding trees, zinc hd the highest negtive index. The second highest negtive index ws for mngnese in high yielding trees, wheres it ws mgnesium in low yielding trees. The order of limiting nutrients by DRIS nlysis ws zinc, mngnese, mgnesium, iron nd clcium in the high yield trees. No DRIS nlyses were conducted for those fields where lef dt were limited to nitrogen, phosphorus nd potssium only.

41 Reltionship between soil nd lef minerl nutrient concentrtion nd yield of selected citrus species 39 Tble 2. Folir levels of nitrogen, iron nd zinc in Februry 1997 in reltion to subsequent yield levels in Vlenci ornge field Yield level-1998 (fruit count) N (%) Zn (ppm) Fe (ppm) Low (<301) Medium ( ) High (>450) Probbility 0.09 NS NS NS not significnt (P = 0.05) Portugl: Soil nd lef nutrient concentrtions tken in Jnury 1997 showed no significnt difference when compred t the three yielding levels of the 1997/1998 crop of Portugl in field Field showed significnt difference (P=0.02) for percent lef phosphorus only, (Tble 3). Soil levels of phosphorus in field showed no significnt difference t the three yield levels but the concentrtion is considered low. The optimum rnge of phosphorus in citrus leves is % (Jorgensen, 1978) nd % (Embleton et l., 1973). The smples from Portugl field in 1997 showed mens of 0.12, 0.14 nd 0.17% (P=0.02) ssocited with trees giving low, medium nd high yields, respectively in the 1997/1998 crop. This suggests tht low yielding trees only were low or deficient in phosphorus. Anlysis of vrince for 1998 lef smples for field showed no significnt differences for ny nutrient t the three yield levels. No regression nlysis ws done s dt were not tken on fruit counts. DRIS nlysis of lef nutrient dt of field in 1997 indicted tht the following nutrients limiting yield, in order of importnce, were clcium, zinc, potssium, mngnese nd iron. When sufficiency rnges were considered (Embleton et l., 1973; Jorgensen, 1978) clcium nd zinc were deficient or low t ll yielding levels. DRIS nlysis of low yielding trees indicted zinc nd clcium s the limiting nutrients. In 1998, the DRIS limiting nutrients, in order of importnce, were clcium, zinc, copper nd mngnese. Appliction of sufficiency rnge stndrds resulted in the sme list of limiting nutrients except for mngnese for which the concentrtion ws close to criticl. The order nd mgnitude of the DRIS negtive indices ws not the sme for Portugl trees tht were high or low yielding, suggesting tht yield ws lower when certin minerl nutrient limittions were more cute. Tble 3. Soil nd lef concentrtions of phosphorus in Portugl trees of field (Jnury 1997) Yield level 1998 (fruit count) P ppm (0-23 cm) P ppm (23-46 cm) P in leves (%) Low (<301) Medium( ) High (>800) Probbility NS NS NS not significnt (P=0.05) Ortnique: Only soil potssium content t the cm depth ws significntly different for the three yield levels (Tble 4) but the reltionship ws not liner. No dequte regression eqution ws derived. When dt for fields nd were combined becuse of similr soil type (Tble 1) soil ph nd lef nitrogen levels becme significnt indictors of yield (Tble 5). Yield level is inversely relted to ph even though the upper limit of men ph is 6.8 (Tble 5). The high ph my hve contributed to unvilbility of zinc seen in Vlenci field (Tble 2). The high nitrogen levels (Tble 5) my be reflecting phosphorus deficiency (Br-Akiv et l., 1968; Rbe nd Lovtt, 1986) or clcium deficiency (Weir, 1969) which would explin the ssocited lower yield level. Tble 4. Men content of soil nutrients in field in reltion to the following yer s yield levels (1997/1998 crop) (Februry 1997) Yield level 1998 (fruit count) K ppm (0 23 cm) K ppm (23-46 cm) P ppm (23-46 cm) Low (<251) Medium ( ) High (>300) Probbility Tble 5. Effect of lef nd soil nutrient chrcteristics on yield level of Vlenci nd Ortnique grown on Sevill/L Ebrnche clys Yield level ph (0-23 cm) ph (23-46 cm) Folir nitrogen (%) Low Medium High Probbility Effect of soil nd lef nutrient levels on percent fruit set: The correltions in Tble 6 re more reveling thn ANOVA or regression nlyses s it is cler tht copper nd iron nd the clcium/mgnesium rtio (indictive of clcium nutrition) were positively relted to percent fruit set in Vlenci nd Portugl. Within the Croni (1975) Limited environment, lef nutritionl dt of Jnury in the yers 1997 nd 1998 reflect the nutritionl sttus tht cn hve direct effect on flowering nd fruit set for the next crop, since no intervening fertilizer ppliction occurred before August (Croni Reserch Sttion, 1999). Only one ppliction of fertilizer ws given in those yers s supplies were received lte. By tht time flowering would hve occurred nd fruit drop stbilized within eight weeks therefter (Andrews, 1996). The correltion nlyses (Tble 6) showed positive reltionship between concentrtion of copper, iron nd clcium/mgnesium rtio in the leves nd fruit set in two cultivrs. Incresed vilbility of minor element concentrtion hs been ssocited with incresed yield under conditions of deficiency (Chpmn, 1968; Alv nd Obrez, 1998) but the effect on percent fruit set hs not been reported. There is generlly decrese in percent fruit set with increse in flowering (Deidd nd Agbbio, 1977; Grci-Luis et l., 1988). However, Becerr nd Gurdiol (1984) hve determined tht, t the sme flowering intensity, high yielding trees hve higher fruit set percent thn low yielding trees. Ortnique lef moisture content mesured in Februry 1997 (Tble 6) could not hve hd direct effect on fruit set some five months lter but my hve been relted to the tree s cpcity for crbohydrte production during tht period nd ffect fruit set indirectly.

42 40 Reltionship between soil nd lef minerl nutrient concentrtion nd yield of selected citrus species Effect of soil nd lef nutrient levels on fruit qulity Nitrogen: The effect of nitrogen ws common for Portugl nd Vlenci nd the reltionship with fruit weight ws generlly negtive (Tble 7), s seen in the derived regression equtions. Significnt nitrogen effect ws not observed in 1997 for Portugl, field 12050, but it ws present in 1998 for lef concentrtion in reltion to fruit weight (Tble 7). The negtive reltionship between lef nitrogen content nd fruit size is consistent with the other reported experiences (Embleton et l., 1973). Phosphorus nd potssium: Soil or lef phosphorus ppered to be importnt in ll three cultivrs for vrious spects of fruit qulity, especilly for peel thickness where the reltionship ws inconsistently positive nd negtive. The effect of potssium on fruit qulity ws wek nd inconsistent. Mgnesium: Mgnesium effect ws limited to fruit weight nd seed number in Vlenci nd to fruit volume nd percent cidity in Portugl in 1998 (Tble 7). This positive effect Tble 6. Person s correltion of citrus percent fruit set with soil nd lef nutrition vribles t Todds Rod estte Cultivr/ Field ID Vlenci Portugl Portugl Vlenci Ortnique Fruit set dte Nutrition vrible Correltion of mgnesium content on juice cidity in the Portugl crop (Tble 7) confirms to the report by Chpmn (1968). The Mg/Zn reltionship with seed number in Vlenci (Tble 7) is probbly due to the more influence of mgnesium thn zinc s folir mgnesium deficiency symptoms re more pronounced in seedy cultivrs (Chpmn, 1968). The removl of folir mgnesium for seed development my be significnt even though Vlenci is not seedy cultivr. Microelements: Effects were observed for the two fields in which folir nlyses for microelements ws done (Tble 7). Except for the effect of mngnese on peel thickness, microelement effects were not consistent by ttributes for the different fields nd yers. The effect of mngnese on peel thickness of Portugl mndrin ws quite pronounced (Tble 7), s the contribution to vribility ws 50.25% in 1997 nd 17.95% in This reltionship hs not been previously reported lthough positive correltion ws found between fruit size (dimeter nd fresh weight) nd flower content of mngnese in Vlenci ornge (Pestn et l., 2001). Some locl growers believe tht fruit with thick peel re fvoured by buyers becuse of less dherence to the endocrp. All fields showed some form of nutrient deficiency tht would ffect yield. There is generl evidence to support the ide tht yield vrition is ssocited with vrition in nutritionl fctors. The reltionship between lef concentrtion of mngnese nd peel thickness in Portugl mndrin suggested by preliminry dt should be investigted further, including the ttrctiveness of peel thickness to the locl mrket. References Ali, A.G., V. Thomsos, K. Morton-Gittens, W. Augustin-Coryt, K. Beckles nd G. Blly, Report No. 3, Agriculture in County St. George Lnd Cpbility Studies Phse II Trinidd nd Tobgo, Ministry of Plnning nd Development, Trinidd nd Tobgo. Alv, A.K. nd T.A. Obrez, By-product iron-humte increses tree growth nd fruit production of ornge nd grpefruit. Hortscience, 33(1): Andrews, L Germplsm improvement in citrus. Croni Reserch Sttion Technicl Report, 31(2): 3-9. Andrews, L Fruit retention in citrus. Croni Reserch Sttion Technicl Report, 33: 175. Tble 7. Effect of soil nd lef nutrient fctors on fruit qulity of Vlenci ornge nd Portugl mndrin Cultivr Field ID Yer Nutrient fctor Regression eqution Adjusted R 2 (%) Probbility Vlenci ornge N Fruit weight = %N 455 %Mg 5.73 P ppm Vlenci ornge N Fruit weight = ph 31.7 %N Portugl mndrin N Fruit weight = %N %P Portugl mndrin N Fruit weight = %N %K Fe/Zn Vlenci ornge Mg Fruit weight = %N 455 %Mg 5.73 P ppm Vlenci ornge Mg Number of seeds = Mg/Zn 76.3 <0.001 Portugl mndrin K/Mg Fruit volume = K/Mg %K Portugl mndrin Mg Acidity (%) = %Mg Mn ppm Portugl mndrin Mn Peel thickness = P ppm Mn ppm 85.5 < %N K ppm Portugl mndrin Mn Peel thickness = Mn ppm refers to the cm soil depth coefficient Probbility 1997 Fe Cu C/Mg Fe Cu C/Mg % K ph 0 23 cm (soil) ph cm (soil) K(ppm) cm % Wter <0.001 K (ppm) cm Log trnsformtion of % set dt 2 % Wter refers to lef moisture content P (ppm) cm

43 Reltionship between soil nd lef minerl nutrient concentrtion nd yield of selected citrus species 41 Andrews, L., R. Phelps nd R.A.I. Brthwite, Reltionship of citrus yield nd virus infection in Trinidd. Journl of Applied Horticulture, 6(2): Andrews, L., G. White nd P. Jordn, The use of field ttributes dtbse for nlysis of citrus yields t Todds Rod. NIHERST, 11th Annul Conference on Agriculturl Reserch nd Development: Br-Akiv, A., V. Hiller nd J. Ptt, Effects of phosphorus nd chicken mnure on yield, fruit qulity nd lef composition of grpefruit trees. Proceedings of the Americn Society for Horticulturl Science, 93: Becerr, S. nd J.L. Gurdiol, Inter-reltionship between flowering nd fruiting in sweet ornge, cultivr Nvelin. Proceedings of the Interntionl Society of Citriculture, 1: Bekele, I. nd L. Andrews, Investigtion into the ccurcy of visul on-tree fruit counts for estimting citrus production. Ntionl Institute of Higher Eduction Reserch Science nd Technology. 9th Seminr on Agriculturl Reserch nd Development. Port of Spin, NIHERST. p 3-9. Beverly, R.B., J.C. Strk, J.C. Ojl nd T.W. Embleton, Nutrient dignosis of 'Vlenci' ornges by DRIS. Journl of the Americn Society of Horticulturl Science, 109(5): Croni Reserch Sttion, Citrus - Reserch nd Production Review nd Plnning Workshop. Croni (1975) Limited. Chpmn, H.D The minerl nutrition of citrus. The Citrus Industry. Volume 2. Eds. W. Reuther, L.D. Btchelor, nd H.J. Webber. Division of Agriculturl Sciences, University of Cliforni, pp Cooper, St. G Citrus production in the Snt Cruz nd Diego Mrtin vlleys in Trinidd. Journl of the Agriculturl Society of Trinidd nd Tobgo, 56: Deidd, P. nd M. Agbbio, Some fctors influencing flowering nd fruit-set of Clementine mndrin. Proceedings of the Interntionl Society of Citriculture, 2: Dey, J.K. nd D.D. Singh, Nutritionl sttus of helthy nd declining citrus (Citrus reticult) orchrds. Indin Journl of Agriculturl Sciences, 68(2): Embleton, T.W. nd C.K. Lbnusks, Plnt nlysis s bsis for the minerl fertiliztion of citrus orchrds. Proceedings of 21st Interntionl Horticulturl Congress, 1: Embleton, T.W., W.W. Jones, C.K. Lbnusks nd W. Reuther, Lef nlysis s dignostic tool nd guide to fertiliztion.in: The Citrus Industry, Volume 3. (W. Reuther, Ed.). University of Cliforni, Division of Agriculturl Sciences. pp Grci-Luis, A., F. Fornes, A. Snz nd J.L. Gurdiol, The regultion of flowering nd fruit set in citrus: reltionship with crbohydrte levels. Isrel Journl of Botny, 37: Jorgensen, K.R Lef nd soil nlysis guide to fertilizing citrus. Queenslnd Agriculturl Journl, 104: Lucie-Smith, M.N Citrus production in Trinidd. Port of Spin, Trinidd, Port of Spin Gzette Limited. Pestn, M., P.J. Corrier, A. de Vrennes, J. Abd nd E.A. Fri, The use of florl nlysis to dignose the nutritionl sttus of ornge trees. Journl Plnt Nutrition, 24(12): Rbe, E. nd C.J. Lovtt, Reltive phosphorus dependency of citrus rootstocks is reflected in lef nitrogen metbolism. Journl Americn Society Horticulturl Science, 111(6): Reuther, W. nd P.F. Smith, A preliminry report on the reltionship of nitrogen, potssium nd mngnese fertiliztion to yield, lef composition nd the incidence of zinc deficiency in ornges. Proceedings Americn Society Horticulturl Science, 56: Richrd, J.E Chemicl chrcteriztion of plnt tissue. In: Soil smpling nd methods of nlysis. (M. Crter, Ed.). Boc Rton, Lewis Publishers pp Rodriguez, O., E. Rojs, nd M. Sumner, Vlenci ornge DRIS norms for Venezuel. Communictions in Soil Science nd Plnt Anlysis 28 (15/16): Vrlkshmi, L.R. nd B.S. Bhrgv, Plnt nutrient sttus nd lef dignostic norms for cid lime (Citrus urntifoli). Indin Journl Agriculturl Sciences, 68(7): Wlworth, J.L. nd M.E. Sumner, The Dignosis nd Recommendtion Integrted System (DRIS). Advnces in Soil Science, 6: Wrdowski, W., J. Soule, J. Whighm nd W. Greirson, Florid citrus qulity tests. SP 99. Ginesville, University of Florid. Weir, C.C A survey of the minerl nutrition of vlenci ornge in Trinidd. Experimentl Agriculture, 1: Weir, C.C Nutrient element blnce in citrus nutrition. Plnt nd Soil, 3:

44 Journl of Applied Horticulture, 8(1): 42-44, Jnury-June, 2006 Studies with thidizuron on the vse life of cut rose flowers Appl Journl Esmeil Chmni*, Donld E. Irving, Dryl C. Joyce nd Mos Arshd** Centre for Ntive Floriculture, School of Agronomy nd Horticulture, The University of Queenslnd, Gtton, Queenslnd 4343, Austrli; * Horticulture Deprtment, Agriculture Fculty, Mohghghegh Ardebil University, Ardebil, Irn, **Horticulture Deprtment, Agriculture Fculty, Mhbd Azd University, Irn Abstrct Effects of posthrvest-pplied thidizuron (TDZ) on the vses life of 7 rose cultivrs (Ros hybrid L.) were investigted. Cut rose flowers were pulse-treted with TDZ for 24 hours t 22 o C. Appliction of 20, 60 nd 100 µm TDZ to Memoire rose did not ffect vse life when compred with the control (0 μm TDZ). Similrly, pulse tretments with 10 µm TDZ did not ffect the vse life of cvs. Chmpgne, Lser, Mgnum, Neon nd Tresor 2000 roses compred with their untreted controls, but did increse the vse life of First Red by 2 dys (+11.5 %). Lterl shoot development ws common side effect of TDZ tretment. Key words: Posthrvest, thidizuron, vse life, Ros hybrid, lterl shoot, pulse tretment, cut flower, longevity Introduction Ros hybrid is mong the most commercilly importnt flowers, with n nnul vlue of $10 billion (Gutermn et l., 2002). Roses re used s cut flowers, flowering pot plnts nd grden plnts. The vse life of cut rose flowers is reltively short (Hung et l., 2002; Mrisen nd Benning, 2001), nd thus, posthrvest technologies for improving the keeping qulity of roses re sought nd pplied. Increse in the posthrvest life of cut flowers following cytokinin tretments hs been reported (Hicklenton, 1991; Lukszewsk et l., 1994; Pull nd Chntrchit, 2001). Bosse nd Vn Stden (1989) reported tht DHZ (dihydrozetin), used s pulse tretment for 6-24 hours t concentrtion of M, significntly delyed crntion flower senescence. An increse in the longevity of crntion flowers by 15 dys ws found t M DHZ. Lukszewsk et l. (1994) showed tht exogenous ppliction of trns-zetin, trns-zetin riboside, 2iP (isopentenyldenine) nd 2iPA (isopentenyldenosine) in holding solutions delyed rose senescence by fold. Their dt lso showed tht zetin nd zetin riboside were most effective on roses t M nd prolonged longevity by 1.3-fold. Thidizuron (N-phenyl-N-1, 2, 3-thidizol-5-ylure) is uretype cytokinin tht is reltively novel compound for treting cut flowers. It is more commonly used t high concentrtion s cotton defolint (Mlik et l., 2002) nd t low concentrtion for regenertion in tissue culture (Singh nd Syml, 2001). TDZ is round times more ctive in inducing cytokinin-like effects thn common cytokinins (Genkov nd Iordnk, 1995). Tretments with TDZ delyed lef yellowing in cut Alstroemeri (Ferrnte et l., 2002), nd ws most effective t 10 µm s pulse tretment or t 1 µm s continuous (vse) tretment. TDZ tretments of less thn 50 µm lso reduced flower shedding nd induced dditionl flower buds during vse life of cut phlox inflorescences (Snkhl et l., 2003). TDZ tretment of cut chrysnthemum nd tulip inhibited lef yellowing but did not enhnce flower qulity (Ferrnte et l., 2003). The present experimenttion ws conducted to determine the effects of TDZ on the vse lifes of seven cut rose cultivrs. Mterils nd methods Rose flower stems were hrvested from greenhouses t the tight bud stge nd trimmed to 40 cm length nd 3 leves. Posthrvest experiments were crried out under vse life evlution room conditions of 22 ± 1 C, % reltive humidity nd 12 h photoperiod with 15µmol m -2 s -1 irrdince from cool white florescent lmps. The Memoire rose cultivr for experiment 1 ws hrvested in December 2003 from ner Tehrn, Irn. For experiment 2, Chmpgne, Neon, Tresor 2000, Lser nd Mgnum cultivrs were hrvested in Februry 2004 from ner Brisbne, Austrli. First Red roses for experiment 3 (My 2004) were hrvested from different grower ner Brisbne. Vse life ws recorded s the time in dys fter hrvest (dy 0) tht flowers reched the end of their longevity due to bent neck or dvnced signs of fding on ll petls (Myk nd Hlevy, 1974; Lio et l., 2000). Experiment 1: TDZ (Sigm Chemicl Co.) ws dissolved in 2 ml liquots of 1M KOH, mde up with distilled wter to concentrtions of 0, 20, 60, nd 100 µm nd neutrlized with 2 ml 1M HCL (Ferrnte et l., 2002). Cut Memoire flowers (white) were then pulse-treted for 24 h t 20 C. Distilled wter ws the control pulse tretment. The post-pulse tretment vse solution ws 200 mg L -1 HQS (hydroxyquinoline sulphte) in distilled wter (Lio et l., 2000). Flower vse life ws ssessed dily nd recorded for ech of 7 single bloom replictes per tretment. A completely rndomised design ws dopted. Dt were nlyzed by one wy ANOVA with Minitb Relese 13.1 for Windows (Minitb Inc.). Experiment 2: Chmpgne (white), Neon (deep pink), Lser (pink), Tresor 2000 (yellow) nd Mgnum (red) were pulsed with 10 µm TDZ for 24 h t 22 C. Distilled wter ws the control pulse tretment. The post-pulse tretment vse solution ws distilled wter contining 10 μl L -1 vilble chlorine supplied

45 Studies with thidizuron on the vse life of cut rose flowers Vse life (dys) b b b 11 Fig. 1. Lterl shoot development (rrowed) during vse life in cut Memoire (left photogrph) nd First Red (right photogrph) roses pulse-treted with 10 µl L -1 TDZ. s dichloroisocynuric cid (DICA) (Joyce et l., 2000) Vse life ws ssessed dily. Ech replicte ws 5 individul stems of ech cultivr for control nd TDZ tretments. A completely rndomised design ws dopted, nd t-tests (Minitb Relese 13.1) were used for dt nlysis for ech cultivr. Experiment 3: First Red roses were treted with 10 µm TDZ, 4% w/v sucrose nd both in combintion s pulses for 24 h t 22 C. Distilled wter ws the control pulse tretment. Vse life ws ssessed dily. Ten replicted stems were used for ech tretment. A completely rndomised design ws dopted, nd dt were nlysed by ANOVA using SAS procedures (SAS Institute 1998). The lest significnt difference (P = 0.05) ws clculted to llow comprisons mong tretment mens. Results nd discussion Pulse-tretment with TDZ t concentrtions of 0, 20, 60 nd 100 µm did not differentilly ffect the vse life of cut Memoire rose. The men (±SD) vselife ws 14.6±0.25 dys. A side-effect of TDZ tretments ws enhnced lterl shoot development (Fig. 1). Pulse-tretment with TDZ t 10 µm did not ffect the vse lives of Chmpgne (18 ± 1.4 dys), Lser (16 ± 1.3 dys), Mgnum (19 ± 0.7 dys), Neon (18 ± 0.3 dys) nd Tresor 2000 (16 ± 0.3 dys). As observed with cv. Memoire, lterl shoot development on TDZ-treted stems ws observed for these 5 cultivrs. Pulse-tretment of First Red roses with 10 µm TDZ did increse the vse life by 1.5 dys (Fig. 1), nd lso encourged lterl shoot development s in Memoire (Fig. 1). Pulse-tretment with 4% (w/v) sucrose lso incresed flower vse life by 1 dy. TDZ pulsing in combintion with sugr pulsing hd synergistic effect on vse life (Fig. 2). Pull nd Chntrchit (2001) reported tht different cultivrs of nthurium hd different vse life responses to benzyldenine tretment. The responses rnged from 20% reduction to 2.5- fold increse in vse life. A lck of response to benzyldenine my hve been due to high nturl cytokinin levels. Perhps in cultivrs with nturlly low levels of cytokinin, TDZ tretment might increse vse life. Further experimenttion is required to exmine this proposition for cut roses. 10 Fig. 2. Effect of TDZ (10 μm), sucrose (4% w/v), nd TDZ plus sucrose in combintion (ll pplied s pulse-tretments) on the vse life of First Red rose. Brs topped with the sme letter re not significntly different (P<0.05). In summry, TDZ pulse-tretments hd no effect on vse life of severl cut roses, but did increse the vse life of cv. First Red, one of the more commercilly importnt cut rose cultivrs. As consistent side effect, TDZ tretments promoted shoot development from lterl vegettive stem buds. Acknowledgments We thnk Aln Lisle for dvice nd ssistnce in nlysing the dt. We lso thnk Mr Brin Shnnon for donting flowers used in one experiment. References Control TDZ Sucrose TDZ + Sucrose Tretments Bosse, C.A. nd J. Vn Stden, Cytokinins in cut crntion flowers. V. Effects of cytokinin type, concentrtion nd model of ppliction on flower longevity. Journl Plnt Physiology, 135: Ferrnte, A., D.A. Hunter, P.H. Wesley nd M.S. Reid, Thidizuron potent inhibitor of lef senescence in Alstroemeri. Posthrvest Biology nd Technology, 25: Ferrnte, A., F. Tognoni, A. Mensuli-Sodi nd G. Serr, Tretment with thidizuron for preventing lef yellowing in cut tulips nd chrysnthemum. Act Hoticulture, 624: Genkov, T. nd I. Iordnk, Effect of cytokinin-ctive phenylure derivtives on shoot multipliction peroxidse nd superoxide dismutse ctivities of in vitro cultured crntion. Bulgrin Journl of Plnt Physiology, 21: Gutermn, I., M. Shlit, N. Mend, D. Piestun, M. Dfny-Yelin, G. Shlev, E. Br, O. Dvydov, M. Ovdis, M. Emnuel, J. Wng, Z. Adm, E. Pichersky, E. Lewinsohn, D. Zmir, A. Vinstein nd D. Weiss, Rose scent: genomics pproch to discovering novel florl frgrnce relted genes. Plnt Cell, 14: Hicklenton, P.R GA 3 nd benzylminopurine dely lef yellowing in cut Alstroemeri stems. HortScience, 26(9): Hung, K.L., L.J. Lio, R.S. Shen, W.S. Chen nd Y.H. Lin, The synergistic effect of mleic cid hydrzide (1,2-dihydro-3,6- pyridzinedione) nd sucrose on vse life of cut roses. Austrlin Journl Experimentl Agriculture, 42: Joyce, D.C., S.A. Mer, S.E. Hetherington nd P. Jones, Effects of cold storge on cut Greville Sylvi inflorescences. Posthrvest Biology nd Technology, 18: Lio, L.J., L.J. Lio, K.L. Hung, W.S. Chen nd Y.M. Cheng, Posthrvest life of cut rose flowers s ffected by silver thiosulfte nd sucrose. Botnicl Bulletin Acdemic Sinic, 41:

46 44 Studies with thidizuron on the vse life of cut rose flowers Lukszewsk, A.J., J. Binco, P. Brthe nd M.T. Le Pge-Degirry, Endogenous cytokinins in rose petl nd the effect of exogenously pplied cytokinins on flower senescence. Plnt Growth Regultion, 14: Myk, S. nd A.H. Hlvey, The ction of kinetin in improving the wter blnce nd delying senescence process of cut rose flowers. Physiologi Plntrum, 32: Mlik, M.N.A., U.D. Shbb, M.I. Mkhdom nd F.I. Chudhry, Use of thidizuron s hrvest id in erly nd lte plnted cotton. Interntionl Journl Agriculture nd Biology, 4: Mrissen, N. nd J. Benning, A nursery comprision on the vse life of the rose First Red : Effects of growth substnces. Act Horticulture, 543: Pull, R.E. nd T. Chntrchit, Benzyldenine nd the vse life of tropicl ornmentls. Posthrvest Biology nd Technology, 21: Snkhl, N., W.A. Mcky nd T.D. Dvis, Reduction of flower bscission nd lef senescence in cut phlox inflorescences by thidizuron. Act Hoticulture, 628: Singh, S.K. nd M.M. Syml, A short pre culture sok in thidizuron or forchlorfenuron improves xillry shoot prolifertion in rose micropropgtion. Scienti Hoticulture, 91:

47 Journl of Applied Horticulture, 8(1): 45-49, Jnury-June, 2006 Growth behviour of pple cctus (Cereus species) in hyper-rid environment Appl Journl Ahmed A. ElObeidy Deprtment of Fruit Horticulture, Fculty of Agriculture, Ciro University, Giz, Egypt. E-mil: Abstrct Introducing new crop with high wter use efficiency into the hyper-rid environment will prticipte in curb rising demnd of wter. Apple cctus (Cereus species) chrcteristics fit with most of the requirements of drought tolernt crop with very high wter-use efficiency. Severl Cereus species were introduced into desert, chrcterized with rre rinfll nd high tempertures. The introduced fruiting species were Cereus hexgonus, C. peruvinus, C. peruvinus monstrose nd C. vlidus. C. pchnoi ws introduced s rootstock. C. peruvinus cuttings survived storge up to eight months. Horizontl position of the cuttings during storge encourged the development of lterl brnches. Plnts were propgted by cuttings, cclimtized nd then trnsplnted into the field in the desert. Growth nd development of the introduced species were ssessed under the new environment. All the introduced species grew successfully except C. vlidus tht ws eliminted during the first summer. C. peruvinus monstrose ws chrcterized with drmtic contrction of the stem in the dry condition. The min stem of C. peruvinus, C. peruvinus monstrose, C. hexgonus nd C. pchnoi grew 9.2, 10.2, 8.1 nd 15 cm/month, respectively. C. peruvinus developed the highest number of sprouts. C. Peruvinus, C. peruvinus monstrose nd C. hexgonus unite with the C. pchnoi to form successful grfts with percentge of success 80, 53 nd 86.5, respectively. C. vlidus filed completely to unite with C. pchnoi. C. peruvinus nd C. peruvinus monstrose were the most promising in the new hyper-rid environment in terms of dptbility nd helthy growth. Key words: Apple cctus, Cereus, drought resistnce, hyper-rid environment, wter use efficiency. Introduction As nturl hzrd, drought imposes differentil vulnerbility on society in the hyper-rid region (Wilhite, 2000). Introducing new crops with high wter use efficiency in the hyper-rid environment will prticipte in curb rising demnd of wter (ElObeidy, 2004). Apple cctus (Cereus sp.) chrcteristics fit with most of the requirements of drought tolernt crop with very high wter-use efficiency. Apple cctus hs physiologicl nd morphologicl methods of exploiting environments tht would soon desiccte other plnt species (Nerd et l., 1988; Wllce nd Gibson 2002). In ddition, pple cctus is chrcterized with crssulcen cid metbolism pthwy tht improve wter-use efficiency (Mld et l., 1999; Cushmn nd Borlnd, 2002). Apple cctus, populr group of ccti from South Americ,consists of bout 60 species. These vigorous-growing ccti re esy to grow in mrginl, infertile, dry lnds where common crops fil (Felger nd Moser, 1976; Wllce nd Gibson 2002). Apple cctus produces unique fruits tht re thornless nd vry in skin colour from violet-red to yellow. The flesh, which is the edible prt of the fruit, is white nd contins smll, edible, nd crunchy seeds (Felger nd Moser, 1976; Nerd et l., 2002). Apple cctus fruits my be eten fresh, dried, or cn be mde into juice (Scheinvr, 1985). Apple cctus fruits cn offer commercil opportunities (Felger nd Moser, 1976). However, t present, it is n unexplored, underutilized fruit (Morton, 1987; Inglese et l., 2002). The im of this study ws to investigte vegettive growth, development, storing bility of cuttings nd grfting success of severl pple cctus species introduced in hyper-rid desert. Mterils nd methods Plnt mteril: Cuttings from four Cereus species were obtined from privte nurseries of Cliforni, USA. The introduced species were C. hexgonus, C. pchnoi, C. peruvinus nd C. vlidus. The subspecies C. peruvinus monstrose ws lso introduced. Forty cuttings with 30 cm length from ech species were used for propgtion. Forty more cuttings of C. peruvinus nd C. peruvinus monstrose were used in the storge experiment. Cutting heling nd storge: Thirty scions of ech of C. hexgonus, C. pchnoi, C. peruvinus, C. peruvinus monstrose, C. vlidus nd C. pchnoi were grfted on C. pchnoi. Cuttings were llowed to hel in dry re t 30 C for ten dys. Other cuttings of C. peruvinus nd C. peruvinus monstrose were stored t 35 C up to eight months. Propgtion by cuttings: Pots (25 cm in dimeter) were filled to bout one-third full of potting mix consists of 1 prt pet moss: 2 prts snd. Ech cutting ws plnted in the center of pot. Pots were completed to two-third full with the potting mix. Pots were wtered nd kept in the greenhouse t 27 C. Propgtion by grfting: Plnts of C. hexgonus, C. peruvinus, C. peruvinus monstrose, C. vlidus nd C. pchnoi, grown in 25 cm dimeter pots, were used s scions. C. pchnoi plnts grown in 25 cm dimeter pots were used s rootstocks. The dimeter of the stems of C. hexgonus, C. pchnoi, C. peruvinus, C. peruvinus monstrose nd C. pchnoi plnts ws 9-12 cm. However the dimeter of the stems of C. vlidus plnts ws 5 cm. With shrp sterile knife, the top of the rootstocks ws cut off 20 cm bove the soil surfce. The bottom of the scion plnts ws cut off 15 cm from the top. The scions nd rootstocks

48 46 Growth behviour of pple cctus (Cereus species) in hyper-rid environment were joined together by lining up the two cylindricl prts nd gently rotted to squeeze out ny ir bubbles. Two rubber bnds of pproprite size were ffixed over the scion nd under the bottom of the pot. Rubbers could exert stedy pressure on the scion, pressing it ginst the stock to support the grft while the cut edges hel. The two rubber bnds were plced over the scion t 90 degrees from ech other, in order to prevent the scion from shifting. The grfts were supported with stkes, then kept in the greenhouse. The rubber bnds were removed fter two months of grfting. Orchrd loction: The orchrd ws locted in Al-Ain desert, United Arb Emirtes (UAE), chrcterized with high tempertures nd rre rinfll. Mximum temperture reches 49 C during the summer nd verge nnul rinfll is 77mm. Soil nd irrigtion wter smples from the orchrd were tken to determine slinity. Trnsplnting: In the fll (15th September), plnts propgted by cuttings were tken from the greenhouse to the shde for ten dys cclimtiztion. The plnts were then trnsplnted into the orchrd. Hole ws dug bout 15 cm wider thn the continer nd couple inches deeper. The plnt ws slipped crefully out of its continer. Hevy gloves nd 40 cm forceps were used to void injuring the technicins or plnts. The plnt ws plced into the hole nd soil ws firmed lightly round. Irrigtion nd fertiliztion: The source of irrigtion is the underground quifer. Drip irrigtion system ws used in the orchrd. Two liters of wter ws dded per plnt during two hours every two dys. Fertilizer 12:4:24 (150g per plnt) ws pplied one month fter trnsplnting into orchrd, then every two months during the growing seson. Dt collection nd nlysis: Dt were collected in two successive sesons (2001/2002 nd 2002/2003) on plnt survivl, stem length nd dimeter nd sprout or lterl brnch development during the growing seson (October My). Other observtions on growth behviour were recorded. In storge experiment, ech cutting ws weighed t the beginning nd every month during the storge period. In grfting experiment the heling nd success of grfts ws observed. Completely rndomised design ws used for sttisticl nlysis nd men comprisons were mde using Duncn's Multiple Rnge test t 5% significnt level (Duncn, 1955). Results nd discussion Four pple cctus species were introduced into hyper-rid desert in the UAE. Orchrd soil found to be sndy with 2200 ppm slinity. Slt concentrtion in the irrigtion wter rnged from 1800 to 2000 ppm. Cuttings of the introduced Cereus species were llowed to dry in wrm, dry re for ten dys to permit the cut surfce to hel nd develop cllus. The cllus helps prevent rotting during propgtion (Hrtmnn et l., 2001). The introduced species grew in the hyper-rid desert successfully except C. vlidus tht ws eliminted during the first summer. All the plnts of C. peruvinus nd C. peruvinus monstrose survived in the new environment. The min stem of C. peruvinus showed 9.2 cm increse in height (Fig. 1) nd 0.9 cm increse in dimeter per month (Fig. 2). Five sprouts developed per plnt (Figs. 3 nd 4). C. peruvinus, found in southestern cost of South Americ, hs lredy ttrcted ttention s potentil fruit-crop (Bckeberg, 1984). The fruit is smooth nd spineless nd vries from yellow to deep red (Weiss et l., 1993). C. peruvinus ws found to be very promising fruiting cctus (Morton, 1987; Inglese et l., 2002). C. peruvinus cuttings survived storge up to eight months. Cuttings lost bout 50% of the originl weight t the end of the storge period (Fig. 5). Horizontl position of the cuttings during storge encourged the development of lterl brnches (Fig. 6). Lterl brnching verged t four brnches per cutting. C. peruvinus monstrose cuttings did not survive more thn three months of storge (Fig. 5). In the field, the min stem of C. peruvinus monstrose incresed 10.2 cm in length nd 1.1 cm in dimeter per month (Figs. 1 nd 2) nd four sprouts developed per plnt. C. peruvinus monstrose exhibited drmtic expnsion nd contrction of the stem s wter vilbility chnged (Fig. 7). Such mechnism enbles the stem to conserve wter nd survive in dry, hot conditions (Wllce nd Gibson, 2002). Over millions of yers, through nturl selection, only the most dpted species survived in desert environment (John, 2001). Increse in stem length (cm/month) C. peruvinus C. peruvinus monstrose C. hexgonus C. pchnoi C. vlidus b c d 0 Species Fig. 1. Rte of the increse in min stem length of the introduced Cereus species. Columns lbeled with the sme letter re not significntly different (P>0.05). Increse in stem dimeter (cm/month) b C. eruvinus monstrose C. peruvinus C. hexgonus C. pchnoi C. vlidus c 0.0 Species Fig. 2. Rte of the increse in min stem dimeter of the introduced Cereus species. Columns lbeled with the sme letter re not significntly different (P>0.05). b e

49 Growth behviour of pple cctus (Cereus species) in hyper-rid environment 47 Number of sprouts developed per plnt b C. peruvinus C. peruvinus monstrose C. hexgonus C. pchnoi C. vlidus 0 Species Fig. 3. Averge number of the developed sprouts per plnt during the first two sesons. Columns lbeled with the sme letter re not significntly different (P>0.05). Generlly, the fmily of Cctcee, contins mny plnts tht re highly dptble to new environment nd ble to tolerte drought, het nd sline soil. Apple cctus, s member in the fmily Cctcee hs devolved to be well dpted to extremely xeric conditions (Museth, 2000; John, 2001). The pthwy of photosynthesis in Cereus is the crssulcen cid metbolism in which stomt open t night (when evportion rtes re usully lower) nd re usully closed during the dy. The CO 2 is converted to n cid nd stored during the night. During the dy, the cid is broken down nd the CO 2 is relesed to RUBISCO for photosynthesis. When conditions re extremely rid, Cereus plnts cn just leve their stomt closed night nd dy. Oxygen given off in photosynthesis is used for respirtion nd CO 2 given off in respirtion is used for photosynthesis (Winter nd Smith, 1996). Importnce of crssulcen cid metbolism species increses in the fce of expnsion of desertifiction round the world (Cushmn nd Borlnd, 2002). C. hexgonus is nother introduced Cereus species with 73% of survivl in the new rid environment. The min stem growth rte of C. hexgonus ws 8.1 nd 0.8 cm/month in height nd dimeter, respectively (Figs. 1 nd 2). Ech C. hexgonus plnt d c developed two sprouts during the first two sesons (Fig. 3). C. hexgonus is known to give ovoid red fruits with soft nd juicy, white to pinkish pulp nd numerous edible smll blck seeds. C. hexgonus ws originted in Brzil (Anderson, 2001). C. vlidus grew 5.1 cm in height nd 0.5 cm in dimeter per month during the first seson (Figs. 1 nd 2). However, it could not survive the high temperture during the summer months. All plnts were dried nd finlly died during the first seson. All C. pchnoi plnts survived the new environment. C. pchnoi grew very fst, verging up to fifteen centimeter month of new growth (Fig. 1). Its stem dimeter incresed 1.1 cm per month (Fig. 2). Three sprouts developed per plnt during the first two sesons (Fig. 3). C. pchnoi ws introduced s rootstock for grfting other species. C. pchnoi is populr s grfting stock for smller, slower growing ccti (Ostolz, 1984). C. peruvinus, C. peruvinus monstrose nd Cereus hexgonus united with C. pchnoi to form grfts with the success of 80, 53 nd 86.5%, respectively. C. Vlidus, however, filed completely to unite with C. Pchnoi (Fig. 8). This filure to form grfts my Weight (%) c b C. peruvinus C. peruvinus monstrose Period of storge (month) Fig. 5. Effect of the storge period on weight of C. peruvinus nd C. peruvinus monstrose cuttings. Vlues lbeled with the sme letter re not significntly different (P>0.05). e bc d e Fig. 4. Sprouts were rmified from the bse of the min stem of C. peruvinus. Fig. 6. Horizontl position of C. peruvinus cuttings during storge encourged the development of lterl brnches.

50 48 Growth behviour of pple cctus (Cereus species) in hyper-rid environment Fig. 7. Drmtic contrction (A) nd expnsion (B) of C. peruvinus monstrose stem s wter vilbility chnges. be due to the incomptibility or the difference in the dimeter between the scion nd the rootstock. Incomptibility between the scion nd the rootstock ffects the heling, nd could ruin the grft. The difference in dimeter between the scion nd the rootstock prevent meeting the cmbil lyers of the scion nd stock resulting filure of heling. The risk is smller if the scion nd the rootstock re bout the sme dimeter (Hrtmnn et l., 2001). However, self grfted C. Pchnoi heled with 93% of successful grfts. Grfting in cctus is relly best used s technique for sving dying cctus from certin deth, speeding up breeding progrms or growing less hrdy species. Often in cctus genetic improvement, hybridiztion, nd clonl selection progrms, the reserchers need more rpid method to prove or disprove the Percentge of successful grfts C. peruvinus C. peruvinus monstrose b c 0 Species Fig. 8. Percentge of succeeded grfts of different Cereus species on Cereus pchnoi. Vlues lbeled with the sme letter re not significntly different (P>0.05). C. hexgonus C. pchnoi C. vlidus merits of fruiting clone. To speed the evlution process for fruit production nd to dpt certin clones to locl conditions, grfting ws done onto mture plnts in the ground (Huffmn, 2003). A grfted cctus cn grow fster, flower sooner nd often be used with cctus species hving sensitive soil requirements (Hrtmnn et l., 2001). The hyper-rid region hs limited renewble freshwter supplies (Wilhite, 2000). So, lrge portion of the hyper-rid region is desert, the mjority of which is uninhbitble becuse of lcking the biologicl production (Al Alwi nd Abdulrzzk, 1993). It is vitl tht the kind of griculture prcticed in hyper-rid region should use s little wter s possible. In ddition, griculturl ctivities should produce mterils tht cn hve economic potentil. Apple cctus species C. peruvinus nd the subspecies C. peruvinus monstrose were the most promising in the new hyper-rid environment in terms of dptbility nd helthy growth. Apple cctus my give the inhbitnts of the hyper-rid nd mrginl lnds wy of mking living. Acknowledgement The uthor would like to thnk Mrs. Susn Nelson-Kluk, Progrm Director, Foundtion Plnt Mterils Service, University of Cliforni, for her ssistnce in collecting the plnt mteril. References Al Alwi, J. nd M. Abdulrzzk, Wter in the Arbin peninsul: Problems nd Perspectives. In: Wter in the Arb World: Perspectives nd Prognoses (Rogers P., Lydon P., eds), Division of Applies Sciences. Hrvrd University, USA. Anderson, E.F The Cctus Fmily, Timber Press, Portlnd Oregon. USA. Bckeberg, C Die Cctcee. Gustv Fisher, Stuttgrt, Germny.

51 Growth behviour of pple cctus (Cereus species) in hyper-rid environment 49 Cushmn, J.C. nd A.M. Borlnd, Induction of crssulcen cid metbolism by wter limittion. Plnt, Cell & Environment, 25: Duncn, D.B Multiple rnge nd multiple F test. Biometrics, 11: ElObeidy, A.A Introducing nd growing some fruiting columnr ccti in new rid environment. Journl Fruit nd Ornmentl Plnt Reserch, 12: Felger, R.S. nd M.B. Moser, Seri Indin food plnts: desert subsistence without griculture. J. Ecol. Food. Nutr., 5: Hrtmnn, H.T., D.E. Kester, R.L. Geneve nd F.T. Dvies, Plnt Propgtion: Principles nd Prctices. Prentice Hll, NJ, USA. Huffmn, M Cctus Grfting Methods. J. PACD: Inglese, P., F. Bsile nd M. Schirr, Cctus per fruit production. In: Ccti: Biology nd Uses (Nobel, P.S., ed), University of Cliforni Press, Cliforni. USA. John, S Desert ecology. An introduction to life in the rid Southwest. University of Uth Press, Slt Lke City, USA. Mld, G., R.A. Bckhus nd C. Mrtin, Altertions in growth nd crssulcen cid metolism (CAM) ctivity of in vitro cultured cctus. Plnt Cell, Tissue nd Orgn Culture, 58: 1-9. Museth, J.D Theoreticl spects of surfce-to-volume rtios nd wter-storge cpcities of succulent shoots. Americn Journl of Botny, 88: Morton, J.F., Cctcee, strwberry per. In: Fruits of wrm climtes (Morton J.F., ed), FL, USA. Nerd, A., J.A. Aronson nd Y. Mizrhi, Introduction nd domestiction of rre nd wild fruit nd nut trees. In: New Crops (Jnick J., Simon J.E., eds.), Timber, Portlnd, OR, USA. Nerd, A., N. Tel-Zur nd Y. Mizrhi, Fruits of vine nd columnr ccti. In: Ccti: Biology nd Uses (Nobel, P.S., ed), University of Cliforni Press, Cliforni. USA. Ostolz, C Trichocereus pchnoi. Cctus nd Succulent Journl, 56: Scheinvr, L Flor Ilustrd Ctrinese Cctcee. Itji. Snt Ctrin, Brzil. Wllce, R.S. nd A.C. Gibson, Evolution nd systemtics. In: Ccti: Biology nd Uses (Nobel, P.S., ed), University of Cliforni Press, Cliforni. USA. Weiss, J., A. Nerd nd Y. Mizrhi, Development of Cereus peruvinus (Apple Cctus) s new crop for the Negev desert of Isrel. In: New crops (Jnick J., Simon J.E., eds.), Wiley, New York, USA. Wilhite, D.A Drought s nturl hzrds. Concepts nd definitions. In: Drought, A Globl Assessment (Wilhite D., ed.), Volume I, Routledge, London nd New York, USA. Winter, K. nd J.A.C. Smith, Crssulcen Acid Metbolism - Biochemistry, Ecophysiology nd Evolution, Springer Verlg, New York. USA.

52 Journl of Applied Horticulture, 8(1): 50-52, Jnury-June, 2006 Assessment of genetic diversity nd reltionships mong some grpe vrieties using ISSR mrkers Appl Journl Mnjush Dhne, S.A. Tmhnkr, S.G. Ptil, G.S. Kribspp*nd V.S. Ro Genetics Group, Aghrkr Reserch Institute, G. G. Agrkr Rod, Pune *Ntionl Reserch Centre for Grpes, Mnjri Frm, Pune Abstrct As result of lrge-scle introduction, the origin nd uthenticity of mny grpe vrieties is uncler nd the subject of some controversy. This hs led to confusion regrding their correct identifiction. Moleculr mrkers hve proved to be useful to nlyze the genetic reltionships s well s diversity between different grpe vrieties. In the present study, 34 grpe vrieties hve been chrcterized using Inter Simple Sequence Repet (ISSR) mrkers. Out of 93 ISSR primers screened initilly, 11 showed good polymorphism. Totl 174 bnds were obtined, out of which 145 were polymorphic. The pir wise similrity indices were clculted from the bnd dt. Cluster nlysis of the vrieties resulted in the formtion of two min clusters, one belonging to Vitis vinifer nd other to V. lbrusc. Vrieties belonging to V. vinifer ppered more diverse nd formed distinct sub-clusters bsed on their colour, flvour nd seeds. Out of 34 vrieties screened, 10 vrieties with green/yellow berries were grouped together in one subcluster nd 15 with red/blck berries in the other. Three vrieties with green/ yellow berries, Itli, Queen of Vineyrd nd Thompson seedless were grouped with the vrieties with red/blck berries. The cluster of lbrusc vrieties showed homogeneity nd hd five vrieties except Dkh, which belongs to vinifer. Concord seprtes initilly from ll other vrieties. Incidentlly, Concord is pure selection from V. lbrusc, while other vrieties like Bnglore Blue, Blck Musct, Ctwb nd Muzzfr Ngr in lbrusc group, my be the hybrids of V. brusc x V. vinifer. The current study thus reveled tht genetic reltionships mong grpe cultivrs could be ssessed using ISSR mrkers. Key words: Diversity, genetic reltionships, ISSR mrkers, grpe vrieties Introduction Grpe is one of the most importnt nd oldest fruit crop throughout the world. In Indi, more thn 90% of grpe produce is utilized for tble purpose nd smll quntity for risins, juice nd wine mking. The long history of viticulture, vegettive propgtion of cultivrs nd the relince on mpelogrphy pose difficulties in ccurte cultivr identifiction. Most of the commercilly cultivted grpe cultivrs re introductions from exotic sources nd the genetic reltionships mong them re not cler which is importnt for plnning breeding progrmme nd conservtion of germplsm. The use of DNA mrkers hs been proposed s n objective nd vible lterntive for mpelogrphy (Thoms et l., 1993). There re number of reports involving use of DNA mrkers for studying genetic reltionships, fingerprinting of clones nd cultivrs s well s prentge studies (Bowers et l., 1993, 1996; Moreno et l., 1995; Cerver et l., 1998; Sefc et l., 1998). Prticulrly, PCR bsed DNA mrkers, provide powerful tools for genetic nlysis becuse of their simplicity nd ese of hndling. Mrkers generted by Inter Simple Sequence Repet mplifiction (ISSR; Zietkiewicz et l., 1994) hve been shown to be useful for detecting polymorphisms nd overcome mny technicl limittions of RFLP nd RAPD nlyses. ISSR nlyses hve been pplied to grpes erlier minly for detecting intrvrietl differences (Moreno et l., 1998) nd distinguishing cultivrs (Herrer et l., 2002). In the present work; we hve used ISSR mrkers to chrcterize seeded grpe vrieties from Indi. Mterils nd methods Plnt mteril: A totl of 32 seeded grpe vrieties were nlysed in the present study. Two seedless vrieties, Thompson Seedless nd Flme Seedless were lso included in the nlysis s stndrd vrieties. All these were obtined from the germplsm mintined t Ntionl Reserch Centre for grpes, Pune. List of vrieties is given in Tble-1 Tble 1. List of vrieties nlysed in present study Green / Yellow berries Red / Blck berries 1 Anb-e-Shhi 18 Blck Musct 2 Angur Kln 19 Blck Prince 3 Bnqui Abyd 20 Ctwb 4 Cheem Shebi 21 Corn Regi 5 Chenin Blnc 22 Concord 6 Gold 23 Convent Lrge Blck 7 Itti 24 Dkh 8 Jos Beli 25 Dimond Jubilee 9 Muller Thergu 26 Musct Hmburg 10 Plomino 27 Muzzfr Ngr 11 Queen of Vineyrd 28 Red Globe 12 Shebi Ali 29 Red Musct 13 Sundekhni 30 Shirz 14 Wlthom Cross 31 Spin Shebi Red / Blck berries Stndrds 15 Bnglore Blue 32 Thompson Seedless 16 Bhrt Prince 33 Flme Seedless 17 Blck Chmp 34 Gulbi DNA isoltion: DNA ws extrcted from young, fully expnded leves by modified CTAB method (Lodhi et l., 1994). The isolted DNA ws processed, quntified nd used for PCR rections.

53 Assessment of genetic diversity nd reltionships mong some grpe vrieties using ISSR mrkers 51 Tble 2. List of ISSR primers used nd polymorphic bnds S. No. Primer Sequence Number of bnds Polymorphic bnds Polymorphism (%) AGA GAG AGA GAG AGA GT ACA CAC ACA CAC ACA CG ACA CAC ACA CAC ACA CYT ACA CAC ACA CAC ACA CYA ACA CAC ACA CAC ACA CYG TGT GTG TGT GTG TGT GRC TGT GTG TGT GTG TGT GRA BDB CAC ACA CAC ACA CA DBD ACA CAC ACA CAC AC VHV GTG TGT GTG TGT GT HVH TGT GTG TGT GTG TG Totl Y=C/T; R=A/G; B=C/G/T; D=A/G/T; H=A/C/T; V=A/C/G ISSR mplifictions nd Gel electrophoresis: ISSR mplifictions were crried out using Primer set#9; obtined from University of British Columbi, Vncouver, Cnd in 25µl rection volume. Rection mixture contined 1X PCR buffer contining 1.5mM MgCl 2, 12.5ng of genomic DNA, 0.5 U Tq DNA Polymerse, 0.1mM of ech dntp, 0.04 mm spermidine, 2% formmide nd 0.3 µm of primer. The therml cycling ws performed in PTC 200 Therml Cycler (MJ Reserch Inc, USA) following the protocol of Ngok nd Ogihr (1997). The PCR rection ws performed t lest three times for ech primer to ensure reproducibility. The mplifiction products were seprted on 1.5% grose gels. The gels were stined with ethidium bromide nd visulized on UV trnsillumintor. Dt Anlysis: Bnds in the mplifiction profiles were recorded s present (1) nd bsent (0). Bsed on the bnd dt, the similrity mtrix ws clculted using Dice coefficient nd the cluster nlysis ws crried out using SAHN module in NTSYS pc 2.1 softwre. Results nd discussion Totl 93 ISSR primers from set #9 were screened initilly for polymorphism nd 11 were finlly selected bsed on the bsis of cler scorble bnd pttern. The list of the primers used nd polymorphic bnds recorded is given in Tble 2. The mplifiction profile obtined with primer UBC 857 is shown in Fig. 1. Totl 174 bnds were obtined with the 11 selected primers; out of which 145 were polymorphic. The bnds rnged from 70 bp to 1.4 kb in size. Primer 891 showed the mximum number of bnds. The number of polymorphic bnds rnged from 6 (UBC 859) to 23 (UBC 890). All these primers contined dinucleotide repets. Primers with (AC) n repets were mximum in number (5/11) followed by (TG) n (3/11). In generl, percentge polymorphism obtined by primers contining (AC) n repets ws lower (75%) s compred to those with (TG) n (89.66%). All the bnds obtined with primers 857, 859 nd 890 were polymorphic. Few genotype specific bnds were observed with primers 856 nd 860 in vriety Blck Prince nd primer 889 in vrieties Concord nd Dimond Jubilee. Bsed on the bnd dt, similrity mtrix ws generted using Dice coefficient nd cluster nlysis ws crried out. The dendrogrm generted by UPGMA lgorithm using NTSYS pc 2.1 softwre is shown in Fig. 2. The similrity coefficient rnged from 0.48 to Two mjor clusters were observed; one consisting of V. lbrusc nd its derivtives nd the other of vrieties from V. vinifer. Vriety Concord seprted initilly from ll others nd showed lest similrity. Cluster I consisted of minly V. lbrusc or lbrusc x vinifer hybrids except Dkh. The cluster II consisted minly of V. vinifer vrieties. In this cluster, Blck Prince nd Red Musct seprted initilly from other vinifer vrieties. Severl subgroups bsed on their colour, flvour nd seeds were observed in this cluster indicting the diverse nture of these vrieties. A distinct sub cluster of 10 vrieties with green / yellow berries could be distinguished. Similrly, 15 vrieties with (red / blck) berries were grouped together. Three vrieties, Itli, Queen of Vineyrd nd Thompson Seedless hving yellowish green berries were lso grouped with these vrieties. Vrieties Anb-e-Shhi, Angur Kln nd Cheem Shebi, ll high yielding nd hrd seeded were grouped together, while three vrieties in the other subgroup nmely Gold, Muller Thergu nd Sundekhni, hve soft seeds nd musct flvour. Four flvoured vrieties; Itli, Bhrt Prince, Musct Hmburg nd Gulbi were grouped together in the sme group. The cluster of lbrusc vrieties showed homogeneity nd hd five vrieties including Dkh, which however, belongs to vinifer. Vrieties Ctwb nd Dkh showed mximum similrity of 89%. Such close reltionship nd the grouping of Dkh in lbrusc group is surprising since Dkh belongs to V. vinifer (Chdh nd Fig. 1. Amplifiction profile obtined with UBC primer 857. M: Moleculr weight mrker, PhiX174DNA/ HeIII digest DNA smples s listed in Tble 1

54 52 Assessment of genetic diversity nd reltionships mong some grpe vrieties using ISSR mrkers Fig. 2. Dendrogrm bsed on ISSR bnd dt nd UPGMA lgorithm showing reltionships mong the grpe vrieties [I]: Vitis lbrusc cluster [II]: Vitis vinifer cluster Rndhw, 1974). Therefore, it ppers to be cse of mistken identity. The seprtion of Concord from V. lbrusc group is lso unexpected since it is one of the oldest lbrusc cultivr. According to Hedrick (1938), Concord hs ll the morphologicl chrcters of the Americn species (V. lbrusc), while, severl other workers hve reported this vriety to be hybrid between V. lbrusc nd V. vinifer (Chdh nd Rndhw, 1974). In our erlier nlysis using RAPD mrkers lso it hd grouped long with other lbrusc vrieties (Tmhnkr et l., 2001). The nlysis of multiple smples collected from different sources will be useful to confirm these results. Although ISSR mrkers re efficient nd relible for the ssessment of genetic reltionships mong grpe vrieties, ppliction of other mrker systems like microstellites (STMS) nd AFLP is necessry for resolving the controversil grouping of some vrieties. Acknowledgements The finncil ssistnce from the Deprtment of Biotechnology (DBT), Ministry of Science nd Technology, New Delhi is grtefully cknowledged. References Bowers, J.E., E.B. Bndmn nd C.P. Meredith, DNA fingerprint chrcteriztion of some wine grpe cultivrs. Am. J. Enol. Viticult., 44: Bowers, J.E., G.S. Dngl, R. Vignni nd C.P. Meredith, Isoltion nd chrcteriztion of new polymorphic simple sequence repet loci in grpes (Vitis vinifer L). Genome, 39: Cerver, M.T., J.A. Cbezs, J.C. Snch, De Mrtinez, F. Tod nd J.M. Mrtinez Zpter, Appliction of AFLPs to the chrcteriztion of grpevine Vitis vinifer L. genetic resources. A cse study with ccessions from Roij (Spin). Theor. Appl. Genet., 97: Chdh, K.L. nd G.S. Rndhw, Grpe vrieties in Indi- Description nd clssifiction, ICAR Technicl Bulletin No.48, ICAR, New Delhi. Hedrick, U.P Cytology of hrdy fruits. The Mcmilln co., New York. Herrer, R., V. Cres, M.J. Wilkinson nd P.D.S. Cligri, Chrcteriztion of genetic vrition between Vitis vinifer cultivrs from centrl Chile using RAPD nd Inter Simple Sequence Repet Mrkers. Euphytic, 124: Lodhi, M.A., G.N. Ye, N.F. Weeden nd B.I. Reisch, A simple nd efficient method for DNA extrction from grpevine cultivrs nd Vitis species. Plnt Mol. Biol. Reporter, 12: Moreno, S., Y. Gogorcen nd J.M. Ortiz, The use of RAPD mrkers for identifiction of cultivted grpe vine (Vitis vinifer L). Scienti Hortic., 62: Moreno, S., J.P. Mrtin nd J.M. Ortiz,1998. Inter-simple sequence repets PCR for chrcteriztion of closely relted grpe vine germplsm. Euphytic,101: Ngok, T. nd Y. Ogihr, Applicbility of inters simple sequence repet polymorphisms in whet for use s DNA mrkers in comprison to RFLP nd RAPD mrkers. Theor. Appl. Genet., 94: Sefc, K.M., F. Regner, J. Glossl nd H. Steinkellner, Genotyping of grpevine nd rootstock cultivrs using microstellite mrkers. Vitis, 37: Tmhnkr, S.A., S.G. Ptil, V.S. Ro, Assessment of the genetic diversity of some importnt grpe genotypes in Indi using RAPD mrkers. Vitis, 40: Thoms, M.R., S. Mtsumoto, P. Cin nd N.S. Scott, Repetitive DNA of grpevine: clsses present nd sequence suitble for cultivrs identifiction. Theor. Appl. Genet., 86: Zietkiewicz, E., A. Rflski nd D. Lbud, Genome fingerprinting by simple sequence repet (SSR)-nchored polymorphism chin rection mplifiction. Genomics, 20:

55 Journl of Applied Horticulture, 8(1): 53-57, Jnury-June, 2006 Impct of polyethylene glycol-induced wter stress on growth nd development of shoot tip cultures from different bnn (Mus spp.) cultivrs Appl Journl Mohsen K.H. Ebrhim 1, Ibrhim A. Ibrhim 2, Hmdy A. Emr 2 nd Ewld Komor3 1 Botny Deprtment, Fculty of Science, Tnt University, Tnt 31527, Egypt. 2 Genetic Engineering nd Biotechnology Reserch Institute, Minufiy University, Egypt, 3 Deprtment of Plnt Physiology, Byreuth University, D Byreuth, Germny, e-mil: ewld.komor@uni-byreuth.de. Abstrct Shoot tip explnts of the Egyptin bnn cultivrs Mghrby, Vlery, Grnd Nin nd Hindy were tested for their tolernce to wter stress. Shoot survivl, shoot growth nd root growth stimultion in presence of polyethylene glycol (PEG) ws strongest in cultivr Hindy followed by Grnd Nin, Mghrby nd Vlery. The ccumultion of soluble sugrs nd proline in shoots ws positively correlted with the pplied polyethylene glycol concentrtion, while the reverse ws true for N, P nd K content. The cultivr Hindy exhibited higher metbolite ccumultion response nd cultivr Mghrby the lest. The effects were most cler on liquid medium wheres solid (gr) medium exerted some dditionl effects incresing the osmotic stress t low PEG concentrtions nd lleviting the PEG effect t high PEG concentrtions. In conclusion, the cultivr Hindy ppered to be the most tolernt to wter stress becuse of strong ccumultion of comptible solutes nd greter stimultion of root development. Keywords: Bnn (Mus spp. L.), medium (solid/liquid), micropropgtion, osmotic stress, polyethylene glycol (PEG), proline, sugrs Introduction Bnn (Mus spp.) fruits represent stple food for bout one billion people ll over the world (FAO, 1992). It hs become one of the strtegic crops in tropicl countries due to its high income potentil for the locl frmers. Since they re seedless, the plnt hs to be propgted vegettively or in vitro. The in vitro propgtion is preferble in modern breeding becuse it llows the production of lrge number of virus-free plnts in reltively short time nd in smll spce (Cronuer nd Krikorin,1984, 1985; Khlil et l., 2002; Sgi et l.,1995). Bnn is cultivted in tropicl climtes wherever stedy yerround wter supply is vilble. Recently it ws introduced into tropicl regions which hve reltively low wter vilbility. This initited the investigtion of bnn plnt response to wter stress during micropropgtion nd under greenhouse conditions. Wter stress cn be induced in the micropropgtion medi by dding comptible osmoticum such s polyethylene glycol (PEG). Despite the numerous studies on micropropgtion of bnn, there is no informtion bout the effect of polyethylene glycol on regenerting or regenerted bnn tip explnts. Since the drought stress tolernce differs mong plnt species nd the cultivrs of given species (Dodd nd Donovn, 1999), the in vitro response of four bnn cultivrs which re importnt for Egyptin horticulture, to polyethylene glycol-induced wter stress ws studied. Mterils nd methods Plnt mteril: Four cultivrs of Mus spp. obtined from the experimentl frm El-Knter El-Khyrei, Klubiy of the Agriculturl Development Systems Project (Giz, Egypt), served s the source mteril for shoot tips during the study period ( ). The cultivrs were Mghrby, Vlery, Grnd Nin, which belong to the semi-dwrf Cvendish group nd Hindy is strin of dwrf Cvendish. All re of triploid Acumint type. Aseptic cultures were estblished from shoot tips which were surfce-sterilized in 3 % NOCl solution (contined 0.1 % Tween 20 s wetting gent) for 20 min. Therefter, the tips were rinsed severl times in sterilized distilled wter to remove ll trces of chlorine. After removl of the outside tissues, picl meristems were verticlly cultured for 4 weeks on Murshige-Skoog bsl medium (Murshige nd Skoog, 1962) supplemented with benzyldenine (3 mg/l) nd gr (6 g/l). The growing explnts were recultured, t 4 week intervls, on fresh medi until the onset of prolifertion (c. 2 months). This culture period ws clled strting phse. In order to obtin sufficient number of explnts, the produced shoots were subcultured four times on solid Murshige- Skoog bsl medi supplemented with benzyldenine (5 mg/l) in so-clled multipliction phse. The obtined plntlets were used for the following experiments (Fig.1). Incubtion with polyethylene glycol: To determine the lethl concentrtion of polyethylene glycol for ech cultivr, PEG-6000 ws dded to solid bsl medi t levels of 0, 5, 15, 25, 35 nd 45 g/l. The produced shoots were cultured for 4 weeks on these medi nd the percentge of survivl ws determined. Polyethylene glycol (0, 10 nd 20 g/l) ws dded to bsl medi which were either supplemented with benzyldeneine (5 mg/l) to produce shoots or α-nphthlene cetic cid (1 mg/l) to produce roots. After 4 weeks, shoot or root growth nd development were determined. This procedure ws pplied in liquid nd in solid medi (0.6 % gr). Shoot strength (or vigour) ws determined ccording to Pottino (1981) on rting scle: 1 = no growth, 2 = below verge, 3 = verge, 4 = bove verge, 5 = excellent.

56 54 Wter stress effects on bnn shoot tip cultures Anlyticl methods: The metbolites were determined from oven-dried shoots. Mixed-cid digestion method ws used in prepring the smple solution used for determintion of N, P nd K (Ebrhim nd Aly, 2002). Totl-nitrogen content (N) ws estimted using the micro-kjeldhl method (Jcobs, 1958). Phosphorus (P) content ws spectrophotometriclly determined by molybdenum-blue method (Pge, 1982). Potssium (K) ws determined ccording to Allen et l. (1974). Sugrs were extrcted in borte buffer ph 8 (0.1 g DW/5 ml buffer), then totl soluble sugrs were determined by the method dopted by Shffer nd Hrtmnn (1921). Proline, ws quntified in ethnol extrct ccording to Btes et l. (1973). All medi were filled into 200 ml Pyrex-glss jrs (25 ml/jr), utoclved for 20 min t 121 o C nd 1.2 kg/cm 2 pressure, then cooled nd kept for 4-15 dys before use. In ll medi, ph ws djusted to 5.7 (before dding gr). Growth ws in growth chmber t 25+3 o C nd 40 μmol m -2 s -1 continuous photosynthetic photon flux provided by cool white fluorescent lmps. Sttisticl nlysis: All experiments were repeted twice nd conducted by using completely rndomised design in fctoril rrngement with t lest 4 replictes. All dt were verged nd sttisticlly nlysed by using two-nd three-wy nlysis of vrince (ANOVA). In cse of percentges, the originl dt were rcsine-trnsformed prior to sttisticl nlysis. The lest significnt difference (LSD) t the 0.05 level ws used to compre between mens directly (Steel nd Torrie, 1980) or indirectly by the multiple rnge test of Duncn (Duncn, 1955). Results Polyethylene glycol is hydrophilic lcohol polymer with high wter solubility nd low toxicity (Fontn et l., 2001). We used it in our study s n inert non-penetrting osmoticum (Almnsouri et l., 2001). Explnt survivl in the multipliction phse ws significntly decresed by high PEG levels (> 15 g/l) (Fig. 2), but the inhibition ws more evident in cse of the cultivrs Mghrby nd Vlery thn Grnd Nin nd Hindy. The ltter survived prtly up to 45 g PEG /l. PEG tretments reduced shoot multipliction nd biomss per shoot, especilly in the cultivrs Mghrby nd Vlery nd less in Grnd Nin nd Hindy (Tble 1). Incresing PEG concentrtion in the rooting medium incresed both root number nd, slightly, root length. This increse ws highest in Hindy, followed by Grnd Nin, Vlery nd Mghrby (Tble 1). This my be interpreted s n dptive response of roots to llevite the reduced wter vilbility. Thus cultivr Hindy ppered to be the most dpted to wter shortge. The comprison of shoot nd root growth t 20 g/l PEG on solid (gr) medium nd in liquid medium reveled tht growth ws better in liquid medium (Tble 2). This finding grees with previous results by Ebrhim (2004) with Cll nd Ebrhim nd Ibrhim (2000) on Mrnt, who ttributed this response to better vilbility for growth substnces nd nutrients nd better ertion in liquid medi. Also gr my dd n osmotic effect in ddition to PEG. The cultivr tolernce to PEG with respect to growth ws gin gretest in the order Hindy>Grnd Nin>Vlery>Mghrby (Tble 2). From the growth response of the 4 bnn cultivrs it ws obvious tht cultivr Hindy could dpt best to wter stress, followed by Grnd Nin nd then Mghrby nd Vlery (with the ltter two virtully identicl). The nlysis of cell contents should show whether the stress tolernce ws correlted with ccumultion of nutrient slts or production of comptible osmotic, especilly soluble sugrs nd proline. N, P nd K content of shoots decresed with incresing levels of PEG, somewht less in cultivr Hindy thn in Grnd Nin or Mghrby (Fig. 3). On solid medium the N, P nd K contents were mrginlly but consistently lower thn in liquid medium. A different, complex picture emerged for sugrs nd proline. Explnts of ll cultivrs in liquid medium exhibited incresing levels of sugrs nd proline with incresing PEG concentrtions. On solid medium there ws the sme trend, but the difference between no PEG tretment nd 10 g/l PEG ws smll or bsent. In ddition the level of sugrs nd proline were higher on solid medium thn in liquid medium when PEG ws bsent, but lower Tble 1. Shoot growth, shoot development, root growth nd root development of in vitro cultured bnn s ffected by the cultivr nd PEG concentrt supple cid (1 mg/l, rooting medium) (Men ±SD) Cultivr PEG (g/l) Shoots/explnt Shoot fresh Roots/shoot Root length weight (g) (cm) ± ± ± ± 0.18 Mghrby Vlery Grnd Nin Hindy ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± 0.19

57 Tble 2. Shoot growth, shoot development, root growth nd root development of in vitro cultured bnn in presence of 20 g/l PEG on solid nd liquid medium. Explnts were cultured with 20 g/l PEG for 4 weeks on Murshige-Skoog bsl medium contining benzyldenine (5 mg/l, multipliction medium) or on Murshige-Skoog bsl medium contining nphthyl cetic cid (1 mg/l, rooting medium) (Men ±SD) Cultivr Medium Shoots/ explnt Shoot frresh weight (g) Mghrby Vlery Grnd Nin Hindy Wter stress effects on bnn shoot tip cultures 55 Shoot vigour Leves/ shoot Roots/ shoot Root length (cm) Solid 1.12 ± ± ± ± ± ± 0.24 Liquid 1.50 ± ± ± ± ± ± 0.28 Solid 1.04 ± ± ± ± ± ± 0.26 Liquid 1.57 ± ± ± ± ± ± 0.32 Solid 1.57 ± ± ± ± ± ±0.24 Liquid 1.93 ± ± ± ± ± ± 0.33 Solid 1.96 ± ± ± ± ± ± 0.22 Liquid 2.22 ± ± ± ± ± ± 0.32 Fig. 1. Regenerting bnn plntlets from meristem tip culture. Successive stges of micropropgtion of Mus spp. (cultivr. Hindy): (A) shoot tips were cultured on Murshige-Skoog bsl medium contining benzyldenine (3 mg/l, strting stge), (B) shoot explnts were proliferted on bsl medium supplemented with benzyldenine (5 mg/l, multipliction stge), nd (C) shoot explnts were rooted on bsl medium contining nphthyl cetic cid (1 mg /l, rooting stge). Biomss (%) Mghrby Vlery Grnd Nin Hindy Fig. 2. Percentge of shoot survivl of in vitro cultured bnn cultivrs in presence of PEG. Explnts were cultured for 4 weeks on Murshige- Skoog bsl medium supplemented with benzyldenine (3 mg/l, strting medium). (The results for Mghrby nd Vlery coincide t high PEGconcentrtions) (Men ±SD). on solid medium thn in liquid medium when PEG ws present (Fig. 4). The different bnn cultivrs showed the sme order in comptible solute content s in tolernce towrds PEG, nmely cultivr Hindy contining the highest levels of nutrients nd comptible solutes followed by cultivr Grnd Nin nd then Mghrby. N content (% mg/g dw) K content (% mg/g dw) P content (% mg/g dw) Fig. 3. N, P nd K-content of shoots of in vitro cultured bnn plntlets in presence of PEG on solid nd liquid medium. Explnts were cultured for 4 weeks on Murshige-Skoog bsl medium contining Discussion Grnd Nin,liquid Hindy,solid Hindy,liquid Mghrby,solid Mghrby,liquid Grnd Nin,solid Effects of PEG on osmotic performnce hd been reported for mny plnts (Bndursk, 2000; Pushpm nd Rngsmy, 2000; Ronde et l., 2000; Liu et l., 2001). Predictbly the growth of the smll, micropropgted bnn plnts of cultivrs Mghrby, Vlery, Grnd Nin nd Hindy ws influenced by ppliction of PEG nd, slightly, by the type of medium, but the mgnitude of the response ws consistently cultivr-dependent. The cultivr