11 Interntionl Conferene on Biology, Environment nd Chemistry IPCBEE vol. (11) (11)IACSIT Press, Singpoore Slisyli Aid Folir nd Soil Applition Effet on Chikpe Resistne to Chilling Stress Simk Immi 1, Soleimn Jmshidi + nd Shhrm Shhrokhi 1 Deprtment of Grdute Agronomy, Miyneh Brnh, Islmi Azd University, Miyneh, Irn Deprtment of Plnt Protetion, Miyneh Brnh, Islmi Azd University, Miyneh, Irn Astrt. To evlute sliyli id (SA) soil nd folige pplition on resistne to hilling stress in two Kuli type hikpe ultivrs, Jm nd ILC, n experiment ws ontted sed on rndomized ompletely design in ftoril with three replitions. Two SA doses (5 nd mmol) were pplied. Soil sprying inresed proline ontent, ut there ws no signifint effet pplying SA s folige sprying on proline ontent. SA pplition redued eletrolyte lekge nd reltive wter ontent. Also, soil pplition ws more effetive on hlorophyll ontent nd plnt height enhnement thn folige pplition. Keywords: indued resistne, SA, Jm, ILC, old stress. 1. Introdution Chikpe (Cier rietinum L.) is one of the most importnt legumes ultivting in ool nd semi-rid regions of Irn where hill nd drought re two limiting ftors in hikpe prodution (Soltni et l., 1999). In old regions, freezing stress predominntly ours during germintion, seedling formtion nd erly vegettive stges of rop growth (Croser, 3). Cold stress is ommon in west Asi nd North Afri whih is inresed its ourrene likelihood y ltering rop system from hikpe spring to winter plnting (Bgheri & Soltni, ). Sliyli Aid is one of the phenoli omponents whih is normlly produed in plnts in very smll quntities (Rskin, 199), whih n e implemented s growth regultors (Aerg, 191). Sliyli id is one of the signling moleules whih effet on plnt growth nd development (Devoto & Turner, 3; Krntev et l., ). Its remrkle effet in mmol onentrtion on itter olive resistne seedlings to hilling stress hs een reported up to 71% (Kvin, ). Also, it ws effetive in.5 μmol on rdish seedlings resistne to hilling stress (Pour-Akr & Noojvn Asghri, 5). Tsgin et l., (3) reported tolerne to freezing in winter whet leves ffeted y SA pplition. Also, Jnd et l. (199) indued resistne of orn to hilling stress dding.5 μmol of SA to nutritive solution in hydroponi system. Syyri et l. (9) proved SA potentil on stored pomegrnte fruits tolerne to hilling stress. The urrent experiment ws performed to evlute SA effiieny on resistne to hilling stress in two Kuli ultivrs of hikpe using two folir nd soil pplition methods.. Mteril nd Methods Chikpe Kuli type seeds (v. Jm nd ILC) were superfiilly sterilized y 1.5%.i. of Croxin nd seeds were plnted in ylindril plsti pots, 15m internl dimeter filled with y lomy silt soil mixed with perlite ( : 1 w/w) nd immeditely irrigted. In -lef stge (fter out five weeks), the hikpe seedlings were tined to equl seedlings. The plnts folige nd soil surfe were spryed y 5 ml of, 5, nd mmol of sliyli id. To void soil ontmintion y SA in folige sprying method, plsti over ws used on soil surfe while sprying. Then, they were kept for h in usul + Corresponding uthor. Tel.: + 9 91337; Fx: + 9 379. E-mil ddress: s.jmshidy@gmil.om 3
greenhouse ondition (5 ± C). For seedlings dpttion to old ondition, ll pots were pled t C for h efore hilling stress. The hilling stress ws performed t C for h. Afterwrd, the plnts were trnsferred to the sme greenhouse ondition nd fter 7h, the plnt height (H), hlorophyll ontent (CC), eletrolyte lekge (EL), reltive wter ontent (RWC) nd proline ontent (PC) were mesured. CC ws determined y holorophyllmeter (SPAD 5DL, Minolt, USA). To ssess memrne permeility, EL ws determined ording to Korkmz et l. (7). Lef diss (5 mm in dimeter) from rndomly hosen two plnts per replite were tken from the middle portion of fully developed youngest lef nd wshed with distilled wter to remove surfe ontmintion. The diss were pled in individul stoppered vils ontining ml of distilled wter. After inuting the smples t room temperture on shker (15 rpm) for h, the eletril ondutivity (EC) of the thing solution (EC 1 ) ws determined. The sme smples were then pled in n utolve t 11 C for min nd seond reding (EC ) ws determined fter ooling the solution to room temperture. The EL ws lulted s EC 1 /EC nd expressed s perent. Lef diss (5 mm in dimeter) from rndomly hosen two plnts per replite were tken from the middle portion of fully developed third ompound lef. Diss were weighed (fresh weight, FW) nd then immeditely floted on distilled wter in petri dish for 5 h in the drk. Turgid weights (TW) of lef diss were otined fter drying exess surfe wter with pper towels. Dry weights (DW) of diss were mesured fter drying t 75 C for h. RWC ws lulted using the following formul: RWC = FW DW TW DW PC ws determined ording to the method desried y Btes et l. (1973). Fresh lef mteril (.5 g) ws homogenized in ml of 3% queous sulfosliyli id nd filtered through Whtmn's No. 1 filter pper. Hlf milliliter of the filtrte ws mixed with 1ml of id-ninhydrin nd 1ml of glil eti id in test tue. The mixture ws pled in wter th for 1 h t C. The retion mixture ws extrted with ml toluene nd the hromophore ontining toluene ws spirted, ooled to room temperture, nd the sorne ws mesured t 5nm with UV/visile spetrophotometer (Jenwy 35, USA). Approprite proline stndrds were inluded for the lultion of proline in the smples. The experiment ws performed sed on ompletely rndomized design in ftoril with three replitions. 3. Results nd disussion 3.1. Plnt height ANOVA of plnt height showed there ws signifint differene (p <.1) etween methods nd SA doses (tle 1). Soil sprying ws more effetive thn folige sprying on plnt height. Also, inresing SA dose from to mmol enhned the plnt height (Fig. 1). Tle 1: ANOVA of under hilling stress hikpe trits ffeted y SA pplition Vrition soures Df Plnt height Fresh weight Dry mtter Men of Squres Chlorophyll ontent Eletrolyte lekge RWC Proline ontent SA dose (D) 5. **.7. 397.7 ** 33. ** 77.31 ** 131. ** Cultivr (C) 1. 11.77 **.1 17. **.779 5.1 ** 19. Method (M) 1 3.73 ** 5. **.3 ** 1. ** 19.75.13 ** 7.95 ** D C.355.31.7.551.9 11.13 ** 5.7 D M 1.93.75..35 73. 9.55 3.5 ** C M 1 1.173.95 **. 5. 11.9.3.51 D C M..57..57.9.9 **.51 Error.\3.35.11.7 55.553.93.1 C.V. (%).51 1.35.37 5.7 9.1.7 5.
Plnt height (m) 11.5 11.5 9.5 9.5 5 SA onentrtion (mmol) Plnt height (m).... 9. 9. 9. Soil sprying Folige sprying SA pplition method Fig. 1: SA different onentrtions effet on hikpe height fter hilling stress Fig. : Different SA pplition method effet on hikpe height fter hilling stress 3.. Fresh d dry weight ANOVA of FW showed there ws signifint differene (p <.1) etween SA pplition methods, ultivrs nd their intertion (tle 1). There ws no signifint differene etween folige nd soil sprying with SA in Jm ultivr. But Soil sprying resulted in more fresh weight thn folige pplition in ILC. Plnt dry weight ws signifintly (p <.1) etween two pplition methods (fig. 3). More dry weight ws hieved when SA pplied s soli sprying thn folige pplition,.5 nd.3 gr/plnt, respetively. fresh weight (gr/plnt) 5 3 1 folige spry Soil spry folige spry Soil spry Jm ILC Fig. 3: SA different pplition methods effet on two hikpe ultivrs fresh weight fter hilling stress 3.3. Chlorophyll ontent ANOVA of CC showed there ws signifint differene (p <.1) etween methods, nd dose of SA nd ultivrs (tle 1). On the whole, CC of Jm ws more thn ILC. Also, plnts reeived SA y soli pplition hd signifintly more CC thn folige spryed plnts. Also, inresing SA dose from to mm enhned hlorophyll ontent (fig. ). Chlorophyll ontent 15 5 5 Chlorophyll ontent 1 1 1 Soil sprying Folige sprying Chlorophyll ontenet 1 1 1 Jm ILC SA onentrtion (mm) SA pplition method Chikpe ultivrs Fig. : SA different onentrtions, pplition methods effet on two hikpe ultivrs hlorophyll ontent fter hilling stress 5
3.. Reltive wter ontent ANOVA of RWC showed there ws signifint differene (p <.1) etween methods, dose of SA, nd ultivrs lso ultivr SA dose nd dose ultivr method intertion (tle 1). RWC of Jm signifintly inresed in oth pplition methods y SA dose inrement. In with folige sprying 5 mmol ws the sme ith mmol. In ll methods, SA pplition ws effetive thn un-spryed plnts (Fig. 5) RWC 9 7 5 3 d d d f ef ef e g mm 5 mm mm mm 5 mm mm mm 5 mm mm mm 5 mm mm folige spry soil spry folige spry soil spry Jm ILC Fig. 5: SA different onentrtions, pplition methods effet on two hikpe ultivrs reltive wter ontent fter hilling stress 3.5. Eletrolyte lekge ANOVA of EL showed there ws signifint differene (p <.1) etween doses of SA (tle 1). Inresing dose of SA, EL signifintly ws deresed (Fig. ). 3.. Proline ontent ANOVA of PC showed there ws signifint differene (p <.1) etween methods, dose of SA, nd ultivrs lso SA dose method intertion (tle 1). On the whole, PC ws inresed signifintly in soil sprying y SA pplition (Fig. 7) Eletrolyte lekge 9 7 5 3 5 Fig. : SA different onentrtion effet on hikpe eletrolyte lekge fter hilling stress proline onent (μmol/gr FW) 1 1 1 1 5 5 folige spry soil spry Fig. 7: SA different dose nd pplition methods effet on hikpe fter hilling stress
. Referenes [1] A. Soltni, K. Ghssemi Golezni, F. R. Khooie, nd M. Moghdm. 1999. A simple model for hikpe growth nd yield. Field Crops Reserh, 1999. : 13-. [] J. S. Croser, H.J. Clrke, K.H.M. Siddique, nd T.N. Khn. Low-temperture stress: implitions for Chikpe (Cier rietinum L.) improvement. Critil Reviews in Plnt Siene, 3. (): 15-19. [3] A. R. Bgheri, nd A. Soltni. Legumes reeding for stresses tolerne. Reserh, Edution nd Extension Orgniztion Pulition: Tehrn.. 55 pp. [] B. Aerg. Plnt growth regultors XLI. Monosustituted enzoi id. Sweden Journl Agriulturl Reserh. 191, 11: 93 5. [5] I. Rskin. Role of sliyli id in plnts. Annul Review of Plnt Physiology, 199. 3: 39-3. [] A. Devoto, J. G. Turner. Regultion of jsmonte-medited plnt responses in Aridpsis. Annul Botny, 3. 9: 39-337. [7] A. Krntev, R. Yordnov, T. Jnd, G. Szli, L. Popov. Tretment with sliyli id dereses the effet of dmium on photosynthesis in miz plnts. Journl of Plnt Physiology,. 15(9): 9-931. [] M. Syyri, M. Blr, S. Klntri., M. Serrno, nd D. Vlero. Effet of sliyli id tretment on reduing hilling injury in stored pomegrntes. Posthrvest Biology nd Tehnology, 9. 53: 15-15. [9] E. Tsgin, O. Atii, nd B. Nlntoglu. Effets of sliyli id nd old nd freezing tolerne in winter whet leves. Plnt Growth Regultors. 3. 1: 31-3. [] B. Kvini, B. Frnso, Shker, H., nd Hddd-hi, Ch. Sliyli id effets on resistne enhnement of emryoni xis in itter olive (Meli zedrh L.) ginst hill nd hrd freezing. Pzhuhesh v Szndegi,. 7: -9. [11] L. Pour-kr, nd Noojn Asghri, M. Sliyli id effet on resistne to hilling stress in rdish seedlings. Siene Mgzine of Trit Mollem University, 5. 3: 9-. 7