Aville online t www.scholrsreserchlirry.com Annls of Biologicl Reserch, 2013, 4 (6):1-9 (http://scholrsreserchlirry.com/rchive.html) ISSN 0976-1233 CODEN (USA): ABRNBW The effect of plnt hormone gierellic cid on germintion indices Secle montnum in vitro nd pot experiments under drought conditions Azdeh Afrign 1, Zhr Jvdni 2, Esfndir Jhnt 3 *, Ruhollh Jhnin 4 nd Adol Azim Bhri 5 1 Deprtment of Renglnd, Science nd Reserch Brnch, Islmic Azd University Tehrn, Irn 2 Deprtment of Horticulture, Guiln University, Irn 3 Young Reserchers Clu, Deshdsht Brnch, Islmic Azd University, Deshdsht, Irn 4 Young Reserchers Clu, Gchsrn Brnch, Islmic Azd University, Gchsrn, Irn 5 Deprtment of Seed Technology, University of Tehrn, Irn ABSTRACT One of the effects of drought is the reduction of germintion, seedling estlishment nd seedling growth. Pretretment improves some spects of seed germintion nd seedling estlishment. To investigtion the effect of pretretment with Gierellic cid on the seed germintion of Secle Montnum in stges of growth in vitro nd pot under drought stress conditions experiment ws conducted in lortory nd greenhouse Tehrn University. Experiments design in fctoril rndomized complete lock with three replictions. Tretments exmintion included two levels of priming t 10 nd 15 C (first fctor), Gierellic cid tretments on three levels: 50, 100 nd 150 ppm for 12 h (second fctor) nd osmotic potentil (PEG 6000) in three 10 -, 12 - nd 14 - times (fctor III), respectively. The results showed tht pretretment with Gierellic cid of seed in drought stress conditions in vitro incresed the germintion percentge, germintion rte, root length nd shoot length nd lso in medium pot, seedling estlishment percentge, seedling estlishment rte, root length nd shoot length s influenced y different hormonl tretments, with Gierellic cid 50 ppm t 10 C nd Gierellic cid 100 ppm t 10 C nd - 10 times surfce in the experimentl vitro nd pot hve hd the most positive effect on germintion nd growth fctors, respectively. The seed priming tretment t 10 C compred with 15 C displyed etter outcome. Key words: secle montnum, Gierellic cid, wter stress, seed germintion nd seedling estlishment INTRODUCTION Cerel is the most importnt source of food in humn life tht it is including 65-70% crohydrte, proteins nd too much B vitmins nd trce minerl. most of Irnin food re supplied y grin nd diry products. Mountin rye (secle montnum) is the plnt of fmily of grsses (pocece). It is the most stle to cold nd hs long clusters nd flt leves. It grows in clerous dry lnd nd seeds of this plnt re used for mking flour nd red. Seed germintion is complex physiologicl process tht it is response to environmentl signls such s wter potentil, light, nitrte nd other fctor.unfvorle environmentl conditions for seed genertion nd seedling emergence in dry nd semi dry lnd is min cuses of emergence nd seed poor estlishment. One of the limiting fctors is drought condition tht cuses stress t the plnt. Drought stress induced y high evportion, high osmotic potentil in sline soils nd soil freezing. Resulting is roken thermodynmic equilirium condition nd reduce ville of wter. Seed germintion nd seedling estlishment phse primrily ws very sensitive to drought stress tht is reduced the germintion nd estlishment of seedling grown [1, 2, 3]. Increse performnce of seed thought the used of different method promoting cn chieve the finl gol the sme incresing seed nd forge proceeds is very effective.among the most common nd efficient methods in this regrd is seed tretment efore plnting which is 1
clled priming [4]. Studies hve shown tht soking seeds with pproprite concentrtions of growth hormone, hs positive effect on germintion, growth nd yield of different plnt species nd stress conditions re norml [5,6]. Gierellic cid is plnt growth hormone tht the most importnt role in the control of seed germintion is fcilitted. In previous study investigted of the effect of gierellic cid levels (ppm 50, 100, 200, 400) on the germintion of 11 species of romtic plnts nd the result showed tht there were significnt differences etween species nd gierellic cid, ppm 400 gretest effect on seeds will germinte [7]. Also investigted of the effects of hormonl priming on germintion nd seedling growth of lettuce, nd then reported tht the concentrtion of GA3 sttisticlly significnt difference [8]. Mountin rye is the nutrients grsses tht use for humn nutrition nd forge production gstronome nd normlly, growth in the erly stges of germintion nd seedling stresses such s drought nd low tempertures encountered Thus, we pre-treted with the hormone gierellic cid seed germintion seed crop in drought conditions improve is possile to increse the strength of the initil seed finlly creted the incresing in the percentge nd rte of seed germintion nd seedling estlishment. The im of this study evlution the gierellic cid tretment in improving germintion under drought stress in vitro nd pot culture. MATERIALS AND METHODS 1 The Mountin rye plnt seeds purchsed from Isfhn Pkn Seed Compny in crop yer t 1391 nd the experiment design to fctoril in rndomized lock with three replictions in greenhouse Tehrn University ws conducted. 2- Lortory studies: First experimentl mterils (petri dishes, filter pper, forceps, etc.) sterilized for 3 hours in n utoclve nd lso plnt seeds plced sterilized in 3% sodium hypochlorite solution for one minute for steriliztion nd then wshed in sterilized distilled wter nd then the seeds treted hormonl with gierellic cid levels t 3, 50, 100 nd 150 ppm (prts per million) t 10 nd 15 C for 12 hours. Then plnt seeds were wshed with distilled wter nd 50 seeds in ech petri dish cultures were grown on two lyers of filter pper nd then plnted seeds in three replictes t 20 C (optiml temperture). Finlly, pply solution of poly ethylene glycol stress concentrtion in three 10 -, 12 - nd 14 - were used in ech petri dish, s this solution ws dded 5 ml nd 7 dys t 20 C for evluted. 3- Pot evlution In this step, the seeds of the plnt crop, such s lortory tests, prepdre nd plced of culture petri dishes in pots (dimeter 10 cm) contining snd ws used for the stress of polyethylene glycol technology ws used t the eginning of culture, 25 ml of solution ws dded to ech pot nd finlly, fter 7 dys of germintion prmeters were mesured. 4 - Prmeters mesured included: germintion percentge, germintion rte, root length nd shoot length (medium petri dish), the percentge rte of seedling estlishment nd seedling estlishment (in medium pot). The during germintion nd the proper temperture germintion ws sed on the sttic informtion. A mountin rye seed t 20 C for 7 dys in germintor ws in drkness. Germintion ws ssessed on dily sis nd re the seeds with rdicl length of 2 mm re considered s seed germintion (AOSA, 1990), nd finlly the forementioned chrcteristics were evluted. 5- Dt nlysis: The seed germintion percentge ws clculted using eqution (1) [9]: 100 (numer of seeds / numer of seeds germint until the lst dy) = germintion percentge (Eqution 1) To clculte the germintion rte germintion (Eqution 2) ws used to Timson index [10]: Germintion Rte (GR) = Σ (G / t ) (Eqution 2) In this eqution, GR is germintion rte nd G is estimted numer seed germintion per ech dy nd t is dys counted. To clculte the rte of seedling estlishment nd seedling estlishment were used of these reltions (etween 1 nd 2), so the reltionship, ws used seedling estlished insted of numer of seeds germinted. Proity Anlysis Sttisticl nlyzes were performed with SAS softwre compre dt using the lest significnt difference (LSD) ws performed with MSTATC softwre nd grphs were plotted y Excel softwre. 2
RESULTS AND DISCUSION The results of the evlution of germrmintion of S. Montnum in drought condition in vitro showed tht don t hve significnt levels the interction of priming temperture * surfce priming * surfce drought. Also the interction of surfce priming * drought stress on germintion percentge nd germintion rte ws significnt t 1% level nd the interction of priming temperture * surfce stress on the rte nd percentge of germintion nd the interction of surfce primin *tempertur showed tempertur effect on length root. The overll temperture of the priming, solution of gierellic cid nd different levels of drought stress on the index germintion evluted hs significnt effect on the t the 1% level (Tle 1-1). Tle (1-1) nlysis of vrince to ssess trits ssocited with plnt shoots S. Montnum ffected y gierellic cid stress conditions in lortory culture Men squres (MS) Sources chnges degrees of Germintion Root During the Germintion rte freedom percentge length shoot Block 2 0.01 ns 0.03 ns 1.04 ** 14.51 Prime temperture 1 13.07 ** 8.24 ** 4.64 ** 177.85 Prime surfces 2 6.44 ** 7.34 ** 21.27 ** 1014.74 ** Drought stress 2 42.65 ** 37.36 ** 20.63 ** 403.29 ** temperture * Surfce priming 2 0.56 ns 0.79 * 0.28 ns 5.85 ns Temperture *Drought stress 2 0.22 ns 0.08 ns 0.62 ** 22.74 * Surfce priming *Drought stress 4 0.02 ns 0.11 ns 1.24 ** 31.40 ** temperture *Surfce priming* 4 0.08 ns 0.112 ns 0.13 ns 4.74 ns stress Error 34 0.25 0.15 0.106 4.63 Coefficient chnges - 12.24 10.17 5.68 5.09 Ns, *, **: Non-significnt, significnt t the 5 nd 1%, respectively Anlysis sed on the comprison of chrcteristics of the interction of gierellic cid on germintion * drought stress levels were difference significntly so tht with incresing levels of gierellic cid nd whit incresing osmotic potentil decresed germintion percentge significntly (Figure 1-1). The results from the nlysis of the verge comprison of germintion percentge of seeds cn e expressed the interction of temperture priming * drought stress levels on germintion percentge hve significnt effects nd with incresing temperture priming nd incresing the osmotic potentil of -10 to -14 times, decresed germintion percentge significntly (Figure 1-2). Germintion percentge c c d fg de ef g ۵٠ ١٠٠ ١۵٠ Interction of gierellic cid levels * levels of stress Figure (1-1): Averge comprison, interctive effects of gierellic cid * surfce of drought stress on seed germintion S. Montnum under drought stress in vitro According to the results, the germintion rte under drought stress in vitro is ffected y interctions of temperture priming * drought stress levels, lso the interction of drought stress levels * gierellic cid level. The effect of seed priming tretments on germintion rte t 10 C ws thn t 15 C nd lso incresing levels of gierellic ppm 50 to ppm 150 nd lso y incresing s well s the stress increses drought stress the germintion rte diminished (Figure 1-3 nd 1-4). Results sed on comprison nlysis reveled tht the interction of priming temperture nd different levels gierellic cid nd concentrtions of different solution hs significnt effect on root length nd mximum length of root ws in priming tretments 10 C gierellic cid ppm 50 with the potentil nd the osmotic 3
concentrtion of -10 times (1-5 nd 1-6). Bsed on the results, the priming effect of gierellic cid on shoot length s well s the positive effect of this tretment on root length ws similr to seed priming tretment t 10 C with solution of ppm 50 nd osmotic concentrtion - 10 time hd the gretest impct (Figure 1-7 nd 1-8 nd 1-9). ntion percentge ١٠-١٢- c Figure (1-2): Averge comprison, interctive effects of temperture priming * drought stress on germintion seeds of S. Montnum under drought stress in vitro Germintion rte c d e f ١٠-١٢- ١۴- Interction of temperture Priming *stress level Figure 1-3: Averge comprison, interctive effects of temperture priming * surfce drought stress on germintion rte of the seeds of S. Montnum under drought stress in vitro Figure (1-4): Averge comprison, interctive effects of gierellic cid * surfce of drought stress on germintion rte of seeds S. Montnum under drought stress in vitro Root length (cm) c ۵٠ ١٠٠ ١۵٠ temperture interction Priming * gierellic cid Figure (1-5): Averge comprison, interctive effects of priming temperture * gierellic cid on root Length S. Montnum under drought stress in vitro 4
Root length (cm) ١٠-١٢- ١۴- Figure (1-6): Averge comprison, the effect of drought stress on root Length S. Montnum under drought stress in vitro c Root length (cm) Priming ١٠ temperture ( ١۵C) Figure (1-7): Averge comprison the verge temperture of the priming effect ( º C) on oth seed length S. Montnum under drought stress in vitro Germintion rte c c e c d e ۵٠ ١٠٠ ١۵٠ Interction of gierellic * stress levels Figure (1-8): Averge comprison, effect of gierellic cid levels (ppm) in shoot length S. Montnum under drought stress in vitro Figure (1-9): Averge comprison, effect of levels of stress (lod) on the shoot length S. Montnum under drought stress in vitro According to nlysis of vrince nlysis in pot culture under conditions of wter stress, the interction priming level * drought stress on root length nd shoot length (5% level), the interction priming temperture * surfce gierellic cid on seedling estlishment rte (5% level) nd temperture effects prime temperture, prime levels nd different levels of drought stress on the components of the germintion (percentge, rte, root length nd shoot length) ws significnt t 1% level (Tle 1-2). The results of the comprison of chrcteristics of seed germintion of S. Montnum the medium vses, nd priming under drought stress ffects different tempertures, with different levels of gierellic cid nd lso reported tht the percentge of seedlings decresed with incresing osmotic solution concentrtion nd generlly the highest estlishment of gierellic cid ppm 50 tretments in the 10 C temperture nd stress - 10 times ws oserved (Figure 1-10, 1-11 nd 1-12). Averge Comprison showed tht the interction of priming temperture * gierellic cid levels on the rte of seedling estlishment in drought conditions hve different significntly nd priming 5
tretments t 10 C nd y gierellic cid 100 nd 150 (ppm) ws more effective in incresing the rte of seedling estlishment hs een in drought conditions (Figure 1-13) nd lso reported tht the increse in osmotic potentil rte of seedling estlishment ws significntly reduced (Figure 1-14). Tle (1-2) nlysis of vrince to ssess chrcteristic ssocited with plnt germintion S. Montnum ffected y gierellic cid in drought stress conditions in pot culture Men squres (MS) Sources chnges degrees of Seedling Rte of seedling Root During the freedom estlishment (%) estlishment length shoot Block 2 0.60 * 0.80 ** 0.10 ns 14.29 * Prime temperture 1 5.22 ** 3.94* * 3.86 ** 20.29 ** Prime surfces 2 9.87 ** 9.39 ** 2.49 ** 163.18 Drought stress 2 29.10 ** 22.27 ** 24.16 ** 757.40 ** temperture * Surfce priming 2 0.007 ns 0.06 ns 0.32 * 7.18 ns Temperture * Drought stress 2 0.52 ns 0.29 ns 0.08 ns 1.40 ns Surfce priming *Drought stress 4 0.63 * 0.45 * 0.19 ns 5.62 ns temperture * Surfce priming* stress 4 0.13 ns 0.07 ns 0.05 ns 0.29 ns Error 34 0.16 0.14 0.07 4.02 Coefficient chnges - 8.68 9.32 5.80 4.78 Ns, *, **: Non-significnt, significnt t the 5 nd 1%, respectively The sed on the ssessment of the root in potting medium under constnt tension throughout the plnt root S. Montnum influenced y the interction of gierellic cid levels * stress levels nd gierellic cid ppm 100 tretment on stress -10 times ws highest nd gierellic cid tretment ppm 50 on stress - 14 times ws lowest root length nd seed priming in temperture t 10 C ws positive effect thn 15 C (1-15 nd 1-16). The length chnge, the study compred n nlysis similr to reported chnges so tht the mximum shoot length in seed priming tretment t 10 C nd solution of gierellic cid 100 nd 150 (ppm) nd potentil t -10 times were oserved (Figure 1-17 nd 1-18). Figure (1-10): Averge comprison, the effect of priming temperture on seed germintion S. Montnum under drought stress conditions in pot Figure (1-11): Averge comprison, the effect of gierellic cid on the germintion of seeds of S. Montnum under drought stress conditions in pot 6
Figure (1-12): Averge comprison, the effect of drought stress on seed germintion S. Montnum under drought stress conditions in pot Figure (1-13): Averge comprison, the interction of priming temperture * gierellic cid surfce on the rte of seedling estlishment of S. Montnum under drought stress conditions in pot Figure (1-14): Averge comprison, the effect of surfce stress on the rte of seedling estlishment of S. Montnum under drought stress conditions in pot Figure (1-15): Averge comprison, the interction of gierellic cid levels * drought stress levels on root length of seeds of S. Montnum under drought stress conditions in pot 7
Figure (1-16): Averge comprison, the effect of temperture priming on the length of the root or seed S. Montnum under drought stress conditions in pot Figure (1-17): Averge comprison, the effect of gierellic cid levels * levels of drought stress on the shoot seeds of S. Montnum under drought stress conditions in pot Figure (1-18): Averge comprison, the effect temperture priming on shoot length S. Montnum under drought stress conditions in pot In drought conditions my e impired wter sorption y seeds, which in the cse of metolic function in seed germintion will occur slowly, the time of root growth nd seed germintion rte of this decrese, To cope with stress disorders nd improve seed germintion, pretreted seed plnt hormone gierellic cid in prticulr will hve positive impct. In this study, the effect of the hormone gierellic cid on seed germintion trits, including the percentge nd rte of germintion, root nd shoot gierellic cid 50 nd 100 (ppm), respectively, in vitro lortory nd greenhouse conditions with the gretest impct on improving seed germintion of stress. Gierellic cid hormone secreted y the emryo seed encoding the genes enzymes involved in seed germintion, especilly the lph-mylse enzyme is ctive nd this is done y incresing the mrna encoding the enzyme tht wy in germintion nd erly seedling estlishment plys role. Bsed on pulished reports, mny reserchers showed tht the effect of gierellic cid on improvement germintion on the stress, tht results re in greement y this study [11, 12, nd 13]. Mny reports suggest the improved germintion ehvior nd relted indictors such s men time to germintion, seed vigor, root length, shoot length, the rte of seed germintion nd initil estlishment of the priming seed [14,15]. Also in this cse study cn e regrding the positive role of gierellic cid on lettuce seed germintion nd promote [16]. On sorghum nd sunflower seeds to increse germintion under drought, slinity nd frost point e. 8
It seems tht the technique of priming to trnscrie erly trnscription of DNA nd incresed RNA nd protein synthesis to the seeds tht grow emryos lso increses, the prts dmged seed repir would dischrge its metolites reduce the rte nd uniformity of seed germintion nd seedling emergence cn improve [16]. CONCLUSION Results from this study showed tht pretreted of S. Montnum seeds y hormone gierellic cid improves germintion under drought stress so tht the priming seed tretment with gierellic cid ppm 50 t 10 C in petri dish culture, germintion percentge, germintion rte, root length nd shoot length incresed significntly in the stress three levels. The results indicte tht the expression of chrcteristic in the pot priming tretments hd significnt effect on germintion componentsso tht the seed tretment with gierellic ppm 100 t 10 C is shown the mximum positive impct in incresing seedling estlishment, seedling estlishment rte, root length nd shoot length. REFERENCES [1] A Soltni, A Ghderi, A Memr, Journl of Agriculturl Sciences nd Nturl Resources, 2008, 14 (5), 8. [2] R Foti, K Aureni, A Tigere, J Gotos, J Gere,, Journl of Arid Environments, 2008, 72, 1127-1130. [3] F Shekri, R Pkmehr, M Rstgoo, M Vzyefi, M Ghoreishi Ns, Journl - Reserch of Agriculturl Sciences, 2011, 13 (4), 29-13. [4] S Bourgne, C Jo, D Jo, Seed Science Reserch, 2000, 10, 153-161. [5] BL Dr, SP Seth, H Sing, RS Mendirtt, Agronomy Journl, 1973, 65, 292-295. [6] R Hurli, FJ Vn stden, MT smith, Annls of Applied Boilogy, 1991, 118, 175-184. [7] A Liop, G Zkynthinos, T Vrzks, I Nickolos Xynis, Journl of Medicinl Plnts Reserch, 2011, 5(17), 4065-4073. [8] H Mhmoudi, H Zrgouni, I Trchoune, O Btour, N Nsri, R Ben Mssoud, M Zghdoudi, w Aidi, J Hung, A Hnnouf, M Lchâl, Z Ouerghi, Africn Journl of Biotechnology, 2012, 11(45), 10373-10380. [9] A Mohseni, R Rezee, M Rmezni, H Moser, Qurterly Journl of Crop Physiology, 2011, 2(2), 44-25. [10] P Msoodi, A Geznchin, A Jjermn, Journl of Agriculturl Science nd Technology, 2009, 22 (1), 67-57. [11] A Pslr, B Rezee, Effect of different tretments on dormncy nd stimulte germintion of Chenopodium, Fifth ntionl conference on new ides in griculture, 2011, pp: 4. [12] FS Mueungu, P Chiduz, LJ Nymugft, WR Clrk Whlley, W.E. Finch- svge. Field Crops Res, 2004, 89, 49-57. [13] S Run, Q Xue, K Tylkowsk, Seed Science technology, 2002, 30, 451-458. [14] SS Lee, JH Kim, SB Honge, SH. Yuu, EH Prk, Koren Journl of crop Science, 1998, 43, 194-198. [15] CA Prer, DJ Cntliff, Jnnick Journl, 1994, 16, 119-141. [16] H Sdeghi, F Khzei, L Yri, S Sheidei, ARPN Journl of Agriculturl nd Biologicl Science, 2011, 6(1), 39-43. 9