Drought mitigation potential of Azospirillum inoculation in canola (Brassica napus)

Transcription

1 Journl o Appli Botny n Foo Qulity 89, (2016), DOI: /JABFQ * Dprtmnt o Botny, PMAS Ari Ariultur Univrsity Rwlpini, Pkistn Drouht mitition potntil o Azospirillum inoultion in nol (Brssi npus) Mimon S 1*, Noshin Ilys, Roomin Mzhr, Ftim Bii, Nzim Btool (Riv Dmr 9, 2015) * Corrsponin uthor Summry Azospirillum is onsir to most tiv Plnt Growth Promotin Rhizotri (PGPR), whih is rsponsil or vrious moiitions in plnts to op with strss onitions. Thror, th prsnt rsrh ws plnn to vlut th t o Azospirillum liporum (GQ ) inoultion on rowth, iohmil, yil ttriuts o nol rown unr rouht onitions. Two irnt mos o inoultion wr us; i.., inoultion o ss irtly n xposur o plnt s in th rhizosphr. Drouht strss ws impos t lowrin st. Azospirillum s inoultion ws hlp mitit strss ts y improvin rmintion prnt up to 12.49%. Root r ws inrs up to 18.5% n 11.38% with s n rhizosphr inoultion in rouht strss rsptivly. Chlorophyll ontnts n wtr potntil wr inrs 12.21%, n 11.0% in ss inoult unr rouht onitions. Suproxi ismuts tivity ws rs up to 24.6% n 12.5% in s n rhizosphr inoult plnts unr wll wtr onitions. S inoultion ws most tiv, s numr o ss pr po n s wiht pr plnt ws siniintly inrs up to 25%, n 14.28% s ompr to th ontrol. In onlusion, Azospirillum n mitit ltrious ts o rouht strss in nol unr wtr iiny onitions. Ky wors: PGPRs, Cnol, rouht Introution Drouht strss is srity o vill wtr in soil, whih is nssry or optimum rowth n rproution o plnts (Lihtous, 2010). Ru wtr supply is on o th most importnt tors whih is rsponsil or rution in riulturl proutivity (Mhmoo t l., 2009; Ashr, 2010). Diiny o wtr uss injurious ts on plnts y ruin rowth, rsin nutrint intk n hnin wtr sttus o plnts (Ali n Ashr, 2011; Shhz t l., 2011). Ru wtr supply lso uss lin in l vlopmnt n n ltr stomtl onutn (Qri t l., 2006). In ition to morpholoy, physioloil prosss, inhiit y wtr strss r photosynthsis, ll turiity n ll rowth (Thir t l., 2007). In short, vry tur o plnt rowth is t y wtr strss inluin ntomy, morpholoy, iohmistry, plnt physioloy n yil (Jons t l., 2003; Hiz t l., 2004). Cnol (Brssi npus L.) oil proutivity stns thir tr soyn n plm oil rops in th worl. It prous s muh s 14.7% o th vtl il oil n hs hih ontnt o unsturt tty is (Ysri t l., 2008). Compr to rl rops, nol is rnk ith tr wht, miz, ri n otton (Croz n Stwrt, 2003). Cnol oil is prmium ookin oil tht hs lss thn 2% rui i n is low in sturt tty is. It lso is rih in mono poly unsturt tty is whih hlps to rs holstrol lvl (Crvloh t l., 2006; Omi i t l., 2010). Drouht is on o th importnt strss tor whih is rsponsil or ruin proution o nol in smi-ri rions o th worl. Almost, 17 to 70% yil rutions hv n ror u to rouht (Nsri t l., 2007). Plnt rowth-promotin rhizotri (PGPR) r l to promot rowth n yil o plnts unr strss onitions. Inoultion o ths miroornisms provis hihr rop yil without intrrin with nturl prosss in th osystm (Thkor, 2006). Durin th lst two s, vrious tri (i.., Azototr sp., Azospirillum sp., Atotr sp., Billus, Psuomons sp.) r us or plnt rowth promotion unr vrious prvilin ioti n ioti strsss (Turn t l., 2006). Azospirillum is on o th vry tiv Plnt Growth Promotin Rhizotri (PGPR) whih t s nrl root olonizr to improv rop rowth n yil up to 5 to 30%. Azosprilium or inxpnsiv n sy pplition whil proviin minrls, n phytophormons s wll s ix nitron, n ru th synthsis o thyln thry Inrsin yil (Ysri t l., 2008). Azospirillum spp inoultion n improv tolrn to wtr strss, improv th rowth o plnts in ri n smiri rions (Ily s n Bno, 2010). Vrious stuis hv oumnt th rol o Azospirillum in improvin rowth n yil o nol (Ysri t l., 2008; Bnihil t l., 2013). Th impt o wtr iit onitions on plnt physioloy hs n stui or mny yrs. Th prsnt stuy ws onut in orr to vlut th t o Azospirillum inoultion on rowth n yil o nol. Furthr, physioloil n iohmil rsponss o nol unr rouht strss n th rol o Azospirillum in mitition o rouht strss in nol wr lso stui. Mtrils n mthos To vlut th t o Azospirillum inoultion on rowth n yil prmtrs o nol plnts unr rouht strss pot xprimnt ws onut in rn hous, Dprtmnt o Botny, Pir Mhr Ali Shh Ari Ariultur Univrsity, Rwlpini. Two nol vritis (i.., Rinow n Con II) wr ollt rom th Ntionl Ariulturl Rsrh Cntr (NARC), Islm. Ss o oth vritis wr sur striliz in 0.1% mruri hlori solution or 5 minuts, thn wsh thorouhly with istill wtr. Inoulum ws prpr y inoultin 24 h ol ultur o Azospirillum in LB mi n kpt or 72 h on shkr, thn ntriu t 10,000 RPM or 10 min. Th pllt ws ilut to 100 ml with istill wtr n th suprntnt ws isr. Optil nsity ws lult to on. Six hour striliz ss wr sok in th inoulum n th ht kill inoulum ws prpr y utolvin th inoulum. Six ss pr pot wr sown rom 19 Novmr 2013 to 10 Frury For rhizosphr inoultion liv ulturs wr into soil tr sowin th striliz ss. Ht kill tri lso wr inoult onto ss. Eh trtmnt ws rplit thr tims. Plnt rouht strss ws impos t th lowrin st y rstritin wtr or tn ys lon with ontrol or h inoultion. Grmintion nlysis For rmintion nlysis unr rouht onitions, inoult n uninoult ss wr pl in st o ptri plts, whih wr

2 Potntil o Azospirillum or rouht 271 provi with moist iltr ppr. Ss wr monitor ily or mn ily rmintion. Th s ws onsir rmint whn its ril mr 5 mm. Un-inoult ss wr sown in nothr st o plts. Followin prmtrs o rmintion wr vlut. Grmintion prnt For h trtmnt, rmintion prnt ws stimt y usin th ollowin ormul: Grmintion % = Totl ss rmint / Totl no o ss plnt 100 Grmintion inx Usin th ollowin ormul (Ist, 2005), rmintion inx ws lult. Grmintion Inx = n/ Whr, = ys tr plntin, n = no o slins mr on y Promptnss inx By usin th mtho o (Norn t l., 2007) Promptnss inx (P.I) ws trmin. P.I = n2 (1.00) + n4 (0.75) + n6 (0.50) + n8 (0.25) Whr (n2 n8) r numr o mrin slins on y 2, 4, 6, 8. th rk rtion in nothr st o tus. Asorn or oth sts o tus ws ror t 560 nm wvlnth (Ginnopolitis n Ris, 1977). SOD μ/ml = Asorn o smpl K vlu ilution tor / Wiht o th smpl Yil prmtrs: Vrious yil prmtrs o nol plnts wr nlys suh s po lnth, numr o ss pr po, numr o pos pr plnt n s wiht pr plnt, stimt tr hrvstin. Trtmnts T0 Wll wtr n ontrol T1 S inoult n wll wtr T2 Rhizosphr inoult n wll wtr T3 Ht kill-inoult n wll wtr T4 Drouht strss n un-inoult T5 S inoult n rouht strss T6 Rhizosphr inoult n rouht strss T7 Ht kill-inoult n rouht strss Sttistil nlysis Th sotwr us or sttistil nlysis ws Sttistix 9.1. A two wy ANOVA with toril lok sin ws rri out or ll trtmnts. Eh trtmnt h thr rplits Slin vior inx Th mtho o Aulki n Anrson (1973) ws us to trmin th Slin vior inx. Slin vior inx = rmintion % Slin lnth (mm) Plnt rowth prmtrs Atr wshin root n shoot lnth wr msur or thr plnts or h trtmnt. Physioloil prmtrs A Sholnr prssur hmr ws us or trmintion o l wtr potntil (Sholnr t l., 1965). Chlorophyll ontnt ws trmin y ollowin th mtho o Bruinsm (1963). Biohmil prmtrs Prolin ontnt ws trmin y usin sptrophotomtr (Bts t l., 1973). Solul sur ws trmin y ollowin Duois (1951). For protin trmintion, n xtrt o plnt smpls ws prpr y homonizin 0.2 o rsh l mtril in 4 ml o soium phospht ur (ph 7) n thn ntriu. In sprt tst tu 0.5 ml o l xtrt n 0.5 ml istill wtr n 3 ml o Coomssi io r y wr mix. Th rtion mixtur ws pl unistur or 5 minuts n th sorn ror t 595 nm (Bror, 1976). Antioxint nzym ssy Suproxi ismuts For trmintion o suproxi ismuts, 10 ml soium phospht ur ws us to rin 0.5 l mtril. Th solution ws llow to sttl n thn 0.1ml o xtrt ws to nothr st o tst tus lon with 0.1 ml riolvin n 3 ml phospht ur. This rtion mixtur ws pl unr luorsnt lmp or 8 min to strt rtin. Th sm rtion mixtur ws prpr or Rsults Grmintion prmtrs Th t o plnt inoultion with Azospirillum liporum (Assion no. GQ255949) on rmintion o two nol vritis ws siniint (p 0.05). Drouht strss rs th rmintion prnt up to 37% s ompr to wll-wtr onitions (T. 1). S inoultion, with tri rsult in 10% inrs in rmintion unr wll-wtr onitions n 12.49% inrs unr wtr strss. Ht kill inoulums h no siniint t on rmintion prnt unr oth wtr rims. Rsults rom th rmintion inx (T. 1) show similr t o rouht strss. Rution in rmintion inx ws improv y trtin th ss with Azospirillum. Atr inoultion unr oth wll wtr n rouht onitions n 8.91% n 12.28% inrs ws osrv in plnts rltiv to thir rsptiv ontrols. Et o th ht kill inoulum rmin insiniint unr oth ontrol n rouht onitions. A siniint inrs in promptnss inx ws nountr in plnts trt with Azospirillum inoulum unr oth wll wtr onitions n unr wtr short (T. 1). Thr ws n inrs o 20.87% in slin viour inx tht ws ror in plnts inoult unr wll wtr onitions s ompr to un-inoult plnts. A similr trn o inoultion ws osrv in plnts rown unr rouht onitions; 12.4% n 4.16% inrs in slin viour inx with oth mos o inoultion ws osrv. Thr ws siniint irn osrv or ll th prmtrs o rmintion in oth nol vritis (T. 1). Et o inoultion on rowth prmtrs Th t o tril inoultion on rowth prmtrs o plnts ws siniint (p 0.05) whn ompr to ontrol plnts in oth s n rhizosphr inoultions. A rs o 10% ws osrv in shoot lnth or rouht xpos plnts s ompr to ontrol plnts (Fi. 1A). With rstrit wtr supply, oth s n rhizosphr inoultions rsult in 7.2% n 4.5% inrs in shoot

3 272 M. S, N. Ilys, R. Mzhr, F. Bii, N. Btool T. 1: Et o Azospirillum inoultion on rmintion prmtrs o two nol vritis unr rouht strss. Trtmnts Slin vior inx Promptnss inx Grmintion inx Grmintion % V1 V2 V1 V2 V1 V2 V1 V2 T0 780±5h ±4i 4.5± ± ± ± ±1 T ±5 650±4h 2.25± ± ±0.2 1± ±0.9 50±1 T ± ±4 6.25±0.2 6± ± ± ± ±0.9 T ± ±4 4.75± ± ± ± ± ±0.9 T ±5k ±4 3.25±0.1i 2.5± ± ± ± ±0.9 T ±5j 700±4 2.5±0.1h 2.25± ±0.2 1± ±0.9 50±1 Whr, To= Un-inoult n wll wtr, T1= Un-inoult n rouht xpos, T2= S inoult n wll wtr, T3= S inoult with ht kill inoulums n wll wtr, T4= S inoult n rouht xpos, T5= S inoult with ht kill inoulums n rouht xpos. An V1=Rinow, V2=Con II. h h Fi. 1A: Et o Azospirillum inoultion on shoot lnth o two nol vritis (v1 n v2) rown unr rouht strss. Whr, To = Un-inoult n wll wtr, T = Un-inoult n rouht xpos, T2 = S inoult n wll wtr, T3 = Rhizosphr inoult n wll wtr, T4 = S inoult with ht kill inoulums n wll wtr, T5 = S inoult n rouht xpos, T6 = Rhizosphr inoult n rouht xpos, T7 = S inoult with ht kill inoulums n rouht xpos. An V1 = Rinow, V2 = Con II. lnth rsptivly. Dt rrin shoot rsh n ry wiht lso show th sm rsults with irnt mos o inoultion (Fi. 1B n 1C). Anlysis o t rom wll wtr n rouht xpos plnts trt with Azospirillum show siniint irn twn inoult n ontrol plnts. S inoultion rsult in 23.68% inrs in root r ompr to ontrols trt unr wll wtr onitions. Evn rhizosphr inoultion rsult in 18.09% inrs in root r ompr to rulrly wtr plnts. A similr rsult ws osrv in rouht xpos plnts, whr 18.5% n 11.38% inrs in r trt with s n rhizosphr inoultions rsptivly. Th inrs o 9.72% in root r ws osrv with ht kill inoulum ompr to ontrol plnts rown unr rouht onitions. Rinow vrity show ttr rspons towrs trtmnts ompr to on II. Physioloil prmtrs Drouht strss us 29.7% rs in wtr potntil or th l in oth nol vritis (Fi. 2A). Inoultion tn to ru th rouht t rsultin in 11.01% n 8.26% inrs in wtr potntil with s n rhizosphr inoultion unr rouht strss onitions. Et o ht kill inoultion ws not siniint un- Fi. 1B: Et o Azospirillum inoultion on shoot rsh wiht o two nol vritis (v1 n v2) rown unr rouht strss. Whr, To = Un-inoult n wll wtr, T = Un-inoult n rouht xpos, T2 = S inoult n wll wtr, T3 = Rhizosphr inoult n wll wtr, T4 = S inoult with ht kill inoulums n wll wtr, T5 = S inoult n rouht xpos, T6 = Rhizosphr inoult n rouht xpos, T7 = S inoult with ht kill inoulums n rouht xpos. An V1 = Rinow, V2 = Con II. r oth wll wtr n rouht onitions. Th t o rouht strss on photosynthti pimnts ws mor pronoun ompr to othr prmtrs. Consirl vrition ws osrv in hlorophyll ontnt o wll wtr n wtr strss plnts. Wtr iit strss rsult in 23.31% rs in th totl hlorophyll ontnt o wtr strss plnts s ompr to wll wtr plnts (Fi. 2B). Plnts with s n rhizosphr inoultion show 20.28% n 9.52% inrs o totl hlorophyll in lvs ompr to ontrols rown on unr wll wtr onitions. A similr trn ws sn in rouht xpos plnts, whr 12.21% n 5.98% inrs ws sn in s n rhizosphr inoult plnts rown unr wtr iit onitions. S inoultion with ht kill inoulum i not show ny siniint irn in plnts rown unr oth wll wtr n wtr iit onitions. Et on omptil soluts A onsirl mount o osmolyts ws umult in wtr strss plnts. In ontrst to wll wtr plnts 23.29% prolin

4 Potntil o Azospirillum or rouht V1 48 V Fi. 1C: Et o Azospirillum inoultion on shoot ry wiht o two nol vritis (v1 n v2) rown unr rouht strss. Whr, To = Un-inoult n wll wtr, T = Un-inoult n rouht xpos, T2 = S inoult n wll wtr, T3 = Rhizosphr inoult n wll wtr, T4 = S inoult with ht kill inoulums n wll wtr, T5 = S inoult n rouht xpos, T6 = Rhizosphr inoult n rouht xpos, T7 = S inoult with ht kill inoulums n rouht xpos. An V1 = Rinow, V2 = Con II. inrs ws osrv in rouht trt plnts (Fi. 3A). S n rhizosphr inoultion llow th plnts to mintin hih prolin lvl up to 10.34% n 8.62% unr wtr strss ompr to plnts without inoultion. A similr inrs o 21.72% in solul sur ontnt ws osrv in rouht xpos plnts (Fi. 3B). Inoultion ts rmin siniint unr limit wtr vilility whr 10.69% n 7.28% inrs ws osrv in sur ontnt o s n rhizosphr inoult plnts rsptivly. Rsults rvl 36.67% rs in solul protins in rouht impos Fi. 1D: Et o Azospirillum inoultion on root sur r o two nol vritis (v1 n v2) rown unr rouht strss. Whr, To = Un-inoult n wll wtr, T = Un-inoult n rouht xpos, T2 = S inoult n wll wtr, T3 = Rhizosphr inoult n wll wtr, T4 = S inoult with ht kill inoulums n wll wtr, T5 = S inoult n rouht xpos, T6 = Rhizosphr inoult n rouht xpos, T7 = S inoult with ht kill inoulums n rouht xpos. An V1 = Rinow, V2 = Con II. Fi. 2A: Et o Azospirillum inoultion on wtr potntil o two nol vritis (v1 n v2) rown unr rouht strss. Whr, To = Un-inoult n wll wtr, T = Un-inoult n rouht xpos, T2 = S inoult n wll wtr, T3 = Rhizosphr inoult n wll wtr, T4 = S inoult with ht kill inoulums n wll wtr, T5 = S inoult n rouht xpos, T6 = Rhizosphr inoult n rouht xpos, T7 = S inoult with ht kill inoulums n rouht xpos. An V1 = Rinow, V2 = Con II. Totl Chlorophyll ontnt (µ/ml) T0 T1 T2 T3 T4 T5 T6 T7 Trtmnts Fi. 2B: Et o Azospirillum inoultion on totl hlorophyll ontnt o two nol vritis(v1 n v2) rown unr rouht strss. Whr, To = Un-inoult n wll wtr, T = Un-inoult n rouht xpos, T2 = S inoult n wll wtr, T3 = Rhizosphr inoult n wll wtr, T4 = S inoult with ht kill inoulums n wll wtr, T5 = S inoult n rouht xpos, T6 = Rhizosphr inoult n rouht xpos, T7 = S inoult with ht kill inoulums n rouht xpos. An V1 = Rinow, V2 = Con II. plnts (Fi. 3C). Inoultion rus th injurious t o rouht with s inoultion rsultin in 10.02% n 9.48% rs in wll wtr n rouht onitions in ontrst to thir rsptiv ontrols. Rsults wr oun to similr with rhizosphr inoultion, with 12.63% rs in wll wtr n 10.70% lin in strss plnts. Rspons o oth vritis ws siniint ollowin inoultion. Rinow show ttr rspons ompr to Con II. An inrs in supr oxi ismuts (Fi. 4A) ws osrv in plnts rown unr rouht strss ompr to normlly irrit plnts. Supr oxi ismuts tivity ws 35.8% inrs in rouht xpos plnts ompr to ontrol plnts. Dt or Azospirillum inoultion show siniint rs in supr oxi ismuts in inoult plnts s ompr to untrt plnts. S n rhizosphr inoultion show 24.6% n 12.5% rs in supr 55 V1 V2

5 274 M. S, N. Ilys, R. Mzhr, F. Bii, N. Btool i n l m k j h Fi. 3A: Et o Azospirillum inoultion on prolin ontnt o two nol vritis (v1 n v2) rown unr rouht strss. Whr, To = Un-inoult n wll wtr, T = Un-inoult n rouht xpos, T2 = S inoult n wll wtr, T3 = Rhizosphr inoult n wll wtr, T4 = S inoult with ht kill inoulums n wll wtr, T5 = S inoult n rouht xpos, T6 = Rhizosphr inoult n rouht xpos, T7 = S inoult with ht kill inoulums n rouht xpos. An V1 = Rinow, V2 = Con II. Fi. 3C: Et o Azospirillum inoultion on solul protin o two nol vritis (v1 n v2) rown unr rouht strss. Whr, To = Un-inoult n wll wtr, T = Un-inoult n rouht xpos, T2 = S inoult n wll wtr, T3 = Rhizosphr inoult n wll wtr, T4 = S inoult with ht kill inoulums n wll wtr, T5 = S inoult n rouht xpos, T6 = Rhizosphr inoult n rouht xpos, T7 = S inoult with ht kill inoulums n rouht xpos. An V1 = Rinow, V2 = Con II. i h h i h Fi. 3B: Et o Azospirillum inoultion on solul sur o two nol vritis (v1 n v2) rown unr rouht strss. Whr, To = Un-inoult n wll wtr, T = Un-inoult n rouht xpos, T2 = S inoult n wll wtr, T3 = Rhizosphr inoult n wll wtr, T4 = S inoult with ht kill inoulums n wll wtr, T5 = S inoult n rouht xpos, T6 = Rhizosphr inoult n rouht xpos, T7 = S inoult with ht kill inoulums n rouht xpos. An V1 = Rinow, V2 = Con II. Fi. 4A: Et o Azospirillum inoultion on supr oxi ismuts o two nol vritis (v1 n v2) rown unr rouht strss. Whr, To = Un-inoult n wll wtr, T = Un-inoult n rouht xpos, T2 = S inoult n wll wtr, T3 = Rhizosphr inoult n wll wtr, T4 = S inoult with ht kill inoulums n wll wtr, T5 = S inoult n rouht xpos, T6 = Rhizosphr inoult n rouht xpos, T7 = S inoult with ht kill inoulums n rouht xpos. An V1 = Rinow, V2 = Con II. oxi ismuts tivity ompr to un-inoult plnts unr wll wtr onitions. A similr trn o inoultion ws osrv in rouht xpos plnts whr 45% n 43% rs ws osrv ollowin s n rhizosphr inoultion rltiv to thir rsptiv ontrols. A siniint irn ws ror in oth vritis. Rspons o Rinow ws muh ttr s ompr to Con II. Yil prmtrs Dt or yil prmtrs irs siniintly twn wll wtr n rouht xpos plnts. Drouht strss us 36.08%, 9.41%, n 41.42% rution in th numr o ss pr po, numr o pos pr plnt n s wiht pr plnt s ompr to ontrol. A siniint irn ws osrv in Azospirillum inoult plnts s ompr to un-inoult plnts. Th st outom o inoultion or numr o ss pr po ws sn in rouhtxpos plnts whr 25% n 16.67% inrs ws osrv with two irnt mos o inoultion (Fi. 5A). Dt rrin to numr o pos pr plnt (Fi. 5B) rvls 9.41% rs in yil in rouht xpos plnts ompr to ontrol plnts. S n rhizosphr inoultion rsult in 6.07% n 4.88% inrs in yil ompr to ontrol plnts rown unr wll wtr onitions. In rouht onitions, only 3.52% n 2.79% inrs ws osrv ollowin s n rhizosphr inoultion. Whil in th s o s wiht pr plnt s n rhizosphr inoultion rsult in 14.14% n 6.06% inrs in wll wtr ompr to ontrol plnts. In rouht xpos plnts, s n rhizosphr inoultion rsult in 14.28% n 7.14% inrs with rspt to ontrol plnts (Fi. 5C).

6 Potntil o Azospirillum or rouht 275 Fi. 5A: Et o Azospirillum inoultion on numr o ss pr po o two nol vritis (v1 n v2) rown unr rouht strss. Whr, To = Un-inoult n wll wtr, T = Un-inoult n rouht xpos, T2 = S inoult n wll wtr, T3 = Rhizosphr inoult n wll wtr, T4 = S inoult with ht kill inoulums n wll wtr, T5 = S inoult n rouht xpos, T6 = Rhizosphr inoult n rouht xpos, T7 = S inoult with ht kill inoulums n rouht xpos. An V1 = Rinow, V2 = Con II. N0 o pos pr plnt To T1 T2 T3 T4 T5 T6 Trtmnts Fi. 5B: Et o Azospirillum inoultion on numr o pos pr plnt o two nol vritis (v1 n v2) rown unr rouht strss. Whr, To = Un-inoult n wll wtr, T = Un-inoult n rouht xpos, T2 = S inoult n wll wtr, T3 = Rhizosphr inoult n wll wtr, T4 = S inoult with ht kill inoulums n wll wtr, T5 = S inoult n rouht xpos, T6 = Rhizosphr inoult n rouht xpos, T7 = S inoult with ht kill inoulums n rouht xpos. An V1 = Rinow, V2 = Con II. Disussion Drouht strss uss siniint rution in rmintion prnt, rmintion inx n slin vior inx whn ompr to ontrol plnts. Dlin in rmintion omponnts o wtr strss is u to lss wtr intk y th s ot n inhiition o wtr intk t th initil st o rowth (Turk t l., 2004; Bhrmi t l., 2012) or rution in xtrnl wtr potntil. Anothr rson or th rution in rmintion is ly in hyrolysis o stor ompouns in th nosprm n otylon, or slow trnsport o wtr to th vlopin mryo xis (Ay z t l., 2000). Unr rouht strss, nzymti tivity slows own n susquntly rs in th rmintion prnt. Rution in wtr sorption rss ll turiity, ivision, n ril lnth (Zizh t l., 2011). Btril inoultion is n tiv strty to nhn rmintion, improv slin mrn n rspon to xtrnl nvironmntl tors (Lutnr t l., 2002). In th prsnt stuy, onsirl inrs in rmintion prmtrs (T. 1) T7 V1 V2 Fi. 5C: Et o Azospirillum inoultion on s wiht pr plnt o two nol vritis (v1 n v2) rown unr rouht strss. Whr, To = Un-inoult n wll wtr, T = Un-inoult n rouht xpos, T2 = S inoult n wll wtr, T3 = Rhizosphr inoult n wll wtr, T4 = S inoult with ht kill inoulums n wll wtr, T5 = S inoult n rouht xpos, T6 = Rhizosphr inoult n rouht xpos, T7 = S inoult with ht kill inoulums n rouht xpos. An V1 = Rinow, V2 = Con II. ws osrv in s inoult plnts ompr to un-inoult plnts. Rsults o s inoultion wr lso mor pronoun ompr to rhizosphr inoultion n us o ht kill inoulum. Similr ts o tril inoultion on th rmintion prmtr hv n rport in rls suh s sorhum (RAJU t l., 1999), miz (Ghrooi t l., 2012), wht (Bnsh t l., 2013) n in sunlowr (Zizh t l., 2011). This improvmnt my u to synthsis o hormons, n inrs tivity o rtin nzyms lik myls whih sp up strh ssimiltion. S vior inx ws lso improv y n inrs in synthsis o uxin y rmintin slins whn inoult with tri (Bhrthi t l., 2004). Th most immit t o rouht strss is rution in rowth (Shlhvt t l., 1995) n numr o morpholoil prosss vrsly t y wtr iit (Dikin n Wriht, 2008). Durin th prsnt stuy thr ws rs in shoot lnth (Fi. 1 A) tht ws osrv in rouht xpos plnts ompr to wll wtr plnts us o rution in ll ivision n lontion. A similr rution in rowth prmtrs ws osrv y Ashr t l. (2013) n Shi t l. (2009). Shoot rsh wiht n ry wiht show siniint rs unr strss onitions. This rution in iomss ws u to tivity o mtoli nzyms in us unr strss onitions (Hon n Ji-Yun, 2007; Xu t l., 2008), hn in mtolism (Chimnti t l., 2006) n inhiition in ll ivision (ARSHAD t l., 2008). Prviously rution in iomss ws rport y Ashr t l. (2013). Plnts with Azospirillum inoultion show n inrs in shoot lnth n iomss (Fi. 1-3 A). S inoultion prou mximum rsults ollow y inoultion o th rhizosphr, n us o ht kill inoultion ompr to ontrols. Phospht soluliztion, proution o uxin, th ixtion o nitron n nhn nutrint intk r likly rsponsil or ttr shoot lnth n shoot wiht (Bshn n Holuin, 1997; Bshn t l., 2004). Dt lookin t l wtr potntil inits onsirl rs in wtr potntil unr rouht strss. Inoultion tns to ru rouht ts. Inrs in wtr potntil ws osrv in s inoult plnts, lthouh it ws not siniintly irnt rom ontrol plnts (Fi. 1 B). Similr rss in wtr potntil unr rouht strss n its mitition y Azospirillum ws rport y Ily s n Bno (2010). Kr i m i n Mohsni (2013) rport similr lin in hlorophyll ontnt in soyn ultivrs rown

7 276 M. S, N. Ilys, R. Mzhr, F. Bii, N. Btool unr rouht strss. Th usl nt hin th rution o hlorophyll ontnt my lin in pimnt synthsis u to isruption o mro-rts o Chl, Chl, or proution o rtiv oxyn spis (ROS) (Smirno, 1993). Thr ws siniint inrs in hlorophyll ontnt o plnts with Azospirillum inoultion unr rouht strss ompr to un-inoult plnts. S inoultion show th st rsult. A onsirl rs in th hlorophyll ontnt o lvs ws osrv unr wtr strss in nol plnts (Kusr t l., 2006). In th prsnt stuy nlysis o t rrin hlorophyll ontnt show trmnous rs in hlorophyll ontnt o rouht xpos plnts ompr to wll wtr onitions (Fi. 2). Proution n umultion o prolin n solul surs is ommon rspons in plnts xpos to wtr iit strss, whih plys rol in prottion o th ll mmrn n mromolul strutur unr strss onitions (Pro t l., 2000). Similr inins wr rport y Din t l. (2011) in nol plnts. Azospirillum inoultion improvs prolin n solul sur ontnts s rsult o wtr inlux inrss in plnt lls (Fi. 1-2 C). Noshn t l. (2011) lso rport th sm rsult ollowin inoultion o nol. Th rson or this inrs ws th rkown o polyshris, whih hlp to stiliz ll turor (Nzrli t l., 2011). Amon omptil soluts solul protins r quit importnt or th mnisttion o rouht strss. Solul protin ontnt ws rs in plnts xpos to rouht strss (Fi. 3 C). Ths inins wr support y Goo n Zplhinski (1994), who osrv rution in solul protin ontnt in Brssi npus unr strss. Th lin in solul protin ontnts ws u to rution in photosynthsis, sn o rw mtrils or protin synthsis tht us lin in or ompltly stop th pross (Mohmmkhni n Hiri, 2008). In th prsnt rsrh, similr lin ws osrv in protin ontnt, ut inoultion o Azospirillum mitit th ts o strss y ruin proportions o plnts with whih it rss without inoultion unr strss onitions. Th wtr iit onition is ssoit with proution o rtiv oxyn spis (ROS) in th hloroplst, mitohonri n proxisom lin to oxitiv strss. Ativtion o ntioxint nzym systms is ommon rspons in plnts to ru oxitiv strss (Foyr n Notor, 2003). Plnts pn on tivitis o supr oxi ismuts (SOD) n onsquntly on th tivitis o othr ntioxint nzyms (Alshr t l., 2002). In th prsnt stuy, trmnous inrs in supr oxi ismuts ws noti in rouht xpos plnts (Fi. 1). Our rsults r onsistnt with thos o Ai n Pkniyt (2010). Dt or inoult plnts show siniint rution in supr oxi ismuts unr oth wll wtr n rouht onitions. S inoultion hs shown ttr rspons s ompr to th rhizosphr n ht kill inoultion with tri. Ht kill inoultion i not show ny siniint irn in oth wtr rims. Btril inoultion rs th lvl o SOD y stimultin th intk o nitron n phosphorous, whih intrt with rohyrts s non-nzymi ntioxints. Ths sustns utiliz lss nry n inrs spily ompr to nzymi ompouns (Di t l., 2009). Drouht imposs ntiv impt on prormn o nol. Howvr this ntiv t on yil pns on st, urtion o strss n ility o th plnt to op with strss (Trotl-Aziz t l., 2000). Drouht strss ru th yil n yil omponnt in th nol plnt (Birunr t l., 2011). Yil prmtrs o oth plnt vritis tst wr ly t y rouht strss, rsultin in rution in yil omponnts lik numr o pos pr plnt, numr o ss pr plnt, n s wiht pr plnt nountr unr wtr iit onitions ompr to wll wtr onitions (Fi. 1-3 E). Rution in photosynthsis unr wtr iit onitions us po ortion, onsquntly rs numr o pos prou (Dipnrok, 2000). Dlin in s wiht is ssoit with rs in th numr o ss pr plnt s wll s numr o ss pr po. Wtr iit h irt impt on siz o sink, rs stor pity o sour n susquntly us rution in s wiht (Shirnir t l., 2013). Ets on s trt in th rhizosphr n with ht kill inoultion wr not vry tiv ompr to irt s trtmnt with th tri. This my u to ility o th Azospirillum to vlop roots, ilitt nutrint n wtr uptk, ispl pthoni tri n hlp in nitron ixtion (Okon n Itzisohn, 1995). From th prsnt rsrh, w onlu tht Azospirillum liporum (Assion no. GQ255950) ws l to mitit th vrs ts o rouht strss y improvin th morpholoil, physioloil n iohmil spts o th plnt. 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