Plant-microbe interaction of Serratia marcescens and Scirpus mucronatus on phytoremediation of gasoline contaminated soil

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1 Intrntionl Journl of ChmTh Rsrh CODEN( USA): IJCRGG ISSN : Vol.6, No.1, pp , Jn-Mrh 2014 Plnt-miro intrtion of Srrti mrsns n Sirpus murontus on phytormition of gsolin ontmint soil Asi Fhil Almnsoory 1,4, *, Mushrifh Iris 2, Siti Rozimh Shikh Aullh 3 n Nurin Anur 3 1 Shool of Environmntl n Nturl Rsour Sins, Univrsiti Kngsn Mlysi, UKM Bngi, Slngor, Mlysi. 2 Tsik Chini Rsrh Cntr, Fulty of Sin n Thnology, Univrsiti Kngsn Mlysi, UKM Bngi, Slngor, Mlysi. 3 Dprtmnt of Chmil n Pross Enginring, Fulty of Enginring n Built Environmnt, Univrsiti Kngsn Mlysi, UKM Bngi, Slngor, Mlysi. 4 Dprtmnt of Biology, Sin ollg, Bsrh Univrsity, Irq. *Corrs. Author: sin.lmnsoory@yhoo.om Astrt: Th im of this stuy ws to invstigt th plnt-miro intrtion of Srrti mrsns isolts in phytormition of gsolin ontmint soil using plnt ulrush of Sirpus murontus. Th xprimnt inlu ontrol plnt n thr phytormition trtmnts, with gsolin s hyroron pollutnt mol t iffrnt gsolin onntrtions (0, 5, 10 n 30 g/kg) in soil, h with 10% (v/v) of S. mrsns. Th ry n wt wights of th plnts slightly inrs on xposur to gsolin for 42 ys. Th highst wt wight in 10 g/kg gsolin ws g t 72 ys whil ry wight mount 35.6 g. In th 5 g/kg n 30 g/kg gsolin onntrtions, th wt wights wr 43.4 g n 10.8 g with 13.1 g n 6.3 g for ry wight rsptivly. Th phytormition trtmnt using S. mrsns in 10 g/kg gsolin show th highst totl ptrolum hyroron (TPH) rmovl ftr 72 ys of xposur of 89.5% ompr to only 55.0% in th orrsponing unplnt ontrol. Th mximum TPH rmovls in th othr two phytormition trtmnts (5 n 30 g/kg gsolin) wr 84.4 n 83.3%, rsptivly, whil th vrg rmovl in th orrsponing ontrol trtmnts wr 56.3 n 54.2% rsptivly. This inits tht th iogrtion pross y S. mrsns ply signifint rol in th trtmnt. Hn, th synrgy intrtion twn S. murontus n th tri n nfiil in rmition of hyroron in gsolin ontmint soil. Kywor: Phytormition, Sirpus muruntus, TPH, gsolin ontmint soil n Srrti mrsns.

2 Asi Fhil Almnsoory t l /Int.J. ChmTh Rs.2014,6(1),pp Introution Th us of plnts n miroorgnisms ssoit in rhizosphr to nhn iogrtion of ptrolum hyroron pollutnts in th soil hs gin inrsing ptn intrntionlly s vil thnology to ln up ontmint soils [1,2]. Miroil tivity is ssntil to nutrint yling in soils, n orgni pollutnts hv fft on soil miroorgnisms [3]. This intrtion twn root xuts n th miro orgnisms populting th rhizosphr hs n shown to nhn grtion of hyroron ontminnts n oul hv potntil for improving phytormition [4, 5, 6]. Growth of plnt n vlopmnt rlt with th toxiity of hyrorons with low molulr wight, furthrmor, hyrophoi proprtis of hyroron ru th ility of plnts n miroorgnisms to sor wtr n nutrints from th soil [7]. Plnts n miros rquir wtr n nutrints from th nvironmnt [8]. Phytormition is ffiint to rmov hyroron ontmint soil sin it n provi th pproprit nvironmnt for oth plnt n ssoit miroorgnisms. Ths proprtis suh s ph, orgni mttr, tion xhng pity n strutur n fft on plnt growth n vlopmnt [6]. Th fft of ptrolum hyroron ontminnts on plnt n miroorgnisms in soil my of hlp in ssssing th rovry potntil of soil [3]. Rhizormition is typ of miroil ssist phytormition, hs shown on of th most sussful mns y whih plnts n influn th grtion of ptrolum hyroron ontminnts [9]. Aoring to [4] n [5] th growth of root oxitiv grtion of ptrolum hyrorons n stimult y promoting soil rtion tht nhns th prtnrship twn root xuts n th miroorgnisms iogrtion. In th rhizosphr, miroil tivity is nhn y th hlth of th plnt whih ffts on th hlth of th rhizosphr n th ntir phytormition systm [8]. Th pplition of sussful rhizormition is pnnt on th pity of ontminnt grrs or plnt growth promoting miros [10]. Th miroil rmovl of ontminnts in th soil through two mthos of iogrtion n miroil uptk for iogrtion tri mit hmil trnsformtion of orgni ompoun whil miroil uptk is th irt rmovl of th ontminnt y soring ompouns to th mmrn surf [8]. Numrs of miroil inrs in th rhizosphr of plnt [11]. On of th most importnt rquisit to rhizormition of hyroron ontminnt soils is tht plnts r l to grmint n om stlish in th prsn of ontminnts [9]. Th ims of this stuy wr to trmin th plnt-miro intrtion of S. mrsns in gsolin ontmint soil to nhn gsolin grtion through goo prtnrship with ulrush of Sirpus muruntus uring phytormition pross. Mtrils n Mthos Exprimntl Dsign for Phytotoxiity Tst Th xprimnt ws onut in grnhous lot t Univrsiti Kngsn Mlysi. Th glss quri wr us for th phytotoxiity tst. Eh qurium with imnsions of m (L W D) ws fill with 30 kg soil s mixtur of 75: 25 (w/w) grn soil: sn. All th quri wr prpr simultnously n run in prlll. Th prpr gsolin onntrtions of 5,10 n 50 g gsolin/kg. Soil with h of th gsolin onntrtions ws plnt with 20 plnts of th slt plnt, S. murontus whih ws lso sown in ontrol xprimnt using soil without gsolin. Thr rplits pr trt qurium n nothr unplnt qurium for ontrol ontminnts (CC), in ition to nothr qurium without th gsolin ontminnt s plnt ontrol (PC). Stnr gsolin otin from lol Ptrons ptrol sttion ws mix with ton (R&M Chmils, U.K.) s solvnt in th rtio of 1:1 (v/v) n prpr t iffrnt onntrtions (5, 10 n 30 g/kg). Aftr sprying th mixtur soil, it ws stirr until homognous n thn lft to plnting. 20 thr-months-ol hlthy S. murontus plnts wr trnsplnt into h qurium ontining gsolin t iffrnt onntrtions. All xprimntl plnts wr wtr with ionis wtr t fix lult volum. Th ulk nsity of th soil ws 100 g soil mixtur pr 26 ml. Smpling ws onut on y 0, 7, 14, 28, 42 n 72. Dtrmintion of physiohmil proprtis To osrv th hngs in wtr qulity, th physiohmil prmtrs of issolv oxygn (DO, mg/l), oxition rution potntil (ORP, mv) n ph wr ror, using n IQ 150 (IQ Sintifi Instrumnts, UK) multi-pro for ph, ORP n DO snsor (GLI Intrntionl, Mol 63, USA).

3 Asi Fhil Almnsoory t l /Int.J. ChmTh Rs.2014,6(1),pp Plnt growth Th growth of S. murontus ws osrv for 72 ys t iffrnt gsolin onntrtions (0, 5, 10 n 30 g/kg). On plnt ws hrvst on h smpling y (Dy 0, 7, 14, 28, 42 n 72) from th thr rplits of h gsolin onntrtions. Th plnts wr rins with tp wtr n sprt into lowr prt (from th stm uri in th soil to th tip of th longst rootlt) n uppr prt (lvs n stm). All prts wr msur grvimtrilly to trmin thir iomss through oth wt n ry wights [12]. All plnt smpls wr ri in n ovn (Mmmrt, Grmny) t 70ºC for 72 h until onstnt mss ws rh to giv th ry wight [4]. Biogumnttion Btri with Soil This isolt S. mrsns ws ollt from prvious stunts n intifi using iohmil n PCR thniqu. Inoulum t 10% (v/v) of ws mix with ulk nsity of wtr in th soil mixtur n to th plnt sit glss sin ontining gsolin ontmint soil. Th ulk nsity of th soil ws 26 ml pr 100 g of th soil mixtur, whil th wight of th soil mixtur in th glss sin ws 30 kg. So, th volum of wtr in th glss sin is 7800 ml. Ths tri grow in th liqui mi ontining Totl Soy Broth (TSB) shk for 24 h t 150 rpm n 37 ºC. Th ition of th tri to th soil mixtur in n mount of 10% (v/v) tri ws to h qurium. Miroil plt ounts Th miroil popultion ws otin from th soil tht ws firmly tth to th rhizosphr zon. Th popultion of liv tri in th rhizosphr of plnts trt with gsolin-ontmint soil ws trmin through sril ilution mtho. Initilly, 10 g of rhizosphr soil ws hrvst n to 100 ml stril istill wtr to otin 10 2 ilutions [12]. This mixtur ws shkn t 150 rpm for 1 h to rls hring miroorgnisms. Susqunt ilutions up to 10 4 fol wr prpr n 100 µl of h of thr ilutions (10 2, 10 3, n 10 4 ) ws plt on stril plts ontining nutrint gr mium y th pltpouring mtho. Th plts wr invrt n inut t 37ºC for 24 h for th tril olonis wr ount n xprss s olony forming units (CFU) pr ml. Colonis ws ount for plts tht hv mor thn 30 n lss thn 300. Th numr of olonis ount ws multipli y th riprol of th ilution n th mount plt n th rsults wr xprss s CFU/mL [4, 14]. Extrtion of totl ptrolum hyroron in soil mixtur Thr rplits of spik mium wr smpl t h smpling prio. Collt smpls wr stor in glss ottls n kpt t 4 o C prior to nlysis. Approximtly 10 g of h smpl ws pl in 100 ml flsk from h qurium on th sm smpling y for ll trtmnts to xtrt totl ptrolum hyroron (TPH). Th TPH in smpls ws xtrt ultrsonilly using solvnt xtrtion mtho [15, 16]. Soil smpls wr ri y mixing with soium sulpht (N 2 SO 4 ) n ltr pl in 100 ml Shott ottl with 50 ml ihloromthn (DCM) (R&M Chmils, U.K.) n th ottl ws gitt in n Ultrsoni Clnr (Thrmo-10D, U.S.A.) for 30 min t 50ºC. Th suprntnt ws filtr through glss wool. Th xtrts wr onntrt n wr lft in th fum hoo for 3 4 ys to llow th solvnt to vport ompltly, ftr whih, 1.5 ml DCM ws n th xtrts wr stor in gs hromtogrphy (GC) vils. Anlysis of totl ptrolum hyroron Th smpl xtrts wr nlys y gs hromtogrphy flm ionistion ttor (GC FID, Agilnt Thnologis, Mol 7890A, UK) with HP-5 5% phnyl mthyl siloxn olumn (30 m 0.32 mm i miron) with hlium s th rrir gs. Th olumn tmprtur ws progrmm to rmin t 50ºC for 1 min, n thn rmp t 15 C pr min to 320ºC for 10 min. Th prntg of TPH grtion on h smpling y ws trmin y iviing th iffrn of th urrnt TPH vlus y th initil TPH vlu in soil. Th prntg of TPH rmovl in soil on h smpling y ws trmin using Eq. (1): TPH TPH = (1) TPH 0 t %Rmovl 100 0

4 Asi Fhil Almnsoory t l /Int.J. ChmTh Rs.2014,6(1),pp Whr TPH 0 = totl ptrolum hyroron on smpling y 0 n TPH t = totl ptrolum hyroron on h smpling y. Rsults n Disussion Dtrmintion of physiohmil prmtrs Th slt physiohmil prmtrs wr ror throughout th phytotoxiity tst for h gsolin onntrtions in two trtmnts plnt n unplnt (Figur 1).Gnrlly, th ph, ORP n DO rs slightly uring th xposur prio. During th 72 ys of th xprimnt, th ph rng twn 6.4 n 7.5 for plnt quri n twn 5.2 n 6.7 for unplnt quri. Th onitions of xprimnts, th roi or nroi, n istinguish with DO n ORP msurmnts. For th plnt n unplnt trtmnts, th DO rng twn 1.2 n 7.2 mg/l, th ORP twn 7.13 n mv. Th rsing DO rings init tht th trtmnt nvironmnt ws noxi onitions. Furthrmor, msurmnts th ORP for ll gsolin onntrtions wr in th roi n noxi rng. [17] Dmonstrt tht th ition of hyrorons to th soil ltr th soil ph n th onitions of th phytotoxiity tst n lss s noxi/nroi, s th onntrtion of issolv oxygn ws vry limit. Plnt rsponss to th gsolin ontminnt Th plnts growing throughout 72 ys in soil mixtur irrigt with gsolin ontmint soil intrt with tri show ovious iffrns in pprn ompr with thos in th orrsponing ontrols (S. murontus without th ontminnt n tri) (Figur 2). No plnt th ws ror with gsolin onntrtions. Howvr, th growth of S. murontus ws inrs with th gsolin onntrtions, initing tht th plnt growth n vlopmnt ws om ttr u to plnts n thir ssoit tri intrt with h othr in plnt trtmnts [6]. Th iomss of S. murontus inrs in th gsolin phytormition trtmnt. Th wt wight iomss of S. murontus t th n of th xposur prio t onntrtion 10g/kg rh ± 1.7 g n ws lmost th wt wight iomss of S. murontus in orrsponing ontrol only (36.4 ±.97 g) (Figur 2 i n ii). This sitution my hv ourr u to th ition of th tri tht t n nhn th growth. Howvr, th plnt iomss inrs until 42 ys in 5 g/kg n 30g/kg for wt mount wr 47.5 g n 15.6 g ut for ry wight t 18.9 g n 6.3 g rsptivly. Svrl prvious stuis show similr rsults. [18] Rport P, rss, n pnsy plnts hv inrs mounts of totl priority ptrolum romti hyroron xtrt from soil y mor thn 17% uring 68 ys. Bs on stuy y [14] lrgr ffiiny in th rmition nhn with th lrgst growth of tri in rhizosphr whn ttmpt plnt Rhizophor mngl L. n osrv lrgr growth of plnts xpos to ontmint simnts y hyroron ftr 3 months of phytormition. Miroil plt ount Th miroil popultion in S. murontus rhizosphr zon ws vlut t iffrnt gsolin onntrtions (0, 5, 10 n 30 g/kg) s shown in (Figur 3). It ws foun tht th gsolin pollutnt nhn th miroil popultion n inrss its ivrsity. Th grtion of hyrorons minly pns on th pilitis of th miroorgnisms in th surrouning rhizosphr [5] During th xprimnt, miroil popultions in th ontrol qurium wr signifintly lowr thn thos in th quri with gsolin onntrtions of 5, 10 n 30 g/kg. Gnrlly, th tril popultion in th trtmnts with iffrnt of gsolin onntrtions inrs until 42 ys, n thn strt to rs to th n of th 72 y prio of xposur. Th popultion of rhizotri in th plnt ontrol qurium without ontmintion (0 mg/l) ws to CFU ml 1 soil t from y 0 to 42, n ws oviously lowr thn tht in th trtmnts with iffrnt gsolin onntrtions. In othr wors, th popultion of tri in quri irrigt with gsolinontmint soil ws lrly highr thn in th ontrol qurium. Similrly, th popultion of tri in trtmnts with th highst gsolin onntrtion of 30g/kg mount to to CFU ml 1 soil t from y 0 to 42, whih ws similr with gsolin onntrtion of 5 n 10g/kg. Th tril popultion in th trtmnt with lowst gsolin onntrtion of 5g/kg mount from to CFU ml 1 soil

5 Asi Fhil Almnsoory t l /Int.J. ChmTh Rs.2014,6(1),pp uring 42 ys of trtmnt. Th growth of tri in 10 g/kg ws highr thn othr onntrtions tht mount 2.4 x 10 4 to CFU ml 1 soil. Figur 1. : Physil prmtr vritions in th phytotoxiity tst with S. murontus using gsolin s th ontminnt g/kg 5g/kg 10g/kg 30g/kg Gsolin onntrtions in soil Wt wight (grm) f f (i) f

6 Asi Fhil Almnsoory t l /Int.J. ChmTh Rs.2014,6(1),pp (ii) Dry wight (grm) g/kg 5g/kg 10g/kg 30g/kg Gsolin onntrtions in soil Figur 2.: Growth rspons prmtrs: (i) wt wight, (ii) ry wight in th phytotoxiity tst of S. murontus using gsolin s th ontminnt. Error rs init th stnr vition (n = 3). Th mns mong iffrnt gsolin onntrtions follow y th sm lttr ( ) wr not signifintly iffrnt t p < Log (CFU/mL) g/kg 5g/kg 10g/kg 30g/kg Dy Figur 3. : Totl ount of th tril popultion uring th 72 ys of th xprimnt omprison of th miroil popultions in th ontrol n with iffrnt gsolin onntrtions of 5, 10 n 30g/kg. TPH Rmovl in Soil To otin mor informtion out grtion of gsolin in th soil mixtur ws onut in (Figur 4) shows grtion n rmovl ovr 72 ys for th thr iffrnt gsolin onntrtions trtmnts plnt n unplnt(5g/kg, 10g/kg n 30 g/kg). Th rmovl ffiiny of gsolin ontminnt in most trtmnts ws signifintly iffrnt twn ll onntrtions n smpling ys (7, 14, 28, 42 n 72). Th mximum TPH grtion rmovl in soil mixtur of 87.5% ourr with gsolin onntrtion of 10g/kg ftr 72 ys of trtmnt, whil th vrg rmovl in its orrsponing ontrol trtmnt ws only 55 %. Similrly, th grtion rts with gsolin onntrtions of 5 n 30 g/kg wr 84.5 n 83.3%, rsptivly, whil th vrg rmovl in th orrsponing ontrol trtmnts ws 56.3 n 54.2%, rsptivly. Th onvrgn of rsults inits th ility of tri S. mrsns prtnrship with plnt S. murontus to nhn grtion of TPH n surviv in ths thr gsolin onntrtions.

7 Asi Fhil Almnsoory t l /Int.J. ChmTh Rs.2014,6(1),pp Sttistil nlysis ws prform twn trtmnts plnt n unplnt t h smpling tim for ll gsolin onntrtions s illustrt in (Figur 4). Th rmovl TPH grtion in ll onntrtions signifintly (p < 0.05) iffrnt twn plnt n unplnt (orrsponing ontrol). Du to th intrtion twn slt tri n plnts th hyrorons wr mtoliz. Th min mhnism to rmovl TPH in phytormition of ontmint soils is ssum to rhizogrtion, th stimultion of tri in th rhizosphr zon to gr n nhn rmovl of TPH [19]. In ontrol soil mixtur my lso th onsqun of iogrtion y inignous miroorgnisms [4] Th phytormition of TPH, spilly in soil on mny stuis oring to [20], th rmovl of isl from soil ontmint with 15,000 mg/kg isl y Sirpus triqutr ws n 72.62% plnt n unplnt, rsptivly. [21] Rport our rsults show tht ivrs plnt spis growing in hyroron ontmint soil with miroil popultions, whih my impt upon th ility of plnts to promot th grtion of spifi typs of hyrorons. Th grtion of TPH y Mirilis jlp n show tht th vrg ffiiny to rmov TPH ovr 127-y ultur prio ws high, up to %, whil th rmovl rt y nturl ttnution ws only % [4]. [22] Show tril inoultion in phytormition nhns plnt rsistn to th ontminnt strss n inrss thir limtion rt n iomss formtion. Similrly hyroron grtion y tri nhn plnt iomss proution n HC grtion [23]. Th synrgisti tion of th plnts n tril inoultion rhizogrtion of hyroron xhiit mor ffiint s ompr to miroil rmition n phytormition [24]. Figur 4.: Dgrtion prntg in soil mixtur xtrtion y S. murontus xpos to gsolin ontmintion t5 g/kg, 10 g/kg n 30 g/kg. Brs init th stnr rror of thr rplits (n = 3). Lttr A: sttistilly signifint gsolin rmovl from soil twn two trtmnts plnt n unplnt ws rprsnt (p < 0.05). Conlusions Th tolrn of plnts to soil ontmintion y gsolin onntrtions ftr 72 monstrt tht S. murontus hs th ility to surviv n provi suitl onitions for rhizotri to gr hyrorons t ll invstigt gsolin onntrtions (5, 10 n 30g/kg). Th ry n wt wight of th plnt slightly inrs on xposur to gsolin t 42 in 5g/kg n 30 g/kg. Th highst wt wight in onntrtion 10 g/kg g t 72 whil ry wight mount 35.6 g. In th 5g/kg n 30 g/kg mount for 43.4g n 10.8g ws wt ut ry wight t13.1g n 6.3g rsptivly. Bs on soil xtrtion, th highst TPH rmovl rt ws 87.5%, in omprison to th rmovl rt y th orrsponing unplnt ontrols of only 55%.Th rmovl rts with gsolin onntrtions of 5 n 30 g/kg wr 84.5 n 83.3%, rsptivly, whil th

8 Asi Fhil Almnsoory t l /Int.J. ChmTh Rs.2014,6(1),pp vrg rmovl in th orrsponing ontrol trtmnts ws 56.3 n 54.2%, rsptivly. This inits tht th iogrtion pross ply rol in th trtmnt with 10% rhizotri. Aknowlgmnt Th uthors woul lik to thnk th Univrsiti of Kngsn Mlysi (UKM) n th Tsik Chini Rsrh Cntr for supporting this rsrh projt unr FRGS grnt (FRGS ). W lso knowlg th Irqi Ministry of Highr Eution n Sintifi Rsrh for proviing otorl sholrship for th first uthor. Rfrns 1. Grhrt, K.E., Hung, X.-D., Glik, B.R., Grnrg, B.M., Phytormition n rhizormition of orgni soil ontminnts: potntil n hllngs.plnt Si. 176, Gskin, S.E., Bnthm, R.H., Rhizormition of hyroron-ontmint soil using Austrlin ntiv grsss. Si. Totl Environ. 408: Lu, V.; Gri, C. n Hrnnz, T Efft of hyroron pollution on th miroil proprtis of sny n ly soil. Chmosphr 66: Png, S., Zhou, Q., Ci, Z., Zhng, Z., Phytormition of ptrolum ontmint soils y Mirilis jlp L. in grnhous plot xprimnt. J. Hzr. Mtr. 168, Lu, S., Tng, Y., Wng, J., Sun, Z., Enhnmnt of pyrn rmov from ontmint soils y Bins mximowizin. Chmosphr 81, Khn, S.; Afzl, M.; Iql, S.; Khn, Q. M Plnt tri prtnrships for th rmition of hyroron ontmint soils. Chmosphr. 90: Kthi, S., n Khn, A. B., Phytormition pprohs to PAH ontmint soil. Inin Journl of Sin n Thnology. 4. ISSN: MPhrson, A Monitoring Phytormition of Ptrolum Hyroron Contmint Soils in Clos n Controll Environmnt. Thsis Sumitt to th Collg of Grut Stuis n Rsrh.Th Dgr of Mstrs of Sin. P MKinnon, G. n Dunn, H.J Phytotoxiity of rnh ylohxns foun in th voltil frtion of isl ful on grmintion of slt grss spis. Chmosphr 90: Zhng, Z., Zhou, Q., Png, S. n Ci, Z Rmition of ptrolum ontmint soils y joint tion of Phritis nil L. n its miroil ommunity. Sin of th Totl Environmnt 408(22): Kirk, J.L., Klironomos, J.N., L, H. n Trvors, J.T Th ffts of prnnil rygrss n lflf on miroil unn n ivrsity in ptrolum ontmint soil. Environmntl Pollution 133: Ogo E.M., Tunu, A. n U R Phytotoxiity ssy of isl ful-spik sustrts rmit with Plurotus turrgium using Z mys. Intrntionl Journl of Appli Rsrh in Nturl Prouts 3 (2): Prsott, L.M., Hrly, J.P., Lortory Exriss in Miroiology, fifth. MGrw-Hill Compnis, Klin DA. 14. Morir, I., Olivir, O., Triguis, J., Sntos, A., Quiroz, A., Mrtins, C., Silv, C., Jsus, R., Phytormition using Rhizophor mngl L. in mngrov simnts ontmint y prsistnt totl ptrolum hyrorons (TPHs). Mirohm. J. 99, Su, Y.-H. n Zhu, Y.-G Uptk of slt PAHs from ontmint soils y ri slings (Oryz stiv) n influn of rhizosphr on PAH istriution. Environmntl Pollution 155(2): Tng, J., Wng, M., Wng, F., Sun, Q. n Zhou, Q Eotoxiity of ptrolum hyroron ontmint soil. Journl of Environmntl Sins 23(5): Tng, J., Lu, Z.; Sun, Q.; Zhu, W Aging fft of ptrolum hyrorons in soil unr iffrnt ttnution onitions. Agriultur, Eosystms n Environmnt 149:

9 Asi Fhil Almnsoory t l /Int.J. ChmTh Rs.2014,6(1),pp List, H. H. n Prutz, I Plnt prformn, ioxygns-xprssing rhizosphr tri n iogrtion of wthr hyrorons in ontmint soil. Chmosphr 62: Ci, Z., Zhou, Q., Png, S., Li, K., Promot iogrtion n miroiologil ffts of ptrolum hyrorons y Imptins lsmin L. with strong nurn. J. Hzr. Mtr. 183, Liu,T.; Hou. J.; Zuo, Y.; Bi, Z. & Jing, J Isoltion n hrtriztion of iosurftnt prouing trium from Dqing oil-ontmint sits. Afr. Journl Miroiol. Rs. 5(21): Phillips, L. A.; Grmi, J. J.; Frrll, R. E. & Grr, C. W Hyroron grtion potntil n tivity of ntophyti tri ssoit with priri plnts. Soil Biology & Biohmistry 40: Murtov, A.Y., Goluv, S., Wittnmyr, L., Dmitriv, T., Bonrnkov, A.,Hirh, F., Mrh, W., Turkovsky, O., Efft of th polyyli romti hyroron phnnthrn on root xution of Sorghum iolor (L.) Monh. Environ. Exp. Bot. 66, Yousf, S., Afzl, M., Rihnur, T.G., Bry, C.L., Sssitsh, A., Hyroron grtion, plnt oloniztion n gn xprssion of lkn grtion gns y nophyti Entrotr luwigii strins. Environ. Pollut. 159: Afzl, M., Yousf, S., Rihnur, T.G., Sssitsh, A Th inoultion mtho ffts oloniztion n prformn of tril inoulnt strins in th phytormition of soil ontmint with isl oil. Int. J. Phytorm. 14: *****