Fungitoxiity of queous n orgni solvent extrts of Dtur metel ginst Asohyt riei *Rukhsn Bjw, Soiy Shfique n Shzi Shfique Deprtment of Myology n Plnt Pthology, University of the Punj, Qui-e-Azm Cmpus, Lhore, Pkistn. *E. mil: rukhsnjw_mppl@yhoo.om Astrt In vitro effiy of queous n methnoli extrts of Dtur metel ws evlute ginst Asohyt riei, (the usl gent of hikpe light) n the sensitivity of olony growth ws stuie in terms of inhiition zone. The inhiitory potentil of ll the extrts ws gretest t lower onentrtion. The queous n methnol extrts of shoot of Dtur metel use 21-34% n -% reution in growth of Asohyt riei wheres the root extrts prove less effetive s they use 15-25% n 11-29% reution in growth of A. riei, respetively. Keywors: Asohyt riei, hikpe, fungitoxiity, Dtur metel, queous n orgni extrts. Introution One of the most importnt legume rops of Pkistn is Chikpe (Cier rietinum L.), whih is mjor soure of protein. It is grown over.963 m h rinfe onitions with nnul proution of.6752 million tons with n verge yiel of 71 kg h -1 (Anonymous, 4), whih is muh lower thn its potentil. Blight isese is mjor limiting ftor for its reue proution (Ilys n Bshir, 1983). Asohyt light is mjor isese of hikpe in most growing res of the worl (Port-Pugili et l., 1997) tht uses -25% yiel loss in hikpe nnully (Iql et l., 5). Vrieties of ontrol mesures re unertken to voi the implitions of yiel losses ue to plnt iseses. In this regr, the iologil inhiitions y ifferent nturl sustnes, suh s essentil oils n plnt extrts hve een investigte wiely ginst fungl tivities. In lst two ees, muh work hs een one on plnt-erive ompouns s environmentlly sfe lterntives to hemils (Duke et l., ; 2; Singh et l., 4). In reent times Alkhil (5) showe tht queous extrts of plnts viz. Allium stivum, Cymopogon proxims, Crum rvi, Azirht ini n Eugeni ryophyllus h strong ntifungl tivity ginst fungi nmely Fusrium oxysporum, Botrytis inere n Rhizotoni solni. Similr effets of Mgnoli grniflor L. extrts ginst Alternri lternt, Helminthosporium spp., Fusrium oxysprum, F. ulmorium n Rhizotoni solni hve lso een reporte y Ahm n Aulgleil (5). More reently Bjw et l. (7) evlute ntifungl potentil of queous n orgni extrts of Aloe ver n reporte tht shoot queous n n-hexne extrts use signifint inhiition in growth n iomss proution of the three teste fungi viz., Alternri lternt, A. itri n A. tenuissim. Thus the present stuy ws unertken to evlute the ntifungl potentil of queous n orgni solvent extrts of Dtur metel on in vitro growth of Asohyt riei Mterils n Methos Prourement n mintenne of trget fungl speies Culture of trget fungl speies of A. riei ws otine from First Fungl Culture Bnk of Pkistn, (FCBP) University of the Punj, Quie-Azm Cmpus Lhore n mintine on mlt extrt gr (MEA) meium. Colletion of plnt mterils Fresh smples of shoot n root of Dtur metel were ollete from University of the Punj, Qui-e-Azm Cmpus Lhore n wshe thoroughly uner tp wter, rie with lotting pper n ut into smll piees. The solule ingreients of the plnt mteril were then extrte y soluiliztion in wter n methnol s ifferent solvents. Preprtion of queous extrt Aqueous extrt of wter solule ingreients of plnt mteril ws prepre
18 Bjw et l. oring to Bjw et l,. (7). A % w/v stok solution of plnt extrt ws ttine y soking the rushe plnt mterils in sterilize istille wter for 48 h t ±2 C. Then mteril ws filtere through muslin loth followe y filter pper. This stok extrt ws store t 4 ºC n use within four ys. Preprtion of orgni solvent extrt The metho of Alkhil (5) ws followe for the preprtion of shoot n root extrt in methnol. The test plnt ws rushe n extrte y merting g of plnt mteril in ml of methnol for 48 h. Mterils were filtere through muslin loth followe y filter pper. Orgni solvent extrt ws evporte uner vuum until its volume ws reue to 2 ml n then ilute y ing pproprite quntity of sterilize istille wter to mke finl volume of ml. These stok extrts were store t 4 ºC n use within four ys. The lower onentrtions of 1, 2, 3 n 4% of oth queous n methnol extrts of shoot n root were prepre y ing pproprite quntity of sterilize istille wter. To mke methnol ontrol, 2 ml of methnol ws e to sterilize istille wter to mke finl volume ml. Antifungl iossys with extrts Mlt extrt gr (MEA) meium ws prepre n oole to ºC. Approprite quntities of stok solution n istille wter were e to MEA meium to get 1, 2, 3 n 4% (v/v) onentrtions of shoot n root extrts in the meium. Control reeive the sme quntity of istille wter. The plnt extrts were thoroughly mixe with the meium. Twenty ml of eh meium ws poure in eh 9 m imeter sterilize Petri plte. Myelil iss of 5 mm imeter were tken with pre-sterilize ork orer from 5-7 ys ol ulture of A. riei n were ple in the entre of eh Petri plte fter soliifition of the MEA meium. Eh tretment ws replite thrie. Pltes were inute in n inutor t 25±2 ºC for 7 ys. Fungl growth ws mesure y verging the three imeters tken t right ngles for eh olony. Perentge growth inhiition of the fungl olonies ws lulte y pplying the following formul: Growth in ontrol - Growth in tretment Growth inhiition (%) = Growth in ontrol Sttistil nlysis All the t were nlyze y nlysis of vrine followe y Dunn s Multiple Rnge Test (Steel n Torrie, 198) using omputer softwre SPSS n COSTAT, respetively. Results n Disussion Effet of queous extrt of D. metel on growth rte of A. riei The results otine from perioi growth ssys of A. riei in vrious onentrtions of shoot n root queous extrt of D. metel showe signifint ntifungl tivity in ll the onentrtions in omprison to ontrol (Fig. 1). The results revele tht shoot extrt ws signifintly the most effetive in suppressing the olony growth s ompre to root extrt. The ssessment of onentrtion effet revele n inrese in growth rte with inrese inution perio. At initil growth stges ll the onentrtions invrily n insignifintly epresse the fungl growth while fter 7 ys inution ll extrt onentrtions epite onsierle epression in growth rte. The mximum ntimyoti tivity ws oserve y 1 & 3% shoot extrt. In ontrst 2 & 4% onentrtions epite less toxiity ginst A. riei. In se of root extrt vrile pttern of ntimyoti tivity ws oserve s 1 & 4% onentrtion of root extrt ws the most effetive in suppressing the growth of A. riei wheres 2 & 3% onentrtions were less epressive (Fig. 3A). Shoot extrt use out 21-33% epression in growth rte of the test orgnism while 15-25% reution ws notie y root extrt. The 1% onentrtion of oth shoot n root queous extrts showe mximum erese in fungl growth whih ws 33% in shoot extrt n 25% y root extrt (Fig. 4A). Effet of orgni extrt of D. metel on growth rte of A. riei Methnoli frtions exhiite more promising results in suppressing the fungl growth thn queous frtions. The perioi t regring fungl growth, expose to vrious onentrtions of methnoli extrts of Dtur metel re presente in Fig. 2. Differenes in growth rte were exhiite with respet to the onentrtions employe. The perioi ssys revele signifint reution in fungl growth rte in ll onentrtions. The frtions of shoot n root orgni extrts i not show ny prtiulr tren in response to inhiition. However, the lowest onentrtion 1% use signifint reution in myelil growth (Fig. 3B).
Fungitoxiity of queous n orgni solvent extrts 19 % 1% 2% 3% 4% A Shoot Extrt of Dtur metel Dy 1 Dy 2 Dy 3 Dy 4 Dy 5 Dy 6 Dy 7 Root Extrt of Dtur metel B Dy 1 Dy 2 Dy 3 Dy 4 Dy 5 Dy 6 Dy 7 Fig. 1: Perioi effet of queous shoot n root extrts of Dtur metel on fungl olony growth of Asohyt riei. Vertile rs show stnr errors of mens of three replites. For eh y vlues with ifferent letters show signifint ifferene (P =.5) s etermine y Dunn's Multiple Rnge Test. The omprison of ifferent onentrtions of shoot n root orgni extrts showe tht the %ge olony growth inhiition ws signifintly greter in shoot extrt in ontrst to root extrt (Fig. 4B). There ws -% reution in fungl growth ue to vrious employe onentrtions of shoot extrt. The 1% shoot extrt use highest reution of out % in fungl growth. Further inrese in extrt onentrtion exhiite signifint ifferene s ompre to 1% extrt. Different onentrtions of root extrt use -29% reution in fungl growth. Mximum reution of 29% ws epite y 1% root extrt. In the present stuy, two types of extrts of D. metel were use ginst A. riei. The results of this stuy lerly reflet tht D. metel hs the potentil to inue toxi effets on myelil growth n prolifertion of fungi. The reltive intensity of this effet, however, vries with the speies involve, s well s the onentrtion of the extrt employe. Erlier Shfique et l., (6) hve reporte tht llelopthi trees viz., Azirht ini, Mngifer ini, Meli zerh n Syzygium umini signifintly suppress the growth of Alternri lternt. Similrly, Bjw et l., (7) rrie out the stuy on ntifungl tivity of queous n n-hexne shoot extrts of Aloe ver ginst few pthogeni speies of Alternri lternt, A. itri n A. tenuissim. They reporte tht the inhiitory effet ws foun to e vrile with the pplie onentrtion n use signifint inhiition in iomss proution of the three test fungi Likewise, Dur et l., (4) reporte tht phytohemil extrtion of D. metel showe ntifungl (MIC 87.5 mg ml -1 ) tivity ginst Aspergillus fumigtus. Extrts in ifferent solvents exhiite vrile ntifungl tivity ginst in vitro growth of A. riei. Methnoli frtions exhiite more promising results in suppressing the fungl growth thn queous frtions. Likewise Dwivei n Duey (1986) reporte tht voltile frtions of two meiinl plnts,
Bjw et l. Azirht ini n Eulyptus gloules were more effetive ginst Mrophomin phseolin thn non-voltile frtions. The vrition in ntifungl tivity of the extrts in ifferent solvents my e ttriute to the ifferent hemil nture of the solvents. It is likely tht ifferent types of hemils were issolve in ifferent solvent tht resulte in vrile tivity of the extrts of sme prt of the plnt in ifferent solvents. There re mny exmples in the literture whih support our finings. Zfr et l., (2) reporte tht hloroform extrt of leves of M. zerh ws tive ginst Fusrium hlmosporum while hexne, ethnol n wter extrts were not. In similr kin of work Bjw et l., (6) reporte the ntimyoti tivity of queous n ihloromethne frtions of Cier rietinum ginst Drehsler tetrmer n D. hwiiensis. % 1% 2% 3% 4% A Shoot Extrt of Dtur metel Dy 1 Dy 2 Dy 3 Dy 4 Dy 5 Dy 6 Dy 7 Root Extrt of Dtur metel B Dy 1 Dy 2 Dy 3 Dy 4 Dy 5 Dy 6 Dy 7 Fig. 2: Perioi effet of orgni shoot n root extrts of Dtur metel on fungl olony growth of Asohyt riei. Vertile rs show stnr errors of mens of three replites. For eh y vlues with ifferent letters show signifint ifferene (P =.5) s etermine y Dunn's Multiple Rnge Test.
Fungitoxiity of queous n orgni solvent extrts 21 % 1% 2% 3% 4% A: Aqueous Extrts 7 Shoot Root B: Methnol Extrts Shoot Fig. 3: Effet of queous n orgni solvent extrts of Dtur metel on in vitro growth of Asohyt riei fter 7 ys of inution. Vertil rs show stnr error of mens of three replites. Vlues with ifferent letters show signifint ifferene s etermine y DMR Test. Root 1% 2% 3% 4% A Aqueous Extrt Shoot Root -5 Derese (%) -15-25 -35 33 21 28 21 25 15 16 22-45 B Methnol Extrt -5 Derese (%) -15-25 -35 24 28 28 11 17-45 Fig. 4: Perentge erese in olony imeter of Asohyt riei ue to ifferent onentrtions of queous n orgni solvent extrts of shoot n root of Dtur metel. Referenes Ahme SM, Aelgleil SAM, 5. Antifungl tivity of extrts n sesquiterpene ltones from Mgnoli grniflor L. (Mgnoliee). Int. J. Agri. Biol., 4: 638 642. Alkhil AA, 5. Antifungl tivity of some extrts ginst some plnt
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