*Shazia Shafique and Sobiya Shafique

Transcription

1 Myopth (28) 6(1&2): Antifungl tivity of n-hexne extrts of Dtur metel ginst Asohyt riei *Shzi Shfique nd Soiy Shfique Institute of Myology nd Plnt Pthology, University of the Punj, Quid-e-Azm Cmpus, Lhore, Pkistn. *E. mil: Astrt The ntimyoti potentil of Dtur metel (syn. Dtur l Nees.) ws investigted in vitro ginst Asohyt riei, the use of hikpe light disese. The pthogen ws exposed to vrious n-hexne onentrtions (1, 2, 3 nd 4% w/v) of shoot nd root extrts of D. metel using poisoning food tehnique. All the employed onentrtions of oth root nd shoot extrts signifintly suppressed the growth of the trget fungl pthogen. There ws 28-34% nd 23-41% redution in olony dimeter of A. riei due to different onentrtions of n-hexne shoot nd root extrts of D. metel. Keywords: Asohyt riei, hikpe, n-hexne extrt, Dtur metel. Introdution Chikpe (Cier rietinum L.) is n importnt legume rop. It is grown under wide gro-eologil onditions nd rnk first mong food legumes in Indin Suontinent nd Mediterrnen sin (Anonymous, 1994). In Pkistn, it is grown under rinfed onditions nd.6752 million tons re produed nnully (Anonymous, 24). Although hikpe prodution is ffeted y vrious ftors ut light disese used y Asohyt riei is the mjor limiting ftor (Ilys nd Bshir, 1983) whih use out 2-25% nnul loss in hikpe yield nd totl rop filure under fvourle epidemi onditions (Iql et l., 25). The exploittion of resistnt vrieties hs een supported s the est ontrol of disese (Iql et l., 22), however, ultivrs with resistne ginst Asohyt light hve not exhiited the desired results due to lk of stility in tolerne euse of pperne of new pthotypes of the pthogen (Iql et l., 22, 25). Chemil ontrol hs een proved effiient nd eonomil in ontrolling light disese. However, inresing puli onern on environmentl issues desires tht lterntive mngement systems e evolved either to redue pestiide dependnt or nturlly ourring ompounds e explored to onstrin the pthogen ttk (Singh et l., 23; Cuthertson nd Murhie, 25). Nturl plnts derived ompounds ontriute lot in fight ginst pthogens (Vyvyn, 22). Severl plnt fmilies like Anthee, Amrnthee, Apiee nd Mgnoliee hve ntifungl nd ytotoxi properties (Mnsill nd Plenzuel, 1999; Neermn, 23). Numerous studies onduted in Pkistn reveled wide spetrum prospets of using extrts of plnts for iologil ontrol of pthogeni fungi (Bjw et l., 21; Bjw et l., 22; Ahmd nd Adelgliel, 25; Brg et l., 27). Vrious plnt extrts e.g., Cier rietinum (Bjw et l., 26), Prthenium hysterophorus (Bjw et l., 24) nd Mgnoli grndiflor (Ahmed nd Adelgleil, 25) et. hve lso een exmined for their ntifungl tivity with the ojetive of exploring environmentlly sfe lterntives of plnt disese ontrol. Thus with the ojetive to ontriute to these studies, the ntifungl tivity of n-hexne extrts of Dtur metel ws investigted ginst Asohyt riei. Mterils nd Methods Culture of A. riei FCBP-72 ws otined from First Fungl Culture Bnk of Pkistn, University of the Punj, Quid-e-Azm Cmpus Lhore nd mintined on mlt extrt gr (MEA) medium. Shoots nd roots of mture Dtur metel plnts were olleted from University of the Punj, Quid-e-Azm Cmpus Lhore. After thorough wshing under tp wter, the mterils were surfe sterilized with 1% sodium hypohlorite solution followed y thorough wshing with sterilized wter. Plnt mterils were rushed nd soked in orgni solvent 2 g/ ml for 48 hours. The extrts were filtered through muslin loth followed y filter pper. Orgni solvent ws evported under

2 32 Shfique nd Shfique vuum to 2 ml nd the remining extrt ws diluted y dding pproprite quntity of sterilized distilled wter to mke 2% extrt. These stok extrts were stored t 4 ºC nd used within four dys. To mke n-hexne ontrol, 2 ml of orgni solvent ws dded to sterilize distilled wter to mke finl volume ml. Two perent MEA medium ws prepred y utolving t 121 ºC nd ooled to ºC. Afterwrds, pproprite quntities of stok solution nd distilled wter were dded to MEA medium to get 1, 2, 3 nd 4% (w/v) onentrtions of shoot nd root extrts in the medium. The plnt extrts were thoroughly mixed with the medium. Control tretments reeived only n-hexne ontrol mixture. Twenty ml of eh medium ws poured in eh 9 m dimeter sterilized Petri pltes. Myelil diss of 5 mm dimeter were tken with sterilized ork orer from 7 dys old ultures of A. riei nd were pled in the entre of eh Petri plte fter solidifition of the medium. Eh tretment ws replited thrie. Pltes were inuted t 25±2 ºC for 7 dys. Fungl growth ws mesured y verging the three dimeters tken t right ngles for eh olony. Perentge growth inhiition of the fungl olonies ws lulted y pplying the following formul: Growth in ontrol - Growth in tretment Grwoth inhiition (%) = Growth in ontrol All the dt were nlyzed y nlysis of vrine followed y Dunn s Multiple Rnge Test (Steel nd Torrie, 198) using omputer softwre COSTAT. Results nd Disussion The sensitivity of olony growth of the usl orgnism of light disese ws evluted using inhiition zone tehnique t vrious onentrtions. The dt regrding the olony growth reveled tht the growth rte ws different in ll the onentrtions employed (Fig. 1A & B). Generlly, root extrt ws more toxi thn shoot extrt to fungl growth. The periodi growth ssys showed tht the olony growth inresed from first dy to seventh dy in ll the tretments, however nominl depression ws oserved fter two nd three dys of inution ut t 7 dys period growth rte ws found to e signifintly depressed s ompred to ontrol. The vrition in ntifungl tivity of the shoot nd root extrts my e ttriuted to the different hemil nture of the ompounds present in these prts (Afshrypuor et l., 1995). Generlly ll the onentrtions signifintly suppressed fungl growth. No prtiulr trend ws oserved in response to frtions of shoot nd root extrts. The mximum ntimyoti tivity ws oserved under 1% shoot extrt. In ontrst, the higher onentrtions (from 2-4%) depited less toxiity ginst A. riei. In se of root extrt inverse pttern of ntimyoti tivity ws oserved s 4% onentrtion of root orgni extrt ws found to e the most effetive in suppressing the growth of A. riei (Fig. 2). Dtur metel is well known for tropne lkloids nd with steroids, yielded severl sphingosine derivtives, one of whih ws hrterised s (4E,8Z)-1-O-(et-D-gluopyr- nosyl)-n-(2'-hydroxyhexdenoyl)-sphing-4,8- dienine from omprehensive spetrl nlysis of its hexette nd known to hve ntifungl properties (Shi et l., 1999). Dur et l. (24) isolted new pyrrole derivtive 1 from the leves of D. metel whih ws hrterised s 2-et-(3, 4-dimethyl-2, 5-dihydro-1H-pyrrol-2-yl)-1'-methylethyl pentnote on the sis of spetrl dt nlyses nd hemil retions whih ws endowed with ntifungl tivity. Erlier Crpinell et l. (23) hve reported tht hexni nd ethnoli extrts from fruit, seed kernels nd senesent leves of Meli zedrh exhiited fungistti tivity ginst Aspergillus flvus, Diporthe phseolorum vr. meridionles, Fusrium oxysporum, Fusrium solni, Fusrium vertiillioides, nd Slerotini slerotiorum. Similrly, Kgle et l. (24) found vrile ntimiroil tivity of D. metel in vitro ginst Rhizotoni solni nd Xnthomons oryze. The omprison of mens in se of different onentrtions of shoot nd root extrts showed tht the perentge olony growth inhiition ws signifintly greter in root extrt in ontrst to shoot extrt (Fig. 3). There ws 22-35% redution in fungl growth due to vrious employed onentrtions of shoot extrt. The 1% shoot extrt used the highest redution of out 35% in fungl growth. Further inrese in extrt onentrtion exhiited signifint differene in ntimyoti tivity s ompred to 1% extrt. Different onentrtions of root extrt used 22-41% redution in fungl growth. Mximum redution of 41% ws depited y 4% root extrt. Erlier, Dur et l. (24) reported tht phytohemil extrtion of D. metel showed ntifungl (MIC 87.5 mg/ml) tivity ginst Aspergillus fumigtus. In onlusion, this study demonstrtes tht n- hexne extrts of D. metel exhiit n effetive ntifungl potentil ginst the test orgnism, A. riei. Finlly, urrent reserh offers the possiility of developing strtegies for ontrolling plnt pthogens with nturl extrts or iotive metolites of mediinl plnts. Myopth (28) 6(1&2): 31-35

3 Antifungl tivity of n-hexne extrts 33 % 1% 2% 3% 4% A Colony dimeter (mm) Shoot Extrt of Dtur metel d d d Dy 1 Dy 2 Dy 3 Dy 4 Dy 5 Dy 6 Dy 7 Colony dimeter (mm) Root Extrt of Dtur metel B Dy 1 Dy 2 Dy 3 Dy 4 Dy 5 Dy 6 Dy 7 Fig. 1: Periodi effet of shoot nd root extrts of Dtur metel on fungl olony growth of Asohyt riei. Vertile rs show stndrd errors of mens of three replites. For eh dy vlues with different letters show signifint differene (P =.5) s determined y Dunn's Multiple Rnge Test. % 1% 2% 3% 4% Colony dimeter (mm) d Shoot Fig. 2: Effet of n-hexne extrts on in vitro growth of Asohyt riei fter 7 dys of inution. Vertil rs show stndrd error of mens of three replites. Vlues with different letters show signifint differene s determined y DMR Test. Root Myopth (28) 6(1&2): 31-35

4 34 Shfique nd Shfique 1% 2% 3% 4% A n-hexne Extrt Shoot Root -5 Derese (%) Fig. 3: Perentge derese in olony dimeter of Asohyt riei due to different onentrtions of n- hexne shoot nd root extrt of Dtur metel. Referenes Afshrypuor S, Mostjern A, Mokhtry R, Vrition of Sopolmine nd Atropine in different prts of Dtur metel during development. Plnt Medi., 61: Ahmed SM, Adelgleil SAM, 25. Antifungl tivity of extrts nd sesquiterpene ltones from Mgnoli grndiflor L. (Mgnoliee). Int. J. Agri. Biol., 4: Anonymous, Food nd Agriulturl Orgniztion of the United Ntions Orgniztion. Prodution Yerook, Rome, Itly, FAO. Anonymous, 24. Agriulturl Sttistis of Pkistn Ministry of Food, Agriulture nd Live-stok. Enomi Division, Islmd. Bjw R, Akhtr N, Jvid A, 21. Antifungl tivity of llelopthi plnt extrts. I. Effet of queous extrts of three llelopthi Astereous speies on growth of Aspergilli. Pk. J. Biol. Si., 4: 3-7. Bjw, R., Riz S, Jvid A, 22. Antifungl tivity of llelopthi plnt extrts. II: In vitro ontrol of Fusrium moniliforme nd F. oxysporum y queous extrts of four llelopthi grsses. In: Proeedings of 3rd Ntionl Conferene of Plnt Pthology, Otoer 1-3, 21, Islmd Bjw R, Shfique S, Anjum T, Shfique S, 24. Antifungl tivity of llelopthi plnt extrts IV: Growth response of Alternri lternt, Fusrium monili-frome nd Drehsler hwiiensis to queous extrt of Prthenium hysterophorus L. Int. J. Agri. Biol., 6: Bjw R, Anjum T, Shfique S, Shfique S, 26. Evlution of ntifungl tivity of Cier rietinum L. Pk. J. Bot., 38: Brg FG, Mri LM, Bouzd RL, Fri, Mtos MO, Moreir FO, Sio E, Coimr ES, 27. Antileishmnil nd ntifungl tivity of plnts used in trditionl mediine in Brzil. J. Ethnophrmol., 111: Crpinell MC, Giord LM, Ferryoli CG, Ferryoli CG, 23. Antifungl effets of different orgni extrts from Meli zedrh L. on phytopthogeni fungi nd their isolted tive omponents. J. Agri. Food Chem., 51: Cuthertson AGS, Murhie AK, 25. Eonomi spry thresholds in need of revision in Northern Irish Brmley orhrds. Bio. News., 32: 19. Dur R, Ali M, Singh H, Gupt J, Shrm GL, 24. A novel ntifungl pyrrole derivtive from Dtur metel leves. Phrmzie., 59: Ilys MB, Bshir M, Evlution of systemi fungiides for the ontrol of grm light. Pk. J. Agri. Si., 21: Iql SM, Ghfoor A, Bkhsh A, Bshir M, 22. Disese resistne sttus of hikpe genotypes ginst Asohyt light. Pk. J. Phytopthol., 14: Iql SM, Bkhsh A, Mlik SR, Hqqni M, 25. Pthogeni vriility in Asohyt riei. Pk. J. Phytopthol., 17 : Myopth (28) 6(1&2): 31-35

5 Antifungl tivity of n-hexne extrts 35 Kgle S, Mrimuthu T, Thyumnvn B, Nndkumr R, Smiyppn R, 24. Antimiroil tivity nd indution of systemi resistne in rie y lef extrt of Dtur metel ginst Rhizotoni solni nd Xnthomons oryze pv. Oryze. Physiol. Mol. Plnt Pthol., 65: 91-. Mnsill H, Plenzuel JA, Minor eudesmnolides from Artemisi nriensis. Phytohemistry, 51: Neermn MF, 23. Sequiterpenes ltones diverse lss of ompounds found in essentil oils possessing ntiteril nd ntifungl properties. Int. J. Aromth., 13: Shi M, Mnikm M, Gupt M, Srivstv A, Ry AB, Chrteristion of ereroside isolted from the leves of Dtur metel. J. Ind. Chem. So. 76: Singh HP, Btish DR, Kohli RK, 23. Allelopthi intertions nd llelohemils: New possiilities for sustinle weed mngement. Cri. Rev. Plnt Si., 22: Steel RGD, Torrie JH, 198. Priniples nd Proedures of Sttistis. MGrw Hill Book Co., In, New York, USA. Vyvyn JR, 22. Allelohemils s leds for new heriides nd grohemils. Tetrhedron, 58: Myopth (28) 6(1&2): 31-35