FUNGITOXIC ACTIVITY OF AQUEOUS AND ORGANIC SOLVENT EXTRACTS OF TAGETES ERECTUS ON PHYTOPATHOGENIC FUNGUS-ASCOCHYTA RABIEI

Transcription

1 Pk. J. Bot., 43(1): 59-64, 211. FUNGITOXIC ACTIVITY OF AQUEOUS AND ORGANIC SOLVENT EXTRACTS OF TAGETES ERECTUS ON PHYTOPATHOGENIC FUNGUS-ASCOCHYTA RABIEI SHAZIA SHAFIQUE, SOBIYA SHAFIQUE, RUKHSANA BAJWA, NOSHEEN AKHTAR AND SANA HANIF Institute of Plnt Pthology, University of the Punj, Quid-e-Azm Cmpus, Lhore, Pkistn. Corresponding uthor: Astrct The In vitro fungitoxic potentil of Tgetes erectus L. ws scrutinized ginst Ascochyt riei, the cusl gent of chickpe light disese. The pthogen ws exposed to vrious concentrtions (1, 2, 3 nd 4% w/v) of queous nd methnol extrcts of flower nd shoot of T. erectus using food poisoning technique. All the employed concentrtions of oth flower nd shoot extrcts significntly suppressed the growth of trget fungl pthogen. There ws 4-35% nd 55-73% reduction in colony dimeter of A. riei due to different concentrtions of queous flower nd shoot extrcts of T. erectus nd 12-5% nd 4-42% due to different concentrtions of methnolic flower nd shoot extrcts of T. erectus, respectively. Introduction One of the most importnt legume crops of Pkistn is chickpe (Cicer rietinum L.), which is mjor source of protein. In Pkistn, it is grown under rinfed conditions nd.6752 million tons re produced nnully (Anon., 24). Although chickpe production is ffected y vrious fctors ut light disese is mjor limiting fctor for its reduced production (Ilys & Bshir, 1983). Ascochyt light is mjor disese of chickpe in most growing res of the world (Port-Pugili et l., 1997) tht cuses 2-25% yield loss in chickpe nnully (Iql et l., 25). Vrieties of mngement strtegies re commenced to void the yield losses due to fungl plnt diseses mong which nturl plnts derived compounds contriute lot in fight ginst pthogens (Vyvyn, 22; Neermn, 23). Mny plnt species re known to hve fungitoxic effects on pthogenic fungl species (Crpinell et l., 23; Bjw et l., 26, 28; Shfique & Shfique, 28). Numerous plnt extrcts e.g., Mgnoli grndiflor (Ahmed & Adelgleil, 25), Aloe ver (Bjw et l., 27), Oryz stiv (Jvid et l., 28) etc., hve een scrutinized for their phytopthogenic ctivity with n intention of exploring environmentlly sfe nd economic lterntives of plnt disese control. Tgetes erectus (mrigold), n ornmentl plnt, which is lso populr edding plnt, cn e used s cover crop. It produces sustnce with pungent odour clled lph-terthienyl tht is suggested useful for inhiiting ttcks from root-knot nemtodes, vine weevils nd other disese promoting orgnisms such s insects, fungi, cteri nd viruses (Khn et l., 1971; Hethelyi et l., 1986; Soule, 1993). Thus the present study ws designed to ssess the ntifungl potentil of queous nd orgnic solvent extrcts of Tgetes erectus on In vitro growth of Ascochyt riei. Mterils nd Methods Procurement nd mintennce of trget fungl species: Culture of trget fungl species of A. riei ws otined from First Fungl Culture Bnk of Pkistn, (FCBP)

2 6 SHAZIA SHAFIQUE ET AL., University of the Punj, Quid-e-Azm Cmpus Lhore nd mintined on mlt extrct gr (MEA) medium. Collection of plnt mterils: Fresh smples of shoot nd flower of Tgetes erectus were collected from University of the Punj, Quid-e-Azm Cmpus Lhore nd wshed thoroughly under tp wter, dried with lotting pper nd cut into smll pieces. The solule ingredients of the plnt mteril were then extrcted y soluiliztion in wter nd methnol s different solvents. Preprtion of queous extrct: Aqueous extrct of wter solule ingredients of plnt mteril ws prepred ccording to Bjw et l., (27). A 2% w/v stock solution of plnt extrct ws ttined y soking the crushed plnt mterils in sterilized distilled wter for 48 h t 25±2 C. Then mteril ws filtered through muslin cloth followed y filter pper Whttmn No. 1. This stock extrct ws stored t 4ºC nd used within four dys. Preprtion of orgnic solvent extrct: The method of Alkhil (25) ws followed for the preprtion of shoot nd flower extrct in methnol. The test plnt ws crushed nd extrcted y mcerting 2 g of plnt mteril in 1 ml of methnol for 48 h. Mterils were filtered through muslin cloth followed y filter pper Whttmn No. 1. Orgnic solvent extrct ws evported under vcuum until its volume ws reduced to 2 ml nd then diluted y dding pproprite quntity of sterilized distilled wter to mke finl volume of 1 ml. The stock extrct ws stored t 4 C nd used within four dys. The lower concentrtions of 1, 2, 3 nd 4% of oth queous nd methnol extrcts of shoot nd flower were prepred y dding pproprite quntity of sterilized distilled wter. To mke methnol control, 2 ml of methnol ws dded to sterilized distilled wter to mke finl volume 1 ml. Lortory iossys: Mlt extrct gr (MEA) medium ws prepred nd cooled to 5 C. Approprite quntities of stock solution nd distilled wter were dded to MEA medium to get 1, 2, 3 nd 4% (v/v) concentrtions of shoot nd flower extrcts in the medium. Control received the sme quntity of distilled wter. The extrcts were thoroughly mixed with the medium. Twenty ml of ech medium ws poured in ech 9 cm dimeter sterilized Petri plte. Mycelil discs of 5 mm dimeter were tken with presterilized cork orer from 5-7 dys old culture of A. riei nd were plced in the centre of ech Petri plte fter solidifiction of the MEA medium. Ech tretment ws replicted thrice. Pltes were incuted in n incutor t 25±2 C for 7 dys. Fungl growth ws mesured y verging the 3 times dimeters tken t right ngles for ech colony. Percentge growth inhiition of the fungl colonies ws clculted y pplying the following formul: Growth in control - Growth in tretment Growth inhiition (%) = 1 Growth in control Sttisticl nlysis: All the dt were nlyzed y nlysis of vrince followed y Duncn s Multiple Rnge Test (Steel & Torrie, 198) using computer softwre SPSS nd COSTAT, respectively.

3 FUNGITOXIC ACTIVITY OF TAGETES ERECTUS ON ASCOCHYTA RABIEI 61 Results Effect of flower extrct of Tgetes erectus on fungl iomss: The results otined from growth ssys of the test species in different concentrtions of queous nd methnolic extrcts of flower of T. erectus re presented in Fig. 1A. A vrile effect of vrious concentrtions ws recorded for the test species. The ntifungl effect of ll the concentrtions of queous extrcts ws significnt ginst the test fungl species except 1% extrct tht proved ineffective in suppressing the fungl growth. Mximum inhiition in fungl growth ws evidenced y 2% queous extrct wheres 3 nd 4% concentrtions exhiited slight nd insignificnt increse in fungl growth ut reveled the significnt ntifungl potentil. In cse of methnol extrct of flower of T. erectus there ws grdul decrese in growth of the test fungl species s the concentrtion of extrct ws incresed from 1 4%. Reltively more toxicity of methnol extrct ws recorded s compred to queous extrct. The 4% methnol extrct concentrtion reveled the gretest inhiitory effect s it cused significnt decrese (5%) in fungl growth (Fig. 2A). Effect of shoot extrct of Tgetes erectus on fungl iomss: The In vitro ntifungl potentil of T. erectus shoot extrcted y queous nd methnol solvent is presented in Fig. 1B. In generl, ll concentrtions of queous extrct were found more inhiitory to the growth of test fungl species thn methnol extrct. All the concentrtions significntly reduced the fungl growth in terms of colony dimeter. Amongst these 4% concentrtion ws the most effective in suppressing the growth up to 73% (Fig. 2B). Methnol extrct lso cused significnt reduction in growth of A. riei s compred to control ut it proved to e comprtively less toxic thn other extrct exmined. However in contrst to queous extrct, 1% concentrtion of Methnol extrct proved ineffective ginst suppressing the fungl growth s it depicted n increse (12.34%) in growth s compred to control. The highest ntifungl ctivity of methnol extrct ws recorded t 4% concentrtion with mximum suppression of fungl growth up to 42% followed y 3 nd 2% concentrtions exhiiting 27 nd 4.5% reduction, respectively (Fig. 2B). Discussion Different extrcts used exhiited vrile ntimycotic ctivity ginst In vitro growth of A. riei. Among the two solvents, shoot queous extrcts were found more inhiitory to test fungl growth thn other extrcts. Aqueous extrcts of shoot of T. erectus reduced the fungl growth y 55-73%. Conversely, mximum reduction of 12-5% ws exhiited y methnolic extrct of flower. The vrition in ntifungl ctivity of the extrcts in different solvents my e ttriuted to the different chemicl nture of the solvents. It is likely tht different types of chemicls were dissolved in different solvents tht resulted in vrile ctivity of the extrcts of sme prt of the plnt in different solvents. There re mny exmples in literture which support these findings. Alkhil (25) studied the effect of queous nd ethnolic extrcts of Allium stivum, Crum crvi, Azidircht indic nd Eugeni cryophyllus ginst Fusrium oxysporum, Botrytis cinere nd Rhizoctoni solni nd found tht queous extrcts were more inhiitory to test fungl species growth thn ethnolic extrct. Similrly, Bjw et l., (27) crried out the study on ntifungl ctivity of queous nd n-hexne shoot extrcts of Aloe ver ginst few pthogenic species viz., Alternri lternt, A. citri nd A. tenuissim. They reported tht the inhiitory effect ws found to e vrile with the pplied concentrtions nd cused significnt inhiition in iomss production of the three test fungi.

4 62 SHAZIA SHAFIQUE ET AL., % 1% 2% 3% 4% A Colony dimeter (mm) Flower Extrct c c Aqueous Extrct Methnol Extrct 7 Shoot Extrct B 6 Colony dimeter (mm) Aqueous Extrct Methnol Extrct Fig. 1. Effect of queous nd orgnic solvent extrcts of Tgetes erectus on In vitro growth of Ascochyt riei fter 7 dys of incution. Verticl rs show stndrd error of mens of three replictes. Vlues with different letters show significnt difference s determined y DMR Test. The extrcts of different prts of T. erectus employed in the present study exhiited vrile ntifungl ctivity ginst the trget pthogenic fungl species A. riei. This vriility in ntifungl potentil of extrcts my e ttriuted to the presence of different types of ntifungl compounds in different prts of the test plnt species. These findings re in line with the work conducted y Zfr et l., (22) who reported tht chloroform extrct of leves of M. zedrch ws ctive ginst Fusrium chlmydosporium while hexne, ethnol nd wter extrcts were not. In similr kind of work Bjw et l.,

5 FUNGITOXIC ACTIVITY OF TAGETES ERECTUS ON ASCOCHYTA RABIEI 63 (26) reported the ntimycotic ctivity of queous nd dichloromethne frctions of Cicer rietinum ginst Drechsler tetrmer nd D. hwiiensis. Likewise, Crpinell et l., (23) reported tht hexnic nd ethnolic extrcts from fruit, seed kernels nd senescent leves exhiited fungisttic ctivity ginst Aspergillus flvus, Diporthe phseolorum vr. meridionles, Fusrium oxysporum, Fusrium solni, Fusrium verticillioides, nd Sclerotini sclerotiorum. The ntifungl ctivity of M. zedrch my e ttriuted to the presence of ntifungl compounds viz., hydroxycoumrin scopoletin, vnillin, 4-hydroxy-3-methoxycinnmldehyde nd (±) pinoresinol (Crpinell et l., 23, 25). The present study confirm tht queous nd orgnic solvent extrcts of vrious prts (flower nd shoot) of T. erectus contin ingredients tht hve ntifungl potentil which re highly effective ginst one of the most destructive pthogens of chickpe. 1% 2% 3% 4% A Flower Extrct Aqueous Methnol Decrese (%) B Shoot Extrct Decrese/Increse ( % ) Fig. 2. Percentge increse/decrese in colony dimeter of Ascochyt riei due to different concentrtions of queous nd orgnic solvent extrcts of flower nd shoot of Tgetes erectus.

6 64 SHAZIA SHAFIQUE ET AL., References Ahmed, S.M. nd S.A.M. Adelgleil. 25. Antifungl ctivity of extrcts nd sesquiterpene lctones from Mgnoli grndiflor L. (Mgnolicee). Int. J. Agric. Biol., 4: Alkhil, A.A., 25. Antifungl ctivity of some extrcts ginst some plnt pthogenic fungi. Pk. J. Biol. Sci., 8 (3): Anonymous. 24. Agriculturl Sttistics of Pkistn Ministry of Food, Agriculture nd Live-stock, Economic Division, Islmd. Bjw, R., A. Jvid, S. Shfique, A. Jvid, K. Jeen nd S. Shfique. 28. Fungisttic ctivity of queous nd orgnic solvent extrcts of rice vrieties on phytopthogenic fungi. Allelopthy Journl, 22(2): Bjw, R., S. Shfique nd S. Shfique. 27. Apprisl of ntifungl ctivity of Aloe ver. Mycopth, 5 (1): 5-9. Bjw, R., T. Anjum, S. Shfique nd S. Shfique. 26. Evlution of ntifungl ctivity of Cicer rietinum L. Pk. J. Bot., 38(1): Crpinell, M.C., C.G. Ferryoli nd C.G. Ferryoli. 25. Antifungl synergistic effect of scopoletin, hydroxycoumrin isolted from Meli zedrch L., fruits. J. Agric. Food Chem., 53: Crpinell, M.C., L.M. Giord, C.G. Ferryoli nd C.G. Ferryoli. 23. Antifungl effects of different orgnic extrcts from Meli zedrch L., on phytopthogenic fungi nd their isolted ctive components. J. Agric. Food Chem., 51: Hethelyi, E., B. Dnos nd P. Tetenyi GC-MS nlysis of the essentil oils of four Tgetes species nd the nti-microil ctivity of Tgetes minut. Flvour nd Frgrnce Journl, 1: Ilys, M.B nd M. Bshir Evlution of systemic fungicides for the control of grm light. Pk. J. Agric. Sci., 21: Iql, S.M., A. Bkhsh, S.R. Mlik nd M. Hqqni. 25. Pthogenic vriility in Ascochyt riei. Pk. J. Phytopthol., 17(2): Jvid, A., S. Shfique, S. Shfique nd T. Riz. 28. Effects of rice extrcts nd residue incorportion on Prthenium hysterophorus mngement. Allelopthy Journl, 22(2): Khn, A.M., S.K. Sxen nd Z.A. Siddiqi Efficcy of Tgetes erect in reducing root infesting nemtodes of tomto nd okr. Indin Phytopth., 24: Neermn, M.F. 23. Sequiterpenes lctones diverse clss of compounds found in essentil oils possessing nticteril nd ntifungl properties. Int. J. Aromth., 13: Port-Pugili, A., P. Infntino, R. Crino nd G. Venor Ascochyt light of chickpe: present sttus nd prospects. Pk. J. Phytopthol., 9(1): Shfique, S. nd S. Shfique. 28. Antifungl ctivity of n-hexne extrcts of Dtur metel ginst Ascochyt riei. Mycopth., 6(1 & 2): Soule, J Tgetes minut: A potentil new her from South Americ. Pp In: New Crops, (Eds.): J. Jnick nd J. E. Simon. Wiley, NY. Steel, R.G.D. nd J.H. Torrie Principles nd Procedures of Sttistics. McGrw Hill Book Co., Inc, New York, USA. Vyvyn, J.R. 22. Allelochemicls s leds for new hericides nd grochemicls. Tetrhedron., 58: Zfr, I., S. Mussrt, H. Frrkh nd B. Sherz. 22. Antifungl properties if some indigenous plnts from Peshwr vlley. Asin. J. Plnt Sci., 1(6): (Received for puliction 2 April 29)