Mycobiology 30(3): 175-179 (2002) Copyright 2002 by The Koren Society of Mycology Inhibitive Activity of Cow Urine nd Cow Dung ginst Sclerotini sclerotiorum of Cucumber A. B. Bsk, Min Woong Lee 1 nd Te Soo Lee 2 Deprtment of Botny, University of Chittgong, Chittgong-4331, Bngldesh 1 Deprtment of Biology, Dongguk University, Seoul 100-715, Kore 2 Deprtment of Biology, University of Inchon, Inchon 402-749, Kore (Received Mrch 7, 2002) A study on comprtive efficcy nd in vitro ctivity of fresh cow urine nd cow dung for controlling Sclerotini rot cused by Sclerotini sclerotiorum of cucumber ws crried out following mycelil growth inhibition test, treted nd untreted scleroti with these orgnic mtters t different dys of. Results showed tht cow urine suppressed more effectively the mycelil growth even fter 5 dys of in comprison to cow dung. The highest inhibition 75.9% of mycelil growth ws recorded in cow dung potto dextrose gr (CUPDA) fter 3 dys of nd lest 22.7% ws in cow dung potto dextrose gr (CUPDA) fter sme dys of. Mycelil growth from scleroti of S. sclerotiorum ws lso influenced by PDA medium mixed with cow urine nd cow dung. After 6 dys of in CUPDA mycelil growth ws only 12.9 mm wheres in CDPDA nd PDA the corresponding growth t the sme time were 65.8 mm nd 80.0 mm. Treted scleroti of the selected fungus with cow urine hd very effective role on suppression of mycelil growth thn tht of untreted one. No mycelil growth ws observed up to 4 dys in treted scleroti with cow urine. After 5 dys only 0.9 mm mycelil growth ws mesured in treted scleroti, while in cse of untreted scleroti the growth ws 42.6 mm. Appliction of cow urine nd cow dung on growing plnts inoculted with the pthogen t different concentrtions lso proved their inhibitive effects. KEYWORDS: Cow urine nd cow dung, Cucumber, Inhibitive ctivity, Sclerotini sclerotiorum Sclerotini sclerotiorum (Lib.) de Bry most common fungl pthogen cusing Sclerotini rot of mny vegetble crops including cucumber, hs been found to cuse severe infection every yer in the crop fields nd glss houses of Kore nd other countries of the world (Jgger, 1920; Kohn, 1979, b; Gmundi nd Spinedi, 1987; Grf nd Schumcher, 1995; KSPP, 1998; Cho et l., 1997; Kim et l., 1999). Due to reltively low temperture nd high reltive humidity in most of the glss houses, the occurrence of the fungl pthogen is very common nd which cn survive or overwinter in the soil s scleroti for long period (Agrios, 1988). Symptoms produced by Sclerotini rot results in not only sudden deth of plnts but lso ppers in stem nd fruit. Mny reserchers recorded the disese s stem rot, stem blight, fruit rot, timber rot nd cottony rot depending on host (Frr et l., 1989). In n intensive survey on six cucurbitceous crops nmelynetted melon, cucumber, pumpkin, summer sqush, wter melon nd orientl melon, Kim et l. (1999) recorded s high s 30~70% disese incidences. Mny reserchers lso identified nd illustrted the cusl orgnism of the Sclerotini rot with chrcteristic typicl symptoms (Willims nd Spooner, 1991; Frr et l., 1989; Purdy, 1979; Cho et l., 1997). Though it is serious pthogen on mny vegetble crops, very little study on *Corresponding uthor <E-mil: bbsk@ctgu.edu> culturl nd chemicl control of the pthogen S. sclerotiorum hs been crried out from different prts of the world (Wlker, 1952; Nyvll, 1989; Agrios, 1988). Recently Bsk nd Lee (2001, b) reported tht fresh cow urine nd cow dung hs positive response in suppression of mycelil growth of Fusrium solni f. sp. cucurbite nd F. oxysporum f. sp. cucumerinum, nd protective effect on Fusrium root rot nd wilt of cucumber by pplying these bio mtters under pot conditions. This reserch crried out to determine the efficcy of cow urine nd dung on Scleroti sclerotiorum under in vivo nd in vitro conditions. Mteril nd Methods Selection of fungus. Sclerotini sclerotiorum ws isolted from the infected smples collected from the different selected glss houses locted t Gimpo city nd Dongguk university frm t Go-yung city, Kore. Inhibitive ctivity on mycelil growth. (1) Cow urine: Inhibition ctivity of cow urine on mycelil growth of S. sclerotiorum ws surveyed on cow urine potto dextrose gr (CUPDA) medium. The ingredients nd their quntities re s follows: peeled nd sliced potto-20 grms, dextrose-20 grms, cow urine- 500 ml nd distilled wter- 500 ml (ph 7.4). Mycelil blocks of 5 mm dimeter of 175
176 Bsk et l. S. sclerotiorum tken from 10 dys old culture were used. Scleroti collected directly from full grown PDA culture medium were lso used in the experiment. In ech Petri dish disc of mycelil block (5 mm) nd scleroti of S. sclerotiorum ws plced t the center. Every fter 24 hours mycelil growth from the disc or scleroti ws mesured nd it ws continued for 5 dys. Percentge of inhibition of mycelil growth ws clculted (Ashrfuzzmn, 1976) (2) Cow dung: For this test, cow dung potto dextrose gr (CDPDA: cow dung- 20 grms, peeled nd sliced potto- 200 grms, dextrose 20 grms, gr- 20 grms nd distilled wter-1000 ml) ws used. The sme method ws followed s described under cow urine. Effect of cow urine on ctivity of scleroti. Scleoti were collected from full grown culture medium nd dipped in cow urine for 24 hour. The treted scleroti were plced on PDA nd mycelil growth from the scleroti ws dily mesured for 5 dys. Inhibitive ctivity of cow urine nd dung on S. sclerotiorum inoculted plnts grown in pots. Inhibitive ctivity of cow urine nd dung ws lso tested on S. sclerotiorum inoculted cucumber plnts grown in pots. S. sclerotiorum ws grown in corn dextrose gr (CDA) medium t 25 o C for 10 dys. In ll 90 cucumber plnts in pots were selected for this experiment. Inocul of S. sclerotiorum from 10 dys old culture were plced t the surrounding soils of the collr prts of 2 months old plnts. Among the plnts, 40 plnts 10 ech were pplied with cow urine nd the other 40 plnts 10 ech with cow dung t concentrtions of un-dilution, 1 : 1, 1 : 2, nd 1 : 4. The remining 10 plnts were not treted with either cow urine or dung. After ppliction of these bio mtters observtion ws tken if ny kinds of wilt symptoms or deth of plnts occurred every fter 24 hours nd this observtion ws continued for 10 dys. Results nd Discussion Tble 1. Effects of cow urine nd cow dung on the mycelil growth from mycelil of Sclerotini sclerotiorum fter different dys of % inhibition of Mycelil growth (mm) in mycelil growth CUPDA CDPDA b PDA CUPDA CDPDA 1 05.6 06.2 13.6 58.6 54.7 2 07.0 08.4 22.0 68.2 61.7 3 09.3 29.7 38.4 75.9 22.7 4 14.9 34.7 45.9 67.5 24.5 5 17.2 40.8 53.8 68.0 24.1 CUPDA : Cow urine potto dextrose gr. b CDPDA : Cow dung potto dextrose gr Effects of cow urine nd cow dung on mycelil growth of S. sclerotiorum vried widely (Tble 1, Fig. 1A nd 1B). Results showed tht cow urine significntly decresed the mycelil growth even fter 5 dys of in comprison to cow dung. After n hour of, fungl growth of CUPDA nd CDPDA were 5.6 nd 6.2 mm, wheres in PDA the mycelil growth ws 13.6 mm. After 5 dys of, in presence of cow urine in medium, the mycelil growth ws recorded s 17.2 mm wheres in PDA the growth ws 53.8 mm. In cse of percentge inhibition of mycelil growth the highest inhibition (75.9%) ws recorded in CUPDA fter 3 dys of nd the lest (22.7%) in CDPDA fter sme dys of (Tble 1). Effect of cow urine nd cow dung medium on direct mycelil growth from scleroti of S. sclerotiorum lso differed gretly (Tble 2, Fig. 1B nd 1E.). No mycelil growth in CUPDA ws recorded within 4 dys of ; while in CDPDA nd PDA some growth of mycelium ws strted. After 6 dys of, mycelil growth ws only 12.9 mm in CUPDA, wheres in CDPDA nd PDA the corresponding growth ws 65.8 mm nd 80.0 mm respectively. In cse of percentge inhibition of mycelil growth the highest vlues (100%) were recorded fter second, third nd fourth dys of nd the lowest (17.8%) fter 6 dys of in CDPDA. Effect of cow urine treted nd untreted scleroti of S. sclerotiorum on verge dimeter of mycelil growth in PDA medium fter different dys of vried widely with respect to dys on nd nture of tretment (Tble 3, Fig. 1C). Results showed tht treted scleroti with cow urine hd very effective role on suppression of mycelil growth thn tht of untreted one. No mycelil growth ws recorded up to 4 dys in treted scleroti with cow urine. After 5 dys only 0.9 mm mycelil growth ws mesured in treted scleroti while in cse of untreted scleroiti the growth ws 42.6 mm. Results of inhibitive of cow urine nd cow dung on inoculted plnts with S. sclerotiorum t different concentrtions re presented in Tble 4, Fig. 1G nd 1H. It ws reveled from the dt tht cow dung hd better inhibitive effect on pthogen inoculted plnts in comprison to plnts treted with cow urine. No plnt infection ws recorded t un-dilution, 1 : 1 nd 1 : 2 concentrtions of cow dung solution when pplied to the pthogens inoculted plnts. However direct ppliction of fresh cow dung t un-diluted condition cused the deth of ll inoculted plnts. Percentge of infected plnts grdully incresed with the decrese of concentrtion rtes of cow urine. In the present study, cow urine nd dung hd different inhibitive ctivities both in vitro nd in vivo conditions. With the increse of concentrtion of these bio mtters the
Inhibitive Activity of Cow Urine nd Cow Dung ginst Sclerotini sclerotium of Cucumber 177 Fig. 1. Effect of cow urine (A-C) nd dung (D-E) on mycelil growth of Sclerotini sclerotiorum nd control. Mycelil growth of the fungus from mycelil block (A), nd from sclerotium (B) in cow urine PDA medium nd from treted sclerotium (C) in PDA medium. Rdil growth of the fungus from mycelil block (D), untreted sclerotium (E) in cow dung PDA medium nd fungl growth in PDA medium (F) s control. Effect of Cow urine (G) nd dung (H) on inoculted plnts with S. sclerotiorum. In both cses, one helthy plnt with the bio mtter tretment (left) nd nother infected plnt (right) without tretment re growing in pots. percentge of plnt infection decresed. So it is indicted clerly tht toxicity of these bio mtters ginst the pthogens depends on their concentrtions. Moreover direct ppliction of cow dung to the pthgen-inoculted plnts cused the deth of plnt. Therefore, cow dung t undiluted concentrtions possessed high toxicity. Results lso showed tht cow dung showed better inhibitive ctivity ginst the selected fungl pthogen thn tht of cow urine. As cow dung is composed of mny digested nd undigested plnt mterils so it is very esy for cow dung to protect plnts creting toxic brriers ginst the inoculted pthogen. Moreover, chemicls or gses present in cow urine my be evported fter ppliction to the plnts. So inhibitive ctivity for cow urine my be lowered in comprison to cow dung. Recently, Bsk nd Lee (2001) reported tht cow urine
178 Bsk et l. Tble 2. Effects of cow urine nd cow dung on mycelil growth from Scleroti of Sclerotini sclerotiorum fter different dys of Incubtion % inhibition of Mycelil growth (mm) in mycelil growth CUPDA CDPDA b PDA CUPDA CDPDA 1 2 02.6 07.2 100.0 63.5 3 13.2 25.0 100.0 47.1 4 42.2 58.0 100.0 29.8 5 07.4 50.3 75.6 90.2 33.5 6 12.9 65.8 80.0 84.0 17.8 CUPDA : Cow urine potto dextrose gr. b CDPDA : Cow dung potto dextrose gr. Tble 3. Effect of cow urine treted nd untreted scleroti of Sclerotini sclerotiorum on verge dimeter of mycelil growth in PDA medium fter different dys of Rdil growth growth (mm) of mycelium on PDA medium of Treted scleroti Untreted scleroti 1 2 02.7 3 10.6 4 25.5 5 0.9 42.6 Tble 4. Effect of cow urine nd cow dung on inoculted plnts with Sclerotini sclerotium t different concentrtions Bio mtters Concentrtion 5 infected plnts in CDA CDA + Cow urine CDA + Cow dung Un-dilution 30.0 0.0 1:1 40.0 0.0 1:2 60.0 0.0 1:4 70.0 40.0 Control 100.0 CDA : Corn dextrose gr. nd cow dung hs n effective role on mycelil growth of F. oxysporum f. sp. cucumerinum nd F. solni f. sp. cucurbite cusing fusrium wilt nd root rot of cucumber, respectively. Both cow urine nd cow dung hve been used in Indo -Bngl sub continent for curing some humn diseses. It is pertinent to mention here tht the role of cow urine nd cow dung in controlling plnt pthogens never hve been studied. So results of the present experiment will provide new technology for controlling plnt pthogens insted of using different chemicl fungicides tht re very hzrdous to both environment nd soil structure. Complete suppression of mycelil growth of S. sclerotiorum my be possible if different herbl plnt extrcts re dded with fresh cow urine nd cow dung before ppliction. In order to estblish the bove results for the commercil use series of different trils should be initited. Acknowledgement Authors re very much grteful to Dr. Chung Duk, President, Kore Science nd Engineering Foundtion (KOSEF) for grnting Postdoctorl Fellowship to the first uthor of this reserch work under which finnce this work ws crried out. Thnks re extended to the President, Dongguk University nd the Hed, Deprtment of Biology, Dongguk University for giving permission to do this work providing ll kinds of lbortory fcilities. References Agrios, G. N. 1988. Plnt Pthology, Third edition, Acdemic Press, p. 803. Ashrfuzzmn, M. H. 1976. Lbortory Mnnul of Plnt Pthology. First edition. Zmn mnzil, Iqbl ngr, Khuln, Bngldesh, p. 95. Bsk, A. B. nd Lee, M. W. 2001. Efficcy of cow dung for controlling root rot nd Fusrium wilt diseses of cucumber plnt. Abstrct published in the 2001 Koren Society of Plnt Pthology nnul meeting nd Interntionl conference, held on 25-30 th October, 2001, Kyongju TEMF hotel, Kore, p. 49. nd. 2001b. 2001. Comprtive efficcy nd in vitro ctivity of cow urine nd cow dung for controlling Fusrium wilt of cucumer. Abstrct published in the 2001 Koren Society of Plnt Pthology nnul meeting nd Interntionl conference, held on 25-30 th October, 2001, Kyongju TEMF hotel, Kore, p. 49. Cho, W. D., Kim, W. G., Jee, H. J., Choi, H. S., Lee, S. D. nd Choi, Y. C. 1997. Compendium of vegetble diseses with color pltes. Ntionl Institute of Agriculturl Science nd Technology. Suwon, Kore, p. 447 Frr, D. F., Bills, G. F., Chmuris, G. P. nd Rossmn, A. Y. 1989. Fungi on plnt nd plnt product in the united sttes, APS press. The Americn Phytopthologicl society, St. Pul, Minnesot, U.S.A. p. 1252. Gmundi, I. J. nd Spinedi, H. A. 1987. Sclerotini Antrctic sp.nov. the teleomorph of the first fungus described from Antrtic. Mycotoxon 29: 81-89. Jggr, I. C. 1920. Sclerotini minor. N.sp. the cuse of decy of lettuce cerley nd other crops. J. Agric. Res. 20: 331-334. Kim, W. G. nd Cho, W. D. 1998. Comprtive chrcteristics of two Sclerotini species ssocited with occurrence of Sclerotini rot on vegetble crops. Proc. And Abs. Of Mycologicl symposium in Asin Reg. Seoul, Kore, Pp. 1-8., nd Jee, H. J. 1999. Occurrence of Sclerotini rot on cucurbitceous vegetble crops in green houses. Koren J. of Mycology 27(3): 198-205. Kohn, L. M. 1979. A monogrphic revision of the genus Scleroti. Mycotoxon 9: 365-444.. 1979b. Delimittion of the economiclly importnt plnt pthogenic Scleroti species. Phytopthology 69: 881-890.
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