Bio-Formulation of Chaetomium cochliodes for Controlling Brown Leaf Spot of Rice

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1 Interntionl Journl of Agriculturl Technology 2014 Vol. 10(2): Aville online ISSN (Online) Fungl Diversity Bio-Formultion of Chetomium cochliodes for Controlling Brown Lef Spot of Rice Soytong, K. * Deprtment of Plnt Production Technology, Fculty of Agriculturl Technology, King Mongkut s Institute of Technology Ldkrng, Bngkok 10520, Thilnd. Soytong, K. (2014). Bio-formultion of Chetomium cochliodes for controlling rown lef spot of rice. Interntionl Journl of Agriculturl Technology 10(2): Astrct Chetomium cochliodes proved to e new ntgonistic fungus ginst rown lef spot of rice vr Pittsnulok 2 cused y Drechsler oryze. It showed good inhiition of mycelil growth of per nd inhiited inoculum production of per. Crude extrcts from Ch cochliodes using hexne, ethyl cette nd methnol t 1,000 ppm could significntly inhiited the inoculum production of rice pthogen per which ED 50 vlue ws ppm when compred to the control (0 ppm). Ch cochliodes ws formulted in different forms for pplying to control rown lef spot of rice. Biologicl products formulted from Ch cochliodes were tested to control rown lef spot of rice cused y D oryze. Result showed tht io-powder formultion gve significntly highest to control lef spot nd highest plnt growth when compred to the non-tret control, followed y pplying crude extrct of Ch cochliodes, enlte nd spore suspension of Ch cochliodes. Moreover, io-powder formultion gve significntly incresed in plnt growth over 44 nd followed y crude extrct of Ch cochliodes, spore suspension of Ch cochliodes nd enlte. Keywords: Chetomium cochliodes, crude extrct, Drechsler oryze Introduction Rice (Oryz stiv L.) is one of economiclly stle crop in the world. During cultivtion of rice, weed, pests nd diseses re invded to destroy the plnts nd low yield. Rice vrieties re reported to infect with severl plnt pthogens eg Trichoconis pdwickii, Curvulri lunt, Fusrium semitectum, Drechsler oryze, Srocldium oryze, Alternri tenuis, Fusrium moniliforme, Nigrospor oryze, Phom spp., Cldosporium spp. nd Pyriculri oryze (Thwt et l, 1997). The importnt diseses occur in susceptile vriety of rice re rice lst cused y Mgnporthe grise (Pyriculri oryze) nd rown lef spot cused y Drechsler oryze (Helminthosporium oryze). Both diseses hve een reported to seriously infected to susceptile vriety of rice leding to low yield over 50. The * Corresponding uthor: Soytong K.; Emil: jksem@gmil.com

2 trditionl chemicl fungicides hve een used for yers nd some cse the pthogens ecome resistnt to those chemicl fungicides. However, there re mny reserchers re reported to use the iologicl control gents to control those diseses. Chetomium cupreum, Chetomium gloosum re reported to e ntgonize Pyriculri oryze cuing rice lst nd Drechsler oryze (Helminthosporium) cusing rown lef spot of rice. Knokmedhkul et l. (2001) stted tht Ch gloosum could produce ntimycocteril nthrquinone-chromnone compound nd disktopiperzine lkloid nd ntifungl Azphilones from Ch cupreum (Phonkerd et l., 2008) including is-spiro-azphilones nd zphilones from Ch cochliodes. Thohinung et l., (2010). The ojective of reserch project ws to evlute io-formultion of Ch. cochliodes to control Drechsler oryze cusing rown lef spot of rice. Mterils nd methods Isoltion of pthogen nd pthogenicity test Rice pthogens were isolted from rice seeds of Chinrt 1, Chinrt 2, Supnuri 2, Supnuri 2, Pitsnulok 2, Koekor 31, Koekor 39,Koekor 41 nd Korkor 47 using moist chmer t room temperture, the signs of pthogens were trnsferred to wter gr (WA) nd then sucultured to potto dextrose gr (PDA) until get pure culture. The most frequency found pthogen would then prove for pthogenicity test followed the Koch s postulte method. Rice seedlings of 15 dys were inoculted with spore suspension of pthogen t the inoculum conrtion of 1x10 6 spore/ml which inoculted to wounded lesions on leves y sterilized needle. The infected res of lesions re re-isolted to get pure culture of pthogen. Identifiction into species level ws morphologiclly done under i-noculr compoud microscope followed instruction of Ellis (1971), Ou (1984), Domsch et l.(1980),von Arx et l. (1986),Soytong nd Quimio (1989), Soytong et l (2001),Pornsuriy et l. (2008) nd Hoog G.S.(2000). Test for control mechnism of Chetomium cochliodes ginst rice pthogen Bi-culturl ntgonistic test ws done using completely rndomized design (CRD) with four replictions. Ch cochliodes. The rice pthogen which proved for pthogenicity ws co-cultured to PDA. The gr plug of ntgonist nd pthogen were trnsferred to PDA in opposite site to ech other, incuted for 18 dys t room temperture. Dt were collected s coloy dimeter (cm) nd spore suspension using hemcytometer. Dt were then trnsformed to 322

3 Interntionl Journl of Agriculturl Technology 2014, Vol. 10 (2): per inhiition (PI) where PI = R1-R2/R1x100, R1 ws colony dimeter or numer of spore in control plte nd R2 ws colony dimeter or numer of spore in co-cultured pltes. The norml spore of pthogen ws lso oserved under inoculr compound microscope. Bioctive compound from Ch cochliodes ws tested for ntifungl ctivity ginst rice pthogen. Ch cochliodes ws cultured in potto dextrose roth (PDB) for 30 dys then filtered nd ir dried t room temperture to get fungl iomss. The dried fungl iomss ws ground nd extrcted in Hexne (1:1, v/v) t sttionry for 3 dys, nd filtered through whtmn filter pper No.4 to get culture filtrte. Culture filtrte ws rifuged using rotry vcuum evportor to get crude hexne. The mrc of fungl iomss ws continued to extrct y ethyl cette (EtOAc) nd methnol (MeOH). Ech crude extrct of hexne, EtOAc nd MeOH ws seprtely dissolved in 2 DMSO (Dimethylsulfoxide) to test ginst rice pthogen. The experiment ws done using two fctors fctoril experiment in CRD with four replictions. Fctor A represented crudecextrct where A1 ws crude hexne, A2 ws crude ethyl cette nd A3 ws crude methnol. Fctor B represented fungl crude extrcts ehich B1 ws 0 ppm, B2 ws 10 ppm, B3 ws 50 ppm, B4 ws 100 ppm, B5 ws 500 ppm nd B6 ws 1,000 ppm. Drechsler oryze ws tested to inhiit y fungl crude extrcts from hexne, ethyl cette dissolved in 2 DMSO (Dimethylsulfoxide) nd methnol incorported with PDA, then sterilized t 121 C, 15 ls for 30 min. The culture gr plug of pthogen ws trnsferred to the middle of PAD incorported with ech crude extrct conrtion, incuted t room temperture. Dt were collected s colony dimeter (cm) nd numer of spores then trnsformed to perge of inhiition = R1-R2/R1x100; R1 = numer of spores in control, R2 = numer of spores in treted pltes X 100. Dt were computed for nlysis of vrince nd tretment mens were compred using Duncn Multiple Rnge Test (DMRT) t P = nd P = The ED 50 ws computed y proit nlysis progrm. Bio-formultion of Chetomium cochliodes testing to control rown lef spot The 15 rice seedlings were trnsferred to soil for 7 dys efore inocultion with spore suspension of pthogen t the conrtion of 1x10 6 spore/ml to wounded lesions in rtificil rice field in greenhouse. The experiment ws performed using rndomized complete lock design (RCBD) with four replictions Tretments were done s follows:- T1 ws inoculted control, T2 ws non inoculted control, T3 ws io-formultion from spore suspension (1x10 6 ) of Ch cochliodes, T4 ws io-powder form of Ch 323

cochliodes pplied t 10 g/1lof wter, T5 ws enlte-chemicl fungicide pplied t recommendtion rte nd T6 ws mixed crude extrct pplied t 10 g/ L of wter.

Dt were collected s disese incidence, plnt height (cm), numer of tillers, fresh nd dried weight of plnt nd computed nlysis of vrince nd tretment mens were

Results Isoltion of pthogen nd pthogenicity test Rice pthogens were isolted from rice seeds of Chinrt 1, Chinrt 2, Supnuri 2, Supnuri 2, Pitsnulok 2, Koekor

It ws found Drechsler oryze which the most frequency ppered s seed orne fungi of rice vr Pitsnulok 2, Koekor 31, Koekor 41 nd Korkor 47.

4 cochliodes pplied t 10 g/1lof wter, T5 ws enlte-chemicl fungicide pplied t recommendtion rte nd T6 ws mixed crude extrct pplied t 10 g/ L of wter. All tretments were pplied t every 15 dys. Dt were collected s disese incidence, plnt height (cm), numer of tillers, fresh nd dried weight of plnt nd computed nlysis of vrince nd tretment mens were compred using Duncn Multiple Rnge Test (DMRT) t P = nd P = Results Isoltion of pthogen nd pthogenicity test Rice pthogens were isolted from rice seeds of Chinrt 1, Chinrt 2, Supnuri 2, Supnuri 2, Pitsnulok 2, Koekor 31, Koekor 39,Koekor 41 nd Korkor 47. It ws found Drechsler oryze which the most frequency ppered s seed orne fungi of rice vr Pitsnulok 2, Koekor 31, Koekor 41 nd Korkor 47. Pure culture showed rown color when mture, septte myceli, porospores with mny septte or cells on one conidi. Chetomium cochliodes showed gry color colony when young nd turned to olivceous greenish gry to rown when mture on potto dextrose gr which relesed purple. Ascocrp ecome rown mture within 6 weeks, 120 x 85 µm, eight scospores per scus, scus clvte, scospore like lemon shpe, 10 x 25 µm. (Fig.1). A B C D Fig. 1. Chetomium cochliodes A : colony on PDA, B :scomt, C : terminl hir, nd D : scospore 324

5 Interntionl Journl of Agriculturl Technology 2014, Vol. 10 (2): Test for control mechnism of Chetomium cochliodes ginst rice pthogen Bi-culturl ntgonistic test Result showed tht Ch cochliodes could inhiit mycelil growth of D oryze which verged colony of 5.56 cm when compred to control plte of 9.00 cm. It could inhiit myceli per in 10 dys. However, Ch cochliodes significntly inhiited spore production of D. oryze per. It could show control mechnism of lysis in hyphe of pthogen (Fig.2). Fig. 2. Hyphe of Drechsler oryze decomposed due to sustnces relesed from Chetomium cochliodes in i-culture ntgonistic test Tle 1. Chetomium cochliodes ginst Drechsler oryze in i-culture ntgonistic test Drechsler oryze Inhiition 2 () C.V. () control Bi-culture Colony (cm) Spore numer Averge of four replictions. Mens followed y common letter re not significntly differed y DMRT t P= Inhiition () = R1-R2/R1x100 where R1ws numer of pthogen spores in control nd R2 ws numer of pthogen spore in i-culture pltes. Bioctive compound test Chetomium cochliodes ws cultured in potto dextrose oth (PDB) of 500 petri dishes (9cm di.) for 30 dys, then get the dried fungl iomss 20.5 g. The fungl iomss ws ground in electricl lender nd soked into hexne 500 ml then plced in shker for 72 h nd filtered through whtmn filter pper 325

6 No 4 to get the filtrte. The filtrte ws then operted in rotry vcuum evportor to get hexne crude extrct of 0.79 g. The mrc ws further extrcted through ethyl cette nd methnol followed the sme method to get ethyl cette crude nd methnol crude extrcts ( Fig. 3). Fig.3. Hexne, ethyl cette nd methnol crude extrcts from Chetomium cochliodes Result showed tht hexne crude extrct from Chetomium cochliodes gve significntly highest inhiition of 54 for the colony growth of Drechsler oryze t the conrtion of 1,000 ppm when compred to the control. Moreover, hexne crude extrct from Ch cochliodes gve significntly highest inhiition of for the spore production of D oryze t the conrtion of 1,000 ppm when compred to the control which the ED 50 ws ppm. Hexne crude extrct from Ch. cochliodes could significntly inhiited colony growth of 54 t conrtion of 1,000 ppm nd inhiited spore production of 90. Moreover, ethyl cette nd methnol crude extrcts could significntly inhiit spore production of nd 93.35, respectively (Tles 2, 3 nd 4). The ED 50 vlues of crude hexne, ethyl cette nd methnol extrcted from Ch. cochliodes could inhiit D oryze t 66.45, nd ppm, respectively (Tle5). All crude extrcts expressed norml spore of D oryze (Fig.5, 6 nd 7) nd those norml spores lost pthogenicity. 326

7 Interntionl Journl of Agriculturl Technology 2014, Vol. 10 (2): Tle 2. Crude extrcts of Chetomium cochliodes testing to inhiit Drechsler oryze t 5 dys Crude extrcts Conrtion (ppm) Colonydimeter (cm) / 1 Growth inhiition () / e c 30.00c Crude hexne de d d e e d Crude ethyl cette c 23.50cd c 25.00c c 29.50c c 29.50c e e Crude methnol e d c 26.00c c 29.50c C.V. () Averge of four replictions. Mens followed y common letter re not significntly differed y DMRT t P= Inhiition () = R1-R2/R1x100 where R1ws colony dimeter of pthogen in control nd R2 ws colony dimeter of pthogen in treted pltes. 327

8 Tle 3. Growth inhiition of crude extrcts from Chetomium cochliodes to Drechsler oryze t 5 dys Crude extrcts Conrtion Mycelil fresh (ppm) weight (g) Inhiition () 3/ h c 65.17f Crude hexne de 78.91cde def 81.78cd def 82.10cd h h g Crude ethyl def 0.00h cette efg 82.05cd efg 82.05cd efg 82.37c h c 65.34f Crude def 79.40cde methnol d 76.79e fg g C.V. )( Averge of four replictions. Mens followed y common letter re not significntly differed y DMRT t P= Inhiition () = R1-R2/R1x100 ;where R1ws mycelil fresh weight of pthogen in control nd R2ws mycelil fresh weight of pthogen in treted plte. 328

9 Interntionl Journl of Agriculturl Technology 2014, Vol. 10 (2): Tle 4. Spore production inhiition of crude extrcts from Chetomium cochliodes to Drechsler oryze t 5 dys Crude extrcts Conrtion (ppm) Numer of spores Inhiition () 2/ j hi Crude hexne de 54.99fg f 62.33de i i j Crude ethyl i fg 63.14de cette h 74.36c i i j c 29.55h Crude methnol d 48.96g ef 57.60ef gh 68.88cd i C.V. () Averge of four replictions. Mens followed y common letter re not significntly differed y DMRT t P= Inhiition () = R1-R2/R1x100 where R1ws numer of pthogen spores in control nd R2 ws numer of pthogen spore in treted plte. 329

10 Tle 5. Effective dose (ED 50 ) of crude extrcts from Chetomium cochliodes to inhiit Drechsler oryze t 5 dys Crude extrcts Crude hexne Crude cette Crude methnol ethyl Conrtion (ppm) Growth imhiition 2 / Mycelil inhiition / Spore production inhiition () e h 0.00j c 65.17f 4.14hi cde 54.99g cd 62.33de cd e 0.00h 0.00j d 8.65g cd 0.00h c 82.05cd c 82.05cd c 82.37c 21.63i 63.14de 74.36c e 0.00h 0.00j e 65.34f e 79.40cde d 76.79e c c h 48.96g 57.60ef 68.88cd ED 50 (ppm) C.V Averge of four replictions. Mens followed y common letter re not significntly differed y DMRT t P= Inhiition () = R1-R2/R1x100 where R1ws colony growth or mycelil growth or numer of pthogen spores in control, R2= colony growth or mycelil growth or numer of pthogen spore in treted plte. Tle 6. Plnt height of rice vr Pitsnulok 2 fter pplying io-formultion of Chetomium cochliodes Tretment 40 dys / Inoculted Control 8.00c Non- Inoculted Control Spore suspention, Ch cochliodes Bio-powder Ch. cochliodes Crude extrct ofch.cochliod es Benlte C.V. () 9.40 Increse d () 2/ 55 dys Increse d ()2/ 70 dys c Increse 85 d ()2/ dys Increse 100 d ()2/ dys e d c c Increse d ()2/ - 1 Averge of four replictions. Mens followed y common letter re not significntly differed y DMRT t P= incresed perge = plnt height in ech tretment plnt height in inoculted tretment /plnt height in ech tretment x

11 Interntionl Journl of Agriculturl Technology 2014, Vol. 10 (2): Tle 7. plnt height nd disese index of rice vr Pitsnulok 2 fter pplying ioformultion of Chetomium cochliodes Tretment 40 dys / incres ed 2/ 55 dys incres ed 70 dys incres ed 85 dys incres ed 100 dys incres ed Dise se index 3 Inoculted c d Control Non c c d Inoculted Control Spore c suspention, Ch. cochliodes Bio-powder c Ch. cochliodes Crude extrct c ofch.cochlio des Benlte C.V. () Averge of four replictions. Mens followed y common letter re not significntly differed y DMRT t P= incresed perge = plnt height in ech tretment plnt height in inoculted tretment /plnt height in ech tretment x Disese Index, 1= lef spot 0, 2=lef spot 1-25,3=lef spot 26-50,4=lef spot nd 5 =lef spot over 75. Tle 8. Plnt growth prmeters of rice vr Pitsnulok 2 fter pplying io-formultion of Chetomium cochliodes Tretment Fresh plnt weig ht (g) incre sed 2/ Fresh root weigh t incre sed 2/ Fresh pnic le weig ht incre sed 2/ Dried plnt weig ht incre sed 2/ Drie d root weig ht incre sed 2/ Dried pnic le weig ht incre sed 2/ Inoculted Control 5 Non Inoculted Control 0 6 Spore suspention, Ch. cochliodes Biopowder Ch. cochliodes Crude extrct ofch.cochl iodes Benlte C.V. () Averge of four replictions. Mens followed y common letter re not significntly differed y DMRT t P= incresed perge = plnt height in ech tretment plnt height in inoculted tretment /plnt height in ech tretment x

12 Fig. 5. Crude Hexne from Chetomium cochliodes t vrious conrtions (A) nd norml spores of Drechsler oryze ; (B) 332

13 Interntionl Journl of Agriculturl Technology 2014, Vol. 10 (2): Fig. 6. Crude ethyl cette from Chetomium cochliodes t vrious conrtions (A) nd norml spores of Drechsler oryze ; (B) 333

14 Fig. 7. Crude Hexne from Chetomium cochliodes t vrious conrtions (A) nd norml spores of Drechsler oryze ; (B) 334

15 Interntionl Journl of Agriculturl Technology 2014, Vol. 10 (2): Bio-formultion of Chetomium cochliodes testing to control rown lef spot Bio-formultion of Ch cochliodes cn e significntly reduced lef spot of rice vr Pitsnulok 2 cused y D. oryze. Result showed tht iopowder of Ch cochliodes, Bio-crude extrct nd enlte treted to rice t 40 dys showing plnt height of 13.41, nd cm, respectively when compred to the control (8 cm). At 70 dys, io-powder, crude extrct of Ch cochliodes nd enlte showed plnt height of 50.83, nd cm, respectively when compred to the control (21 cm). At 85 dys, io-powder, crude extrct of Ch cochliodes nd enlte showed plnt height of 30.91, nd cm, respectively when compred to the control (33.91 cm). At 100 dys, iopowder, crude extrct of Ch cochliodes nd enlte showed plnt height of 70.83, nd cm, respectively when compred to the control (47.66 cm). For numer of tillers t 40 dys, spore suspension of Ch cochliodes, enlte, io-powder nd crude extrct of Ch cochliodes showed numer of tillers of 3.58, 3.23, 3.16 nd 3.16 tillers, respectively when compred to the control (3.0 tillers). At 55 dys, enlte, crude extrct nd io-powder of Ch cochliodes showed numer of tillers of 13.00, nd tillers, respectively when compred to the control (6.5 tillers). For numer of tillers t 70 dys, io-powder, crude extrct of Ch cochliodes nd enlte showed numer of tillers of 18.66, nd tillers, respectively when compred to the control (7.33 tillers). For numer of tillers t 85 dys, io-powder, crude extrct of Ch cochliodes nd enlte showed numer of tillers of 19.08, nd tillers, respectively when compred to the control (7.50 tillers). For numer of tillers t 100 dys, io-powder, crude extrct of Ch cochliodes nd enlte showed numer of tillers of 19.16, nd tillers, respectively when compred to the control (10.58 tillers). Fresh weight of plnt, io-powder, spore suspension of Ch cochliodes nd enlte nd crude extrct of Ch cochliodes gve fresh weight of , , nd g, respectively when compred to the control (83.75 g). Rice pnicle weight showed tht spore suspension, enlte, io-powder of Ch cochliodes gve fresh pnicle weight of 48.75, nd g respectively when compred to the control (28.75 g). Spore suspension of Ch cochliodes, enlte nd crude extrct of Ch cochliodes gve pnicle dried weight of 85.75, nd g when compred to the control (31.25 g). Root dried weight, It showed tht spore suspention, crude extrct nd io-powder of Ch cochliodes showed root dried weight of 52.50, nd g respectively when compred to the control (9.50 g). Bio-poweder, spore suspension nd crude extrct of Ch cochliodes gve significntly controlled lef spot of rice cuse y D oryze when compred to inoculted with pthogen nd enlte chemicl fungicide ws 335

16 significntly etter disese control thn those ll io-formultion of Ch cochliodes (Tle 6). Discussion Drechsler oryze ws isolted from lef spot of rice from different vrieties. It elongs to Deuteromycotin, Hyphles, Demticee which showing Imperfect stge. The fungus morphology is similr to report of Bred de Hn (1900) which four cell conidi, X µm, colony deep rown to lckish rown s similr report y Ellis (1971) nd Ou (1984) Chetomium cochliodes elongs to Ascomycotin, Pyrenomycetes, Chetomiles, Chetomicee which showing perfect stge. It produces peritheci, cylindricll sci nd 8-scospore per scus which reported y Soytong ( 2535), Soytong et l. (2001), Treit nd Moore (1954) nd Johnston nd Booth (1983). As result, Ch. cochliodes proved to e ntgonized D. oryze cusing lef spot of rice vr Pitsnulok 2. Ch. cochliodes could inhiit colony growth nd spore production of D. oryze s in i-culture ntgonistic test which similr report of Soytong (1992). Bi-culture test showed lysis of pthogen spore which served s control mechnism seport y Soytong (2548). Crude hexne of Ch cochliodes inhiited spore production of pthogen t conrstion of 1,000 µg/ml (93.85 ) which ED 50 ws ppm. This similr report y Bisws et l. (2002) showed tht crude extrct of Chetomium sp could Drechsler sorokinin cusing spot lotch of whet (Bisws et l., 2002). As result, io-powder, crude extrct nd spore suspension of Ch cochliodes which similr to Soytong et l. (2001) who pplied iopowder formulted from Chetomium gloosum nd Chetomium cupreum cn e inhiited severl plnt pthogens especilly D. oryze cusing rown spot of rice nd Soytong (2535) reported tht rice seed coted with Ch gloosum, Ch cupreum nd Ch cochliodes could inhiit rice lst cused y P. oryze. References Bker, K. F. (1987). Evolving concepts of iologicl control of plnt pthogen. Annul Review of Phytopthology 25: Bisws, S. K. Srivstv, K. D. Bisws, D. R. nd Aggrwl, R. (2002). Effect of folir spry of Chetomium gloosum on totl protein, nitrogen nd cron contents of whet. Annls of Plnt Protection Sciences 10: Counce, P. A., Keisling, T. C. nd Mitchel, A. J. (2000). A uniform, ojective, nd dptive system for expressing rice development. Crop Science 40: Greyson, R. I. (1994). The Development of Flowers. Oxford University Press, U.S.A. Hyuncheol, O., Swenson, D. C., Gloer, J. B., Wicklow, D. T. nd Dowd, P. F (1998). Chetochlsin A: A new ioctive metolite from Chetomium rsiliense. Tetrhedron Letters 39:

17 Interntionl Journl of Agriculturl Technology 2014, Vol. 10 (2): Smith, C. W. nd Dildy, R. H. (2002). Rice: Origin, History, Technology, nd Production. John Wiley nd Sons, Inc., USA. Kndnsoom, P. nd Sitichi, T. (1967). Vrietl difference in helminthosporium lef spot nd some prolems of control mesure in Thilnd. Proceedings of symposium on rice diseses nd their control y growing resistnt vrieties nd other mesures. Agriculture, Forestry nd Fisheries Reserch Council, Ministry of Agriculturl nd Forests, Tokyo, Jpn. pp Knokmedhkul, S., Nsomji, P., Loungsysouphnh, S., Soythong, K., Isoe, M., Kongseree, K., Prpi, S. nd Suksmrn, A. (2006). Antifungl zphilones from the fungus, Chetomium cupreum CC3003. Journl of Nturl Products 69: Lmp, C. A., Fores, S. J. nd Cde, J. W. (1990). Grss of temperte Austrli. Inkt Press, Sydney. Mhmud, K. nd Khnzd, A. K. (1989). Seed-orne fungl disese of rice in sindh, Pkistn. pp Mnndhr, P. N., Thpliyl, P. N. nd Sinclir, J. B. (1986). Potentil iocontrol fungi for selected soyen fungl pthogens. Biologicl Control nd Cultursl Tests. pp. 36. Moldenhuer, K. A. K, nd Gions, J. H. (2003). Rice morphology nd development. pp Nyvll, R. f. (1999). Field crop diseses. Low Stte University Press, USA. 1: pp. 21. Odette, L. S. nd Ellis, J. J. (1974). Helminthosporium Drechsler nd Bipolris Toxin. Mycotoxins nd Other Fungl Relted Food Prolems. pp Ou, S. H. (1985). Rice Diseses. Commonwelth Mycologicl Institute. pp Pornsuriy, C., Lin, F. C., Knokmedhkul, S. nd Soytong, K. (2008). New record of Chetomium species isolted from soil under pinepple plnttion in Thilnd. Journl of Agriculturl Technology 4: Sennyke, N., De Dtt, S. K., Nylor, R. E. L., nd Thompson, W. J. (1991). Lowlnd rice picl development: stges nd cultivr differences detected y electron microscope. Agronomy Journl 83: Soytong, K. (1989). Appliction of Chetomium cupreum to control rice lst. Thi Phytopthology 9: Soytong, K. (1992). Antgonism of Chetomium cupreum to Pyriculri oryze. Journl of plnt protection in the tropics 9: Soytong, K. (1992). Biologicl control of rice lst disese y seed coting with ntgonistic fungi. Songklnkrin Journl Science Technology 14: Soytong, K. nd Knokmedhkul, S., Kukongviriypn, V., nd Isoe, M. (2001). Appliction of Chetomium species (Ketomium) s new rod spectrum iologicl fungicide for plnt disese control: review rticle. Fungl Diversity 7:1-15. Soytong, K. nd Quimio, T. H. (1989). Antgonism of Chetomium gloosum to the rice lst pthogen, Pyriculri oryze. Ksetsrt Journl Nturl Science 23: Tkeok, Y. M., Shimizu, M. nd Wd, T. (1993). Pnicle. Science of the Rice Plnt, Volume I. Nounkyo, Tokyo. pp The Interntionl Rice Resherch Institute (IRRI). (1985). The flowering response of the rice plnt to photoperiod. The Interntionl Rice Reserch Institute, Philippines. Policy Reserch Center, Jpn. Vergr, B. S. nd Chng, T. T. (1985). The flowering response of the rice plnt to photoperiod: review of the literture. Interntionl Rice Reserch Institute. Thohinung, S., Knokmedhkul, S., Knokmedhkul, K., Kukongviriypn, V., Tusskorn, O., Soytong, K. (2010). Cytotoxic 10-(indol-3-yl)-[13]cytochlsns from the fungus Chetomium eltum ChE01. Fculty of Science, Khon Ken University 33: Tveit, M. nd M. B., Moore. (1954). Isolte of Chetomium tht protect ots from Helminthosporium victorie. Phytophtology 44: (Received 13 Jnury 2014; ccepted 28 Ferury 2014) 337

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