稻曲病菌菌核降解微生物的筛选与作用机制分析
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摘要
菌核是稻曲病菌天然的越冬菌源和来年病害发生的重要初侵染源,控制田间菌核越冬数量能够从源头上遏制和减轻稻曲病的发生。为此,本研究对来自稻田土壤、菌核表面、以及青海高原的能够降解菌核的真菌进行了筛选,得到了6个有较好降解作用的生防菌株。在实验室条件下它们能够在30~70 d内彻底降解稻曲病菌菌核。真菌形态特征和r DNA-ITS序列分析表明,这些菌株分别为淡色生赤壳菌Bionectria ochroleuca、粘鞭霉Gliomastix polychroma、烟曲霉Aspergillus fumigatus、草酸青霉Penicillium oxalicum、粉红粘帚霉Gliocladium spp.和疣孢漆斑菌Myrothecium verrucaria。进一步超微结构观察发现,不同生防菌对菌核的降解机制可分为以重寄生作用和直接降解为主2种基本模式。春季田间撒施试验表明,其中2种生防菌的防治效果可达33.9%和46.8%。
Sclerotia of Villosiclava virens, the pathogen of rice false smut, can overwinter and germinate in the next year to produce ascospores as the primary inocula of rice, so the measures decreasing the sclerotial number in paddy field will be practically useful in control of the disease. We screened sclerotium-degrading fungi against V. virens from paddy field soil, the sclerotium-attached fungi, and the fungi from Tibetan plateau. Tests indicated that Bionectria ochroleuca, Gliomastix polychroma, Aspergillus fumigates, Penicillium oxalicum, Gliocladium roseum, and Myrothecium verrucaria could degrade completely the sclerotia within 30-70 d. Ultrastructural examinations showed that the degradation mechanisms included myco-parasitism and saprophytic modes, respectively. Two of the fungi spreading in paddy field in the spring could decrease the false smut balls by 33.9% and 48.6%, respectively.
Sclerotia of Villosiclava virens, the pathogen of rice false smut, can overwinter and germinate in the next year to produce ascospores as the primary inocula of rice, so the measures decreasing the sclerotial number in paddy field will be practically useful in control of the disease. We screened sclerotium-degrading fungi against V. virens from paddy field soil, the sclerotium-attached fungi, and the fungi from Tibetan plateau. Tests indicated that Bionectria ochroleuca, Gliomastix polychroma, Aspergillus fumigates, Penicillium oxalicum, Gliocladium roseum, and Myrothecium verrucaria could degrade completely the sclerotia within 30-70 d. Ultrastructural examinations showed that the degradation mechanisms included myco-parasitism and saprophytic modes, respectively. Two of the fungi spreading in paddy field in the spring could decrease the false smut balls by 33.9% and 48.6%, respectively.
引文
[1]胡东维,王疏.稻曲病菌侵染机制研究的现状与展望[J].中国农业科学,2012,45(22):4604-4611.
[2]Singh R A,Dube K S.Occurrence of true sclerotia in Claviceps oryzae-sativae causal organism of false smut of rice[J].Current Science,1976,45(21):772-773.
[3]Yaegashi H,Fujita Y,Sonoda R.Severe outbreak of false smut of rice in 1988[J].Plant Protection Tokyo,1989,43(6):311-314.
[4]Rush M C,Shahjahan A K M,Jones J P,et al.Outbreak of false smut of rice in Louisiana[J].Plant Disease,2000,84(1):100.
[5]Atia M M M.Rice false smut(Ustilaginoidea virens)in Egypt[J].Journal of Plant Diseases and Protection,2004,111(1):71-82.
[6]Sakurai M.On the causal fungus of rice false smut[J].Annals of the Phytopathological Society of Japan,1934,3(1):70-71.
[7]Ikegami H.Studies on the false smut of rice.IX.occurrence and development of sclerotia of the rice false smut fungus[J].Research Bulletin of the Faculty of Agriculture,Gifu University,1963,(18):47-53.
[8]严位中,王永华,钱均端.稻曲病菌核的初步研究[J].云南农业大学学报,1993,8(3):206-207.
[9]Tanaka E,Ashizawa K,Sonoda R,et al.Villosiclava virens gen.nov.,comb.nov.,teleomorph of Ustilaginoidea virens,the causal agent of rice false smut[J].Mycotaxon,2008,106(1):491-501.
[10]Koiso Y,Li Y,Iwasaki S,et al.Ustiloxins,antimitotic cyclic peptides form false smut balls on rice panicles caused by Ustilaginoidea virens[J].Journal of Antibiotics(Tokyo),1994,47(7):765-773.
[11]邓启得.菌核在稻曲病菌生活史中的作用研究[D].杭州:浙江大学,2015.
[12]Honkura R,Miura Y.Some notes on the spore disperse and the period of infection of rice false smut pathogenic fungus[J].Annual Report of the Society of Plant Protection of North Japan,1988,39:88-91.
[13]Fujita Y,Sonoda R,Yaegashi H.The fruiting body formation in the sclerotia of Ustilaginoidea viren and the effect by light[J].Annual Report of the Society of Plant Protection of North Japan,1990,41:205.
[14]缪巧明.稻曲病菌核的研究[J].云南农业大学学报,1994,9(2):101-104.
[15]Singh R A,Dubey K S.Sclerotial germination and ascospore formation of Claviceps oryzae-sativae in India[J].Indian Phytopathology,1984,37(2):168-170.
[16]邓根生,陈嘉孚,杨治华,等.稻曲病菌厚垣孢子在侵染循环中作用的研究[J].中国农学通报,1990,6(2):20-22.
[17]樊荣辉,王永强,刘兵,等.稻绿核菌无性孢子形成过程及厚垣孢子萌发率测定[J].菌物学报,2010,29(2):188-192.
[18]兰时乐,陈海荣,肖宏英,等.稻曲病菌拮抗菌的筛选及拮抗活性测定[J].植物保护,2004,30(2):69-71.
[19]李竞生,张敏,膨化贤,等.枯草芽孢杆菌xi-55对稻曲病菌的拮抗活性测定[J].中国农学通报,2008,24(7):375-377.
[20]尹小乐,陈志谊,刘永峰,等.稻曲病拮抗细菌的筛选与评价[J].江苏农业学报,2011,27(5):983-989.
[21]Yamamoto S,Yoshino J.Storage period of Sclerotia of Ustilaginoidea virens[J].Annual Report of the Kanto-Tosan Plant Protection Society,1955,2:10.
[22]李世东,刘杏忠.食菌核葚孢霉——一种有潜力的菌核病生防菌[J].中国生物防治,2000,16(4):177-182.
[23]Gracia-Garza J A,Reeleder R D,Paulitz T C.Degradation of sclerotia of Sclerotinia sclerotiorum by fungus gnats(Bradysia coprophila)and the biocontrol fungi Trichoderma spp.[J].Soil Biology and Biochemistry,1997,29(2):123-129.
[24]Sarrocco S,Mikkelsen L,Vergara M,et al.Histopathological studies of sclerotia of phytopathogenic fungi parasitized by a GFP transformed Trichoderma virens antagonistic strain[J].Mycological Research,2006,110(2):179-187.
[25]Zeng W,Wang D,Kirk W,et al.Use of Coniothyrium minitans and other microorganisms for reducing Sclerotinia sclerotiorum[J].Biological Control,2012,60(2):225-232.
[26]Sennoi R,Singkham N,Jogloy S,et al.Biological control of southern stem rot caused by Sclerotium rolfsii using Trichoderma harzianum and arbuscular mycorrhizal fungi on Jerusalem artichoke(Helianthus tuberosus L.)[J].Crop Protection,2013,54(1):148-153.
[27]高必达,钟杰.稻曲病菌侵染过程研究进展[J].湖南农业科学,2011,(2):93-97.
[28]刘永芳,陈增建.稻曲病初次侵染源及侵染途径的研究[J].天津农业科学,1995,1(1):9-11.
[29]杜爱玲.鸡腿菇炭角菌病害发生机理及其防治研究[D].杨凌:西北农林科技大学,2006.
[30]Lumsden R D,Ridout C J,Vendemia M E,et al.Characterization of major secondary metabolites produced in soilless mix by a formulated strain of the biocontrol fungus Gliocladium virens[J].Canadian Journal of Plant Pathology,1992,38(9):1274-1280.
[31]何迎春,高必达.立枯丝核菌的生物防治[J].中国生物防治,2000,16(1):31-34.
[32]李晶晶,张拥华,陈巧云,等.粉红粘帚霉67-1菌株寄生核盘菌菌核的酶学动态研究[J].菌物研究,2007,5(2):107-109.
[2]Singh R A,Dube K S.Occurrence of true sclerotia in Claviceps oryzae-sativae causal organism of false smut of rice[J].Current Science,1976,45(21):772-773.
[3]Yaegashi H,Fujita Y,Sonoda R.Severe outbreak of false smut of rice in 1988[J].Plant Protection Tokyo,1989,43(6):311-314.
[4]Rush M C,Shahjahan A K M,Jones J P,et al.Outbreak of false smut of rice in Louisiana[J].Plant Disease,2000,84(1):100.
[5]Atia M M M.Rice false smut(Ustilaginoidea virens)in Egypt[J].Journal of Plant Diseases and Protection,2004,111(1):71-82.
[6]Sakurai M.On the causal fungus of rice false smut[J].Annals of the Phytopathological Society of Japan,1934,3(1):70-71.
[7]Ikegami H.Studies on the false smut of rice.IX.occurrence and development of sclerotia of the rice false smut fungus[J].Research Bulletin of the Faculty of Agriculture,Gifu University,1963,(18):47-53.
[8]严位中,王永华,钱均端.稻曲病菌核的初步研究[J].云南农业大学学报,1993,8(3):206-207.
[9]Tanaka E,Ashizawa K,Sonoda R,et al.Villosiclava virens gen.nov.,comb.nov.,teleomorph of Ustilaginoidea virens,the causal agent of rice false smut[J].Mycotaxon,2008,106(1):491-501.
[10]Koiso Y,Li Y,Iwasaki S,et al.Ustiloxins,antimitotic cyclic peptides form false smut balls on rice panicles caused by Ustilaginoidea virens[J].Journal of Antibiotics(Tokyo),1994,47(7):765-773.
[11]邓启得.菌核在稻曲病菌生活史中的作用研究[D].杭州:浙江大学,2015.
[12]Honkura R,Miura Y.Some notes on the spore disperse and the period of infection of rice false smut pathogenic fungus[J].Annual Report of the Society of Plant Protection of North Japan,1988,39:88-91.
[13]Fujita Y,Sonoda R,Yaegashi H.The fruiting body formation in the sclerotia of Ustilaginoidea viren and the effect by light[J].Annual Report of the Society of Plant Protection of North Japan,1990,41:205.
[14]缪巧明.稻曲病菌核的研究[J].云南农业大学学报,1994,9(2):101-104.
[15]Singh R A,Dubey K S.Sclerotial germination and ascospore formation of Claviceps oryzae-sativae in India[J].Indian Phytopathology,1984,37(2):168-170.
[16]邓根生,陈嘉孚,杨治华,等.稻曲病菌厚垣孢子在侵染循环中作用的研究[J].中国农学通报,1990,6(2):20-22.
[17]樊荣辉,王永强,刘兵,等.稻绿核菌无性孢子形成过程及厚垣孢子萌发率测定[J].菌物学报,2010,29(2):188-192.
[18]兰时乐,陈海荣,肖宏英,等.稻曲病菌拮抗菌的筛选及拮抗活性测定[J].植物保护,2004,30(2):69-71.
[19]李竞生,张敏,膨化贤,等.枯草芽孢杆菌xi-55对稻曲病菌的拮抗活性测定[J].中国农学通报,2008,24(7):375-377.
[20]尹小乐,陈志谊,刘永峰,等.稻曲病拮抗细菌的筛选与评价[J].江苏农业学报,2011,27(5):983-989.
[21]Yamamoto S,Yoshino J.Storage period of Sclerotia of Ustilaginoidea virens[J].Annual Report of the Kanto-Tosan Plant Protection Society,1955,2:10.
[22]李世东,刘杏忠.食菌核葚孢霉——一种有潜力的菌核病生防菌[J].中国生物防治,2000,16(4):177-182.
[23]Gracia-Garza J A,Reeleder R D,Paulitz T C.Degradation of sclerotia of Sclerotinia sclerotiorum by fungus gnats(Bradysia coprophila)and the biocontrol fungi Trichoderma spp.[J].Soil Biology and Biochemistry,1997,29(2):123-129.
[24]Sarrocco S,Mikkelsen L,Vergara M,et al.Histopathological studies of sclerotia of phytopathogenic fungi parasitized by a GFP transformed Trichoderma virens antagonistic strain[J].Mycological Research,2006,110(2):179-187.
[25]Zeng W,Wang D,Kirk W,et al.Use of Coniothyrium minitans and other microorganisms for reducing Sclerotinia sclerotiorum[J].Biological Control,2012,60(2):225-232.
[26]Sennoi R,Singkham N,Jogloy S,et al.Biological control of southern stem rot caused by Sclerotium rolfsii using Trichoderma harzianum and arbuscular mycorrhizal fungi on Jerusalem artichoke(Helianthus tuberosus L.)[J].Crop Protection,2013,54(1):148-153.
[27]高必达,钟杰.稻曲病菌侵染过程研究进展[J].湖南农业科学,2011,(2):93-97.
[28]刘永芳,陈增建.稻曲病初次侵染源及侵染途径的研究[J].天津农业科学,1995,1(1):9-11.
[29]杜爱玲.鸡腿菇炭角菌病害发生机理及其防治研究[D].杨凌:西北农林科技大学,2006.
[30]Lumsden R D,Ridout C J,Vendemia M E,et al.Characterization of major secondary metabolites produced in soilless mix by a formulated strain of the biocontrol fungus Gliocladium virens[J].Canadian Journal of Plant Pathology,1992,38(9):1274-1280.
[31]何迎春,高必达.立枯丝核菌的生物防治[J].中国生物防治,2000,16(1):31-34.
[32]李晶晶,张拥华,陈巧云,等.粉红粘帚霉67-1菌株寄生核盘菌菌核的酶学动态研究[J].菌物研究,2007,5(2):107-109.