一株能拮抗3种土传病害病原真菌的长枝木霉
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摘要
为了丰富生防木霉菌(Trichodermaspp.)种资源,在园林植物八宝景天(Sedum spectabile)根际土壤中分离得到一株优势木霉。根据该菌株菌落的形态特征、形态学显微观察、菌株rDNA-ITS序列分析以及邻接法同源性比对等结果判定木霉菌株种类。采用平板对峙法将该菌株与3种土传病害病原菌[(Sclerotinia sclerotiorum)、立枯丝核菌(Rhizoctonia solani)和细链格孢菌(Alternaria alternata)]进行对峙培养。并用该木霉诱导山新杨(Populus davidiana×P.albavar.Pyramidalis)组培苗,观察离体叶片抵抗细链格孢菌侵染的能力。结果表明,此木霉为长枝木霉(Trichoderma longibrachiatum),命名为Tl-70。该菌株对3种土传病害病原真菌均有较明显的拮抗作用。对核盘菌的抑菌率最高为77.71%,显著高于对细链格孢菌和立枯丝核菌的抑制率(P<0.05);对立枯丝核菌的抑菌率为58.56%,显著高于对细链格孢菌的抑制率(P<0.05);对细链格孢菌的抑菌率最低,为53.32%。并且,该木霉能提高山新杨组培苗叶片拮抗细链格孢菌侵染的能力。说明此菌株是具有潜力的生防菌株。
To enrich the microbial resources of Trichoderma spp.for biocontrol,a dominant Trichoderma strain was isolated from the rhizosphere soil of a landscape plant,Sedum spectabile Boreau.The isolated Trichodermastrain was identified using colonial-and micro-morphological observations,ribosomal DNA(rDNA)-internal transcribed spacer(ITS)sequence alignment,and homology analysis using a Neighbour-Joining method.An agar disk diffusion method was used to investigate the antagonism between the isolated Trichoderma strain and three other soil-borne pathogenic fungi,Sclerotinia sclerotiorum,Rhizoctonia solani,and Alternaria alternata.The tissue-cultured plantlets of Populus davidiana×P.alba var.Pyramidalis were further inoculated with the isolated Trichoderma strainand the resistance of the inoculated seedling leaves against A.alternata was evaluated in vitro.The results demonstrated that the isolated Trichodermastrain was Trichoderma longibrachiatum and it exhibited evident and differential antagonistic effects against each of the three tested soilborne pathogenic fungi.We designated the isolated Trichoderma strain as Tl-70.The antifungal activity of Tl-70 against S.sclerotiorum was a 77.71%inhibition,which was the highest and significantly higher than that against R.solani or A.alternata(P<0.05).The antifungal activity against R.solani was a 58.56%inhibition,and significantly higher than that against A.alternata(P<0.05,53.32%),which was the lowest of the three tested strains.Moreover,inoculation with Tl-70 enhanced the disease resistance of the leaves of tissuecultured Populus davidiana×P.albavar.Pyramidalis seedlings against the pathogen A.alternata.Our results suggest that Tl-70 is a potential biocontrol strain.
To enrich the microbial resources of Trichoderma spp.for biocontrol,a dominant Trichoderma strain was isolated from the rhizosphere soil of a landscape plant,Sedum spectabile Boreau.The isolated Trichodermastrain was identified using colonial-and micro-morphological observations,ribosomal DNA(rDNA)-internal transcribed spacer(ITS)sequence alignment,and homology analysis using a Neighbour-Joining method.An agar disk diffusion method was used to investigate the antagonism between the isolated Trichoderma strain and three other soil-borne pathogenic fungi,Sclerotinia sclerotiorum,Rhizoctonia solani,and Alternaria alternata.The tissue-cultured plantlets of Populus davidiana×P.alba var.Pyramidalis were further inoculated with the isolated Trichoderma strainand the resistance of the inoculated seedling leaves against A.alternata was evaluated in vitro.The results demonstrated that the isolated Trichodermastrain was Trichoderma longibrachiatum and it exhibited evident and differential antagonistic effects against each of the three tested soilborne pathogenic fungi.We designated the isolated Trichoderma strain as Tl-70.The antifungal activity of Tl-70 against S.sclerotiorum was a 77.71%inhibition,which was the highest and significantly higher than that against R.solani or A.alternata(P<0.05).The antifungal activity against R.solani was a 58.56%inhibition,and significantly higher than that against A.alternata(P<0.05,53.32%),which was the lowest of the three tested strains.Moreover,inoculation with Tl-70 enhanced the disease resistance of the leaves of tissuecultured Populus davidiana×P.albavar.Pyramidalis seedlings against the pathogen A.alternata.Our results suggest that Tl-70 is a potential biocontrol strain.
引文
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[2]Olson H A,Benson D M.Induced systemic resistance and the role of binucleate Rhizoctoniaand Trichoderma hamatum 382in biocontrol of Botrytisblight in geranium.Biological Control,2007,42(2):233-241.
[3]Druzhinina I S,Seidl-Seiboth V,Herrera-Estrella A,Horwitz B A,Kenerley C M,Monte E,Mukherjee P K,Zeilinger S,Grigoriev I V,Kubicek C P.Trichoderma:The genomics of opportunistic success.Nature Reviews Microbiology,2011,9(10):749-759.
[4]李江遐,蒋立科.木霉菌在坪用黑麦草上的应用效果初报.草业科学,2006,23(1):103-105.Li J X,Jiang L K.Primary report of application effects of Trichoderma on perennial ryegrass.Pratacultural Science,2006,23(1):103-105.(in Chinese)
[5]Hanada R E,Pomella A W V,Soberanis W,Loguercio L L.Biocontrol potential of Trichoderma martiale against the black-pod disease(Phytophthora palmivora)of cacao.Biological Control,2009,50(2):143-149.
[6]Wijesinghe C J,Wijeratnam R S W,Samarasekara J K R R,Wijesundera R L C.Biological control of Thielaviopsis paradoxa on pineapple by an isolate of Trichoderma asperellum.Biological Control,2010,53(3):285-290.
[7]Chen L,Yang X,Raza W,Luo J,Zhang F,Shen Q.Solid-state fermentation of agro-industrial wastes to produce bioorganic fertilizer for the biocontrol of Fusarium wilt of cucumber in continuously cropped soil.Bioresource Technology,2011,102(4):3900-3910.
[8]杨合同.木霉分类与鉴定.北京:中国大地出版社,2009:160-306.Yang H T.Trichoderma Classification and Identification.Beijing:China Land Press,2009:160-306.(in Chinese)
[9]Sant D,Casanova E,Segarra G,Aviles M,Reis M,Isabel-Trillas M.Effect of Trichoderma asperellumstrain T34on Fusarium wilt and water usage in carnation grown on compost-based growth medium.Biological Control,2010,53(3):291-296.
[10]闫峰,徐凤花,顾金刚,姜瑞波.木霉属真菌的生物降解及生物转化作用研究进展.微生物学杂志,2009,29(3):77-80.Yan F,Xu F H,Gu J G,Jiang R B.Research progress of biodegradation and biotransformation with Trichoderma.Journal of microbiology,2009,29(3):77-80.(in Chinese)
[11]金潇,颜冬云,秦文秀.有机磷农药的微生物降解技术.湖南农业科学,2011(5):93-97.Jin X,Yan D Y,Qin W X.Technology of microbial degradation for organophosphorus pesticides.Hunan Agricultural Sciences,2011(5):93-97.(in Chinese)
[12]杨振兴,田从魁,党晨原,常方,倪晋仁.真菌对重金属Pb(Ⅱ),Cd(Ⅱ),As(Ⅲ)和Cr(Ⅵ)耐受性的比较研究.北京大学学报:自然科学版,2015,51(4):667-675.Yang Z X,Tian C K,Dang C Y,Chang F,Ni J R.Comparative study on Pb(Ⅱ),Cd(Ⅱ),As(Ⅲ),Cr(Ⅵ)resistance characteristics of fungus.Acta Scientiarum Naturalium Universitatis Pekinensis,2015,51(4):667-675.(in Chinese)
[13]姚彦坡,王雅玲,吕国忠.木霉对草坪上2种重要土传病害生防效果的研究.草业科学,2007,24(8):96-99.Yao Y P,Wang Y L,Lyu G Z.Research on preventing the turfgrass plant diseases with Trichoderma harzianum preparation.Pratacultural Science,2007,24(8):96-99.(in Chinese)
[14]赵晓燕,吴晓青,李哲,赵忠娟,陈泉,陈凯,李纪顺,杨合同.13个省、自治区滩涂湿地木霉菌分布初探.山东科学,2015,28(3):34-37.Zhao X Y,Wu X Q,Li Z,Zhao Z J,Chen Q,Chen K,Li J S,Yang H T.Trichodermadistribution of coastal beach and wetland of thirteen provinces and autonomous regions.Shandong Science,2015,28(3):34-37.(in Chinese)
[15]张寅媛,刘英,白龙.干旱胁迫对4种景天科植物生理生化指标的影响.草业科学,2014,31(4):724-731.Zhang Y Y,Liu Y,Bai L.Effect of drought stress on physiological indexes of 4Crassulaceae species.Pratacultural Science,2014,31(4):724-731.(in Chinese)
[16]汤聪,刘念,郭微,蔡鑫,苏建华,刘萍.广州地区8种草坪式屋顶绿化植物的抗旱性.草业科学,2014,31(10):1867-1876.Tang C,Liu N,Guo W,Cai X,Su J H,Liu P.The drought stress tolerance of several lawn-style roof green species in Guangzhou area.Pratacultural Science,2014,31(10):1867-1876.(in Chinese)
[17]赵成爱,余梅燕,吴现芳,杨琳琳.八宝景天乙醇提取物对菜青虫的生物活性.农药,2012,51(1):62-63,70.Zhao C A,Yu M Y,Wu X F,Yang L L.Bioactivity of Sedum spectabile Boreau extracts against Pieris rapae.Agrochemicals,2012,51(1):62-63,70.(in Chinese)
[18]Taylor J W,Jacobson D J,Kroken S,Kasuga T,Geiser D M,Hibbett D S,Fisher M C.Phylogenetic species recognition and species concepts in fungi.Fungal Genetics and Biology,2000,31(1):21-32.
[19]李兴龙,李彦忠.土传病害生物防治研究进展.草业学报,2015,24(3):204-212.Li X L,Li Y Z.Research advances on biological control of soil-borne disease.Acta Prataculturae Sinica,2015,24(3):204-212.(in Chinese)
[20]蒋家珍,赵美琦,薄怀霞,肖杰,李学锋.生物农药在草坪病害防治中的应用研究.草业科学,2004,21(11):93-95.Jiang J Z,Zhao M Q,Bao H X,Xiao J,Li X F.Study on application of biological fungicide to several turf diseases.Pratacultural Science,2004,21(11):93-95.(in Chinese)
[21]张茹,李金华,柴兆祥,王蒂.甘肃河西马铃薯根际生防木霉菌对接骨木镰刀菌的拮抗筛选及鉴定.草业学报,2009,18(2):138-145.Zhang R,Li J H,Chai Z X,Wang D.Screening of biocontrol Trichodermafrom potato rhizosphere of Western Gansu against Fusarium sambucinum and their identification.Acta Prataculturae Sinica,2009,18(2):138-145.(in Chinese)
[22]刘志华,范海娟,张荣沭,季世达.一种植物根际木霉菌分离方法:中国专利201510354447.3.2015-09-30Liu Z H,Fan H J,Zhang R S,Ji S D.A strategy for seperating plant rhizospheric Trichoderma.China,Patent application receipted by State Intellectual Property Office of the People's Republic of China,2015,Accepted:201510354447.3.(in Chinese)
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