橡胶树毛色二孢叶斑病病原菌的鉴定及其生物学特性研究
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摘要
近期在海南省三亚市的橡胶树种植区发现一种新的橡胶树叶斑病,利用组织分离法和单孢分离法获得纯化菌株HNSY003。通过形态学特征观察、rDNA-ITS和EF1-α序列分析及柯赫氏法则验证,确定该病原菌为假可可毛色二孢(Lasiodiplodia pseudotheobromae)。这是假可可毛二孢所致橡胶树叶斑病的首次发现。生物学特性测定结果表明,病菌最适生长条件为:温度32℃,pH 5,碳源为D-果糖,氮源为酵母浸膏。不同光照条件对菌丝生长影响不显著。
A new leaf spot disease of rubber tree was found in Sanya, Hainan. The pathogen strain HNSY003 was isolated and purified by methods of tissue isolation and single spore isolation. The pathogen was identified as Lasiodiplodia pseudotheobromae by morphological observation, sequence analyses of rDNA-ITS and EF1-α, and its pathogenicity was confirmed by Koch's postulate. The fungal species is reported the first time as the pathogen of robber tree leaf spot. The results of biological characteristics test showed that the optimum conditions for the pathogen growth were as follows: temperature 32 ℃, pH 5, D-fructose as the carbon source, yeast extract as the nitrogen source. Different light conditions had little effect on the growth rate of the pathogen hypha.
A new leaf spot disease of rubber tree was found in Sanya, Hainan. The pathogen strain HNSY003 was isolated and purified by methods of tissue isolation and single spore isolation. The pathogen was identified as Lasiodiplodia pseudotheobromae by morphological observation, sequence analyses of rDNA-ITS and EF1-α, and its pathogenicity was confirmed by Koch's postulate. The fungal species is reported the first time as the pathogen of robber tree leaf spot. The results of biological characteristics test showed that the optimum conditions for the pathogen growth were as follows: temperature 32 ℃, pH 5, D-fructose as the carbon source, yeast extract as the nitrogen source. Different light conditions had little effect on the growth rate of the pathogen hypha.
引文
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[2]祁栋灵,王秀全,张志扬,等.中国天然橡胶产业现状及其发展建议[J].热带农业科学,2013,33(2):79-87.
[3]Jayasinghe C K,Jayasinghe C K.Check list of rubber pathogens in Sri Lanka[M].Sri Lanka:National Science Foundation Colombo,2001.
[4]Organisation de Cooperation et de Developpement Economiques.Donnees OCDE sur lenvironnement:Compendium2004 Edition complete ISBN 9264007989[J].Sourceocde Environnement et Développement Durable,2006,2005:i-313(313).
[5]华南热带作物学院.热带作物病虫害防治学[M].第2版.北京:农业出版社,1993:66-70.
[6]刘秀娟,杨业铜,冷怀琼.我国植胶垦区橡胶树炭疽病菌的种型鉴定[J].热带作物学报,1987,8(1):93-101.
[7]Hoolliday P,黄雅志.橡胶树根腐病原菌及其防除措施[J].云南热作科技,1983(4):57-62.
[8]希尔顿,周郁文,李芳.橡胶树绯腐病[J].热带作物译丛,1964(3):19-23.
[9]黎辉,朱智强.海南西培农场橡胶树条溃疡病发生规律及防治经验总结[J].热带农业工程,2011,35(2):15-16.
[10]方忠达.植病研究方法[M].北京:中国农业出版社,1998:122-127.
[11]吴如慧,李增平,孙先伊晴,等.橡胶树臭根病菌的鉴定及其生物学特性研究[J].热带作物学报,2018,39(5):940-947.
[12]White T J,Bruns T,Lee S,et al.Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics[M]//Innis M A,Gelfand D H,Sninsky J J,et al(eds).PCR protocols:A guide to methods and applications.New York:Academic Press,1990:315-322.
[13]Alves A,Crous P W,Correia A,et al.Morphological and molecular data reveal cryptic speciation in Lasiodiplodia theobromae[J].Fungal Diversity,2008,28:1-13.
[14]Tamura K,Stecher G,Peterson D,et al.MEGA6:molecular evolutionary genetics analysis version 6.0[J].Molecular Biology and Evolution,2013,30(12):2725.
[15]Alves A,Crous P W,Correia A,et al.Morphological and molecular data reveal cryptic speciation in Lasiodiplodia theobromae[J].Fungal Diversity,2008,28:1.
[16]Punithalingam E.CMI descriptions of pathogenic fungi and bacteria No.519[M].Wallingford,UK:CAB International,1976:59.
[17]张中义,冷怀琼,张志铭,等.植物病原真菌学[M].成都:四川科技出版社,1991:449.
[18]王佳宁,赵洪海,尉莹莹,等.蓝莓毛色二胞枝枯病的病原菌[J].菌物学报,2016,35(6):657.
[19]Alves A,Crous P W,Correia A,et al.Morphological and molecular data reveal cryptic speciation in Lasiodiplodia theobromae[J].Fungal Diversity,2008,28:1-13.
[20]Trakunyingcharoen T,Cheewangkoon R,To-Anun C.Phylogenetic study of the Botryosphaeriaceae species associated with avocado and para rubber in Thailand[J].Chiang Mai Journal of Science,2015,42:104.