卵叶山蚂蝗丛枝植原体的分子鉴定
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摘要
2016年在生长于海南省澄迈县的卵叶山蚂蝗上发现了丛枝、小叶等疑似植原体感染症状。利用植原体16S rRNA基因的通用引物R16mF2/R16mR1,对卵叶山蚂蝗丛枝样品进行了PCR检测,并对检测到的植原体16S rRNA基因片段(1.4 kb)进行克隆测序、序列比对、虚拟限制性片段长度多态性分析和系统进化树构建分析。结果表明,卵叶山蚂蝗丛枝植原体(beggarweed witches'-broom phytoplasma,MK956144)为来檬植原体(Candidatus Phytoplasma aurantifolia,U15442)的相关株系,属于花生丛枝植原体组A亚组(16SrII-A),这是首次发现植原体感染卵叶山蚂蝗的报道。
Phytoplasma suspected symptoms of witches'-broom and little leaves were found on beggarweed(Desmodium ovalifoliumin Wall) in Chengmai County,Hainan Province in 2016. DNA specific fragments of approximately 1.4 kb were amplified from symptomatic samples of beggarweed in PCR assays by using universal phytoplasma specific primers R16 mF2/R16 mR1. Sequence comparison,phylogeny and virtual RFLP analysis of 16 S r RNA gene sequences were used in this study. The beggarweed witches'-broom phytoplasma(MK956144) was identified to be associated with strain related to 'Candidatus Phtoplasma aurantifolia'(U15442) and belong to 16 SrII-A subgroup,in the peanut witches'-broom phytoplasma group.This is the first report of phytoplasmas strain infecting beggarweed.
Phytoplasma suspected symptoms of witches'-broom and little leaves were found on beggarweed(Desmodium ovalifoliumin Wall) in Chengmai County,Hainan Province in 2016. DNA specific fragments of approximately 1.4 kb were amplified from symptomatic samples of beggarweed in PCR assays by using universal phytoplasma specific primers R16 mF2/R16 mR1. Sequence comparison,phylogeny and virtual RFLP analysis of 16 S r RNA gene sequences were used in this study. The beggarweed witches'-broom phytoplasma(MK956144) was identified to be associated with strain related to 'Candidatus Phtoplasma aurantifolia'(U15442) and belong to 16 SrII-A subgroup,in the peanut witches'-broom phytoplasma group.This is the first report of phytoplasmas strain infecting beggarweed.
引文
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[9] Yang Y, Jiang L, Che H Y, et al. Molecular identification of a 16SrII-A group-related phytoplasma associated with cinnamon yellow leaf disease in China. Journal of Phytopathology, 2016, 164(1):52-55.
[10] Yang Y, Jiang L, Che H Y, et al. Phytoplasma in Association With Rubber Tree(Heveabrasiliensis)Stem Fasciation in China. Plant Disease, 2016, 100(12):2520.
[11] Yang Y, Jiang L, Che H Y, et al. Identification of a novel subgroup 16SrII-U phytoplasma associated with papaya little leaf disease. International Journal of Systematic and Evolutionary Microbiology, 2016, 66(9):3485-3491.
[12] Gurr G M, Johnson A C, Ash G J, et al. Coconut Lethal Yellowing Diseases:A Phytoplasma Threat to Palms of Global Economic and Social Significance. Frontiers in Plant Science, 2016, 7:21.
[13] Laimer M, Lemaire O, Herrbach E, et al. Resistance to viruses, phytoplasmas and their vectors in the grapevine in Europe. A review Journal of Plant Pathology, 2009, 91(1):7-23.
[14] Lee I M, Hammond R W, Davis R E, et al. Universal amplification and analysis of pathogen 16S r DNA for classification and identification of mycoplasma like organisms.Phytopathology, 1993, 83(8):834-842.
[15] Zhao Y, Wei W, Lee I M, et al. Construction of an interactive online phytoplasma classification tool, iPhyClassifier,and its application in analysis of the peach X-disease phytoplasma group(16SrⅢ). International Journal of Systematic and Evolutionary Microbiology, 2009(59):2582-2593.
[16] Davis R E, Zhao Y. Criteria for phytoplasma 16Sr group/subgroup delineation and the need of a platform for proper registration of new groups and subgroups. International Journal of Systematic and Evolutionary Microbiology, 2016, 66(5):2121-2123.
[17] Yang Y, Jiang L, Tian Q, et al. Detection and identification of a novel subgroup 16SrII-V phytoplasma associated with Praxelis clematidea phyllody disease. International Journal of Systematic and Evolutionary Microbiology, 2017, 67(12):5290-5295.
[2]杨毅,车海彦,曹学仁,等.植原体基因组学研究进展.植物保护, 2014, 40(6):1-6.
[3]周俊义,刘孟军,侯保林,等.枣疯病研究进展.果树科学,1998, 15(4):354-359.
[4]田国忠,张志善,李志清,等.我国不同地区枣疯病发生动态和主导因子分析.林业科学, 2002, 38(2):83-91.
[5]徐启聪,田国忠,王振亮,等.中国各地不同枣树品种上枣疯病植原体的PCR检测及分子变异分析.微生物学报,2009, 49(11):1510-1519.
[6]任国兰.泡桐丛枝病的研究现状与进展.河南农业大学学报,1996, 30(4):358-364.
[7]田国忠,黄钦才,袁巧平,等.感染MLO泡桐组培苗代谢变化与致病机理的关系.中国科学(B辑), 1994, 24(5):484-490.
[8] Wu Y F, Hao X G, Li Z N, et al. Identification of the phytoplasma associated with wheat blue dwarf disease in China. Plant Disease, 2010, 94(8):977-985.
[9] Yang Y, Jiang L, Che H Y, et al. Molecular identification of a 16SrII-A group-related phytoplasma associated with cinnamon yellow leaf disease in China. Journal of Phytopathology, 2016, 164(1):52-55.
[10] Yang Y, Jiang L, Che H Y, et al. Phytoplasma in Association With Rubber Tree(Heveabrasiliensis)Stem Fasciation in China. Plant Disease, 2016, 100(12):2520.
[11] Yang Y, Jiang L, Che H Y, et al. Identification of a novel subgroup 16SrII-U phytoplasma associated with papaya little leaf disease. International Journal of Systematic and Evolutionary Microbiology, 2016, 66(9):3485-3491.
[12] Gurr G M, Johnson A C, Ash G J, et al. Coconut Lethal Yellowing Diseases:A Phytoplasma Threat to Palms of Global Economic and Social Significance. Frontiers in Plant Science, 2016, 7:21.
[13] Laimer M, Lemaire O, Herrbach E, et al. Resistance to viruses, phytoplasmas and their vectors in the grapevine in Europe. A review Journal of Plant Pathology, 2009, 91(1):7-23.
[14] Lee I M, Hammond R W, Davis R E, et al. Universal amplification and analysis of pathogen 16S r DNA for classification and identification of mycoplasma like organisms.Phytopathology, 1993, 83(8):834-842.
[15] Zhao Y, Wei W, Lee I M, et al. Construction of an interactive online phytoplasma classification tool, iPhyClassifier,and its application in analysis of the peach X-disease phytoplasma group(16SrⅢ). International Journal of Systematic and Evolutionary Microbiology, 2009(59):2582-2593.
[16] Davis R E, Zhao Y. Criteria for phytoplasma 16Sr group/subgroup delineation and the need of a platform for proper registration of new groups and subgroups. International Journal of Systematic and Evolutionary Microbiology, 2016, 66(5):2121-2123.
[17] Yang Y, Jiang L, Tian Q, et al. Detection and identification of a novel subgroup 16SrII-V phytoplasma associated with Praxelis clematidea phyllody disease. International Journal of Systematic and Evolutionary Microbiology, 2017, 67(12):5290-5295.