| 猕猴桃溃疡病菌biovar 3群体MLVA分型技术的建立与应用 |
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| 引用本文： | 赵志博,杜淑凤,李月,杨文,樊荣,王勇,龙友华.猕猴桃溃疡病菌biovar 3群体MLVA分型技术的建立与应用[J].植物病理学报,2019,49(4):445-455. |
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| 作者姓名： | 赵志博 杜淑凤 李月 杨文 樊荣 王勇 龙友华 |
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| 作者单位： | 贵州大学农学院,贵阳550025 |
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| 基金项目： | 贵州省科技计划项目(黔科合平台人才[2017]5788号);贵州大学引进人才项目(贵大人基合字[2017]55号);贵州省科技计划项目(黔科合支撑[2017]2566-1);贵阳市科技计划项目(筑科合同[2017]26-1);修文县猕猴桃产业发展项目(修猕合同[2017]02) |
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| 摘 要： | 丁香假单胞菌猕猴桃致病变种生物型3(Pseudomonas syringae pv. actinidiae biovar 3,Psa3)是猕猴桃溃疡病菌的世界流行群体,但仅在中国存在复杂的遗传多样性。开发适于Psa3群体分型的MLVA(multilocus variable-number tandem-repeat analysis)技术是探索中国Psa3起源与流行学特性的基础。本研究对7个Psa3菌株进行了全基因组测序,结合已公布的86个全基因组数据,进行比较分析发现,中国Psa3至少存在7个亚群;在各亚群间存在多态性的24个串联重复序列中,其中10个可以通过琼脂糖凝胶电泳区分开且变异指数合适,据此建立了适于Psa3的MLVA技术。采用该技术对分别来自贵州和陕西的62和9个Psa3菌株进行群体分型,分型结果与全基因组分析高度一致,证明该MLVA体系分型准确。MLVA分型结果表明:贵州主产区修文县Psa3有3个MLVA群体,其中亚群4的组内分化明显,代表最早发生的群体;而亚群 1和3的结构单一,且多在新果园发现,是新传入群体。总之,本研究建立了一套可用于Psa3群体分型的MLVA技术,将有助于解析中国各猕猴桃产区Psa3群体结构,以及探索中国Psa3的起源、传播和流行学特征。
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| 关 键 词： | 猕猴桃细菌性溃疡病 群体结构 基因组分型 MLVA 流行 |
| 收稿时间： | 2018-10-29 |
Establishment and application of MLVA typing for Pseudomonas syringae pv. actinidiae biovar 3 populations |
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| ZHAO Zhi-bo,DU Shu-feng,LI Yue,YANG Wen,FAN Rong,WANG Yong,LONG You-hua.Establishment and application of MLVA typing for Pseudomonas syringae pv. actinidiae biovar 3 populations[J].Acta Phytopathologica Sinica,2019,49(4):445-455. |
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| Authors: | ZHAO Zhi-bo DU Shu-feng LI Yue YANG Wen FAN Rong WANG Yong LONG You-hua |
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| Institution: | College of Agriculture, Guizhou University, Guiyang 550025, China |
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| Abstract: | Pseudomonas syringae pv. actinidiae biovar 3 (Psa3), the causative agent of a severe global pandemic bacterial canker disease of kiwifruit, showed vast diversity only in China. To gain insights into the population differences within Chinese Psa3 and explore the origin and transmission routes of the pathogen, an MLVA (multilocus VNTR analysis) scheme was established based on ten VNTR (variable-number tandem-repeat) loci in this study. Across the seven Chinese Psa3 subpopulations inferred from the whole-genome tree based on 7 newly sequenced Psa3 isolates in this study and 86 Psa3 genomic data available in NCBI database, 24 VNTR loci showed polymorphic alleles and were subsequently used in MLVA study. According to the discriminatory power and reproducibility by agarose gel electrophoresis, 10 of them were involved in the MLVA scheme for Psa3. Furthermore, the MLVA clustering for 62 and 9 Psa3 isolates from Guizhou and Shaanxi provinces of China, respectively, was largely congruent with aforementioned genomic clustering result, indicating the reliability of the MLVA scheme. In Xiuwen county, the largest kiwifruit cultivation area in Guizhou province, there were three MLVA clades, destined as clade 1, 3 and 4, respectively. The clade 4 showed more within-group diversity, representing the older population spreading in Xiuwen county, whereas the others showed very limited diversity and mostly infected the trees in new orchards, representing recent invasion events in this region. In summary, a reliable MLVA scheme has been established,which is successfully used for genotyping of Psa3 isolates in Guizhou province, and would be used for the thorough analysis of Chinese Psa3 isolates to explore the origin and transmission routes of the pathogen. |
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| Keywords: | bacterial canker of kiwifruit population structure genomic typing MLVA pandemic |
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