| 中国4省猕猴桃细菌性溃疡病菌的生物型检测和遗传多样性分析 |
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| 引用本文： | 代玉立,兰成忠,甘 林,刘晓菲,龚国淑,杨秀娟.中国4省猕猴桃细菌性溃疡病菌的生物型检测和遗传多样性分析[J].植物保护,2022,48(6):58-68. |
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| 作者姓名： | 代玉立 兰成忠 甘 林 刘晓菲 龚国淑 杨秀娟 |
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| 作者单位： | (1. 福建省农业科学院植物保护研究所, 福建省作物有害生物监测与治理重点实验室, 福建省作物有害生物绿色防控工程研究中心, 福州 350013; 2. 四川农业大学农学院, 成都 611130) |
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| 基金项目： | 国家重点研发计划（2021YFC2600402）；福建省农业科学院“5511”协同创新工程（XTCXGC2021011，XTCXGC2021017）；福建省农业科学院农作物重大有害生物灾变机制与绿色防控科技创新团队（CXTD2021002-1）；福建省农业科学院科技创新平台专项（CXPT202105） |
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| 摘 要： | 由丁香假单胞菌猕猴桃致病变种Pseudomonas syringae pv. actinidiae (Psa)侵染引起的猕猴桃细菌性溃疡病(kiwifruit bacterial canker)是全球猕猴桃生产上最具毁灭性的细菌病害。为探明福建、安徽、四川和陕西4省Psa菌株的生物型和遗传多样性,用5对PCR特异性引物PsaJ-F/-R、PsaK-F/-R、Tac-F/-R、Con002-F/-R和avrRps4-F1/-R2检测Psa菌株的生物型;用4对PCR引物27F/1492R、PsaF1/PsaR2、gapA-Fps/Rps和rpoD+364s/-1222ps分别扩增16S rRNA、ITS、gapA和rpoD基因,进行多基因联合分析Psa菌株的遗传多样性。结果表明,特异性引物Tac-F/-R从47株Psa菌株中均能扩增出一条545 bp的特异条带,其他4对引物未扩增出任何条带,说明供试Psa菌株的生物型均为biovar 3。多基因联合分析表明,4省Psa存在丰富的遗传多样性,4个群体共检测出27个单倍型,单倍型多样性为0.955。安徽、福建、四川和陕西群体的单倍型数差异较大,分别为1、8、12个和12个。4个群体的多态性位点数、核苷酸多样性和平均核苷酸差异数差异极显著(P<0.01),其中福建群体的多态性最丰富,而安徽群体的多态性最低。AMOVA分析表明,3.6%的遗传变异来源于种群间,而96.4%的遗传变异来源于种群内,说明种群内变异是遗传变异的主要来源。遗传分化分析表明,安徽省Psa群体与其他3个群体间的遗传分化极高(Fst>0.175),福建、四川和陕西群体间的遗传分化水平较低(Fst<0.017)。研究结果有利于了解福建省Psa的来源,为阻断Psa的传播和猕猴桃细菌性溃疡病的长期可持续控制提供了理论参考。
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| 关 键 词： | 猕猴桃细菌性溃疡病菌 生物型 遗传多样性 多基因联合分析 猕猴桃 |
| 收稿时间： | 2022/6/17 0:00:00 |
| 修稿时间： | 2022/7/22 0:00:00 |
Detection of Pseudomonas syringae pv. actinidiae biovars from four provinces in China and genetic diversity analysis |
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| DAI Yuli,LAN Chengzhong,GAN Lin,LIU Xiaofei,GONG Guoshu,YANG Xiujuan.Detection of Pseudomonas syringae pv. actinidiae biovars from four provinces in China and genetic diversity analysis[J].Plant Protection,2022,48(6):58-68. |
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| Authors: | DAI Yuli LAN Chengzhong GAN Lin LIU Xiaofei GONG Guoshu YANG Xiujuan |
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| Institution: | (1. Fujian Key Laboratory for Monitoring and Integrated Management of Crop Pests, Fujian Engineering Research Center for Green Pest Management, Institute of Plant Protection, Fujian Academy of Agricultural Sciences, Fuzhou 350013, China; 2. College of Agronomy, Sichuan Agricultural University, Chengdu 611130, China) |
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| Abstract: | Kiwifruit bacterial canker, caused by Pseudomonas syringae pv. actinidiae (Psa), is the globally most devastating bacterial disease affecting kiwifruit production. To determine the biovars and genetic diversity of Psa strains from four provinces (Anhui, Fujian, Sichuan and Shaanxi), the biovars of Psa strains were detected by PCR with five biovar-specific primer pairs (PsaJ-F/-R, PsaK-F/-R, Tac-F/-R, Con002-F/-R and avrRps4-F1/-R2). The genes encoding 16S rRNA, ITS, gapA and rpoD were amplified using four primer pairs (27F/1492R, PsaF1/PsaR2, gapA-Fps/-Rps and rpoD+364s/-1222ps), respectively, and a combined multiple gene analysis of the genetic diversity of Psa strains was performed. The results showed that an expected 545 bp fragment was amplified specifically from 47 Psa strains using the specific primer Tac-F/-R, but no amplicons were obtained using the other four biovar-specific primer pairs, indicating that the biovar of the tested Psa strains was all biovar 3. Combined analysis of multiple genes indicated that abundant genetic diversity and a total of 27 haplotypes existed in the four populations, with a haplotype diversity of 0.955. There was a significant difference in the number of haplotypes among populations from Anhui, Fujian, Sichuan and Shaanxi, with a haplotype number of 1, 8, 12 and 12, respectively. A significant difference (P<0.01) in the number of polymorphic sites, nucleotide diversity and average number of nucleotide differences was observed among the four populations. The most abundant polymorphism was observed in Fujian population, whereas low polymorphism was detected in Anhui population. Analysis of molecular variance (AMOVA) demonstrated that only 3.6% of genetic variation occurred among populations, while 96.4% of genetic variation derived from within populations, suggesting that the major source of genetic variation was derived from within populations. Genetic differentiation analysis showed that a highly genetic differentiation (Fst>0.175) occurred between Psa populations from Anhui and those of the other three populations. However, a low genetic differentiation was observed among populations from Fujian, Sichuan and Shaanxi (Fst<0.017). These results are beneficial for understanding the source of Psa in Fujian province, and providing theoretical reference for blocking the spread of Psa and long-term sustainable control of kiwifruit bacterial canker. |
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| Keywords: | Pseudomonas syringae pv actinidiae biovar genetic diversity combined analysis of multiple genes kiwifruit |
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