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汤亚飞, 李正刚, 佘小漫, 于 琳, 蓝国兵, 何自福*.广东番茄巨芽病植原体的分子鉴定[J].植物保护,2024,50(1):203-210.
广东番茄巨芽病植原体的分子鉴定
Molecular identification of phytoplasma associated with tomato big bud disease in Guangdong province
投稿时间:2022-08-12  修订日期:2022-09-29
DOI:10.16688/j.zwbh.2022487
中文关键词:  番茄巨芽病  植原体  16S rRNA  分子鉴定
英文关键词:tomato big bud disease  phytoplasma  16S rRNA  molecular identification
基金项目:广东省重点领域研发计划(2020B0202090002); 国家自然科学基金(32072392); 广东省现代农业产业技术体系创新团队建设项目(2021KJ134); 科技创新战略专项资金(R2019PY-JX005); 广东省农业科学院“十四五”学科团队建设项目(202105TD)
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汤亚飞, 李正刚, 佘小漫, 于 琳, 蓝国兵, 何自福* 广东省农业科学院植物保护研究所, 农业农村部华南果蔬绿色防控重点实验室, 广东省植物保护新技术重点实验室, 广州 510640 hezf@gdppri.com 
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中文摘要:
      2022年, 对在广东省湛江市廉江市田间发现的疑似番茄巨芽病病株, 利用分子生物学方法对其相关植原体进行了鉴定。以番茄病株叶片总DNA为模板, 利用植原体16S rRNA基因通用引物R16mF2/R16mR1进行PCR扩增, 获得了广东番茄巨芽病植原体(TBB-GD-2022)16S rRNA基因片段(1 430 bp, GenBank登录号为ON102780)。16S rRNA基因序列相似性分析显示, TBB-GD-2022与16SrⅡ组植原体菌株的相似性较高, 为96.82%~100%, 其中与隶属于16SrⅡ-V亚组的6个植原体株系相似性为100%。系统进化分析显示, TBB-GD-2022与16SrⅡ组各植原体株系聚类在一个大分支, 并与16SrⅡ-V亚组成员聚类在一个小分支, 亲缘关系较近。16S rRNA 基因相似系数分析表明, TBB-GD-2022与16SrⅡ-V亚组的参照株系‘Praxelis clematidea’ phyllody phytoplasma (GenBank登录号:KY568717) 的相似系数为1.00。上述研究结果表明, 广东番茄巨芽病植原体隶属16SrⅡ-V亚组成员。本文首次报道在广东发现番茄巨芽病, 通过其16S rRNA序列分析进一步确定了其相关植原体的分类地位, 为该病害的防控提供了科学依据。
英文摘要:
      In 2022, the suspected tomato big bud diseased plants were found in Lianjiang city, Zhanjiang city, Guangdong province. The related phytoplasma was detected and identified using molecular technology. The 16S rRNA gene fragment (1 430 bp, GenBank accession no. ON102780) of phytoplasma associated with tomato big bud disease in Guangdong province (TBB-GD-2022) was amplified from total DNA of diseased tomato plants using universal primers R16mF2/R16mR1. Sequence similarity based on 16S rRNA gene sequence showed that TBB-GD-2022 had higher similarity (96.82%-100%) with phytoplasmas from 16SrⅡ group, while had highest similarity (100%) with six phytoplasmas belonged to 16SrⅡ-V subgroup. Phylogenetic analysis showed that TBB-GD-2022 formed a large evolutionary branch with 16SrⅡ group phytoplasmas, and a small evolutionary branch with 16SrⅡ-V subgroup phytoplasmas. iPhyClassifier analysis for 16S rRNA gene showed that the similarity coefficient between TBB-GD-2022 and ‘Praxelis clematidea’ phyllody phytoplasma (GenBank accession no. KY568717) of the reference strain from 16SrⅡ-V subgroup was 1.00. These results showed that phytoplasma associated with tomato big bud disease in Guangdong province belonged to the 16SrⅡ-V subgroup. In this paper, tomato big bud disease was discovered firstly in Guangdong, and the taxonomic status of its related phytoplasma was further determined. The results provide the scientific basis for controlling tomato big bud disease in Guangdong province.
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