| 汤亚飞1#, 林 祺1, 2#, 佘小漫1, 李正刚1, 于 琳1, 蓝国兵1, 何自福1*.广东花生丛枝病植原体的分子鉴定[J].植物保护,2022,48(5):83-90. |
| 广东花生丛枝病植原体的分子鉴定 |
| Molecular identification of peanut witches’-broom phytoplasma in Guangdong province |
| 投稿时间：2021-07-26 修订日期：2021-10-26 |
| DOI：10.16688/j.zwbh.2021409 |
| 中文关键词: 花生丛枝病 植原体 分子鉴定 16S rRNA SecY基因 序列分析 |
| 英文关键词:peanut witches’-broom disease phytoplasma molecular identification 16S rRNA SecY gene sequence analysis |
| 基金项目:广东省重点研发计划(2019B020217003-05); 广东省现代农业产业共性关键技术研发创新团队建设项目(2020KJ134); 广东省农业科学院科技创新战略专项资金(高水平农科院建设)(R2019PY-JX005); 广东省农业科学院“十四五”学科团队建设项目(202105TD) |
| 作者 | 单位 | E-mail | | 汤亚飞1#, 林 祺1, 2#, 佘小漫1, 李正刚1, 于 琳1, 蓝国兵1, 何自福1* | 1. 广东省农业科学院植物保护研究所, 广东省植物保护新技术重点实验室, 广州 510640
2. 华南农业大学植物保护学院, 广州 510642 | hezf@gdppri.com |
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| 中文摘要: |
| 2020年在广东省湛江市遂溪县田间发现表现明显丛枝?小叶, 类似植原体感染症状的花生病株?本研究利用分子生物学技术对其病原进行鉴定?以花生病叶的总DNA为模板, 利用植原体16S rRNA和SecY基因通用引物进行PCR扩增, 获得广东花生丛枝病植原体(PnWB-GDSX-2020)16S rRNA基因片段(1 430 bp, GenBank登录号为MZ427281)和SecY基因片段(1 709 bp, GenBank登录号为MZ437794)?序列一致性和系统进化分析显示, PnWB-GDSX-2020的16S rRNA序列与16SrⅡ-A?16SrⅡ-D和16SrⅡ-V亚组植原体一致性最高, 亲缘关系最近; 进一步利用iPhyClassifier对16S rRNA序列进行在线虚拟RFLP分析, 结果显示, PnWB-GDSX-2020的虚拟RFLP 图谱与16SrⅡ-V亚组的参照株系‘Praxelis clematidea’ phyllody phytoplasma (GenBank登录号:KY568717) 酶切图谱一致, 相似系数为1.00?因此, PnWB-GDSX-2020属于16SrⅡ-V亚组成员?所获得的PnWB-GDSX-2020 Sec Y基因序列与花生丛枝植原体的一致性最高, 亲缘关系最近?本文确定了广东花生丛枝病相关植原体的分类地位, 为当地病害诊断?检测以及防控提供科学依据? |
| 英文摘要: |
| In 2020, peanut plants exhibiting symptoms of witches’-broom and fewer leaves like phytoplasma infection were found in fields in Suixi county, Zhanjiang city, Guangdong province. In this study, the pathogen was identified using molecular detection technology. The 16S rRNA fragment (1 430 bp, GenBank acc. no. MZ427281) and SecY gene fragment (1 709 bp, GenBank acc. no. MZ437794) of phytoplasma associated with peanut witches’-broom in Guangdong province (PnWB-GDSX-2020) were amplified from total DNA of diseased peanut leaves using phytoplasma universal primers of 16S rRNA and SecY gene. Sequence similarity and phylogenetic analysis based on 16S rRNA sequence showed that PnWB-GDSX-2020 shared the highest similarity and closest relationship with phytoplasmas from 16SrⅡ-A, 16SrⅡ-D and 16SrⅡ-V subgroup. The iPhyClassifier online virtual RFLP analysis on the 16S rRNA sequence showed that the virtual RFLP map of PnWB-GDSX-2020 was consistent with that of ‘Praxelis clematidea’ phyllody phytoplasma (GenBank acc. no. KY568717), the reference strain from 16SrⅡ-V subgroup, with a similarity coefficient of 1.00. Based on the 16S rRNA sequence analysis results, PnWB-GDSX-2020 belonged to 16SrⅡ-V subgroup of phytoplasma. The SecY gene sequence of PnWB-GDSX-2020 also had the highest similarity and the closest relationship with peanut witches’-broom phytoplasma. This is the first study for determining the classification status of phytoplasma associated with peanut witches’-broom disease in Guangdong province. The results provide the scientific basis for diagnosis, detection, prevention and control of this disease. |
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