| 何孟兰1, 蔡银枫1, 吴道慧1, 苏 帆1, 万琼莲2, 王柱华1, 陈相聪1, 吴艳芬1, 乔 开1, 蔡 红1*.基于线粒体 COⅠ基因及形态特征的喜树丛枝植原体主要媒介昆虫种类鉴定[J].植物保护,2023,49(1):262-270. |
| 基于线粒体 COⅠ基因及形态特征的喜树丛枝植原体主要媒介昆虫种类鉴定 |
| Identification of main vector insects of the phytoplasma based on mitochondrial COⅠ gene and morphological characteristics |
| 投稿时间：2021-10-19 修订日期：2022-01-02 |
| DOI：10.16688/j.zwbh.2021576 |
| 中文关键词: CO I基因 植原体媒介昆虫 分子鉴定 形态鉴定 拟帕小绿叶蝉 |
| 英文关键词:COⅠ gene vectors insects of phytoplasma molecular identification morphological identification Empoasca (Matsumurasca) paraparvipenis |
| 基金项目:国家自然科学基金（31960535） |
| 作者 | 单位 | E-mail | | 何孟兰1, 蔡银枫1, 吴道慧1, 苏 帆1, 万琼莲2, 王柱华1, 陈相聪1, 吴艳芬1, 乔 开1, 蔡 红1* | 1. 云南农业大学植物保护学院, 昆明 650201
2. 玉溪师范学院化学生物与环境学院, 玉溪 653100 | caihong0623@126.com |
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| 中文摘要: |
| 植原体病害是喜树上危害最严重的病害之一, 传播喜树植原体病害的主要媒介昆虫是喜树上的一种小绿叶蝉, 为进一步分析媒介昆虫传播植原体机制以及植原体在昆虫体内的侵染循环过程, 为植原体病害的科学防控提供参考, 媒介昆虫的种类及分类地位必须先行明确。通过网捕法采集云南文山喜树上的小绿叶蝉标本, 在体视显微镜下根据各龄期外部形态以及雄性外生殖器特征进行种类鉴定, 结合PCR扩增线粒体 COⅠ 基因序列片段进行测序和比对, 利用Kimura 2-parameter 模型计算出平均遗传距离并用最大似然法构建系统发育树, 进一步明确其分类地位, 再根据前人制作的属及亚属分类检索表明确其分类地位。共检视并解剖小绿叶蝉雄虫标本100余头, 提取20余头DNA进行测序, 得到16条709 bp大小的 COⅠ 基因序列片段, 结合NCBI上相关的叶蝉 COⅠ 序列, 建立基因进化树。综合分析其外部形态、雄性外生殖器特征以及 COⅠ 序列比对, 明确了喜树丛枝植原体主要媒介昆虫是拟帕小绿叶蝉 Empoasca ( Matsumurasca ) paraparvipenis Zhang et Liu -CA, 分类地位为小绿叶蝉属 Empoasca Walsh, 1862, 松村叶蝉亚属 Empoasca (Matsumurasca) Anufriev。 |
| 英文摘要: |
| The phytoplasma disease is one of the most serious diseases of Camptotheca acuminata, and Empoasca sp . is the main vector insect of the C. acuminata phytoplasma diseases. In order to provide a reference for further analysis of the transmission mechanism of phytoplasma by vector insects and the phytoplasma infection cycle process, and provide a reference for scientific prevention and control of phytoplasma diseases, the species of vector insects and the taxonomic status of insects should be clarified. The specimens of Empoasca sp . on C. acuminate were collected by net sweeping in Wenshan, Yunnan, and the stereomicroscope was used to identify the species according to the external morphology and male external genitalia. The mitochondrion COⅠ gene sequence fragments was amplified by PCR and sequenced. The average genetic distance was calculated by Kimura 2-parameter model and phylogenetic trees were constructed using the maximum likelihood method. The classification status was clarified according to the identification keys at the genus and sub-genus levels. More than 100 specimens of male Empoasca sp. were observed and dissected; more than 20 DNA samples were extracted for sequencing; sixteen 709-bp COⅠ sequences were obtained, and the phylogenetic tree was established with the reference Cicadellidae COⅠ sequences from NCBI. Through analyzing the external morphology, male external genitalia characteristics and comparing COⅠ sequences, the main vector insect of C.acuminata witches’-broom phytoplasma was identified as Empoasca (Matsumurasca) Zhang et Liu -CA, and its taxonomic status was the subgenus Empoasca ( Matsumurasca ) Anufriev in the genus Empoasca Walsh, 1862. |
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