寄生中国倒挂金钟的草莓滑刃线虫鉴定

doi:10.1016/j.jia.2024.07.002

摘要/Abstract

摘要：

【目的】四川省成都市倒挂金钟（Fuchsia × hybrid Voss.）发生严重的叶斑病，从病叶中分离到大量滑刃线虫属线虫，对其形态学、分子特征和寄生性等进行详细研究，以明确其种类及对倒挂金钟等作物的寄生性，为草莓滑刃线虫的准确鉴定及倒挂金钟叶斑病防控提供科学依据。【方法】将倒挂金钟的发病叶片剪碎，用浸泡法分离叶片中的滑刃线虫，通过胡萝卜愈伤组织对该线虫进行纯化培养，分离纯化培养的线虫制作永久玻片，在光学显微镜和扫描电子显微镜下观察雌虫和雄虫的形态特征，并进行形态特征测量。采用两对引物1096F/1912R及1813F/2646R分别扩增18S小亚基部分区段（SSU）、引物D2A和D3B扩增28S大亚基D2-D3区段（LSU D2D3）及引物18S和26S扩增内转录间隔区（internal transcribed spacer, ITS），经序列测定，采用贝叶斯法构建该线虫种群与滑刃线虫属其他线虫种群的SSU、LSU D2D3和ITS系统进化树，分析其系统发育关系。在室内采用人工接种的方法，测定该线虫对倒挂金钟、草莓和蜈蚣凤尾蕨的寄生性。【结果】形态学观察和特征测量结果表明，从四川成都倒挂金钟病叶中分离到的滑刃线虫雌虫和雄虫形态特征与草莓滑刃线虫（Aphelenchoides fragariae）的形态特征基本一致；序列比对发现该线虫的SSU、LSU D2D3和ITS均与草莓滑刃线虫的序列相似性最高，在3个rDNA系统发育树中，该线虫也均与草莓滑刃线虫以100%的支持率形成一个单系。结合形态学和分子生物学结果，将分离自四川成都倒挂金钟病叶上的滑刃线虫鉴定为草莓滑刃线虫。寄生性研究结果表明，该线虫不仅可寄生和危害倒挂金钟，也可寄生和危害草莓和蜈蚣凤尾蕨这两种草莓滑刃线虫的典型寄主。【结论】引起我国四川成都倒挂金钟叶斑病的病原为草莓滑刃线虫，系首次发现草莓滑刃线虫危害倒挂金钟，也系国内首次通过结合形态学和分子特征对草莓滑刃线虫进行确定。

Abstract:

The strawberry crimp nematode (Aphelenchoides fragariae) is a serious pathogen of ornamental crops and an important quarantine object in approximately 50 countries and regions including China. One nematode population within the genus Aphelenchoides was discovered from diseased leaves of fuchsia plants (Fuchsia × hybrid Voss.) in Chengdu city, Sichuan province of China. Morphological and morphometric data were obtained using light microscopy and scanning electron microscopy. After detailed examination, the species was identified as A. fragariae. Three rDNA sequences of this species, including partial rRNA small subunit, D2-D3 expansion domains of the rRNA large subunit and internal transcribed spacer, were amplified and sequenced. Bayesian trees inferred from these three rDNA sequences were constructed, revealing that this species is placed in a high support monophyletic clade with A. fragariae but clearly separated from all other Aphelenchoides species. Moreover, host-suitability tests showed that the Aphelenchoides population not only can harm and reproduce in F. hybrid, but also in Fragaria ananassa and Pteris vittata (two common hosts of A. fragariae). In conclusion, the study confirmed A. fragariae identity of the nematode from F. hybrid in Chengdu city based on morphology, molecular analysis and host-suitability tests. To our knowledge, this is the first molecular and morphological confirmation of A. fragariae in China, and F. hybrid was first discovered to be attacked by A. fragariae.

Key words: strawberry crimp nematode , morphology , molecular characterization , host-suitability test , fuchsia

. 寄生中国倒挂金钟的草莓滑刃线虫鉴定[J]. Journal of Integrative Agriculture, DOI: 10.1016/j.jia.2024.07.002.

Qiuling Huang, Yan Liao, Chunhui Huang, Huan Peng, Lingchiu Tsang, Borong Lin, Deliang Peng, Jinling Liao, Kan Zhuo. Integrative identification of Aphelenchoides fragariae (Nematoda: Aphelenchoididae) parasitizing Fuchsia hybrid in China[J]. Journal of Integrative Agriculture, DOI: 10.1016/j.jia.2024.07.002.

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