Specific and sensitive detection of the guava fruit anthracnose pathogen (Colletotrichum gloeosporioides) by loop-mediated isothermal amplification (LAMP) assay.
Specific and sensitive detection of the guava fruit anthracnose pathogen (Colletotrichum gloeosporioides) by loop-mediated isothermal amplification (LAMP) assay.
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通过环介导等温扩增 (LAMP) 检测特异性、灵敏地检测番石榴果实炭疽病病原体（胶孢炭疽病菌）。
DOI：
10.1139/cjm-2019-0099
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发表时间：
2019
影响因子：
2.8
通讯作者：
Junxi Jiang
中科院分区：
文献类型：
--
作者：
C. Lan;Jinai Yao;Xiujuan Yang;H. Ruan;Deyi Yu;Junxi Jiang
番石榴炭疽病是由胶孢炭疽菌（Colletotrichum gloeosporioides）引起的一种病害，是限制番石榴生产的主要因素。及时准确地检测病原体对于制定疾病管理策略非常重要。本文建立了一种特异、灵敏的环介导等温扩增（LAMP）方法。使用靶向β-微管蛋白2（TUB 2）基因的引物开发了gloeosporioides。最佳反应条件为64 ℃反应60 min。gloeosporioides分离物、炭疽菌Colletotrichum ssp.分离物和其他属的分离物。阳性结果仅在C. gloeosporioides，而没有观察到交叉反应的其他物种。在25 μL反应液中，LAMP检测限为10 fg基因组DNA。LAMP法成功地检测了C. gloeosporioides在番石榴果实收集在外地。结果表明，建立的LAMP方法是一种简便、经济、快速、灵敏、特异的诊断番石榴炭疽病的方法。gloeosporioides，并可能用于疾病管理。
Anthracnose of guava, caused by the fungus Colletotrichum gloeosporioides, is a major factor limiting guava production worldwide. Timely and accurate detection of the pathogen is important for developing disease a management strategy. Herein, a loop-mediated isothermal amplification (LAMP) assay for the specific and sensitive detection of C. gloeosporioides was developed using primers targeting the β-tubulin 2 (TUB2) gene. The optimal reaction conditions were 64 C for 60 min. The specificity of the method was tested against C. gloeosporioides isolates, Colletotrichum ssp. isolates, and isolates of other genera. Positive results were obtained only in the presence of C. gloeosporioides, whereas no cross-reaction was observed for other species. The detection limit of the LAMP assay was 10 fg genomic DNA in a 25 μL reaction. The LAMP assay successfully detected C. gloeosporioides in guava fruits collected in the field. The results indicated that the developed LAMP assay is a simple, cost-effective, rapid, highly sensitive and specific tool for diagnosis of guava anthracnose caused by C. gloeosporioides, and might be useful for disease management.
影响因子：
2.7
作者：
Goto, Motoki;Honda, Eiichi;Hanaki, Ken-Ichi
通讯作者：
Hanaki, Ken-Ichi