Molecular characterisation and diagnostics of the red ring nematode, Bursaphelenchus cocophilus, from Mexico

IF 16.4 1区 化学 Q1 CHEMISTRY, MULTIDISCIPLINARY
S. Subbotin, Ignacio Cid del Prado-Vera
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引用次数: 0

Abstract

The red ring nematode, Bursaphelenchus cocophilus, vectored by the South American palm weevil, Rhynchophorus palmarum, is the causal agent of red ring disease in coconut and other palms in countries of Central and South America. The populations of B. cocophilus collected in the states of Guerrero and Tabasco, Mexico, were molecularly characterised using the D2-D3 expansion segments of 28S rRNA, ITS rRNA and COI gene sequences. Mexican B. cocophilus populations were molecularly different from other Central and South American populations. Comparative analysis of available rRNA sequences obtained from several countries showed that B. cocophilus consists of molecularly different populations and its species structure is likely congruent with that of the beetle vector. Conventional PCR, real-time PCR and recombinase polymerase amplification (RPA) with lateral flow dipstick (LF) assays have been developed for the identification of the red ring nematode in this study. The specificity of the ITS rRNA primers in the assays were examined using B. cocophilus and other 15 species of family Aphelenchoididae. Detection sensitivity levels, determined by using a dilution series of B. cocophilus extracts, was 0.13 nematode per reaction tube for real-time PCR and 0.25 nematode per reaction tube for conventional PCR and LF-RPA assays. The application of the LF-RPA assay has great potential for diagnosing infestation of this species with a minimal laboratory infrastructure.
墨西哥红环线虫 Bursaphelenchus cocophilus 的分子特征和诊断方法
红环线虫(Bursaphelenchus cocophilus)由南美洲棕榈象鼻虫(Rhynchophorus palmarum)传播,是中美洲和南美洲国家椰子和其他棕榈树红环病的病原体。利用 28S rRNA、ITS rRNA 和 COI 基因序列的 D2-D3 扩展片段,对在墨西哥格雷罗州和塔巴斯科州采集到的椰子象鼻虫种群进行了分子鉴定。墨西哥嗜球菌种群与其他中美洲和南美洲种群存在分子差异。对从几个国家获得的现有 rRNA 序列进行比较分析表明,嗜椰子虫由分子上不同的种群组成,其物种结构可能与甲虫载体的物种结构一致。本研究开发了常规 PCR、实时 PCR 和重组酶聚合酶扩增(RPA)与侧流浸渍棒(LF)测定法,用于鉴定红环线虫。研究人员使用嗜球虫和其他 15 种 Aphelenchoididae 科物种检验了 ITS rRNA 引物在检测中的特异性。通过使用嗜球虫提取物稀释系列确定的检测灵敏度水平为:实时 PCR 为每反应管 0.13 条线虫,传统 PCR 和 LF-RPA 检测为每反应管 0.25 条线虫。LF-RPA 分析法的应用潜力巨大,只需少量实验室基础设施即可诊断该物种的侵染情况。
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来源期刊
Accounts of Chemical Research
Accounts of Chemical Research 化学-化学综合
CiteScore
31.40
自引率
1.10%
发文量
312
审稿时长
2 months
期刊介绍: Accounts of Chemical Research presents short, concise and critical articles offering easy-to-read overviews of basic research and applications in all areas of chemistry and biochemistry. These short reviews focus on research from the author’s own laboratory and are designed to teach the reader about a research project. In addition, Accounts of Chemical Research publishes commentaries that give an informed opinion on a current research problem. Special Issues online are devoted to a single topic of unusual activity and significance. Accounts of Chemical Research replaces the traditional article abstract with an article "Conspectus." These entries synopsize the research affording the reader a closer look at the content and significance of an article. Through this provision of a more detailed description of the article contents, the Conspectus enhances the article's discoverability by search engines and the exposure for the research.
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