Cost-effective multiplex PCR assay for simultaneous detection of bacterial leaf blight, blast and brown planthopper resistance genes in rice

IF 16.4 1区化学 Q1 CHEMISTRY, MULTIDISCIPLINARY Accounts of Chemical Research Pub Date : 2024-01-25 DOI:10.1007/s13562-023-00870-0

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Abstract

Rice production is drastically affected by biotic stresses, namely, bacterial leaf blight (BB), blast (BL) and brown planthopper (BPH). As marker-assisted selection for uniplex gene screening in breeding population is often tedious, time-consuming and expensive, multiplex PCR is practically a reasonable choice for the testing of genes in the pyramided lines as it offers significant time- and cost-saving advantages. Therefore, a cost-effective multiplex PCR-based screening for simultaneous detection of three BB (Xa21, xa13, and Xa4), two BL (Pi1and Pi54) and two BPH resistance genes (Bph20 and Bph21) has been developed. This multiplex system can detect different combinations of BB, BL and BPH resistance. In addition to this most cost-effective method, a five-gene (Xa21, xa13, Pi54, Bph20 and Bph21) multiplex assay in a single PCR tube for three biotic stresses, BB, BL and BPH, has been developed. These standardized multiplex assays have been incorporated into our breeding programs for accelerated results. These assays have displayed no deviation and are in perfect correlation with the individual marker screenings. As one of the most efficient and cost-saving technologies, this technology will have a great impact in the seed industry.

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同时检测水稻细菌性叶枯病、稻瘟病和褐斑病抗性基因的经济高效的多重 PCR 分析法

摘要水稻生产受到细菌性叶枯病（BB）、稻瘟病（BL）和褐斑病（BPH）等生物胁迫的严重影响。在育种群体中进行标记辅助选择的单基因筛选通常比较繁琐、耗时且昂贵，而多重 PCR 具有显著的省时、省钱优势，因此实际上是检测金字塔型品系基因的合理选择。因此，我们开发了一种经济高效的基于多重 PCR 的筛选方法，可同时检测三个 BB（Xa21、xa13 和 Xa4）、两个 BL（Pi1 和 Pi54）和两个 BPH 抗性基因（Bph20 和 Bph21）。这种多重系统可以检测 BB、BL 和 BPH 抗性的不同组合。除了这种最具成本效益的方法外，还开发了一种在单个 PCR 管中检测 BB、BL 和 BPH 三种生物胁迫的五基因（Xa21、xa13、Pi54、Bph20 和 Bph21）多重检测方法。这些标准化的多重检测方法已被纳入我们的育种计划，以加快结果的产生。这些检测结果没有任何偏差，与单个标记筛选结果完全吻合。作为最高效、最节省成本的技术之一，这项技术将对种子行业产生巨大影响。

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来源期刊

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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