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潜在入侵植物病害的快速检测技术研究进展

花匠小妙招
2024-11-18 03:52

摘要： 在国家重点基础研究发展计划(973)“农林危险生物入侵机理与控制基础研究”(2002CB111400)项目的支持下，针对小麦矮腥黑穗病、梨火疫病及大豆疫霉根腐病等潜在外来入侵植物病害，围绕快速检测的技术问题，采用分子生物学技术与方法，研制出了小麦矮腥黑粉菌(TCK)快速分子检测的特异性SCAR标记；建立了以梨火疫病菌基因组和质粒DNA序列为基础的一步双重PCR检测方法及以生物富集PCR技术为核心的受侵组织梨火疫病菌快速检测程序；研究证明中国不同地区的大豆疫霉菌群体在与大豆品种的互作中发生了广泛的遗传变异，建立了一种简单、有效的土壤中大豆疫霉菌的分离方法及以大豆疫霉菌ITS序列为基础的快速分子检测体系，对大豆疫霉游动孢子和卵孢子的检测理论精度可达0.5 个孢子。在上述研究的基础上，成功研制出了梨火疫病、小麦矮腥黑穗病及大豆疫霉根腐病快速检测试剂盒。

关键词: “973”计划 , 作物入侵病害 , 快速检测 , 小麦矮腥黑穗病 , 梨火疫病 , 大豆疫霉根腐病

Abstract: For developing rapid detection technologies of alien invasive diseases of TCK, pear fire blight and soybean Phytophthora root rot, associated research were carried out with the support of National Program on Key Basic Research Projects (973 Program): “Invasive Mechanisms and Control Basic Research of Agricultural and Forest Alien Invasive Species” (No.2002CB111400). SCAR marker for rapid and sensitive detection of TCK was obtained. Sensitive molecular detection method of Erwinia amylovora with “One step and duplex PCR” based on chromosomal and pEA29 plasmid sequences and detection protocol of identification in asymptomatic samples infected by E. amylovora based on Bio-PCR habe been developed. It is indicated that wide genetic variations were occurred in the long history of interaction between Phytophthora sojae populations and soybean varieties in different areas in China. A simple and effective method of isolating P. sojae from soils and molecular detection system of soybean Phytophthora root rot based on ITS sequences were established. The protocol was determined to efficiently detect pathogen at a theoretical rate of half spore. The Kits for detecting pear fire blight, TCK and soybean Phytophthora root rot based on our research results have been exploited.

Key words: National Basic Research and Development Program (973 program) , Crop invasive diseases , Rapid molecular detection , TCK , Pear fire blight , Soybean Phytophthora root rot