基于特异性引物分子标记技术的大豆根腐病主要病原菌分子判别体系研究

Study on Molecular Discrimination System for the Pathogens of Soybean Root Rot Based on the Molecular Marking Technology Using the Specific Primer

【目的】探索大豆根腐病主要病原菌分子检测方法,为复合侵染病害的进一步深入研究及大豆根腐病的科学防治奠定基础。【方法】采用病原菌rDNA-ITS区段序列分析对各病原菌种进行分子验证,采用病原菌特异性引物对大豆根腐病各病原菌种进行分子标记,以特异性引物分子标记技术建立多病原的分子判别体系。【结果】通过序列比对使收集到的7种大豆根腐病致病菌(立枯丝核菌、瓜果腐霉菌、尖孢镰孢霉、禾谷镰孢霉、茄腐镰孢霉和茄腐镰孢霉蓝色变种、燕麦镰孢霉)的鉴定结果得到分子验证;采用6种针对各病原菌种的引物(茄腐镰孢霉和茄腐镰孢霉蓝色变种为同一引物)进行的扩增实验均有特异性条带;经扩增条件优化和灵敏度检测初步建立针对大豆根腐病多种病原菌的分子判别体系。【结论】采用特异性引物分子标记技术基础上的病原菌分子判别体系可用于多病原复合侵染大豆根腐病的病原菌种类的分子判别,并为该病分子检测技术的建立奠定基础。

[ Objective] Exploring molecular detection methods aimed at the main pathogens of soybean root rot, and, in order to lay the foundation for further research on the composite infection diseases and the scientific controlling for soybean root rot. [ Method ] Using sequence analysis with the pathogen＇ s rDNA - ITS section for molecular verifying aimed at various pathogens, take the molecular marker aimed at various pathogens by the pathogen＇ s Specific primer, and establish the molecular discriminant system aimed at the pathogens based on molecular markers with the specific primer. [ Result ] Through Aligning sequences, make 7 species pathogens （Rhizoctoni solani, Pythium aphanidenatum, Fusarum oxysporum, F. graminearum. , F. avenum. ,F. solani . and F. solanivar coeruleum） of soybean root rot which the collected received molecular validating. In addition to , F. solani . and F . solanivar coeruleum are the same kind of primers. All have shown specificity strips In the amplification experiments that be done Used 6 primers which the respectively aimed at various pathogens. After optimization of PCR conditions and sensitivity test, set up a moleculardiscriminant system aimed at the main pathogens of soybean root rot. [ Conclusion] The molecular discriminant system of pathogens based on molecule marker technology used with the specific primer, be possible used in molecular discriminant aimed at multiple pathogens of soybean root rot, and lay the foundation for establish the molecular detection techniques aimed at this disease.