根据太子参尖孢镰刀菌的测序结果,用设计的一对特异性引物,构建其分子定量检测方法,并进行不同连作年限太子参根际土壤中该尖孢镰刀菌的菌群定量分析,研究结果表明,结果表明:(1)扩增的阳性条带106 bp,特异性、敏感性结果显示,该PCR方法对太子参镰刀菌DNA的最低检测量为0.000012 ng/μL,而禾谷镰刀菌、茄病镰刀菌、黄曲霉和解淀粉芽孢杆菌的扩增结果均为阴性。(2)运用此方法对该太子参镰刀菌的定量结果表明,尖孢镰刀菌随太子参连作年限的增加而逐年增多。该检测方法具有快速、准确、灵敏度高、易于操作等优点,并且实现了由定性向定量的跨越,可以为太子参根际微生态中该种病原菌的动态变化研究提供方法及依据,也为阐释太子参重茬连作时伴随爆发大规模病害提供了参考。
The specific primers of Fusarium oxysporum f.sp.were designed and the molecular quantitative detection method was built to be used to quantify and analyze the Fusarium oxysporum f.sp.in rhizosphere soil under monoculturally cropped Pseudostellaria heterophylla for different years.The results showed that ( 1 ) the amplified positive strip was 106 bp;the minimum detectable amount of Fusarium oxysporum f.sp.DNA was 0.000 012 ng/μL with this method and the amplification results of Fusarium graminearum,Fusarium solani,Aspergillus flavus,bacillus amyloliquefaciens were all negative,(2) the quantifying results of Fusarium oxyspo-rum f.sp.with this method suggested that the number of Fusarium oxysporum f.sp.increased with the monocul-ture years.This detecting method is of accuracy,high sensitivity and easy accessibility,which realizs the leap from qualitative to quantitative.It offers the methods and foundation for analysis of this pathogen in the rhizo-sphere micro ecology of Pseudostellaria heterophylla,and illuminates the reason for large-scale occurrence of the disease in successively cropped Pseudostellaria heterophylla.