土壤根肿病菌休眠孢子PCR快速检测方法的建立

Establishment of PCR Rapid Detection of Plasmodiophora brassica Resting Spores in Soils

根据GenBank中根肿病菌的保守序列,设计了一对引物,该引物能从供试的根肿病菌根组织中特异性扩增出一条498bp的预期条带。采用改良土壤DNA提取方法对人工配制病土中根肿病菌休眠孢子DNA提取,通过检测不同稀释度和不同孢子量的DNA检测结果证明,该检测方法能检测出土壤根肿病菌含量大于104个孢子/g孢子浓度的带菌土壤,且该方法具有快速、准确、简便、有效的特点。

According to GenBank sequence of Plasmodlophora brassicae,a pair of primers were designed; the primers could specifically am- plify an expected 498 bp band from the infected root tissues. Through improving extraction method, disease soils prepared with resting spores by artificial method could be extracted for total DNA. In the case of different dilutions, the amount of different Spores of total DNA test results showed that the detection method could detect that containing more than 10^4 spores / g in soils, and the method was rapid, accurate, simple and effective.