黄芪根腐病菌锐顶镰刀菌的实时荧光定量PCR快速检测方法OA

Fusarium acuminatum(FA)is one of the important pathogens causing root rot of Astragalus membranaceus var.mongholicus(AMM).To rapidly detect and accurately quantify FA in AMM plants and soil,a quantitative real-time PCR(qPCR)detection method was established and its practicability in detecting the pathogen in AMM plants and soil samples was verified.The results showed that primer pair Fae F4/Fae R4 designed based on the elongation factor-1α gene(EF-1α)sequence of FA was highly specific.For the FA in AMM plants and soil,the sensitivities of qPCR detection method were 2.56×10-3 ng/μL of DNA concentration and 1×102 conidia/g,respectively;the correlation coefficients of the constructed standard curves were 0.999 7 and 0.983 8;the amplification efficiencies were 1.07 and 0.87.Repeatability evaluation found that the method was stable and reliable.With the established method,the pathogen FA could be detected in inoculated-AMM samples as early as 1 hour after inoculation,and the contents increased with time.The detection rate of FA was 100%in suspected diseased AMM samples.FA was tested positive in all soil samples and the number of conidia in diseased soil samples was significantly greater than that in healthy soil at the same cultivation years.In conclusion,the qPCR detection method established in this study could be used to rapidly and quantitatively detect FA in AMM plants and soil,providing a reliable technical support for the accurate detection and timely prevention and control of AMM root rot.