The study aimed at establishing a rapid and accurate real-time fluorescence quantitative PCR(RT-qPCR)method for the detection of Fusarium solani,the pathogenic fungus of Astragalus membranaceus var.mongholicus(AMM)root rot.Based on the elongation factor-1α(EF-1α)sequence of F.solani,a pair of specific primers FP.F/FP.R was designed and verified for its specificity.The RT-qPCR detection method was established and then used to detect the DNA content of F.solani in AMM plants and soil samples for verifying its detection effect.The results showed that the designed specific primer FP.F/FP.R was highly specific,and the sensitivity of the established RT-qPCR method was 0.0192 ng/μL.Repeatability evaluation showed that the method was stable and reliable.With the mothed,the pathogen F.solani was detected in F.solani-inoculated and suspected diseased AMM plants as well as soil samples,the positive detection rate being 100%for all samples in the three groups.Meanwhile,the changes of F.solani content in AMM plants and soil samples were consistent with the severity of root rot.The results indicated that the RT-qPCR method could be used in quantitative detection of F.solani in AMM plants and soil to provide technical support for the early diagnosis of and warning against root rot,and the dynamic monitoring of pathogens.
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