Establishment of a KASP detection method for the root rot pathogen Heterobasidion parviporum on imported wood
Heterobasidion parviporum is a strong devastating pathogen and great harmful to coniferous forests.It is not distributed in our country until now,but there is transmission risk of being introduced through imported wood.In order to establish a KASP detection method for H.parviporum,multiple sequence comparison was performed on 223 genes from Heterobasidion and its similar genus,and then a SNP site was screened on H.parviporum GAPDH gene,which can be used as marker in this study.The primers Hpar-FAM-F1,Hpar-HEX-F2 and Hpar-R were designed based on design rules for H.parviporum KASP molecular detection.Test results show that it could successfully identify H.parviporum from 22 similar species including H.irregulare,H.occidentale and others by genotyping,indicating that the marker has high specificity.DNA concentration of H.parviporum was ten-fold diluted from initial concentration 100 ng/μL in sensitivity testing.The reaction result of gradient 10-4 and 10-5 were close to that of double diluted water,and therefore,the minimum concentration of the marker detected was up to 0.1 ng/μL.Model positive sample tests were performed in two forms,one was basidiocarp powder mixed with wood and another was adding DNA of H.parviporum in wood.Results show that model positive samples and the reference species displayed typical genotyping on axis.The KASP marker could accurately identify H.parviporum on coniferous wood,and the KASP method can be used to detect H.parviporum on imported wood.
万方数据
维普
