Genetic Diversity Analysis Of Inonotus Obliquus By SRAP And RSAP Markers

Inonotus obliquus (Fr.) Pilat is a kind of rare and valuable medicinal fungi and is in danger of extinction because of excessive pick in recent years. Although the artificial cultivation of Inonotus obliquus have succeed, but the biological efficiency is very low, while the study on breeding of new varieties is still blank. In this paper, SRAP and RSAP markers were used to analyze the genetic diversity of20Inonotus obliquus strains from5different countries and areas:China, Finland, Russia, Japan and North Korea hoping to provide reference for scientific protection and utilization of Inonotus obliquus resources. The main results are as follows:1. The reaction system of SRAP-PCR and RSAP-PCR suitable for Inonotus obliquus were established. The main components in optimal25μl SRAP-PCR amplification reaction mixture contain10×buffer2.5μl, DNA template20ng, primer0.3μmol·L-1, dNTPs150μmol·L-1, Taq DNA polymerase1.0U, Mg2+1.5mmol·L-1. The optimal RS AP-PCR system contains10×buffer2.5μl, DNA template25ng, primer0.4μmol·L-1, dNTPs300μmol·L-1, Taq DNA polymerase2.0U, Mg2+2.5mmol·L-1.2.12pairs of SRAP primers and9pairs of RSAP primers of high polymorphism and good repeatability from153pairs of SRAP primers and45pairs of RSAP primers were used to analyzed the finger-print of20Inonotus obliquus stains. A total of215locis were amplified using12pairs of SRAP primers, of which203locis were polymorphic with a high polymorphism of94.5%and at an average of17.6locis per primer. A total of154locis were amplified using9pairs of RSAP primers, of which141locis were polymorphic with a high polymorphism of90.8%and at an average of17.1locis per primer. The results suggested that SRAP and RSAP were both applicable in the genetic diversity analysis of Inonotus obliquus.3. The p-distance based on SRAP and RSAP marker of the20Inonotus obliquus strains were from0.030to0.453and0.044to0.497, distinctively, which indicate there are some genetic differences between different strains. The cluster results by SRAP and RSAP marker were about the same, both divided20Inonotus obliquus strains into6groups. The results showed that the genetic differences between strains and geographic distribution have a strong correlation, when10Inonotus obliquus strains from Japan were devided into3groups, Inonotus obliquus strains from Russia and Finland came to one group,2Inonotus obliquus strains from North Korea and5Inonotus obliquus strains from Jilin, China were in the same group, the1Inonotus obliquus strain from Heilongjiang, China occupied one group alone.