Salvia Quality Of Est-ssr Molecular Markers And Functional Gene Expression Studies

ObjectiveTo explore the molecular mechanisms of differences in quality of Radix Salvia Miltiorrhizae from different producing areas and provide a foundation for future studies of molecular mechanisms of genuine Radix Salvia Miltiorrhizae.Medthod(1) The analysis of characteristic chromatograms of Radix Salvia Miltiorrhizae from different producing areas were carried out and the content of Salvianic acid A, Cryptotanshinone and Tanshinone IIA in the herb were investigated by HPLC method.(2) After one year of cultivation under the same environment, the expression of genes including SmAACT, SmIPPI and SmCMK from Salvia miltiorrhiza bunge were determined by Real-time PCR. And the correlation bettween the gene expression and the content of Tanshinone was analyzed by SAS software.(3) The genetic diversity of Salvia miltiorrhiza bunge from different producing areas were analyzed by EST-SSR molecular markers developed in this paper.Results(1) The dissertation succeeded in establishing a method of HPLC characteristic chromatograms for analysis of Radix Salvia Miltiorrhizae. And the similarity analysis result showed that the value of Radix Salvia Miltiorrhizae from Henan, Shanxi, Sichuan and Hubei were above 0.9.(2) The expression level of genes of SmAACT and SmCMK showed the positive correlation between the content of Cryptotanshinone and Tanshinone IIA.(3) The study succeeded in establishing method for EST-SSR markers of Salvi miltiorrhiza and above 386 SSR loci were found in Salvia miltiorrhiza bunge. Among them, the occurrences of different repeat units were mainly the di-type(63.0%). The CT/AG was the most frequent motif type (21.8%) and the GAA/TCC was the most frequent motif in trinucleotide repeats (5.7%).Conclusion(1) The HPLC analysis results showed great differences of quality of Radix Salvia Miltiorrhizae from different producing areas, which verified partially the rationality of genuine producing areas of Radix Salvia Miltiorrhizae recorded in literature.(2) There was some correlation between the expression levels of genes of SmAACT and SmCMK and the content of Cryptotanshinone and Tanshinoneâ…¡A, which suggestioned that the differences of gene expression partly attributed to the quality differences of Radix Salvia Miltiorrhizae from different producing areas.(3) The quality differences of Salvia miltiorrhiza bunge had relationship with its genetic diversity.