Genetic diversity and phylogeny analysis of 3-hydroxy 3-methylglutaryl-CoA reductase gene (SmHMGR) in Danshen (Salvia miltiorrhiza Bunge)

Abstract

HMGR, 3-hydroxy-3-methylglutaryl-CoA reductase, is a major rate-limiting enzyme in mevalonate (MVA) pathway for isoprenoids and subsequent tanshinone biosynthesis in the Chinese traditional bulk herbal medicine Danshen, Salvia miltiorrhiza, mainly for cardiovascular disorders. In this paper, the genomic SmHMGR genes of 38 cultivated populations of S. miltiorrhiza collected in China were for the first time sequenced to reveal the genetic diversity and phylogeny. The SmHMGR gene was shown to be intron-free, 1650~1659 bp in complete CDS with the majority being 1656 bp, and two unique populations (W-FJLY-V-1 and W-SCHY-W-1) being 1659 and 1650 bp respectively. A total of 103 SNP variation sites were detected with a variation rate of 6.22%, most of which occurred in S. miltiorrhiza f. alba population W-SCHY-W-1; a total of 25 amino acid variation sites were found, of which 19 was in W-SCHY-W-1. The same four populations, W-SCHY-W-1, V-HBAG-V-1, V-JLCC-V-1 and S-NM-V-1 could be discriminated from the remaining 34 by both the SNP fingerprints and the deduced amino acid variation sites. Other or composite DNA markers are needed for better identification. The SmHMGR gene of white flower S. miltiorrhiza f. alba population W-SCHY-W-1 is especially rich in variations and worthy of further studies. Phylogenetic trees based on both the gene and the deduced amino acid sequences showed a very similar two-clade topological structure. This research enriched the content and the genetic means for the molecular identification, genetic diversity and phylogenetic studies of the cultivated S. miltiorrhiza populations, and laid a solid foundation for further related and in-depth investigations.