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To identify Salvia shandongensis and its relatives at molecular level, the psbA-trnH intergenic region of three species including Salvia shandongensis, Salvia miltiorrhiza and S. miltiorrhiza f. alba were amplified and sequenced. Sequences were assembled with CodonCode Aligner. The K2P genetic distances between Salvia shandongensis and its relatives were calculated and UPGMA tree was performed by MEGA5.0. The results indicated that the lengths of psbA-trnH regions of Salvia shandongensis were about 391 bp, while the lengths of psbA-trnH regions of Salvia miltiorrhiza and S. miltiorrhiza f. alba were about 386 bp. The psbA-trnH sequences showed considerable variations between species and thus were revealed as a promising candidate for barcoding of Salvia shandongensis and its relatives. The intra-specific genetic distances of Salvia shandongensis were 0, while the intra-specific genetic distances of Salvia miltiorrhiza and S. miltiorrhiza f. alba were 0.002 and 0.001 respectively. Additionally, the genetic distance of Salvia shandongensis and Salvia miltiorrhiza ranged from 0.034 to 0.04, and the genetic distance of Salvia shandongensis and S. miltiorrhiza f. alba ranged from 0.005 to 0.008, the intra-specific genetic distances of Salvia shandongensis were much smaller than that of Salvia miltiorrhiza and S. miltiorrhiza f. alba; clustering results showed that there were obvious differences between Salvia shandongensis, Salvia miltiorrhiza and S. miltiorrhiza f. alba, which was consistent with morphological characteristics. This study not only firstly provides the scientific basis for establishing the taxonomy position in molecular level and revealing their genetic relationships of S. shandongensis, S. miltiorrhiza and S. miltiorrhiza f. alba; but also provides DNA molecular identification scientific basis for the development of new medicinal plant resources of Salvia shandongensis. Our results suggest that the psbA-trnH intergenic spacer region can be used as a barcoding to identify Salvia shandongensis, Salvia miltiorrhiza and S. miltiorrhiza f. alba.

Sclareol is a member of labdane type diterpenes mostly used as fragrance ingredient. To enable microbial production of sclareol, synthetic pathways were constructed by incorporating labdenediol diphosphate synthase (LPPS) and terpene synthase (TPS) of the plant Salvia sclarea into Saccharomyces cerevisiae. It was found that sclareol production could be benefited by overexpression of key enzyme for precursor biosynthesis, construction of fusion protein for substrate channeling, and removal of signal peptides from LPPS and TPS. Under optimal shake flask culture conditions, strain S6 produced 8.96 mg/L sclareol. These results provided useful information for development of heterologous hosts for production of terpenoids.

Objective: To study the ethylene responsive element binding protein genes of Salvia miltiorrhiza through bioinformatics and characterization of its tissue expression in regenerated plantlets., Method: The ethylene responsive element binding protein genes were obtained by cDNA microarray analyze. BLAST was used for alignment, ORF finder software was used to find open reading frame, Prosite database was used to analyze the protein. Semi-quantitative RT- PCR method was used to detect the gene expression level., Result: One ethylene responsive element binding protein was obtained, named as SmERF. SmERF had an open reading frame of 699 bp with 5'-URT 87 bp and 3'-URT 166 bp. The putative protein SmERF contains a highly conserved ERF/AP2 domain. Semiquantitative RT- PCR illustrated that SmERF was expressed in all tissues such as root, stem and leaf in regenerated shoots, while the expression level was higher in root than in stem and leaf., Conclusion: It was the first time to obtain ERF gene in S. miltiorrhiza and set a good foundation for its further functional study.

Molecular systematic techniques were applied to reveal the genetic diversity of medicinal plants and their related species in Salvia. The internal transcribed spacer (ITS) as well as 5.8S rDNA sequences of 27 samples of Salvia were amplified using PCR method and sequenced. Mega 3.1 was used to analyze the genetic diversity within genus. The complete sequences of ITS plus 5.8S rDNA are about 612-617 bp. A phylogenetic tree generated by Neighbor-Joining method partly supported the morphological classification within Salvia, but incompatible results were also obtained in the treatment of phylogenetic positions of some species such as Salvia trijuga, Salvia flava var. flava and Salvia flava var. megalentha. The ITS regions of present Salria species showed considerable variation between subgenera in contrast with the conservative 5.8S rDNA sequences. The native Salvia species might have a different origin from the foreign species. The phylogenetic positions of subgenera and sections inferred by ITS analysis were comparable with that of traditional classification, while the phylogeny within sections is still doubtful due to limited information in ITS sequence and need to be further proved by other evidence. ITS analysis in this study supports the rationality of using species from Drymosphace section as substitute drug resources of Dan shen, but also reveals significant genetic differences between high mountain Dan shen species such as Salvia przewalskii with traditional Dan shen origins.

Objective: To explore the technique of induction of polyploidy in Salvia bowleyana by colchicine treatment., Method: The three kinds of explant of bud, leaf and calli were induced by colchicine treatment., Result: The induction effects were better when the calli was treated by colchicines (15 mg x L(-1)) and the leaf was pre-cultured for one week. The doubling rate was 33.33%, while the majority were wholy doubled plants, and the leaves were thicker and broader, the color was darker, the root was thicker and the stoma size was obviously bigger than the diploid plants. The number of chromosome were 8 to 64. Isoenzyme analysis showed that the enzyme activities between the polyploid and the diploid plants were quite different., Conclusion: Induction of polyploidy by colchicine treatment is efficacious. The part of the doubled plants were identified as homologmous tetraploids.