Not less than 14 annotated TDFs have been assigned into secondary metabolic path

Not less than 14 annotated TDFs had been assigned into secondary metabolic pathways via searching the KEGGPATHWAY database and they were primarily concerned in biosynthesis of phenylpropanoids, alkaloids, terpenoids and steroids. The expression amounts of 9 TDFs have been positively connected to tanshinones and phenolic compounds production and have been also co regulated with phenolic compounds and tanshinones biosynthesis related genes by YEL. They were genes encoding lipoxygenase, jasmonate zim domain protein, pyruvate decarboxylase, catalase, cinnamyl alcohol dehydrogenase like protein, HD domain class transcription element, dihydroflavonol reductase and two unknown genes. The sequence data while in the present work KSP protein inhibitor not merely supplied us candidate genes involved in biosynthesis of tanshinones and phenolic compounds but also gave us additional insight into secondary metabolism in Salvia. Supplies and Methods Plant components The red roots of S. castanea Diels f. tomentosa Stib and S. miltiorrhiza along with the white roots of S. miltiorrhiza expanding for three month during the wild have been obtained from the health-related plants garden of Northwest A&F University in Shaanxi province. S. miltiorrhiza seedlings have been cultured in MS liquid medium under natural temperature and photoperiod for 120 days, and then the hydroponic roots were harvested.
A few different plants each for S1, S2, S3, and S4 were collected for analysis of metabolic profiles and cDNA AFLP. Docetaxel The plants have been authenticated by Professor Yuejin Zhang of Northwest A&F University. The root samples have been frozen in liquid nitrogen immediately, and then stored at 280uC until RNA isolation. Hairy root culture and treatment S. miltiorrhiza hairy roots had been derived after the infection of plantlets with Agrobacterium rhizogenes bacterium. Experiments in this study were carried out in a 250 mL shake flask on an orbital shaker running at 110 120 rpm and at 25uC in darkness. Each flask was filled with 50 mL liquid medium and inoculated with 0.3 g fresh hairy roots from a 3 week old shakeflask culture. The liquid medium was made of hormone free MS medium with 30 g/L sucrose but without ammonium nitrate as previously described. The polysaccharide fraction of yeast extract was prepared by ethanol precipitation as described. The treatment with yeast elicitor liquid were conducted on day 18 post inoculation of the hairy root culture. The equal volume of distilled water was added to the hairy root culture as the control. Three independent biological replications had been performed. HPLC analysis of tanshinones and phenolic compounds HPLC analysis was performed using a Waters system with a binary pump and Photodiode Array Detector as previously described. A SunFire C18 column was used. Data have been acquired and processed using Empower 2 software.