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Salvia (Salvia miltiorrhiza Bunge) is a medicinal plant, roots and rhizomes of medicine with blood to regulate menstruation, dispel transvestite students new sedative, cooling blood consumption carbuncle, swelling and pain and other effects, is widely used in coronary heart disease, cerebrovascular disease, hepatitis, cirrhosis, chronic kidney failure, irregular menstruation and upset, insomnia and other illnesses. Salvia active ingredients can be divided into two categories: one is the water-soluble phenolic acids, including salvianolic acid, rosmarinic acid, rosemary acid methyl ester and protocatechuic aldehyde, etc.; other is a fat-soluble diterpene quinone tanshinone Ⅰ, tanshinone Ⅱ A, cryptotanshinone and dihydro tanshinone etc.. The tanshinone substances mostly diterpenoid quinone compound. Geranyl geraniol based pyrophosphate synthase (Geranylgeranyldiphosphate synthase, GGPPS) is one of the rate-limiting enzyme in the synthesis of higher plants diterpenoid substance regulate metabolic branch point precursor it geranylgeranyl geranyl pyrophosphate (Geranylgeranyl diphosphate, GGPP) diterpenoid synthesis of the substance. This study, degenerate primers were cloned by chromosome walking Salvia GGPP synthase gene (is named: SmGGPPS) bioinformatic analysis of the sequence and the promoter region of the gene regulation information, real-time quantitative PCR method expression of this gene in different organs, under the influence of different environmental factors are discussed; the using factors can upregulate SmGGPPS gene expression processing Salvia plants and Salvia diterpenoid substance synthesis under the influence of these factors changes detected by HPLC technology . In this study, to further clarify Salvia the diterpenoid compound (Tanshinone class) biosynthesis mechanism and its regulation to provide certain information. The main findings are as follows: using degenerate primers and combination of chromosome walking cloned DNA and cDNA level from Salvia Salvia geranyl Mang geranylgeranyl pyrophosphate synthase gene (SmGGPPS). The cDNA contains 2758 bp, contains an intron sequence; open reading frame 1086 bp, encoding 361 amino acids (amino acid sequence is named: SmGGPPS). GGPP synthase gene SmGGPPS homologous nucleic acid level and encoded amino acid levels and other plants have been reported have a high consistency (64%). 2.SmGGPPS encoding amino acids containing a mutational pentenyl pyrophosphate synthase (Trans-isoprenyl diphosphatesynthases) domain belonging to turn isopentenyl pyrophosphate synthase protein family members. GGPP synthase family of the two aspartic acid-rich characteristic the sequence: DDXXXXD DDXXD. The bioinformatic analysis shows containing the polypeptide sequence of the N-terminal chloroplast transit peptide fragment, chloroplast protein is hydrophilic, and the helical structure is its main structural unit. 3 the Salvia SmGGPPS gene 5 'flanking sequences (1095 bp fragment), contains the promoter core components related acting elements and tissue-specific expression outside induced damage, drought, heat, GA, SA, ABA and pathogens such as cis-acting elements. Expression the speculated SmGGPPS Keegan light, high temperatures, may be affected by pathogens disseminated, injury, plant hormones (GA, ABA and SA), salt solution factor regulation. 4 gene expression analysis, The Salvia SmGGPPS gene specifically expressed in leaves: dark, high temperature, pathogens, mechanical damage, salt solution (200 mM NaCl solution), gibberellin (50 mM GA , 3 ), abscisic acid (100 mM ABA) and salicylic acid (5 mM SA), etc. Several factors can induce upregulation of the gene. 5.NaCl solution (500 mM), high temperature, GA 3 (50 mM) and SA (10 mM) and other four factors can promote the accumulation of tanshinone compounds in Salvia ingredients, including SA (10 mM ) the effect of the highest, is about three times that of the control group; ABA (100 mM) and dark can induce the Salvia GGPP synthase gene expression raised, but the promoting effect of accumulation of tanshinone class compounds is not obvious.
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