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1

Molecular cloning and expression analysis of a new putative gene encoding 3-hydroxy-3-methylglutaryl-CoA synthase from Salvia miltriorrhiza

100%

Zhang L. , Yan X. , Wang J. , Li S. , Liao P. , Kai G.

Acta Physiologiae Plantarum

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2011

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tom 33

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nr 3

EN

A new putative gene encoding 3-hydroxy-3-methylglutaryl coenzyme A synthase (designated as SmHMGS, GenBank Accession No. FJ785326), which catalyses the condensation of acetyl-CoA and acetoacetyl-CoA to form 3-hydroxy-3-methylglutaryl-CoA as an early step in the mevalonic acid pathway, was isolated from young leaves of Salvia miltiorrhiza by rapid amplification of cDNA ends (RACE) for the first time. The full-length cDNA of the putative SmHMGS was 1,655 bp containing a 1,381 bp open reading frame (ORF) encoding a polypeptide of 460 amino acids. Comparative and bioinformatic analyses revealed that SmHMGS showed extensive homology with HMGSs from other plant species. Phylogenetic tree analysis indicated that SmHMGS belonged to the plant HMGS super family and had the closest relationship with HMGS from Hevea brasiliensis. Tissue expression pattern analysis revealed that the putative SmHMGS was constitutively expressed in all the tested tissues and strong in leaf, moderate in stem, weak in root, which was in contrast to SmHMGR reported before. The putative SmHMGS was found to be an elicitor-responsive gene, which could be induced by exogenous elicitors, including salicylic acid (SA) and methyl jasmonate (MJ). These results will help in understanding the role of HMGS in tanshinones biosynthesis in S. miltiorrhiza.

2

Molecular characterization and expression of 1-deoxy-D-xylulose5-phosphate reductoisomerase (DXR) gene from Salvia miltiorrhiza

100%

Yan X. , Zhang L. , Wang J. , Liao P. , Zhang Y. , Zhang R. , Kai G.

Acta Physiologiae Plantarum

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2009

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tom 31

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nr 5

EN

1-Deoxy-D-xylulose 5-phosphate (DXP) reductoisomerase (DXR; EC 1.1.1.267) catalyzes the first committed step of the 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway for isoprenoid biosynthesis in plants. The present study describes the cloning and characterization of a cDNA encoding DXR from Salvia miltiorrhiza (designated as SmDXR, GenBank Accession No. FJ476255). Comparative and bioinformatic analyses revealed that SmDXR showed extensive homology with DXRs from other plant species. Phylogenetic tree analysis indicated that SmDXR belongs to the plant DXR superfamily and has the closest relationship with DXR from Lycopersicon esculentum. Tissue expression pattern analysis revealed that SmDXR expressed strongly in leaves, followed by roots and stems, implying that SmDXR was a constitutively expressed gene. This is the first report on the mRNA expression profile of genes encoding key enzymes involved in tanshinone biosynthetic pathway in Salvia plants. The expression profiles revealed by RT-PCR under different elicitor treatments such as methyl jasmonate (MJ) and salicylic acid (SA) were compared for the first time, and the results revealed that SmDXR was an elicitor-responsive gene, which could be induced by SA in leaves and inhibited by exogenous MJ in three tested tissues. The functional color assay in Escherichia coli showed that SmDXR could accelerate the biosynthesis of lycopene, indicating that SmDXR encoded a functional protein. The characterization, expression profile and functional analysis of SmDXR gene will be helpful for further study in the role of SmDXR in tanshinones biosynthetic pathway and metabolic engineering to increase tanshinones production in S. miltiorrhiza.

3

Effects of endogenous salicylic acid synthesized through PAL and ICS pathway on baicalin and baicalein accumulation in Scutellaria baicalensis Georgi

84%

Su H. , Yu C. , Shang J. , Yan X. , Liao P. , Zhu Y. , Gong Y. , Zeng B.

Acta Physiologiae Plantarum

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2016

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tom 38

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nr 07

4

Molecular cloning, characterization and expression analysis of a new gene encoding 3-hydroxy-3-methylglutaryl coenzyme A reductase from Salvia miltiorrhiza

84%

Liao P. , Zhou W. , Zhang L. , Wang J. , Yan X. , Zhang Y. , Zhang R. , Li L. , Zhou G. , Kai G.

Acta Physiologiae Plantarum

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2009

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tom 31

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nr 3

EN

The 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) catalyzes the conversion of HMG-CoA to mevalonate (MVA), which is the first committed step in MVA pathway for isoprenoid biosynthesis in plants. In this study, a full-length cDNA encodingHMGRwas isolated from Salvia miltiorrhiza by rapid amplification of cDNA ends (RACE) for the first time, which was designated as SmHMGR (Gen-Bank Accession No.EU680958). The full-length cDNA of SmHMGR was 2,115 bp containing a 1,695 bp open reading frame (ORF) encoding a polypeptide of 565 amino acids. Bioinformatic analyzes revealed that the deduced SmHMGR had extensive homology with other plant HMGRs contained two transmembrane domains and a catalytic domain. Molecular modeling showed thatSmHMGRis a newHMGR with a spatial structure similar to other plant HMGRs. Phylogenetic tree analysis indicated that SmHMGR belongs to the plant HMGR super-family and has the closest relationship with HMGR from Picrorhiza kurrooa. Expression pattern analysis implied that SmHMGR expressed highest in root, followed by stem and leaf. The expression of SmHMGR could be up-regulated by salicylic acid (SA) and methyl jasmonate (MeJA), suggesting that SmHMGR was elicitorresponsive. This work will be helpful to understand more about the role of HMGR involved in the tanshinones biosynthesis at the molecular level.

5

Molecular mechanism of elicitor-induced tanshinone accumulation in Salvia miltiorrhiza hairy root cultures

67%

Kai G. , Liao P. , Xu H. , Wang J. , Zhou C. , Zhou C. , Zhou W. , Qi Y. , Xiao J. , Wang Y. , Zhang L.

Acta Physiologiae Plantarum

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2012

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tom 34

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nr 4

EN

To develop an optimal bioprocess for the production of tanshinone which is mainly used for the treatment of cardiocerebral vascular disease, the tanshinone biosynthetic pathway regulation must be better understood. In this paper, expression of tanshinone biosynthetic pathway related genes as well as tanshinone accumulation in Salvia miltiorrhiza hairy root cultures were investigated, in response to biotic and abiotic elicitors, respectively. Our results showed tanshinone accumulation in S. miltiorrhiza hairy roots was highly regulated by the coordination of the expression of several genes involved in tanshinone biosynthesis pathway. Our results showed a positive correlation between gene expression and tanshinone accumulation, suggesting that tanshinone accumulation may be the result of the coexpression up-regulation of several genes involved in tanshinone biosynthesis under the treatment of various elicitors. Meantime, SmHMGR, SmDXS2, SmFPPS, SmGGPPS and SmCPS were identified as the potential key enzymes in the pathway for targeted metabolic engineering to increase accumulation of tanshinone in S. miltiorrhiza hairy roots. This is the first report integrating comprehensively the transcript and metabolite biosynthesis of tanshinone in S. miltiorrhiza hairy roots.