Characterization and expression profiling of 4-hydroxyphenylpyruvate dioxygenase gene (Smhppd) from Salvia miltiorrhiza hairy root cultures.
A novel 4-hydroxyphenylpyruvate dioxygenase gene (designated as Smhppd) was cloned from hairy roots of Salvia miltiorrhiza Bung. The full-length cDNA of Smhppd was 1,736 bp long with an ORF (open reading frame) that putatively encoded a polypeptide of 481 amino acids, with a predicted molecular mass of 52.54 kDa. The deduced amino acid sequence of the Smhppd gene shared high homology with other known HPPDs. Analysis of Smhppd genomic DNA revealed that it contained two exons and one intron. The analysis of Smhppd promoter region was also presented. Southern-blot analysis revealed that the Smhppd was a low-copy gene in S. miltiorrhiza. Real-time quantitative PCR analysis indicated that Smhppd was constitutively expressed in roots, stems and leaves of S. miltiorrhiza, with the high expression in roots. In addition, Smhppd expression level under different stress condition was also analyzed during the hairy root culture period, including signaling components for plant defence responses, such as methyl jasmonate and salicylic acid, as well as an abiotic elicitor, Ag(+) and a biotic elicitor, yeast extract. This study will enable us to further understand the role Smhppd plays in the synthesis of active pharmaceutical compounds in S. miltiorrhiza at molecular level.
Xiao
Ying
Y
Department of Pharmacognosy, School of Pharmacy, Second Military Medical University, Shanghai, China.
Di
Peng
P
Chen
Junfeng
J
Liu
Ying
Y
Chen
Wansheng
W
Zhang
Lei
L
D015111:4-Hydroxyphenylpyruvate Dioxygenase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D024506:5' Flanking Region / Q000235:genetics; D000595:Amino Acid Sequence; D001483:Base Sequence; D015139:Blotting, Southern; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D019143:Evolution, Molecular; D020869:Gene Expression Profiling; D015971:Gene Expression Regulation, Enzymologic; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant; D018745:Genome, Plant / Q000235:genetics; D053858:Metabolic Networks and Pathways; D008969:Molecular Sequence Data; D009928:Organ Specificity / Q000235:genetics; D010802:Phylogeny; D018517:Plant Roots / Q000201:enzymology* / Q000235:genetics*; D027561:Salvia miltiorrhiza / Q000201:enzymology* / Q000235:genetics*; D016415:Sequence Alignment; D046509:Tissue Culture Techniques
D018076:DNA, Complementary; D015111:4-Hydroxyphenylpyruvate Dioxygenase