Molecular cloning and characterization of the promoter of SmGGPPs and its expression pattern in Salvia miltiorrhiza.
Geranylgeranyl diphosphate (GGPP) synthase is an important branch point enzyme in terpenoid biosynthesis. It regulates the formation of diterpenoid, such as tanshinones. We cloned a gene for GGPP synthase SmGGPPs involved in diterpenoid biosynthesis from Salvia miltiorrhiza. At 2,767 bp long, this gene comprises an intron and two exons that encode a polypeptide of 364 amino acid residues. Then the 5' flanking sequence of SmGGPPs was characterized by bioinformatics method. Deletion analysis of the promoter of SmGGPPs using tobacco plant displayed that the promoter was induced by heat and cold. To further search these cis-elements involved in induction regulation in the 5' flanking sequence of SmGGPPs, many putative cis-elements were predicted with the PlantCARE and PLACE databases. A group of putative cis-acting elements are involved in induction regulation, including G-Box, WRKY, MYC and ATCT motifs. Real-time PCR analysis revealed that SmGGPPs is mainly expressed in the leaves and can also be induced by various factors, such as NaCl, wounding, high temperature, darkness, pathogen, methyl jasmonate, abscisic acid, salicylic acid, and gibberellins. This study provides useful information for further study of SmGGPPs and its regulator effect on the biosynthetic process of tanshinones so that researchers can improve the tanshinone contents in S. miltiorrhiza.
Key Laboratory of the Ministry of Education for Medicinal Resources and Natural Pharmaceutical Chemistry, National Engineering Laboratory for Resource Development of Endangered Crude Drugs in Northwest of China, College of Life Sciences, Shaanxi Normal University, Xi'an, 710062, Shaanxi Province, People's Republic of China.
Wenping
W
Hua
Jie
J
Song
Cuiqin
C
Li
Zhezhi
Z
Wang
D024506:5' Flanking Region / Q000235:genetics; D001483:Base Sequence; D053898:Biosynthetic Pathways / Q000235:genetics; D003001:Cloning, Molecular; D003080:Cold Temperature; D004224:Diterpenes / Q000378:metabolism; D051231:Farnesyltranstransferase / Q000235:genetics* / Q000378:metabolism; D020869:Gene Expression Profiling; D015971:Gene Expression Regulation, Enzymologic; D018506:Gene Expression Regulation, Plant; D005966:Glucuronidase / Q000378:metabolism; D018869:Heat-Shock Response / Q000235:genetics; D008969:Molecular Sequence Data; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction; D027561:Salvia miltiorrhiza / Q000201:enzymology* / Q000235:genetics*; D014026:Nicotiana / Q000235:genetics
D004224:Diterpenes; D051231:Farnesyltranstransferase; D005966:Glucuronidase