Molecular cloning and characterization of the promoter of SmGGPPs and its expression pattern in Salvia miltiorrhiza
Molecular cloning and characterization of the promoter of SmGGPPs and its expression pattern in Salvia miltiorrhiza
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丹参SmGGPPs启动子的分子克隆、特性及其表达模式
DOI：
10.1007/s11033-011-1388-8
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发表时间：
2012-05-01
影响因子：
2.8
通讯作者：
Wang, Zhezhi
中科院分区：
文献类型：
--
作者：
Hua, Wenping;Song, Jie;Wang, Zhezhi
香叶基香叶基二磷酸合成酶（GGPP）是萜类化合物生物合成中的一个重要分支点酶。它调节二萜类化合物的形成，如丹参酮。我们克隆了一个基因的GGPP合酶SmGGPPs参与二萜类化合物的生物合成丹参。该基因长2，767 bp，包含一个内含子和两个外显子，编码364个氨基酸残基的多肽。利用生物信息学方法对SmGGPPs的5 '侧翼序列进行了分析。对SmGGPPs基因启动子的缺失分析表明，SmGGPPs基因启动子受热、冷诱导。为了进一步寻找SmGGPPs 5 '侧翼序列中参与诱导调控的顺式元件，利用PlantCARE和PLACE数据库预测了许多推测的顺式元件。一组推定的顺式作用元件参与诱导调节，包括G-Box、WRKY、MYC和ATCT基序。实时荧光定量PCR分析表明，SmGGPPs主要在叶片中表达，并且受多种因素的诱导，如NaCl、伤害、高温、黑暗、病原菌、茉莉酸甲酯、脱落酸、水杨酸、赤霉素等。本研究为进一步研究SmGGPPs及其对丹参酮生物合成过程的调控作用提供了有用的信息，为进一步提高丹参中丹参酮的含量提供了理论依据。丹参。
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.