Two jasmonate-responsive factors, TcERF12 and TcERF15, respectively act as repressor and activator of tasy gene of taxol biosynthesis in Taxus chinensis
Two jasmonate-responsive factors, TcERF12 and TcERF15, respectively act as repressor and activator of tasy gene of taxol biosynthesis in Taxus chinensis
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红豆杉中两个茉莉酸反应因子TcERF12和TcERF15分别作为紫杉醇生物合成tasy基因的阻遏子和激活子
DOI：
10.1007/s11103-015-0382-2
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发表时间：
2015-11-01
影响因子：
5.1
通讯作者：
Fu, Chunhua
中科院分区：
文献类型：
--
作者：
Zhang, Meng;Li, Shutao;Fu, Chunhua
茉莉酸甲酯（Methyl jasmonate，MeJA）是紫杉醇生物合成基因，特别是紫杉醇合成酶基因的最有效的诱导剂之一。然而，MeJA调节β-内酰胺酶的潜在机制仍不清楚。在本研究中，获得了一个550-bp的5 '侧翼序列，并确认为启动子的cytokine基因。缺失分析表明，包含一个GCC盒的片段从-150到-131是关键的茉莉酸（JA）响应元件，命名为JRE。以JRE为诱饵，发现了两个结合蛋白，即TcERF 12和TcERF 15。序列比对和系统发育分析表明，TcERF 12与AtERF 3相关，TcERF 15与ORA 59相关，是典型的GCC盒结合乙烯应答因子。两者在0.5h内对MeJA的反应分别为10倍和4.5倍。当这两种TcERF在红豆杉细胞中过表达时，TcERF 12过表达细胞中的ERF基因表达降低了2.1倍，而TcERF 15过表达细胞中的ERF基因表达增加了2.5倍。结果表明，TcERF 12和TcERF 15分别是负性和正性调节子，在JA信号转导到BMP 1基因中，通过结合BMP 1启动子的JRE中的GCC盒。这些结果为进一步研究紫杉醇生物合成的调控机制提供了依据。
Methyl jasmonate (MeJA) is one of the most effective inducers of taxol biosynthetic genes, particularly the tasy gene. However, the mechanism underlying the regulation of tasy by MeJA is still unknown. In this study, a 550-bp 5'-flanking sequence was obtained and confirmed as the promoter of the tasy gene. Deletion analysis revealed that the fragment containing a GCC-box from -150 to -131 was the crucial jasmonate (JA)-responsive element, designated as JRE. Using JRE as bait, two binding proteins, namely TcERF12 and TcERF15, were discovered. Sequence alignment and phylogenetic analysis showed that TcERF12 was related to the repressor AtERF3, while TcERF15 was more related to the activator ORA59; these are typical GCC-box-binding ethylene-responsive factors. Both could significantly respond to MeJA for 10 and 4.5 times, respectively, in 0.5 h. When the two TcERFs were overexpressed in Taxus cells, tasy gene expression decreased by 2.1 times in TcERF12-overexpressing cells, but increased by 2.5 times in TcERF15-overexpressing cells. Results indicated that TcERF12 and TcERF15 were negative and positive regulators, respectively, in the JA signal transduction to the tasy gene by binding the GCC-box in the JRE of the tasy promoter. Our results promote further research on regulatory mechanisms of taxol biosynthesis.