Vascular preferential activity of the Pennisetum purpureum cinnamyl alcohol dehydrogenase promoter in transgenic tobacco plants.
Vascular preferential activity of the Pennisetum purpureum cinnamyl alcohol dehydrogenase promoter in transgenic tobacco plants.
复制标题
DOI：
10.1016/j.plaphy.2018.04.010
复制
发表时间：
2018-08
期刊：
影响因子：
--
通讯作者：
Tian-Xiu Zhong;Ran Tang;J. Song;Cheng-Cheng Fu-Cheng;Yang Liu;Cong-Cong Zhou-Cong;Xiang-Qian Zhang;Shu Chen;Xin-Ming Xie
中科院分区：
文献类型：
--
作者：
Tian-Xiu Zhong;Ran Tang;J. Song;Cheng-Cheng Fu-Cheng;Yang Liu;Cong-Cong Zhou-Cong;Xiang-Qian Zhang;Shu Chen;Xin-Ming Xie
木质素生物合成基因启动子与调节分子信号和响应各种逆境的调节蛋白之间的相互作用知之甚少。在本研究中，我们对象草中木质素生物合成途径肉桂醇脱氢酶(CAD)基因的启动子区域进行了研究。对PpCAD启动子转录水平的定量分析表明，PpCAD启动子在维管组织特别是木质部中优先表达。组织化学和荧光分析证实了PpCAD启动子的血管优势表达，因为在表达1154bpPpCAD启动子-GUS融合结构的转基因烟草植株中，β-葡萄糖醛酸苷酶(GUS)活性在基茎中最高。此外，5‘-缺失的PpCAD启动子分析表明，1154-bpPpCAD启动子片段具有最高的转录活性，而2054-BP片段具有多种诱导活性，对赤霉素(GA)、茉莉酸甲酯(MeJA)、脱落酸(ABA)和伤害做出反应。−2 48~−2 43bp和−1416~−1411bp区域含有W-盒顺式元件，用凝胶迁移率改变分析检测到。对GA反应元件(−561~−555bp和−1077~−1071bp)、MeJA反应元件(−1146~−1142bp)和ABA反应元件(−1879~−1874bp)的结合效应也进行了验证。根据我们的结果，我们认为与PpCAD启动子活性相关的木质素沉积通过涉及GA、MeJA和ABA的分子信号来适应环境。
Little is known about the cross talk between the lignin biosynthesis gene promoters and the regulatory proteins that modulate molecular signaling and respond to various stresses. In this study, we characterized the promoter region of the lignin biosynthesis pathway cinnamyl alcohol dehydrogenase (CAD) gene in elephant grass,Pennisetumpurpureum. Quantification of the transcript levels of thePpCADpromoter revealed it is preferentially expressed in vascular tissue, especially xylem. Histochemical and fluorometric assays confirmed the vascular-preferential expression of thePpCADpromoter, as the highest β-glucuronidase (GUS) activity was found in the basal stem in transgenic tobacco plants expressing a 1154-bpPpCADpromoter-GUS fusion construct. Moreover, 5′-deletedPpCADpromoter analyses showed that the 1154-bpPpCADpromoter fragment had the highest transcriptional activity, whereas the 2054-bp fragment had multifarious inducible activity responding to gibberellin (GA), methyl jasmonate (MeJA), abscisic acid (ABA), and wounding. The regions from −248 to −243 bp and −1416 to −1411 bp contained W-boxcis-elements, which were detected by electrophoretic mobility shift assay (EMSA). The binding effects of the GA-responsive elements (from −561 to −555 bp and −1077 to −1071 bp), MeJA-responsive element (from −1146 to −1142 bp), and the ABA-responsivecis-element (from −1879 to −1874 bp) were also validated by EMSA. Based on our results, we suggest that lignin deposition associated withPpCADpromoter activity adapts to the environment through molecular signaling involving GA, MeJA, and ABA.