Functional characterization of OsHAK1 promoter in response to osmotic/drought stress by deletion analysis in transgenic rice
Functional characterization of OsHAK1 promoter in response to osmotic/drought stress by deletion analysis in transgenic rice
复制标题
通过转基因水稻缺失分析来表征 OsHAK1 启动子响应渗透/干旱胁迫的功能
DOI：
10.1007/s10725-019-00504-3
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发表时间：
2019-07-01
影响因子：
4.2
通讯作者：
Qian, Qian
中科院分区：
文献类型：
--
作者：
Chen, Guang;Hu, Jiang;Qian, Qian
水稻基因HAK 1（OsHAK 1）在植物经历干旱胁迫时被激活。在此，对位于HAK 1翻译起始密码子上游的3037 nt序列进行缺失分析，以鉴定哪个启动子区域对于其对水分胁迫的响应性在功能上是重要的。将4个5′端截短的启动子序列（Dp 1820、Dp 1524、Dp 1069和Dp 556）与完整的启动子序列（Dp 3037）沿着融合到GUS基因上，分别转化水稻。在不存在任何强加的非生物胁迫的情况下，GUS活性在携带Dp 3037和Dp 1820构建体的植物中是普遍存在的并且具有相似的强度，但是在携带Dp 1524、Dp 1069或Dp 556的那些植物中是检测不到的。当通过向培养基中添加聚乙二醇使植物经受水分胁迫时，Dp 3037的载体强烈表达GUS，但Dp 1820的载体没有反应，这意味着位于核苷酸-3037和-1821之间的序列负责渗透胁迫诱导。该关键序列的计算机分析揭示了许多推定的顺式作用元件的存在，包括ABRE（脱落酸响应）和DRE（脱水响应）。核苷酸-1820和-1525之间的296 nt片段包含CAAT盒和TATA盒序列。Dp 3037序列代表了调节干旱响应转基因表达的潜在合适候选序列。
The rice gene HAK1 (OsHAK1) is activated when the plant experiences drought stress. Here, a deletion analysis of the 3037 nt sequence lying upstream of the HAK1 translation initiation codon was carried out to identify which promoter region(s) are functionally important for its responsiveness to moisture stress. Four 5′ truncated sequences of the promoter (Dp1820, Dp1524, Dp1069 and Dp556) along with the intact sequence (Dp3037) were fused to GUS, and the resulting constructs transformed separately into rice. In the absence of any imposed abiotic stress, GUS activity was both ubiquitous and of similar intensity in plants carrying the Dp3037 and Dp1820 constructs, but was undetectable in those carrying either Dp1524, Dp1069 or Dp556. When the plants were subjected to moisture stress by the addition of polyethylene glycol to the culture medium, carriers of Dp3037 strongly expressed GUS, but carriers of Dp1820 did not respond, implying that a sequence(s) lying between the nucleotides − 3037 and − 1821 were responsible for osmotic stress inducibility. An in silico analysis of this key sequence revealed the presence of a number of putative cis-acting elements, including ABREs (abscisic acid-responsive) and DREs (dehydration-responsive). The 296 nt segment between the nucleotides − 1820 and − 1525 harbored both a CAAT-box and a TATA-box sequence. The Dp3037 sequence represents a potentially suitable candidate for regulating the expression of drought-responsive transgenes.