Zhang Hongying, Jing Ruilian, Mao Xinguo
College of Tobacco Science, Henan Agricultural UniversityZhengzhou, China.
The National Key Facility for Crop Gene Resources and Genetic Improvement, Institute of Crop Science, Chinese Academy of Agricultural SciencesBeijing, China.
Front Plant Sci. 2017 Jul 13;8:1198. doi: 10.3389/fpls.2017.01198. eCollection 2017.
Drought, salinity, and cold are the major factors limiting wheat quality and productivity; it is thus highly desirable to characterize the abiotic-stress-inducible promoters suitable for the genetic improvement of plant resistance. The sucrose non-fermenting 1-related protein kinase 2 () family genes show distinct regulatory properties in response to abiotic stresses. The present study characterized the approximately 3000-bp upstream sequence (the 313 bp upstream of the ATG was the transcription start site) of the promoter under abscisic acid (ABA) and abiotic stresses. Four different-length 5' deletion fragments of promoter were fused with the reporter gene and transformed into . Tissue expression analysis showed that the promoter region from position -1481 to -821 contained the stalk-specific elements, and the region from position -2631 to -1481 contained the leaf- and root-specific elements. In the ABA-treated seedlings, the deletion analysis showed that the promoter region from position -821 to -2631 contained ABA response elements. The abiotic stress responses of the promoter derivatives demonstrated that they harbored abiotic-stress response elements: the region from position -821 to -408 harbored the osmotic-stress response elements, whereas the region from position -2631 to -1481 contained the positive regulatory motifs and the region from position -1481 to -821 contained the leaf- and stalk-specific enhancers. Further deletion analysis of the promoter region from position -821 to -408 indicated that a 125-bp region from position -693 to -568 was required to induce an osmotic-stress response. These results contribute to a better understanding of the molecular mechanisms of in response to abiotic stresses, and the promoter seems to be a candidate for regulating the expression of abiotic stress response genes in transgenic plants.
干旱、盐度和低温是限制小麦品质和产量的主要因素;因此,非常需要鉴定适合用于植物抗性基因改良的非生物胁迫诱导型启动子。蔗糖非发酵1相关蛋白激酶2(SnRK2)家族基因在响应非生物胁迫时表现出不同的调控特性。本研究对脱落酸(ABA)和非生物胁迫下SnRK2.4启动子约3000 bp的上游序列(ATG上游313 bp为转录起始位点)进行了表征。将SnRK2.4启动子的四个不同长度的5'缺失片段与GUS报告基因融合并转化到拟南芥中。组织表达分析表明,-1481至-821位的SnRK2.4启动子区域包含茎特异性元件,-2631至-1481位的区域包含叶和根特异性元件。在ABA处理的幼苗中,缺失分析表明,-821至-2631位的SnRK2.4启动子区域包含ABA响应元件。SnRK2.4启动子衍生物的非生物胁迫响应表明它们含有非生物胁迫响应元件:-821至-408位的区域含有渗透胁迫响应元件,而-2631至-1481位的区域包含正调控基序,-1481至-821位的区域包含叶和茎特异性增强子。对-821至-408位启动子区域的进一步缺失分析表明,-693至-568位的125 bp区域是诱导渗透胁迫响应所必需的。这些结果有助于更好地理解SnRK2.4响应非生物胁迫的分子机制,并且SnRK2.4启动子似乎是调控转基因植物中非生物胁迫响应基因表达的候选者。