College of Life Science, Henan Agricultural University, Zhengzhou 450002, China.
Library of Henan Agricultural University, Zhengzhou 450002, China.
Int J Mol Sci. 2019 Jul 6;20(13):3326. doi: 10.3390/ijms20133326.
Drought adversely affects crop growth and yields. The cloning and characterization of drought- or abscisic acid (ABA)-inducible promoters is of great significance for their utilization in the genetic improvement of crop resistance. Our previous studies have shown that maize sulfite oxidase (SO) has a sulfite-oxidizing function and is involved in the drought stress response. However, the promoter of the maize gene has not yet been characterized. In this study, the promoter (, 1194 bp upstream region of the translation initiation site) was isolated from the maize genome. The in-silico analysis of the promoter identified several -elements responsive to the phytohormone ABA and drought stress such as ABA-responsive element (ABRE) and MYB binding site (MBS), besides a number of core -acting elements, such as TATA-box and CAAT-box. A 5' RACE (rapid amplification of cDNA ends) assay identified an adenine residue as the transcription start site of the . The activity was detected by β-glucuronidase (GUS) staining at nearly all developmental stages and in most plant organs, except for the roots in transgenic . Moreover, its activity was significantly induced by ABA and drought stress. The 5'-deletion mutant analysis of the in tobacco plants revealed that a 119-bp fragment in the (upstream of the transcription start site) is a minimal region, which is required for its high-level expression. Moreover, the minimal was significantly activated by ABA or drought stress in transgenic plants. Further mutant analysis indicated that the MBS element in the minimal region (119 bp upstream of the transcription start site) is responsible for ABA and drought-stress induced expression. These results improve our understanding of the transcriptional regulation mechanism of the gene, and the characterized 119-bp promoter fragment could be an ideal candidate for drought-tolerant gene engineering in both monocot and dicot crops.
干旱会对作物生长和产量产生不利影响。克隆和鉴定抗旱或脱落酸(ABA)诱导启动子对于其在作物抗性遗传改良中的应用具有重要意义。我们之前的研究表明,玉米亚硫酸氧化酶(SO)具有亚硫酸盐氧化功能,并参与干旱胁迫响应。然而,玉米 基因的启动子尚未得到鉴定。在本研究中,从玉米基因组中分离出该基因的启动子(,翻译起始位点上游 1194 个碱基对)。对 启动子的计算机分析鉴定了几个响应植物激素 ABA 和干旱胁迫的 -元件,如 ABA 响应元件(ABRE)和 MYB 结合位点(MBS),以及一些核心 - 作用元件,如 TATA 盒和 CAAT 盒。5' RACE(快速扩增 cDNA 末端)分析确定了一个腺嘌呤残基作为 的转录起始位点。通过 β-葡萄糖醛酸酶(GUS)染色在转基因 的几乎所有发育阶段和大多数植物器官中检测到 的活性,但根除外。此外,它的活性被 ABA 和干旱胁迫显著诱导。烟草植物中 的 5'缺失突变分析表明, (转录起始位点上游)中的 119 个碱基对片段是一个最小区域,需要其高水平表达。此外,该最小 在转基因植物中被 ABA 或干旱胁迫显著激活。进一步的突变分析表明,最小 区域中的 MBS 元件(转录起始位点上游 119 个碱基对)负责 ABA 和干旱胁迫诱导的表达。这些结果提高了我们对 基因转录调控机制的理解,并且鉴定的最小 119 个碱基对启动子片段可以成为单子叶和双子叶作物中耐旱基因工程的理想候选者。