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两个启动子区域赋予番茄 SlDREBA4 在热激下的激活。

Two promoter regions confer heat-induced activation of SlDREBA4 in Solanum lycopersicum.

机构信息

College of Horticulture and Landscape, Yunnan Agricultural University, Kunming, 650224, PR China.

College of Horticulture and Landscape, Yunnan Agricultural University, Kunming, 650224, PR China.

出版信息

Biochem Biophys Res Commun. 2020 Apr 9;524(3):689-695. doi: 10.1016/j.bbrc.2020.01.153. Epub 2020 Feb 4.

Abstract

Dehydration-responsive element binding (DREB) transcription factors activate the expression of downstream functional genes in combination with a dehydration-responsive element (DRE), and thereby improve the resistance of plants to abiotic stresses such as heat. However, the upstream regulatory mechanism of DREB genes under heat is unclear. A DREBA4 subfamily transcription factor (SlDREBA4), which is heat-responsive and improves heat resistance, was isolated from Solanum lycopersicum 'Microtom'. In this study, promoter truncation experiments were performed to verify changes in β-glucuronidase (GUS) enzyme activity and GUS gene expression levels in transgenic plants with different lengths of promoter fragments under heat and to identify specific regions in the promoter that respond to heat. Our results showed that the GUS reporter gene was constitutively expressed in tissues of the full-length promoter transgenic 'Microtom' plants, with higher expression in conducting tissues of root, stem, and leaf, as well as sepals of flowers and fruits. Under heat treatment, GUS enzyme activity and GUS gene expression levels in tissues of the full-length promoter transgenic plants increased. Promoter deletion analysis identified two positive regulatory regions (-1095 to -730 bp and -162 to -38 bp) responsible for the promoter's response to heat. These results indicated that the heat shock element (HSE) and MYC recognition sequences may cooperate in heat-induced activation of SlDREBA4 promoter.

摘要

脱水响应元件结合(DREB)转录因子与脱水响应元件(DRE)结合,激活下游功能基因的表达,从而提高植物对非生物胁迫(如热)的抗性。然而,DREB 基因在热胁迫下的上游调控机制尚不清楚。从番茄 'Microtom' 中分离出一种热响应且提高耐热性的 DREBA4 亚家族转录因子(SlDREBA4)。本研究通过启动子截短实验,验证了不同长度启动子片段转基因 'Microtom' 植物在热胁迫下β-葡萄糖醛酸酶(GUS)酶活性和 GUS 基因表达水平的变化,并鉴定了启动子中对热响应的特定区域。结果表明,全长启动子转基因 'Microtom' 植物组织中 GUS 报告基因持续表达,根、茎、叶的导组织以及花萼和果实中的表达水平较高。在热处理下,全长启动子转基因植物组织中的 GUS 酶活性和 GUS 基因表达水平均增加。启动子缺失分析鉴定出两个负责启动子对热响应的正调控区域(-1095 至-730 bp 和-162 至-38 bp)。这些结果表明,热休克元件(HSE)和 MYC 识别序列可能在 SlDREBA4 启动子的热诱导激活中协同作用。

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