Isolation and characterization of dehydration-responsive element-binding factor 2C (MsDREB2C) from Malus sieversii Roem.
Isolation and characterization of dehydration-responsive element-binding factor 2C (MsDREB2C) from Malus sieversii Roem.
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DOI：
10.1093/pcp/pct087
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发表时间：
2013-09
影响因子：
4.9
通讯作者：
K. Zhao;Xinjie Shen;Hua-zhao Yuan;Yun Liu;Xiong Liao;Qi Wang;Linlin Liu;Fang Li;Tianhong Li
中科院分区：
文献类型：
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作者：
K. Zhao;Xinjie Shen;Hua-zhao Yuan;Yun Liu;Xiong Liao;Qi Wang;Linlin Liu;Fang Li;Tianhong Li
DREB2(脱水反应元件结合因子2)类转录因子在植物逆境相关的调控网络中起着关键作用。在本研究中，我们从海棠中分离并鉴定了一个DREB2同源物，命名为MsDREB2C(GenBank登录号：JQ790526)。在酵母菌株YGR-2中，MsDREB2C定位于核并反式激活了报告基因。实时荧光定量聚合酶链式反应分析表明，MsDREB2C在干旱、盐、寒冷、高温和ABA的诱导下呈结构性表达。过量表达MsDREB2C的转基因拟南芥植株表现出根和叶的生长和脯氨酸水平的增加，水分损失和气孔开度的减少。在对照和非生物胁迫条件下，转基因拟南芥植株中脱水反应元件下游功能基因的转录水平高于野生型植株。此外，在对照和致病条件下，MsDREB2C的结构性表达抑制了病程相关(PR)基因的表达和过氧化物酶的活性。结果表明，转基因植株具有较强的耐旱性、耐热性和耐寒性，但对PST DC3000(紫丁香假单胞菌)的抗性较强。番茄DC3000)侵染比对照植株高。对MsDREB2C启动子在转基因烟草植株中的β-葡萄糖醛酸酶表达分析表明，MsDREB2C主要在维管组织和种子中表达。缺失分析确定了植物对干旱(-831到-680)、脱落酸(-831到-680和-335到-148)、盐(-831到-335)、冷(-1,317到-831和-335到-148)和热(-335到-148)的调控区域。
DREB2 (dehydration-responsive element-binding factor 2)-type transcription factors play a critical role in the stress-related regulation network in plants. In this study, we isolated and characterized a DREB2 homolog from Malus sieversii Roem., designated MsDREB2C (GenBank accession No. JQ790526). MsDREB2C localized to the nucleus and transactivated reporter genes in yeast strain YGR-2. Quantitative real-time PCR analysis demonstrated that MsDREB2C was constitutively expressed and significantly induced by drought, salt, cold, heat and ABA. Transgenic Arabidopsis plants overexpressing MsDREB2C exhibited increased root and leaf growth and proline levels, and reduced water loss and stomatal aperture. The transcriptional level of genes that function downstream of dehydration-responsive elements was greater in the transgenic Arabidopsis plants than in wild-type plants under control and abiotic stress conditions. Furthermore, constitutive expression of MsDREB2C repressed the expression of pathogenesis-related (PR) genes and the activity of peroxidase in transgenic plants under control and pathogenic conditions. As a result, transgenic plants were more tolerant to drought, heat and cold, but more sensitive to Pst DC3000 (Pseudomonas syringae pv . tomato DC3000) infection than control plants. β-Glucuronidase expression analysis of the MsDREB2C promoter in transgenic tobacco plants showed that MsDREB2C was mainly expressed in the vascular tissues and seeds. Deletion analysis identified the regulatory regions responsible for the plant's response to drought (-831 to -680), ABA (-831 to -680 and -335 to -148), salt (-831 to -335), cold (-1,317 to -831 and -335 to -148) and heat (-335 to -148).