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一种来自甘薯的新型Cys2/His2锌指蛋白基因IbZFP1,参与转基因拟南芥的耐盐和耐旱过程。

A novel Cys2/His2 zinc finger protein gene from sweetpotato, IbZFP1, is involved in salt and drought tolerance in transgenic Arabidopsis.

作者信息

Wang Feibing, Tong Wenjie, Zhu Hong, Kong Weili, Peng Rihe, Liu Qingchang, Yao Quanhong

机构信息

Shanghai Key Laboratory of Agricultural Genetic Breeding, Biotech Research Institute, Shanghai Academy of Agricultural Sciences, Shanghai, 201106, China.

Yunnan Academy of Tobacco Agricultural Sciences, Kunming, 650021, China.

出版信息

Planta. 2016 Mar;243(3):783-97. doi: 10.1007/s00425-015-2443-9. Epub 2015 Dec 21.

Abstract

IbZFP1, encoding a Cys 2/His 2 zinc finger protein gene from sweetpotato, enhances salt and drought tolerance in transgenic Arabidopsis by regulating ABA signaling pathway, proline biosynthesis, stress responses and ROS scavenging. In plants, Cys2/His2 zinc finger proteins play important roles in regulating the growth and development or responses to abiotic stresses. In this study, a novel Cys2/His2 zinc finger protein gene, named IbZFP1, was isolated from drought-tolerant sweetpotato [Ipomoea batatas (L.) Lam.] line Xu55-2. Subcellular localization analysis in onion epidermal cells indicated that IbZFP1 was localized to the nucleus. Expression analysis in yeast showed that the full length of IbZFP1 exhibited transcriptional activation. Expression of IbZFP1 was induced by NaCl, polyethylene glycol and abscisic acid (ABA). Overexpression of IbZFP1 significantly enhanced salt and drought tolerance in transgenic Arabidopsis plants. Real-time quantitative PCR (qRT-PCR) analysis showed that overexpression of IbZFP1 up-regulated the genes involved in ABA signaling pathway, proline biosynthesis, stress responses, and ROS scavenging under salt and drought stresses. Meanwhile, Western blot and enzymatic analyses showed that the activities of 9-cis-epoxycarotenoid dioxygenase, pyrroline-5-carboxylate synthase, superoxide dismutase, catalase, ascorbate peroxidase, and peroxidase were also increased. Further component analyses indicated that the significant increase of ABA, proline, soluble sugar and total chlorophyll content and the significant reduction of H2O2 and malonaldehyde content were observed under salt and drought stresses. In addition, the rates of electrolyte leakage and water loss were reduced in transgenic plants. The overall results demonstrate the explicit role of IbZFP1 in conferring salt and drought tolerance in transgenic Arabidopsis plants. The IbZFP1 gene has the potential to be used to enhance the tolerance to abiotic stresses in plants.

摘要

IbZFP1编码一种来自甘薯的Cys2/His2锌指蛋白基因,通过调节脱落酸(ABA)信号通路、脯氨酸生物合成、应激反应和活性氧清除,增强转基因拟南芥的耐盐性和耐旱性。在植物中,Cys2/His2锌指蛋白在调节生长发育或对非生物胁迫的响应中发挥重要作用。在本研究中,从耐旱甘薯[Ipomoea batatas (L.) Lam.]品系徐55-2中分离出一个新的Cys2/His2锌指蛋白基因,命名为IbZFP1。洋葱表皮细胞中的亚细胞定位分析表明,IbZFP1定位于细胞核。酵母中的表达分析表明,IbZFP1的全长具有转录激活作用。IbZFP1的表达受NaCl、聚乙二醇和脱落酸(ABA)诱导。IbZFP1的过表达显著增强了转基因拟南芥植株的耐盐性和耐旱性。实时定量PCR(qRT-PCR)分析表明,在盐胁迫和干旱胁迫下,IbZFP1的过表达上调了参与ABA信号通路、脯氨酸生物合成、应激反应和活性氧清除的基因。同时,蛋白质免疫印迹和酶活性分析表明,9-顺式环氧类胡萝卜素双加氧酶、吡咯啉-5-羧酸合成酶、超氧化物歧化酶、过氧化氢酶、抗坏血酸过氧化物酶和过氧化物酶的活性也有所增加。进一步的成分分析表明,在盐胁迫和干旱胁迫下,ABA、脯氨酸、可溶性糖和总叶绿素含量显著增加,H2O2和丙二醛含量显著降低。此外,转基因植株的电解质渗漏率和水分流失率降低。总体结果表明IbZFP1在赋予转基因拟南芥植株耐盐性和耐旱性方面具有明确作用。IbZFP1基因具有增强植物对非生物胁迫耐受性的潜力。

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