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通过CRISPR/Cas9介导番茄植株中SlMAPK3诱变降低耐旱性

Reduced Drought Tolerance by CRISPR/Cas9-Mediated SlMAPK3 Mutagenesis in Tomato Plants.

作者信息

Wang Liu, Chen Lin, Li Rui, Zhao Ruirui, Yang Meijing, Sheng Jiping, Shen Lin

机构信息

College of Food Science and Nutritional Engineering, China Agricultural University , Beijing 100083, China.

School of Agricultural Economics and Rural Development, Renmin University of China , Beijing 100872, China.

出版信息

J Agric Food Chem. 2017 Oct 4;65(39):8674-8682. doi: 10.1021/acs.jafc.7b02745. Epub 2017 Sep 25.

Abstract

Drought stress is one of the most destructive environmental factors that affect tomato plants adversely. Mitogen-activated protein kinases (MAPKs) are important signaling molecules that respond to drought stress. In this study, SlMAPK3 was induced by drought stress, and the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (CRISPR/Cas9) system was utilized to generate slmapk3 mutants. Two independent T1 transgenic lines and wild-type (WT) tomato plants were used for analysis of drought tolerance. Compared with WT plants, slmapk3 mutants exhibited more severe wilting symptom, higher hydrogen peroxide content, lower antioxidant enzymes activities, and suffered more membrane damage under drought stress. Furthermore, knockout of SlMAPK3 led to up- or down-regulated expressions of drought stress-responsive genes including SlLOX, SlGST, and SlDREB. The results suggest that SlMAPK3 is involved in drought response in tomato plants by protecting cell membranes from oxidative damage and modulating transcription of stress-related genes.

摘要

干旱胁迫是对番茄植株产生不利影响的最具破坏性的环境因素之一。丝裂原活化蛋白激酶(MAPK)是对干旱胁迫作出响应的重要信号分子。在本研究中,SlMAPK3受干旱胁迫诱导,利用成簇规律间隔短回文重复序列(CRISPR)/CRISPR相关蛋白9(CRISPR/Cas9)系统生成了slmapk3突变体。使用两个独立的T1转基因系和野生型(WT)番茄植株进行耐旱性分析。与WT植株相比,slmapk3突变体在干旱胁迫下表现出更严重的萎蔫症状、更高的过氧化氢含量、更低的抗氧化酶活性,并且遭受了更多的膜损伤。此外,SlMAPK3的敲除导致干旱胁迫响应基因(包括SlLOX、SlGST和SlDREB)的表达上调或下调。结果表明,SlMAPK3通过保护细胞膜免受氧化损伤和调节胁迫相关基因的转录参与番茄植株的干旱响应。

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