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拟南芥SWEET1/2/3基因突变体构建及功能鉴定

文献类型: 中文期刊

作者: 辛红佳 1 ; 李鹏程 1 ; 滕守振 1 ; 李圣彦 1 ; 汪海 1 ; 郎志宏 1 ;

作者机构: 1.中国农业科学院生物技术研究所

关键词: CRISPR/Cas9;拟南芥SWEET1/2/3;葡萄糖敏感性;角果发育

期刊名称: 中国农业科技导报

ISSN: 1008-0864

年卷期: 2020 年 02 期

页码: 39-49

收录情况: 北大核心 ; CSCD

摘要: SWEET(sugars will eventually be exported transporters)基因广泛存在于植物、动物和微生物中,在细胞膜或者细胞器膜形成跨膜孔道,协助糖类物质完成跨膜运输。拟南芥SWEET1/2/3基因属于SWEET基因家族CladeⅠ分支。通过表达谱数据分析发现,SWEET1基因在花器官优先表达,SWEET2基因在营养生长和生殖生长期均有表达,SWEET3基因在花中表达。三个基因在体外鉴定具有葡萄糖转运活性,但由于缺少功能缺失的突变体和功能冗余,它们的生理功能仍不清楚。通过CRISPR/Cas9系统在SWEET1/2/3基因中创建了靶向突变,鉴定获得了sweet1、sweet1/2、sweet3和sweet1/2/3突变体。突变体在营养生长过程中表型与野生型相同,但它们的角果长度显著短于野生型的角果。在高浓度葡萄糖培养基上,sweet1、sweet1/2和sweet1/2/3突变体对葡萄糖敏感,其特征是根更短,高度严重降低,表明SWEET1/2/3基因在葡萄糖信号中具有功能。对葡萄糖信号通路中关键基因的进一步分析发现,HXK1、KIN10和KIN11在野生型和突变体之间转录和翻译水平没有显著差异。结果表明,拟南芥SWEET1/2/3基因在葡萄糖信号传导和调节角果的发育中起着重要作用。

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