木薯糖转运蛋白基因MeSWEET17b的克隆及表达分析

薛晶晶<sup>1; 2</sup>; 韦卓文<sup>1</sup>; 罗秀芹<sup>1</sup>; 安飞飞<sup>1; 2*</sup>

引用本文:	薛晶晶, 韦卓文, 罗秀芹, 安飞飞.木薯糖转运蛋白基因MeSWEET17b的克隆及表达分析[J].广西植物,2024,44(12):2212-2221.[点击复制]
	XUE Jingjing, WEI Zhuowen, LUO Xiuqin, AN Feifei.Cloning and expression analysis of sugar transporter protein gene MeSWEET17b in cassava[J].Guihaia,2024,44(12):2212-2221.[点击复制]

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*木薯糖转运蛋白基因MeSWEET17b的克隆及表达分析*
薛晶晶^1,2, 韦卓文¹, 罗秀芹¹, 安飞飞^1,2*
1. 中国热带农业科学院热带作物品种资源研究所/农业农村部木薯种质资源保护与利用重点实验室, 海口571101;2. 中国热带农业科学院三亚研究院, 海南三亚 572025

摘要:

木薯(Manihot esculenta)是热带亚热带地区重要的粮食作物。糖转运蛋白SWEETs能够促进糖在细胞间的流动,在植物生长发育过程中起着重要作用。为明确 SWEET 家族基因在木薯生长发育过程中的功能,该研究以木薯‘KU50'为实验材料,采用基因克隆、生物信息学分析、亚细胞定位、体外酵母检测及RT-qPCR等方法研究木薯MeSWEET17b的基因特性。结果表明:(1)MeSWEET17b基因开放阅读框为726 bp,编码242个氨基酸,定位于细胞膜; MeSWEET17b蛋白与AtSWEET16、AtSWEET17 亲缘关系较近,含有7个跨膜结构域,属于疏水性蛋白。(2)体外酵母检测表明MeSWEET17b主要转运果糖。(3)RT-qPCR结果表明,MeSWEET17b在叶柄和茎杆中表达趋势基本一致,成熟期相对表达量最高,在叶片中相对表达量较低,在块根膨大期相对表达量最高,随着块根生长发育,相对表达量急速下降。(4)对‘KU50'水培苗进行高盐(8 g·L^-1 NaCl)、干旱(100 mmol·L^-1甘露醇)、氧化(10% H₂O₂)和低温(15 ℃ 24 h,后降至4 ℃ 24 h)等非生物胁迫处理。RT-qPCR结果显示,干旱胁迫下,MeSWEET17b在叶片和茎杆中的相对表达量变化差异最大; 盐胁迫下,MeSWEET17b在叶片和须根中的相对表达量变化最显著; 氧化和低温胁迫下,MeSWEET17b在须根中和叶柄中的相对表达量随着处理时间的延长呈极显著上升趋势。该研究为进一步研究糖转运蛋白SWEETs在木薯中的作用机制提供依据。

关键词: 木薯, 糖转运蛋白, 亚细胞定位, 糖转运能力, 非生物胁迫

DOI：10.11931/guihaia.gxzw202310038

分类号:Q943

文章编号:1000-3142(2024)12-2212-10

基金项目:中国热带农业科学院国家热带农业科学中心科技创新团队(CATASCXTD202301)。

Cloning and expression analysis of sugar transporter protein gene MeSWEET17b in cassava

XUE Jingjing^1,2, WEI Zhuowen¹, LUO Xiuqin¹, AN Feifei^1,2*

1. Tropical Crops Genetic Resources Institute, Chinese Academy of Tropical Agricultural Sciences / Key Laboratory of Conservation and Utilization for Cassava Germplasm Resources, Haikou 571101, China;2. Sanya Research Institute of Chinese Academy of Tropical Agricultural Sciences, Sanya 572025, Hainan, China 1. Tropical Crops Genetic Resources Institute, Chinese Academy of Tropical Agricultural Sciences / Key Laboratory of Conservation and Utilization for Cassava Germplasm Resources, Haikou 571101, China; 2. Sanya Research Institute of Chinese Academy of Tropical Agricultural Sciences, Sanya 572025, Hainan, China

Abstract:

Cassava(Manihot esculenta)is an important food crop in tropical and subtropical regions. Sugar transporter protein SWEETs facilitate the flow of sugar between cells and play an important role in plant growth and development. In order to clarify the function of SWEET family genes in cassava, cassava‘KU50' was used as material in this study, and the gene properties of MeSWEET17b were studied by gene cloning, bioinformatics analysis, subcellular localization, in vitro yeast detection and RT-qPCR, etc. The results were as follows:(1)The open reading frame of MeSWEET17b was 726 bp, encoding 242 amino acids, and located in the plasma membrane. MeSWEET17b had the closer genetic relationship with AtSWEET16 and AtSWEET17, containing 7 transmembrane domains, and belonging to hydrophobic protein.(2)MeSWEET17b mainly transport fructose through the in vitro yeast detection.(3)The results of RT-qPCR showed that the expression trend of MeSWEET17b in stem was basically consistent with in petiole, and the expression was the highest at maturity. The relative expression of MeSWEET17b was relatively low in leaves, and the highest in the expansion stage of tuberous root, while decreased rapidly with the growth of tuberous roots.(4)The ‘KU50' seedlings were subjected to abiotic stress treatments such as high salt(8 g·L^-1 NaCl), drought(100 mmol·L^-1 mannitol), oxidation(10% H₂O₂)and cold(15 ℃ for 24 h, then dropped to 4 ℃ for 24 h). RT-qPCR showed that the relative expressions of MeSWEET17b in leaf and stem had the greatest difference under drought stress. The relative expressions of MeSWEET17b in leaf and fibrous root changed most significantly under salt stress; under oxidation and cold stress, the relative expressions of MeSWEET17b in fibrous root and petiole increased significantly with the extention of treating time. This study provides a theoreticac reference for further studying the function mechanism of sugar transporter protein SWEETs in cassava.

Key words: cassava(Manihot esculenta), sugar transporter protein, subcellular localization, sugar transport capacity, abiotic stress