茶树CsMGTs基因的克隆及其镁转运功能分析

唐磊; 肖罗丹; 黄伊凡; 肖斌; 龚春梅

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茶叶科学 ›› 2021, Vol. 41 ›› Issue (6) : 761-776.

研究报告

茶树CsMGTs基因的克隆及其镁转运功能分析

唐磊, 肖罗丹, 黄伊凡, 肖斌, 龚春梅^*

作者信息 +

Cloning and Magnesium Transport Function Analysis of CsMGTs Genes in Tea Plants (Camellia sinensis)

TANG Lei, XIAO Luodan, HUANG Yifan, XIAO Bin, GONG Chunmei^*

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文章历史 +

摘要

镁离子（Mg²⁺）作为叶绿素的中心成分,是植物细胞中含量最丰富的二价阳离子,也是多种酶的激活剂,特别是茶氨酸合成酶的活性依赖Mg²⁺,常被作为茶树专用肥的特征成分,因此对茶树的生长发育和茶叶的品质形成至关重要。MRS2/MGT家族镁转运蛋白在维持植物体内Mg²⁺的吸收转运、胞内平衡和耐逆性等方面起着重要的作用。为探究茶树MRS2/MGT家族镁转运蛋白基因的功能,以茶树品种陕茶1号为材料,克隆了4条MRS2/MGT镁转运蛋白基因,分别命名为CsMGT1、CsMGT2、CsMGT2.1和CsMGT3。生物信息学分析表明,这4个转运蛋白在C末端均含有2个跨膜结构域和1个保守的GMN三肽基序;系统发育分析显示,CsMGT1属于Clade C成员,而CsMGT2、CsMGT2.1和CsMGT3属于Clade B成员,所编码蛋白与木本植物柑橘MGT家族的亲缘关系最近。实时荧光定量PCR（qRT-PCR）分析表明,CsMGT1、CsMGT2、CsMGT2.1和CsMGT3在茶树的根、茎、叶、花中呈组成型表达,且根和叶对Mg²⁺浓度均表现出不同程度的响应。鼠伤寒沙门氏菌镁转运缺失突变株MM281功能互补试验表明,CsMGT1和CsMGT2均具有Mg²⁺转运功能,且CsMGT1的Mg²⁺转运功能优于CsMGT2,而CsMGT2.1和CsMGT3几乎没有镁离子转运功能。本研究结果丰富了茶树CsMGTs家族的生物学功能,为进一步阐明茶树通过镁转运功能机制实现对镁离子的利用奠定了前期研究基础。

Abstract

Magnesium (Mg²⁺), as the central atom of chlorophyll, is the most abundant divalent cation in plant cells. Magnesium is also an activator of various enzymes, especially theanine synthase, whose activity depends on Mg²⁺ concentration and is often used as a characteristic component of special fertilizer for tea plants. Therefore, it is very important for both the growth and development of tea plants and the formation of tea quality. The MRS2/MGT magnesium transporter family plays an important role in maintaining the absorption and transport of Mg²⁺, intracellular balance and stress tolerance in plants. In order to explore the functions of the MRS2/MGT magnesium transport genes in tea plants, this study cloned four MRS2/MGT magnesium transporter genes (namely CsMGT1, CsMGT2, CsMGT2.1 and CsMGT3, respectively) from tea cultivar ‘Shaancha 1'. Bioinformatics analysis shows that the four proteins all contain two transmembrane domains and a conserved GMN tripeptide motif at the C-terminal. Phylogenetic analysis shows that CsMGT1 is a member of Clade C, while CsMGT2, CsMGT2.1 and CsMGT3 belong to Clade B, and the four encoded proteins are most closely related to the woody Poncirus trifoliata MGT family. Real-time fluorescence quantitative PCR (RT-qPCR) shows that CsMGT1, CsMGT2, CsMGT2.1 and CsMGT3 were constitutively expressed in the roots, stems, leaves and flowers of tea plants, and the roots and leaves all showed different degrees of response to Mg²⁺. Functional complementation tests in a magnesium deletion mutant strain MM281 of Salmonella typhimurium show that both CsMGT1 and CsMGT2 possessed Mg²⁺ transport function, and the Mg²⁺ transport function of CsMGT1 was superior to that of CsMGT2, while CsMGT2.1 and CsMGT3 had almost no Mg²⁺ transport function. The results of this study enriched the biological functions of tea plant CsMGTs family, and laid a foundation for further utilization of magnesium through magnesium transporters in tea plants.

导出引用

唐磊, 肖罗丹, 黄伊凡, 肖斌, 龚春梅. 茶树CsMGTs基因的克隆及其镁转运功能分析[J]. 茶叶科学. 2021, 41(6): 761-776

TANG Lei, XIAO Luodan, HUANG Yifan, XIAO Bin, GONG Chunmei. Cloning and Magnesium Transport Function Analysis of CsMGTs Genes in Tea Plants (Camellia sinensis)[J]. Journal of Tea Science. 2021, 41(6): 761-776

中图分类号： S571.1 Q946.91+2

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