茶树捕光色素蛋白复合体基因CsLhcb2的鉴定及低温响应分析

胡志航; 秦志远; 李静文; 杨妮; 陈益; 李彤; 庄静

doi:10.13305/j.cnki.jts.2023.02.007

茶叶科学 >

2023 , Vol. 43 >Issue 2: 183 - 193

DOI: https://doi.org/10.13305/j.cnki.jts.2023.02.007

研究报告

茶树捕光色素蛋白复合体基因CsLhcb2的鉴定及低温响应分析

胡志航 ,
秦志远 ,
李静文 ,
杨妮 ,
陈益 ,
李彤 ,
庄静

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1.南京农业大学园艺学院,茶叶科学研究所,农业农村部华东地区园艺作物生物学与种质创制重点实验室,江苏南京 210095;
2.南京农业大学,作物遗传与种质创新利用全国重点实验室,江苏南京 210095

胡志航,男,博士研究生,主要从事茶树遗传育种与分子生物学研究。

收稿日期: 2022-12-09

修回日期: 2023-03-02

网络出版日期: 2023-05-05

基金资助

国家自然科学基金（31870681）、江苏省政策引导类计划（SZ-LYG202126）、江苏高校优势学科建设项目（PAPD）

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Identification of the Light-harvesting Chlorophyll-protein Complex Gene CsLhcb2 and Its Response to Low Temperature in Tea Plants

HU Zhihang ,
QIN Zhiyuan ,
LI Jingwen ,
YANG Ni ,
CHEN Yi ,
LI Tong ,
ZHUANG Jing

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1. Ministry of Agriculture and Rural Affairs Key Laboratory of Biology and Germplasm Enhancement of Horticultural Crops in East China, Tea Research Institute, College of Horticulture, Nanjing Agricultural University, Nanjing 210095, China;
2. National Key Laboratory of Crop Genetics & Germplasm Enhancement and Utilization, Nanjing Agricultural University, Nanjing 210095, China

Received date: 2022-12-09

Revised date: 2023-03-02

Online published: 2023-05-05

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摘要

茶树是我国重要的经济作物之一,其生长和发育过程中会遭受不同逆境影响,导致茶叶产量和品质下降。捕光色素蛋白复合体（Light-harvesting chlorophyll-protein complex）主要影响植物光合效率,在植物适应环境胁迫方面也发挥着重要作用。为研究茶树捕光色素蛋白复合体的特性,从龙井43克隆获得编码捕光色素蛋白复合体基因CsLhcb2,分析该基因编码蛋白序列特征、进化树、理化性质、亚细胞定位、二级结构、三级结构及其在4 ℃低温处理的表达情况。结果表明,CsLhcb2开放阅读框为798 bp,共编码265个氨基酸;该基因含有典型的Chloroa-b-bind保守结构域;与15种植物的LHCB2氨基酸序列进行多重比对,氨基酸序列的相似度达91.32%。进化树分析显示,CsLHCB2与曼陀罗、东南景天、葡萄亲缘关系较近,与麻竹和毛竹亲缘关系较远。CsLHCB2分子量为28 662.77,理论等电点为5.69,属于亲水性蛋白;亚细胞定位预测结果显示CsLHCB2主要定位在叶绿体中。荧光定量结果显示,CsLhcb2可能参与茶树低温胁迫的过程。常温处理条件下,一个光周期（24 h）内CsLhcb2的相对表达量呈现先上升,后下降趋势,龙井43和舒茶早在光照处理1 h达峰值,白叶1号在光照处理6 h达峰值;4 ℃低温条件下,3个茶树品种中CsLhcb2的表达均在光照处理12 h达到峰值,舒茶早中CsLhcb2表达量较高,分别为龙井43和白叶1号的1.18倍和1.98倍。研究结果为进一步研究茶树捕光色素蛋白复合体在茶树低温响应中的作用提供了参考。

关键词： 茶树; 捕光色素蛋白复合体; 低温胁迫; 表达分析

本文引用格式

胡志航 , 秦志远 , 李静文 , 杨妮 , 陈益 , 李彤 , 庄静 . 茶树捕光色素蛋白复合体基因CsLhcb2的鉴定及低温响应分析[J]. 茶叶科学, 2023 , 43(2) : 183 -193 . DOI: 10.13305/j.cnki.jts.2023.02.007

Abstract

Tea is one of the important cash crop in China. Its growth and development will be affected by different adversity, leading to the decline of tea quality and yield. Light-harvesting chlorophyll-protein complex mainly affects the photosynthetic efficiency of plants, and also plays important roles in adaptation to environmental stresses. In order to study the characteristics of the light-harvesting chlorophyll-protein complex in tea plants, the gene CsLhcb2 encoding the light-harvesting protein complex was cloned from tea cultivar ‘Longjing 43’, and the sequence characteristics, phylogenetic tree, physical and chemical properties, subcellular localization, secondary structure, tertiary structure and its expression profiles under low temperature treatment were analyzed. The results show that the open reading frame of CsLhcb2 gene is 798 bp, encoding 265 amino acids. This gene contains a typical of Chloroa-b-bind conservation domain. The similarity of CsLHCB2 amino acid sequence with 15 plant species was 91.32%. Phylogenetic tree analysis shows that the CsLHCB2 protein of tea plant was closely related to Datura stramonium, Sedum alfredii and Vitis vinifera, and far from Dendrocalamus latiflorus and Phyllostachys edulis. The relative molecular weight of CsLHCB2 protein is 28 662.77 and the theoretical isoelectric point is 5.69, which belongs to hydrophilic protein. Subcellular localization prediction results show that CsLHCB2 protein is mainly located in chloroplasts. Quantitative RT-PCR results show that CsLhcb2 gene may participate in the process of low temperature stress in tea plants. Under normal temperature treatment, the relative expression level of CsLhcb2 gene showed a trend of increasing first and then decreasing within a photoperiod (24 h). ‘Longjing 43’ and ‘Shuchazao’ reached their peak value at 1 h after light treatment, and 'Baiyeyihao' reached their peak value at 6 h after light treatment. Under low temperature of 4 ℃ treatment, the expression of Lhcb2 of the three tea cultivars all reached the peak at 12 h of light treatment, among which the expression level of CsLhcb2 in ‘Shuchazao’ was the highest, which was 1.18 and 1.98 times higher than that of ‘Longjing 43’ and ‘Baiyeyihao’, respectively. The results provided a reference for further research on the role of light-harvesting chlorophyll-protein complex response to low temperature in tea plants.

Key words： Camellia sinensis; light-harvesting chlorophyll-protein complex; low temperature stress; expression analysis

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