铝诱导的茶树根系转录组变化分析

黄丹娟; 谭荣荣; 陈勋; 王红娟; 龚自明; 王友平; 毛迎新

doi:10.13305/j.cnki.jts.2019.05.002

茶叶科学 >

2019 , Vol. 39 >Issue 5: 506 - 520

DOI: https://doi.org/10.13305/j.cnki.jts.2019.05.002

铝诱导的茶树根系转录组变化分析

黄丹娟 ,
谭荣荣 ,
陈勋 ,
王红娟 ,
龚自明 ,
王友平 ,
毛迎新

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1. 湖北省农业科学院果树茶叶研究所,湖北武汉 430064;
2. 湖北省农业科学院植保土肥研究所,湖北武汉 430064

黄丹娟,女,助理研究员,主要从事茶树栽培生理研究,E-mail: huangdjtea@163.com。

收稿日期: 2019-05-21

修回日期: 2019-06-28

网络出版日期: 2019-10-15

基金资助

国家重点研发计划（2016YFD0200900）、中央引导地方科技发展专项（2018ZYYD009）、湖北省农科院青年科学基金（2018NKYJJ15）、国家茶叶产业技术体系（CARS-19）、湖北省农业科技创新中心团队（2016-620-000-001-032）

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Transcriptome Analysis of Root Induced by Aluminum in Tea Plants (Camellia sinensis)

HUANG Danjuan ,
TAN Rongrong ,
CHEN Xun ,
WANG Hongjuan ,
GONG Ziming ,
WANG Youping ,
MAO Yingxin

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1. Institute of Fruit and Tea, Hubei Academy of Agricultural Sciences, Wuhan 430064, China;
2. Institute of Plant Protection, Soil and Fertilizers, Hubei Academy of Agricultural Sciences, Wuhan 430064, China

Received date: 2019-05-21

Revised date: 2019-06-28

Online published: 2019-10-15

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摘要

探讨茶树响应铝（Aluminum,Al）的基因调控网络和表达模式,确定一些关键候选基因,为茶树耐Al分子机制研究奠定基础。测定了0、0.2、1、2、4βmmol·L^-1 5个Al³⁺浓度处理7βd的福鼎大白茶根系抗氧化酶活性和Al含量变化,并提取0βmmol·L^-1（R0）、1βmmol·L^-1（R1）和4βmmol·L^-1（R4）3个浓度下的茶树根系总RNA,通过Illumina Hiseq Xten平台进行高通量转录组测序。结果表明,随着Al³⁺浓度的升高,根系POD（Peroxidase,过氧化物酶）活性逐渐下降,APX（Ascorbic acid peroxidase,抗坏血酸过氧化物酶）活性则逐渐升高。SOD（Superoxide dismutase,超氧化物歧化酶）活性在Al³⁺浓度为1βmmol·L^-1时最高,CAT（Catalase,过氧化氢酶）活性在各处理间无显著差异。根系中Al含量随着Al³⁺浓度的升高呈先上升后下降趋势,在Al³⁺浓度为1βmmol·L^-1时达到最高。经筛选得到R1 VS R0,R4 VS R0,R4 VS R1的DEGs（Differentially expressed genes）分别为1β894、2β439个和1β384个,显著上调（下调）的差异表达基因分别有733（1β161）、846（1β593）个和628（756）个。GO富集分析表明,3个处理组在生物学途径中富集最多的类别均为刺激响应。在分子功能和细胞组件方面,R1 VS R0和R4 VS R0富集最多的类别均为核酸结合转录因子活性和细胞外围,R4 VS R1富集最多的类别为氧化还原酶活性相关基因和膜区域。KEGG富集分析表明,R1 VS R0、R4 VS R0、R4 VS R1分别显著富集了29、41条和19条Pathway,它们包括转录因子、转运蛋白、植物-病原菌互作、苯丙烷生物合成途径等,鉴定到多个参与调控活性氧代谢、有机酸或金属转运蛋白、转录因子及细胞壁结构修饰等生理过程的基因在Al诱导后上调或抑制表达,显示这些基因与茶树耐Al分子机制密切相关。

关键词： 茶树; Al; 根系; 差异表达基因; RNA测序

本文引用格式

黄丹娟 , 谭荣荣 , 陈勋 , 王红娟 , 龚自明 , 王友平 , 毛迎新 . 铝诱导的茶树根系转录组变化分析[J]. 茶叶科学, 2019 , 39(5) : 506 -520 . DOI: 10.13305/j.cnki.jts.2019.05.002

Abstract

The aim of this study was to investigate the gene regulation network and expression pattern of the response to aluminum (Al) in tea plants, and to identify the key candidate genes for understanding molecular mechanism of Al tolerance in tea plants. The roots’ antioxidant enzyme activities and Al content of Fuding Dabaicha cultivar were detected under 0βmmol·L^-1, 0.2βmmol·L^-1, 1βmmol·L^-1, 2βmmol·L^-1 and 4βmmol·L^-1 Al³⁺ concentrations for 7βd. The total RNA of roots under 0βmmol·L^-1 (R0), 1βmmol·L^-1 (R1) and 4βmmol·L^-1 (R4) Al³⁺ concentrations were extracted for high-through transcriptome sequencing by Illumina Hiseq Xten platform. The results showed that with the increase of Al concentration, POD activity decreased while APX activity increased gradually. SOD activity reached the highest peak at the Al concentration of 1βmmol·L^-1. However, CAT activity showed no significant difference among five treatments. The Al content first increased and then decreased with the increase of Al³⁺ concentration, and reached the highest peak at the Al³⁺ concentration of 1βmmol·L^-1.The DEGs of R1 VS R0, R4 VS R0, R4 VS R1 were 1β894, 2β439 and 1β384 respectively with 733 (1β161), 846 (1β593) and 628 (756) DEGs significantly up-regulated (down-regulated). GO enrichment analysis shows that the most enrichment biological pathway of three samples were all stimulus responses. In terms of molecular function and cell components, R1 VS R0 and R4 VS R0 were mostly enriched in nucleic acid binding transcription factor activity and cell periphery, while R4 vs R1 were mostly enriched in redox enzyme activity and membrane. KEGG enrichment analysis illustrates that they were significantly enriched in 29, 41, and 19 pathways, respectively, including transcription factors, transporters, plant-pathogen interactions, and phenylpropanoid biosynthesis pathways. It was found that genes involved in physiological processes such as reactive oxygen metabolism, organic acids or metal transporters, transcription factors and cell wall structure modification were up-regulated or inhibited after Al induction, suggesting that these genes were closely related to the molecular mechanism of Al tolerance in tea plants.

Key words： Camellia sinensis; aluminum; root; differential expressed genes; RNA sequencing

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