利用cDNA-AFLP技术分析炭疽菌危害诱导茶树的差异表达基因

doi:10.13305/j.cnki.jts.20200117.001

茶叶科学 ›› 2020, Vol. 40 ›› Issue (1): 26-38.doi: 10.13305/j.cnki.jts.20200117.001

利用cDNA-AFLP技术分析炭疽菌危害诱导茶树的差异表达基因

魏日凤¹, 赖建东^1,4, 彭成彬¹, 张承康², 连玲丽^3,*, 刘伟^1,2,*

1. 福建农林大学园艺学院,福建福州 350002;
2. 宁德师范学院生命科学学院,福建宁德 352100;
3. 福建农林大学生命科学学院,福建福州 350002;
4. 湖北科技学院,湖北咸宁 437100

收稿日期:2019-04-02 修回日期:2019-06-04 出版日期:2020-02-15 发布日期:2020-02-04
通讯作者: *lianll2002@163.com，liuwei0591@126.com
作者简介:魏日凤，女，实验师，主要从事茶叶品质与提升方面的研究。
基金资助:
福建省“2011 协同创新中心”中国乌龙茶产业协同创新中心专项（闽教科[2015]75 号）、福建省科技厅高校产学项目（2016N5010）、宁德师范学院科研发展资金项目（2016FZ30）、宁德师范学院协同创新中心资金项目（2017Z02）

cDNA-AFLP Reveals Differential Gene Expression Profiles of Tea Plant (Camellia sinensis cv. Maoxie) Induced by Colletotrichum sp.1 Infection

WEI Rifeng¹, LAI Jiandong^1,4, PENG Chengbin¹, ZHANG Chengkang², LIAN Lingli^3,*, LIU Wei^1,2,*

1.College of Horticulture, Fujian Agriculture and Forestry University, Fuzhou 350002, China;
2. Ningde Normal University, Ningde 352100, China;
3. College of Life Science, Fujian Agriculture and Forestry University, Fuzhou 350002, China;
4. Hubei University of Science and Technology, Xianning 437100, China

Received:2019-04-02 Revised:2019-06-04 Online:2020-02-15 Published:2020-02-04

摘要/Abstract

摘要： 应用cDNA-AFLP技术筛选炭疽菌（Colletotrichum sp.1）危害诱导茶树毛蟹品种（Camellia sinensis cv. Maoxie）的差异表达基因,为探究茶树抗炭疽菌的分子机制奠定基础。利用256对选择性扩增引物组合分析病菌接种后0 h和48 h的叶片cDNA,经筛选、测序和BLAST比对获得136条差异表达片段（TDFs）。同源基因功能分析结果表明,128条TDFs与Nr数据库已有基因同源,其功能以胁迫响应、生物调控与信号转导为主;其中51条TDFs与TPIA数据库中冷害、干旱、盐碱条件下的差异表达基因高度同源。进一步对27条TDFs进行qRT-PCR验证,结果显示,有24条TDFs的表达情况与cDNA-AFLP的结果一致,WRKY转录因子、乙烯响应转录因子、过氧化物酶和超氧化物歧化酶等抗逆相关基因的表达显著上调。本试验通过筛选获得差异表达的抗逆相关基因,为后续的基因功能研究奠定基础。

关键词: 茶树, 炭疽菌, cDNA-AFLP, 差异表达基因

Abstract: Differentially expressed genes related to Camellia sinensis cv. Maoxie against Colletotrichum sp.1 infection were screened by cDNA-AFLP (cDNA-amplified fragment length polymorphism) to provide reference for further illustrating the molecular mechanism underlying tea resistance to anthracnose. A total of 256 pairs of selective primers were applied to amplify the cDNA of leaves collected at 0 h and 48 h after Colletotrichum infection, and 136 differentially expressed transcript-derived fragments (TDFs) were obtained via screening, sequencing and BLAST analysis. The analysis of the homologous genes revealed that 128 TDFs were homologous with the known genes in Nr database. Most of them were related to stress responses, biological regulation and signal transduction, and 51 TDFs were highly homologous to differential expressed genes of tea plant under cold, drought or salt conditions in TPIA database. Further qRT-PCR analysis of 27 TDFs showed that the expression profiles of 24 TDFs were consistent with those of cDNA-AFLP results, among which several resistance-related genes including WRKY transcription factor, ethylene-responsive transcription factor, peroxidase and superoxide dismutase were significantly up-regulated. The differential expressed genes found in this study would lay a foundation for further analysis of their functions.

Key words: Camellia sinensis, Colletotrichum sp.1, cDNA-AFLP, differential expression genes

中图分类号:

魏日凤, 赖建东, 彭成彬, 张承康, 连玲丽, 刘伟. 利用cDNA-AFLP技术分析炭疽菌危害诱导茶树的差异表达基因[J]. 茶叶科学, 2020, 40(1): 26-38. doi: 10.13305/j.cnki.jts.20200117.001.

WEI Rifeng, LAI Jiandong, PENG Chengbin, ZHANG Chengkang, LIAN Lingli, LIU Wei. cDNA-AFLP Reveals Differential Gene Expression Profiles of Tea Plant (Camellia sinensis cv. Maoxie) Induced by Colletotrichum sp.1 Infection[J]. Journal of Tea Science, 2020, 40(1): 26-38. doi: 10.13305/j.cnki.jts.20200117.001.

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利用cDNA-AFLP技术分析炭疽菌危害诱导茶树的差异表达基因

cDNA-AFLP Reveals Differential Gene Expression Profiles of Tea Plant (Camellia sinensis cv. Maoxie) Induced by Colletotrichum sp.1 Infection

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