基于转录组测序的海巴戟果实中乙烯调控相关基因表达分析

doi:10.16036/j.issn.1000-2650.2020.05.004

四川农业大学学报 ›› 2020, Vol. 38 ›› Issue (05): 528-537.doi: 10.16036/j.issn.1000-2650.2020.05.004

基于转录组测序的海巴戟果实中乙烯调控相关基因表达分析

胡丹焱¹, 张正雪², 刘娟¹, 蓝增全³, 吴田^1,*

1.西南林业大学国家林业和草原局西南风景园林工程技术研究中心/园林园艺学院,昆明 650224;
2.西南林业大学生态与水土保持学院,昆明 650224;
3.西南林业大学西南绿色发展研究院,昆明 650224

收稿日期:2020-07-06 出版日期:2020-10-28 发布日期:2021-02-05
作者简介:胡丹焱,硕士研究生。^*责任作者：吴田,博士,副教授,主要从事植物分子生物学研究,E-mail：wutianpotato@swfu.edu.cn。
基金资助:
国家林业和草原局项目“诺丽果高产优质生产及加工技术推广示范”（[2019]27）; 国家留学基金（[2019]75）

Expression Analysis of Ethylene Regulates Related Genes in Morinda citrifolia Fruit Based on Transcriptome Sequencing

HU Danyan¹, ZHANG Zhengxue², LIU Juan¹, LAN Zengquan³, WU Tian^1,*

1. Southwest Forestry University, Southwest Landscape Architecture Engineering Research Center of National Forestry and Grassland Administration, School of Landscape Architecture and Horticulture Sciences, Kunming 650224, China;
2. Southwest Forestry University, College of Ecology and Soil Water Conservation, Kunming 650224, China;
3. Southwest Forestry University, Southwest Institute of Green Development, Kunming 650224, China

Received:2020-07-06 Online:2020-10-28 Published:2021-02-05

摘要/Abstract

摘要： 【目的】为了明确乙烯相关基因在海巴戟（Morinda citrifolia L.）果实采后后熟过程中的功能。【方法】以刚采摘的成熟海巴戟果实（0 d）和室温（25 ℃）后熟2 d的海巴戟果实为研究对象,通过转录组测序（RNA-Seq）和实时荧光定量PCR技术（qRT-PCR）,分析海巴戟果实的转录组数据、乙烯相关基因及其表达水平。【结果】海巴戟果实转录组经组装获得35 721个转录本,26 202个注释基因,筛选后获得3 754个差异基因（P≤0.05,LogFC≥2）。从差异基因中随机选取10个上调基因和10个下调基因进行荧光定量验证,结果表明转录组测序结果可靠。在乙烯合成途径和乙烯信号转导通路中发现4个乙烯合成相关基因-metK、ACS10、ACS12、ACO5和7个乙烯信号转导相关基因-ETR、CTR1、MAPK6、EIN2、EIN3、EBF1、ERF1,这11个基因在乙烯合成途径和乙烯信号转导通路中表达量较高且差异显著,选取其中表达量较高的metK、ACO5、EIN3和EBF1共4个基因进行荧光定量,进一步明确了这4个基因在海巴戟果实后熟过程中的重要作用。【结论】在乙烯合成途径和乙烯信号转导通路中发现的11个相关基因与海巴戟果实后熟和乙烯积累密切相关,可以从metK、ACO5、EIN3和EBF1入手,以继续进行海巴戟果实后熟机理研究。

关键词: 转录组测序, 海巴戟, 果实后熟, 乙烯调控

Abstract: 【Objective】 In order to define the function of ethylene related genes in Morinda citrifolia fruit during post-harvest ripening. 【Method】 The study chose two types M. citrifolia fruit of different mature degrees as the research object, the one is newly picked mature M. citrifolia fruit (0 day), and the other is M. citrifolia fruit that matured naturally on the indoor temperature (25 ℃) for 2 days, through the transcriptome sequencing, RNA-Seq and qRT-PCR technique, analyze the transcriptome data, ethylene related gene and its expression level of M. citrifolia fruit. 【Result】 The transcriptome of M. citrifolia fruit was assembled by trinity software to obtain 35 721 transcripts and 26 202 annotated genes, and 3 754 differentially expressed genes were screened (P≤0.05 and│LogFC│≥2). Ten up-regulated genes and ten down-regulated genes were randomly selected from the differentially expressed genes for qRT-PCR verification, and the results showed that the transcriptome sequencing results were reliable. Based on the effect of ethylene on fruit ripening, in the study, four ethylene synthesis related genes - metK, ACS10, ACS12, ACO5 and seven ethylene signal transduction related genes-ETR, CTR1, MAPK6, EIN2, EIN3, EBF1 and ERF1 were found from the ethylene synthesis pathway and ethylene signal transduction pathway, these ten genes have high expression levels and significant differences in ethylene synthesis pathway and ethylene signal transduction pathway, four genes with high expression levels, metK, ACO5, EIN3 and EBF1, were selected for fluorescence quantification, which further clarified that these four genes play an important role in the post-harvest ripening. 【Conclusion】 The 11 related genes found from the ethylene synthesis pathway and ethylene signal transduction pathway are closely related to the post-ripening and ethylene accumulation of M. citrifolia fruit, and among them, metK, ACO5, EIN3 and EBF1 can be used as key candidate genes, to continue the study on post ripening mechanism of M. citrifolia fruit.

Key words: transcriptome sequencing, Morinda citrifolia, post-ripening of fruit, ethylene regulated

中图分类号:

Q786

胡丹焱, 张正雪, 刘娟, 蓝增全, 吴田. 基于转录组测序的海巴戟果实中乙烯调控相关基因表达分析[J]. 四川农业大学学报, 2020, 38(05): 528-537.

HU Danyan, ZHANG Zhengxue, LIU Juan, LAN Zengquan, WU Tian. Expression Analysis of Ethylene Regulates Related Genes in Morinda citrifolia Fruit Based on Transcriptome Sequencing[J]. Journal of Sichuan Agricultural University, 2020, 38(05): 528-537.

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基于转录组测序的海巴戟果实中乙烯调控相关基因表达分析

Expression Analysis of Ethylene Regulates Related Genes in Morinda citrifolia Fruit Based on Transcriptome Sequencing

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