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林业科学 ›› 2014, Vol. 50 ›› Issue (8): 68-75.doi: 10.11707/j.1001-7488.20140810

• 论文与研究报告 • 上一篇    下一篇

基于RNA-Seq的油茶种子α-亚麻酸代谢途径及相关基因分析

江南1,2, 谭晓风1, 张琳1, 曾艳玲1   

  1. 1. 中南林业科技大学 经济林培育与保护省部共建教育部重点实验室 经济林育种与栽培国家林业局重点实验室 长沙 410004;
    2. 湖南工业大学 株洲 412008
  • 收稿日期:2013-10-11 修回日期:2014-02-24 出版日期:2014-08-25 发布日期:2014-07-31
  • 基金资助:

    国家自然科学基金面上项目(31070603);国家自然科学基金项目(31100497);湖南省科技计划项目(2012NK3065)。

Gene Analysis of α-Linolenic Acid Metabolism of Camellia oleifera Seeds Based on RNA-Seq

Jiang Nan1,2, Tan Xiaofeng1, Zhang Lin1, Zeng Yanling1   

  1. 1. Key Laboratory of Cultivation and Protection for Non-Wood Forest Trees of Ministry of Education Key Laboratory of Non-Wood Forest Products of State Forestry Administration Central South University of Forestry and Technology Changsha 410004;
    2. Hunan University of Technology Zhuzhou 412008
  • Received:2013-10-11 Revised:2014-02-24 Online:2014-08-25 Published:2014-07-31
  • Contact: 谭晓风

摘要:

以油茶果实膨大期和油脂合成高峰期的种子为材料,进行转录组测序和表达谱数据库构建,并对油茶种子α-亚麻酸代谢途径相关基因进行分析。转录组测序获得油茶种子α-亚麻酸代谢非冗余基因Unigenes序列共112条,与α-亚麻酸代谢密切相关的不饱和脂肪酸合成途径非冗余基因Unigenes序列94条,包含12种调控α-亚麻酸代谢主要酶基因。表达谱数据分析显示,果实膨大期和油脂合成高峰期的α-亚麻酸合成代谢途径相关基因具有不同的表达模式,参与油茶种子α-亚麻酸的合成及形成多种不饱和脂肪酸的物质转化。采用实时荧光定量PCR方法验证油茶种子LOX,AOS,ACOX,AOC,OPR,AACT,PLA 2,HPL,DOX,MFP,FAD2,FAD3 等关键基因在油茶种子油脂合成不同时期的相对表达量,结果表明各基因表达规律与表达谱数据分析结果一致。克隆、调控这些相关基因将会为提高α-亚麻酸含量的油茶育种提供依据。

关键词: 油茶, 转录组, 表达谱, α-亚麻酸代谢, 基因表达

Abstract:

In an effort to better understand the factors that regulate α-linolenic acid metabolism in Camellia oleifera seeds, the transcriptome library and expression profile of C. oleifera seeds both in the fruit expanding period and at the peak stage of lipid biosynthesis have been constructed and the functional genes related to α-linolenic acid metabolism have been analyzed in detail. Study on transcriptome showed that there were one hundred and twelve non-redundant gene unigenes related to α-linolenic acid metabolism and ninety-four unigenes related to biosynthesis of unsaturated fatty acids of C. oleifera seeds, including twelve kinds of key enzyme genes regulating α-linolenic acid metabolism. The results of expression profile analysis showed that these genes had different expression patterns. These genes regulate the synthesis of α-linolenic acid and its conversion process to other unsaturated fatty acid at different developmental stage. The real-time quantitative RT-PCR analysis revealed relative expression regularity of the key enzyme genes such as LOX, AOS, ACOX, AOC, OPR, AACT, PLA2, HPL, DOX, MFP, FAD2, FAD3 in α-linolenic acid metabolism in fruit expanding period and the oil synthesis peak stage, which agreed with the results of expression profile analysis. The findings would provide a scientific basic for Camellia breeding of good quality with increasing content of α-linolenic acid.

Key words: Camellia oleifera, transcriptome, expression profile, α-linolenic acid metabolism, gene expression

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