基于RNA-Seq技术的湿加松特定时期产脂差异基因筛选及表达分析  

Screening and Expression Analysis of Different Genes for Oleoresin Production in the Specific Period of Pinus elliottii×P.caribaea by RNA-Seq Technology

作　　者：周晨晨 李义良 王哲[1,2] 赵奋成 钟岁英 林昌明[3] 谭志强 李福明 吴惠姗 郭文冰 廖仿炎 王建革[4] ZHOU Chen-Chen;LI Yi-Liang;WANG Zhe;ZHAO Fen-Cheng;ZHONG Sui-Ying;LIN Chang-Ming;TAN Zhi-Qiang;LI Fu-Ming;WU Hui-Shan;GUO Wen-Bing;LIAO Fang-Yan;WANG Jian-Ge(Guangdong Provincial Key Laboratory of Silviculture,Protection and Utilization,Guangzhou 510520;Guangdong Academy of Forestry,Guangzhou 510520;Taishan Hongling Seed Orchard,Taishan 529200;Jiangxi Agricultural University/College of Landscape and Art,Nan-chang 330045)

机构地区：[1]广东省森林培育与保护利用重点实验室,广州510520 [2]广东省林业科学研究院,广州510520 [3]广东省台山市红岭种子园,台山529200 [4]江西农业大学园林与艺术学院,南昌330045

出　　处：《植物研究》2021年第3期419-428,共10页Bulletin of Botanical Research

基　　金：广东省省级科技计划项目(2017B020232001);广东省林业科技创新专项资金项目(2018KJCX017)。

摘　　要：湿加松(Pinus elliottii×P.caribaea)是我国南方重要的脂材兼用树种,为挖掘调控湿加松松脂产量的相关基因,在湿加松的产脂高峰期,选取生长量相近但产脂量显著差异的湿加松植株,采集次生木质部组织进行高通量测序,并结合萜类物质合成、转运通路进行分析。结果显示,测序共产生594 108 382条clean reads,经de novo拼接获得190 514条unigenes,有304个unigene与萜类物质的合成有关。经过差异表达分析,得到414个差异表达unigenes,其中183个上调表达,231个下调表达。利用qRT-PCR技术对筛选的基因进行表达量验证分析,发现细胞色素P450、ABC转运蛋白、双功能松脂醇落叶松醇还原酶(PLR)在高脂树中表达量升高,异戊烯基转移酶表达量在低脂树中表达量较高。本研究丰富了湿加松及松科植物的转录本信息,为进一步研究湿加松松脂产量差异的分子机理奠定了重要基础。The experiment was conducted to explore the differential expression of genes which regulate oleoresin synthesis and transport in Pinus elliottii×P. caribaea. At the peak of oleoresin production,P. elliottii×P. caribaea with similar growth but significant differences in oleoresin production of plants were selected,and secondary xylem tissues were collected for high-throughput sequencing platform. The results showed that 594 108 382 clean reads were generated by sequencing. 190 514 unigenes were obtained by de novo splicing,and 304 unigenes were related to the synthesis of terpenoids. The 414 unigenes were obtained,differential expression analysis,including 183 unigenes up-regulated and 231 unigenes down regulated. The genes encoding CYPs,ABC transport proteins,PLR,Strictosidine synthase activity increased in high oleoresin-yielding,while the genes encoding prenyltransferase activity increased in low oleoresin-yielding. This study enriched the transcript information of P. elliottii× P. caribaea,and provided reference value for further study of the molecular mechanism of the difference in P. elliottii×P. caribaea and Pines plant resin production.

关 键 词：湿加松 转录组测序 产脂基因 QRT-PCR 

分 类 号：S791.244[农业科学—林木遗传育种] S791.246[农业科学—林学]