基于银杏花芽3个分化时期转录组测序的相关基因筛选与表达分析  

Screening and Expression Analysis of Related Genes Based on Transcriptome Sequencing of Ginkgo Flower Buds at Three Differentiation Stages

作　　者：何崇单 蔡雨蒙 李萌[1,2] 董金金 刘伟[2] 王俊燚 王义强[1,2] HE Chongdan;CAI Yumeng;LI Meng;DONG Jinjin;LIU Wei;WANG Junyi;WANG Yiqiang(l Hunan Provincial Key Laboratory for Forestry Biotechnology,Central South University of Forestry and Technology,Changsha 410004,Chin;Key Lab of Non-wood Forest Nurturing and Protection of National Ministry of Education Biotechnology Laboratory,Central South University of Forestry and Technology,Changsha 410004,China)

机构地区：[1]中南林业科技大学林业生物技术湖南省重点实验室,长沙410004 [2]中南林业科技大学经济林培育与保护教育部重点实验室,长沙410004

出　　处：《园艺学报》2018年第8期1479-1490,共12页Acta Horticulturae Sinica

基　　金：国家自然科学基金项目(31570682);国家重点研发计划项目(2017YFD0600701)

摘　　要：鉴定银杏花芽分化调控的关键基因,揭示银杏花芽分化调控的主要分子机制,为缩短银杏童期和选育银杏早花品种提供理论指导。本研究中采用高通量测序技术对银杏花芽分化3个时期(花芽未分化期、花芽分化始期、花芽分化盛期)的样品进行转录组测序,并分析数字表达谱,筛选开花调控相关基因并进行荧光定量PCR(RT-qPCR)表达验证。转录组测序共产生27.52 Gb原始数据,注释到8大功能数据库(GO、COG、KEGG、KOG、NR、Pfam、Swiss-Prot、egg NOG)上的unigene总数为35 179个。通过GO分类和KEGG Pathway富集性分析,将unigene分别归于55个GO类别和126个代谢途径。差异表达基因分析显示,花芽未分化期较花芽分化始期有2 253个基因上调,2 032个基因下调;花芽分化始期较花芽分化盛期有1 770个基因上调,1 901个基因下调;花芽未分化期较花芽分化盛期有1 865个基因上调,2 042个基因下调。发掘出大量的开花相关的基因涉及5个开花调控途径(光周期途径、春化途径、赤霉素途径、自主途径和年龄途径)。筛选出gene.Gb_17618(GI序列)、gene.Gb_19790(FT/TFL1序列)、gene.Gb_16301(AG序列)、gene.Gb_28337(花发育MADS-box序列)、gene.Gb_01884(SOC1序列)和gene.Gb_41704(CO序列)等6个银杏花芽分化差异表达关键基因序列,荧光定量PCR检测表达水平与转录组结果一致。This study identified of Ginkgo biloba flower bud differentiation key genes,to reveal the main molecular mechanism of ginkgo flower bud differentiation regulation,and provide theoretical guidance for shortening the juvenile phase and breeding early flowering varieties. High-throughput sequencing technology was used to sequence the transcriptomes of three different stages of flower bud differentiation,and digital expression profiles were analyzed to screen ginkgo flowering regulatory related genes. Genes were verified by q PCR. A total of 27.52 Gb of raw data was generated by transcriptome sequencing,and the total number of unigenes annotated in the 8 functional databases（GO,COG,KEGG,KOG,NR,Pfam,Swiss-Prot,and egg NOG）was 35 179. The unigenes were classified into 55 GO categories and 126 metabolic pathways. Analysis of differentially expressed genes revealed that 2 253 genes were up-regulated and 2 032 genes were down-regulated during the flower bud undifferentiated stage vs. initial stage of flower bud differentiation. A total of 1 770 genes were up-regulated and 1 901 genes were down-regulated during the initial stage of flower bud differentiation vs. flower bud differentiation stage. The flower bud undifferentiated stage vs. flower bud differentiation stage,1 865 genes were up-regulated and 2 042 genes were down-regulated. A large number of genes related to flowering were discovered, involving five flowering regulatory pathways（ Photocycle pathway,Vernalization pathway,Gibberellin pathway,Autonomous pathway,and Age pathway）. Six key genes,such as gene.Gb_17618（GI unigene）,gene.Gb_19790（FT/TFL1 unigene）,gene.Gb_16301（AG unigene）,gene.Gb_28337（MADS-box unigene）,gene.Gb_01884（SOC1 unigene）,and gene.Gb_41704（CO unigene）,were selected. Their RT-q PCR expression was consistent with that of the transcriptome.

关 键 词：银杏 转录组测序 开花调控 GI FT/TFL1 SOC1 

分 类 号：S664.3[农业科学—果树学]