低温条件下蓖麻种子萌发期转录组分析  

Transcriptome Analysis of Castor Seeds at Germination Stage under Low Temperature

作　　者：白雪[1,2] 曹帅 向殿军 李兴 刘鹏[1,3] Bai Xue;Cao Shuai;Xiang Dianjun;Li Xing;Liu Peng(College of Agriculture, Inner Mongolia University for Nationalities, Tongliao, 028042;Allukorqin Agricultural Bureau, Chifeng, 024200;Inner Mongolia Industrial Engineering Research Center of Universities for Castor, Inner Mongolia Key Laboratory of Castor Breeding, Inner Mongolia Collaborate Innovation Cultivate Center for Castor, Inner Mongolia University for Nationalities, Tongliao, 028043)

机构地区：[1]内蒙古民族大学农学院,通辽028042 [2]阿鲁科尔沁旗农业局,赤峰024200 [3]内蒙古民族大学,内蒙古自治区高校蓖麻产业工程技术研究中心,内蒙古自治区蓖麻育种重点实验室,内蒙古自治区蓖麻产业协同创新培育中心,通辽028043

出　　处：《分子植物育种》2019年第12期3834-3844,共11页Molecular Plant Breeding

基　　金：国家自然科学基金项目(31860389);内蒙古自然科学基金项目(2018MS03023);内蒙古自治区高校蓖麻产业工程技术研究中心开放课题(BMYJ2015-06);内蒙古自治区草原英才创新团队——蓖麻分子育种创新研究团队项目共同资助

摘　　要：蓖麻(Ricinus communis L.)是一种冷敏感作物,解析其响应低温胁迫的关键调控基因,对于选育耐低温蓖麻品种具有重要的理论意义。以蓖麻耐低温品种‘通蓖5号’为材料,构建15℃(低温)和25℃(适温)萌发条件下完全展开的子叶cDNA文库,利用Illumina测序技术进行转录组测序(RNA-Seq),筛选差异表达基因(differentially expressed genes, DEGs)。结果表明:借助RNA-Seq技术共筛选到1 530个DEGs,其中低温相对于适温表达上调的DEGs有848个,表达下调的DEGs为682个;对1 530个DEGs进行GO功能分类和富集分析显示,共有953个DEGs被注释到GO功能三大分类的57个亚类中,涉及生物学过程、细胞成分及分子功能的DEGs比例分别为88.46%、69.67%和25.71%;KEGG功能分类显示,低温相对于适温表达上调的DEGs有243个,被富集到201个代谢通路中,其中显著富集的通路包括细胞周期、激素信号转导、DNA复制、细胞周期-酵母、减数分裂酵母和孕酮介导的卵母细胞成熟等20个通路,差异基因数富集最多的代谢通路为细胞周期;利用qRT-PCR对上调表达且显著富集的8个DEGs进行了表达分析,证实了转录组测序结果的准确性。该研究将为揭示蓖麻种子低温条件下萌发的分子机制提供理论依据。Castor (Ricinus communis L.) is a cold sensitive crop. It is of great theoretical significance to analyze its key regulatory genes in response to low temperature stress for breeding low temperature tolerant varieties of castor. A fully expanded cotyledon cDNA library was constructed under germination conditions at 15℃ (low temperature) and 25 ℃(suitable temperature) using castor variety 'Tongbi 5' as the material. To screen differentially expressed genes (DEGs) of castor, the Illumina sequencing technique was used to carry out transcriptional sequencing (RNA-Seq). The results showed that 1 530 DEGs were screened compared to optimum suitable temperature by RNA-Seq, including 848 DEGs associated with up-regulation and 682 DEGs associated with down?regulation. Go functional classification and enrichment analysis indicated that 953 of 1 530 DEGs were annotated into 57 subclasses of the three major classification. The proportion of DEGs involved in biological process, cell composition and molecular function was 88.46%, 69.67% and 25.71%, respectively. KEGG functional classification indicated that there were 243 up-regulated DEGs at low temperature relative to suitable temperature, which were enriched into 201 metabolic pathways. Twenty significant enrichment pathways included cell cycle, DNA replication, cell cycle-yeast, meiotic yeast and progesterone mediated oocyte maturation, and the cells cycle were metabolic pathways with the largest number of differentially enriched genes. The qRT-PCR was used to analyze the expression of 8 DEGs with up-regulation and significant enrichment, which confirmed the accuracy of the results of transcriptome sequencing. This study would provide theoretical basis for revealing the molecular mechanism of castor seed germination at low temperature.

关 键 词：蓖麻(Ricinus communis L.) 低温 萌发期 转录组 

分 类 号：S565.6[农业科学—作物学]