为探究草地早熟禾无融合生殖胚囊发育过程中基因表达谱的变化，本研究利用高通量Illu-mina Hiseq测序技术对'清水'草地早熟禾无融合生殖子房发育的无孢子起始细胞发生时期(8DBA)和无孢子生殖胚囊发育时期(6DBA、4DBA和2DBA)共4个阶段的小穗进行转录组测序。结果显示，不同差异分组(2DBA vs 4DBA、2DBA vs 6DBA和2DBA vs 8DBA)共有13575个DEGs，包括7191个上调基因和6384个下调基因。GO富集分析发现，3个差异分组的DEGs显著富集在过氧化氢的响应、蛋白质重折叠、营养水库活动、错误折叠蛋白结合和双糖生物合成等过程中。KEGG通路分析表明，3个差异分组的DEGs显著富集在代谢途径、次生代谢产物的生物合成、淀粉和蔗糖代谢及内质网中的蛋白加工等通路上，在2DBA vs 8DBA分组中发现植物激素信号转导途径的富集。4个时期占比最多的转录因子家族是NAC、AP2/ERF-ERF和MYB-related等。将在 4个发育阶段特异性高表达的基因OPR1、ASN2、SAMS2、HSP81-2、PXG4和FPK2作为与'清水'草地早熟禾无融合生殖胚囊发育相关的候选基因。qRT-PCR分析表明，除WOX2在4DBA上调，与RNA-seq的表达存在差异外，其余16个DEGs的表达量变化趋势与高通量测序结果基本一致。
Transcriptomic Analysis of the Development of Apomixis Embryo Sac in Poa pratensis(Kentucky Bluegrass)
In this research for changes of gene expression profile in apomixis embryo sac development of Poa pratensis,transcriptomic sequencing analysis was applied on spikelet of Poa pratensis cultivar'Qin-gshui'in four stages of apomixis ovary development,apospore initiation cell stage(8DBA)and apospore embryo sac development stages(6DBA,4DBA,and 2DBA),by Illu-mina Hiseq sequencing technology.Results revealed a difference with 13575 DEGs for differential groups(2DBA vs 4DBA,2DBA vs 6DBA,and 2DBA vs 8DBA),including 7191 up-regulated genes and 6384 down-regulated genes.GO enrichment analysis showed that DEGs of three differential groups significantly clustered in processes of hydrogen per-oxide response,protein refolding,nutrition reservoir activity,misfolded protein binding,and disaccharide biosynthesis.DEGs of three differential groups were found significantly enriched on pathways such as meta-bolic pathways,biosynthesis of secondary metabolites,starch and sucrose metabolism,and protein process-ing in endoplasmic reticulum,from KEGG pathway analysis.Enrichment of plant hormone signal transduc-tion pathway was found in the 2DBA vs 8DBA group.The most parts of TFs in four stages are NAC,AP2/ERF-ERF,and MYB-related,etc.Genes specifically high expressed in four development stages,OPR1,ASN2,SAMS2,HSP81-2,PXG4,and FPK2,were selected as candidates of apomixis embryo sac development in P.pratensis'Qingshui'.qRT-PCR analysis indicated that trend of expression changes in 16 DEGs was consistent with results of high-throughput sequencing,except the up-regulation of WOX2 on 4DBA,whose expression was different to RNA-seq.This study provide evidence for further function research on vital genes related to development of apomixis embryo sac in P.pratensis.
万方数据
维普
