目的 探知宁夏枸杞Lycium barbarum雄性不育发生涉及的相关基因。方法 以雄性不育品种“宁杞5号”和正常可育品种“宁杞1号”单双核花粉时期花药为材料，构建cDNA文库，Illumina HiSeq2500高通量平台进行转录组测序。结果 组装得到14 154条Unigenes序列，在错误发现率（false discovery rate，FDR）＜0.05且差异倍数（fold change，FC）≥2的条件下，获得3012个差异表达基因（differentially expressed genes，DEGs），其中，有2327个DEGs在NCBI的非冗余蛋白质数据库（non-redundant protein databasse，NR）、基因本体数据库（gene ontology，GO）、直源同源群集数据库（clusters of orthologous groups，COG）、基因功能和代谢途径数据库（Kyoto encyclopedia of genes and genomes，KEGG）、真核同源群数据库（euKaryotic orthologous Groups，KOG）、去冗余的蛋白序列数据库（swiss prot protein database，Swiss-Prot）等数据库中有注释信息。相对于“宁杞1号”，在“宁杞5号”中有1019个DEGs上调，1993个DEGs下调。GO分析表明，1116个DEGs被注释到生物学过程、细胞组分和分子功能中；此外，693个DEGs富集到KEGG数据库的50条代谢通路。结合PubMed文献库检索，筛选到细胞色素P450类蛋白（cytochrome P450 98A3-like，CYP450）、受体蛋白激酶（receptor-like protein kinase，RPK）、果胶裂解酶（pectate lyase-like，PLL）、查耳酮合酶（chalcone synthase，CHS）、花药特异蛋白（anther-specific protein，ASP）等23个参与雄性不育相关基因，选取其中9个基因进行qRT-PCR分析，结果与转录组测序基因表达趋势一致。结论 为进一步通过转基因技术验证基因功能提供了候选基因，为解析宁夏枸杞雄性不育发生的分子机制提供了研究基础。

Objective To investigate the genes involved in the occurrence of male sterility in Lycium barbarum. Methods The single Microspore and bi-cellular pollen stage anthers of male sterile variety “Ningqi No.5” and normal fertile variety “Ningqi No.1” were used as materials to construct a cDNA library and sequenced by IlluminaHiSeq2500 high throughput platform transcriptome. Results A total of 14 154 Unigenes sequences were assembled. Under the condition of false discovery rate (FDR) ＜ 0.05 and fold change (FC)≥2, there were 3012 differentially expressed genes. Compared with “Ningqi No.1”, 1019 genes were up-regulated and 1993 down-regulated in 'Ningqi No.5'. Among them, 2327 genes had annotation information in non-redundant protein databasse (NR), gene ontology (GO), clusters of orthologous groups (COG), Kyoto encyclopedia of genes and genomes (KEGG), euKaryotic orthologous Group (KOG), swiss prot protein database (Swiss-Prot) and other databases. A total of 1116 differentially expressed genes were annotated into biological processes,cellular components and molecular functions, and 693 differentially expressed genes were enriched into 50 metabolic pathways in KEGG database. Combined with the search of PubMed literature database, 23 genes related to male sterility, such as cytochrome P450 proteins (CYP450), receptor protein kinase (RPK), pectin lyase (PLL), chalcone synthase (CHS) and anther specific protein (ASP) were screened, and 10 of them were selected for RT-qPCR analysis. The results were consistent with the trend of gene expression in transcriptome.Conclusion The results provide candidate genes for further verification of gene function by transgenic technology, and provide research basis for analyzing the molecular mechanism of male sterility in L. barbarum.

R286.12

宁夏枸杞产业发展中心项目“宁夏枸杞核心种质资源差异性分析及功能基因挖掘”；宁夏回族自治区重点研发计划重点项目（2022BBF02010）