目的 为了探知宁夏枸杞Lycium barbarum花药发育过程及挖掘花药发育相关基因。方法 以单双核花粉时期和成熟花粉时期的花药为研究材料，通过高通量Illumina HiSeq2500测序平台进行转录组分析。结果 组装共获得128 151条Unigene，将Unigene与NCBI的非冗余蛋白质数据库（non-redundant protein databasse，NR）、去冗余的蛋白序列数据库（Swiss prot protein database，Swiss-Prot）、基因本体数据库（gene ontology，GO）、直源同源群集数据库（clusters of orthologous groups，COG）、真核同源群数据库（euKaryotic orthologous groups，KOG）、基因功能和代谢途径数据库（Kyoto encyclopedia of genes and genomes，KEGG）数据库比对，共有40 140条得到注释。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，单双核花粉时期与成熟花粉时期的差异表达基因共有46个。KEGG注释发现有4个差异表达基因参与甘油酯代谢、亚油酸代谢、苯丙氨酸代谢和苯丙烷类生物合成4个途径中。富含亮氨酸重复蛋白、细胞周期蛋白、类受体蛋白激酶、E3泛素蛋白连接酶等11个差异表达基因参与了花药发育过程。随机选取其中5个差异表达基因进行qRT-PCR验证，表达结果与转录组数据基本一致，表明其数据真实可靠。结论 为进一步解释宁夏枸杞花药发育的相关分子机制奠定了基础，也为后续研究宁夏枸杞花药发育的生物学特征提供了有益的基因资源。

Objective To investigate the anther development process of Lycium barbarum, and explore the genes related. Methods The anther transcriptome analysis was performed by the high-throughput Illumina HiSeq2500 sequencing platform using anthers of single microspore and bi-cellular pollen stage and mature pollen stage as research materials. Results A total of 128,151 unigene were obtained after assembly, and 40,140 unigenes were annotated by searching databases of NR, Swiss-Prot, GO, COG, KOG and KEGG. Under the condition of FDR (false discovery rate) ＜ 0.05 and FC (fold change) ≥ 2, a total of 46 differentially expressed genes were found between single microspore and bi-cellular pollen stage and mature pollen stage. KEGG annotations identified four differentially expressed genes involved in four pathways: glycerol ester metabolism, linoleic acid metabolism, phenylalanine metabolism and phenylpropanoid biosynthesis.There were eleven differentially expressed genes were involved in the anther development process, such as leucine-rich repeat protein (LRR), cyclin (Cyclin), receptor-like protein kinase (RPK), E3 ubiquitin protein ligase (MBR1), and so on. Five of the different expression genes among the eleven genes were randomly selected for qRT-PCR verification, and the expression results were basically consistent with the transcriptome data, indicating that the data were true and reliable. Conclusion This study lays the foundation for further explaining the molecular mechanisms related to anther development in L. barbarum., and also provides useful genetic resources for subsequent studies on the biological characteristics of anther development in L. barbarum.

R282.12

宁夏自然科学基金资助项目（2020AAC03097）；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（重大）重点项目（2019BBF02022）