雄性不育枸杞花药比较转录组分析

Comparative Transcriptome Analysis of Male Sterile Anther in Wolfberry (Lycium barbarum L.)

为筛选宁夏枸杞(Lycium barbarum L.)雄性不育花药发育相关的差异表达基因,本研究开展了雄性不育品种‘宁杞5号’和可育品种‘宁杞1号’成熟花粉时期花药转录组分析,共获得差异表达基因1759个,在‘宁杞5号’中有753个基因上调表达,1006个基因下调表达。KEGG富集表明,这些差异表达基因主要富集到氨基酸的生物合成、植物激素信号传导、苯丙素生物合成、淀粉和蔗糖代谢、脂肪酸代谢等相关通路。对有注释信息的差异表达基因进行NR和Uniprot数据库检索,筛选到28个花药发育相关基因,其中有20个在‘宁杞5号’中下调表达,8个上调表达。选取其中的5个差异表达基因进行RT-qPCR验证,结果与转录组数据趋势一致。甘油醛-3-磷酸脱氢酶(Glyceraldehyde-3-phosphate dehydrogenase,GAP)、束状阿拉伯半乳糖蛋白3(Fasciclin-like arabinogalactan protein 3,FLAs)和花药特异表达启动子LAT52系统发育树分析表明,3个基因分别与同科的物种在同一个进化分支上且同源性较高。这些差异表达基因可能在宁夏枸杞‘宁杞5号’成熟花粉时期花药发育及雄性不育的发生中发挥重要作用。研究结果为进一步研究宁夏枸杞花药发育及其调控机制、雄性不育发生的分子机制提供了参考依据。

In order to screen differentially expressed genes related to anther development of Lycium barbarum L.,the anther transcriptome analysis of male sterile variety‘Ningqi No.5’and fertile variety‘Ningqi No.1’at mature pollen stage were carried out.1759 differentially expressed genes were obtained.Among them,753 differentially expressed genes were up-regulated and 1006 were down-regulated in‘Ningqi No.5’.KEGG enrichment showed that these differentially expressed genes were mainly enriched in amino acid biosynthesis,plant hormone signal transduction,phenylpropanoid biosynthesis,starch and sucrose metabolism,fatty acid metabolism and other related pathways.The differentially expressed genes with annotated information were searched by NR and Uniprot database.28 genes related to anther development were screened,of which 20 were down-regulated and 8 up-regulated in‘Ningqi No.5’.Five differentially expressed genes were verified by RT-qPCR,which showed the results were basically consistent with the transcriptome data.Phylogenetic tree analysis of Glyceraldehyde-3-phosphate dehydrogenase(GAP)、Fasciclin-like arabinogalactan protein 3(FLAs)and anther specific expression promoter LAT52 showed they were in the same evolutionary branch and had the highest homology with the species of the same family.These differentially expressed genes may play an important role in anther development and male sterility in mature pollen stage of‘Ningqi No.5’.The results provide a reference basis for further study on the anther development and regulation of Lycium barbarum L.and the molecular mechanism of male sterility.