目的:利用Illumina/Solexa Hiseq2000测序平台对女贞果实进行转录组测序.方法:将测序得到的大量数据进行拼接与组装,并应用生物信息学方法对所得序列进行功能注释、功能分类、代谢途径分析和简单重复序列位点查找等研究.结果:共获得32856614条clean reads,含4.93 Gb的有效数据.对clean reads应用Trinity软件进行组装,共获得163092条unigene,总长度、平均长度和N50分别为108.57 Mb、665.68 bp和1345 bp.公共数据库(Nr,Nt和 SwissProt数据库)的BLAST比对分析显示,有83903条unigene获得基因注释,进一步的功能分类表明,在GO数据库中有16876条unigene可被分别注释到细胞组分、分子功能和生物学过程三大类别中.将unigene与COG数据库进行比对,可获得25个功能类别,其中较多unigene涉及果实中物质的合成与转运相关的营养功能.KEGG数据库作为搜索对象,可将unigene定位到21类代谢途径中,涉及重要次生代谢产物黄酮类和萜类生成的unigene数目分别为258条和929条.SSR位点查询发现,在14270条unigene中共可找到15925个SSR位点.结论:本研究在转录组水平对女贞果实进行了系统研究,这为进一步开展女贞果实的分子标记开发和重要次级代谢产物的生物合成奠定了基础.%Objective:This study used Illumina/Solexa Hiseq2000 platform to sequence the transcriptome of Ligus-trum lucidum fruits.Methods:A large number of data obtained by sequencing were spliced and assembled,and bioinformatics methods were used to study the functional annotation,functional classification,metabolic pathway analysis and simple sequence repeat site search.Results:The results showed that 32 856 614 clean reads were acquired including 4.93 Gb valid data.The Trinity software was used to assemble the clean reads,receiving 163 092 unigenes with a total read length of 108.57 Mb,an average read length of 665.68 bp and N50of 1345 bp.The 83 903 unigenes were annotated using BLAST searches against the public database(Nr,Nt and SwissProt database).Further functional classification indicates that 16 876 unigenes were classified into cellular components,molecular functions and biological processes in GO database.Aligning unigenes with COG database,25 functional categories could be obtained,among which lots of unigenes involved in nutritional function of fruit to synthesize and transport substances.As the search object,unigene could be located into 21 kinds of metabolic pathways in KEGG database,in which the number of unigene involved in the production of important secondary metabolites flavonoids and terpenoids was 258 and 929,respectively.The analysis for SSR loci showed that 15 925 SSR loci could be detected from 14 270 unigenes.Conclusion:In this study,the L.lucidum fruits was studied systematically at the transcriptome level,which laid a foundation for the further development of molecular markers and biosynthesis of important secondary metabolites from L.lucidum.
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