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以‘泽西’越橘(Vaccinium corymbosum‘Jersey’)不同发育阶段的果实为试材,构建De novo转录组测序文库并进行数据分析,利用Web Logo 3、CLC Sequence Viewer 6等软件和荧光定量PCR技术对所获数据中的R2R3-MYB转录因子进行生物信息学分析和表达分析。结果表明,转录组测序共获得有效数据6.01 Gb,通过组装得到Unigene序列60 481个。功能注释结果显示,有39 612个Unigene可以注释到GO和COG等数据库上;20 869个Unigene无匹配信息。利用GO和COG分类工具可将这些序列分别划分为55个和25个功能类别,涉及信号转导和转录调控等功能。利用所获数据分析越橘R2R3-MYB基因,结果表明共得到21个R2R3-MYB。序列分析显示,这21个基因均含有完整的R2R3-MYB区域,在此区域内2(G)和4(W)等30个氨基酸保守不变。进化树分析结果显示,越橘R2R3-MYB基因分为11个亚组。其中,S4、S5和S6亚组中有6个基因涉及到花青苷生物合成。荧光定量PCR结果分析显示,这6个基因在果实发育的7个阶段均能表达,但表达模式不同,这6个基因可能对越橘果实着色具有一定作用。
The fruits of different developmental stages of Vaccinium corymbosum’Jersey ’were used as experimental material to construct De novo transcriptome sequencing library and analyze the data. Using software such as Web Logo 3, CLC Sequence Viewer 6 and real-time quantitative PCR The bioinformatics analysis and expression analysis of the R2R3-MYB transcription factor in the obtained data were performed. The results showed that the total number of validated data was 6.01 Gb and 60 481 Unigene sequences were obtained by sequencing. Functional annotation results showed 39 612 Unigene annotated databases such as GO and COG; 20 869 Unigene no matching information. These sequences can be divided into 55 and 25 functional categories using GO and COG classification tools, respectively, involving functions such as signal transduction and transcriptional regulation. The data of bilberry R2R3-MYB was analyzed. The results showed that a total of 21 R2R3-MYB were obtained. Sequence analysis showed that these 21 genes all contain the complete R2R3-MYB region, in which 30 amino acids such as 2 (G) and 4 (W) are conserved. Phylogenetic tree analysis showed that the bilberry R2R3-MYB genes were divided into 11 subgroups. Among them, six of the S4, S5 and S6 subgroups involved anthocyanin biosynthesis. Fluorescent quantitative PCR results showed that the six genes were expressed in seven stages of fruit development, but the expression patterns were different. These six genes may play a role in the coloring of bilberry fruit.