基于转录组测序的紫叶李叶色相关基因分析

Analysis of Leaf Color Related Genes Based on Transcriptome Sequencing in Prunus cerasifera Ehrhar f.atropurpurea Rehd.

紫叶李是一种具有较高观赏价值的落叶乔木,叶色是紫叶李重要的观赏性状之一。为探究影响紫叶李花色素苷合成的关键酶基因,本研究应用Illumina NovaSeq6000测序平台,对性状分别为红叶、绿叶和普通叶的紫叶李进行了转录组测序。实验以红叶紫叶李为实验组,绿叶和普通叶分别做对照组,每个样品各3个生物学重复,共获得195548106条Clean reads。通过对差异表达基因分析,筛选到红叶相对绿叶的差异表达基因5376个,其中上调表达基因2686个,下调表达基因2690个;红叶相对普通叶的差异表达基因6197个,其中上调表达基因3193个,下调表达基因3004个。通过GO功能分类,可分为生物过程、细胞组分和分子功能3大类,其中红叶紫叶李和绿叶紫叶李比对注释到44个功能组,红叶紫叶李和普通叶紫叶李比对注释到47个功能组。通过代谢通路KEGG pathway分析表明,红叶紫叶李和绿叶紫叶李比对有897个差异表达基因参与了120条已知的代谢通路,红叶紫叶李和普通叶紫叶李比对有1108个差异表达基因参与了121条已知的代谢通路,其中苯丙素合成通路、类黄酮合成通路及黄酮和黄酮醇合成通路3条花色素苷相关代谢通路的部分基因表达存在显著的差异,并通过实时荧光定量,验证花色素苷合成相关差异表达基因在3种不同叶色紫叶李中的表达模式。本研究丰富了紫叶李的转录组信息,为进一步揭示紫叶李叶片着色的相关分子机制、生长相关基因克隆和表达等研究提供科学依据。

Prunus cerasifera Ehrhar f.atropurpurea Rehd.is a kind of deciduous tree with high ornamental value.Leaf color is one of the important ornamental characters of P.cerasifera.In order to explore the key enzyme genes affecting anthocyanin synthesis of P.cerasifera,in this study,transcriptome sequencing of P.cerasifera with green leaf and red leaf phenotype was performed using Illumina NovaSeq6000 sequencing platform.In this experiment,195548106 clean reads were obtained with red P.cerasifera as the experimental group,green leaf and common leaf as the control group,and each sample had 3 biological replicates.5376 differentially expressed genes in red and green leaves were screened,including 2686 up-regulated genes and 2690 down-regulated genes.There were 6197 differentially expressed genes in red leaves,including 3193 up-regulated genes and 3004 down-regulated genes.According to GO functional classification,it can be divided into three categories:biological process,cell component and molecular function.Among them,red leaf of P.cerasifera and green leaf of P.cerasifera were compared to 44 functional groups,and red leaf of P.cerasifera and common leaf of P.cerasifera were compared to 47 functional groups.KEGG pathway analysis showed that there were 897 differentially expressed genes involved in 120 known metabolic pathways in the ratio of red leaf of P.cerasifera to green leaf of P.cerasifera,and 1108 differentially expressed genes involved in 121 known metabolic pathways in the ratio of red leaf of P.cerasifera to common leaf of P.cerasifera.There were significant differences in the expression of some genes in the three anthocyanin-related metabolic pathways:phenylpropanin synthesis pathway,flavonoid synthesis pathway,and flavonoid and flavonol synthesis pathway.The expression patterns of anthocyanin synthesis-related differentially expressed genes in three different leaf of P.cerasifera were verified by real-time fluorescence quantitative analysis.This study enriched the transcriptome information of P.cerasifera,and provided a scientific basis for further study on the molecular mechanisms related to leaf coloring and the cloning and expression of growth-related genes.