转录组和代谢组联合分析桑葚发育过程中可溶性糖和有机酸代谢的变化  

Transcriptome and metabolome combined analysis metabolism change of soluble sugars and organic acids in mulberry fruit during development stages

作　　者：张若彤 李蒙 齐一鸣 王晓萍 孙志超 ZHANG Ruotong;LI Meng;QI Yiming;WANG Xiaoping;SUN Zhichao(Institute of Sericulture,Chengde Medical University,Chengde 067000,Hebei,China)

机构地区：[1]承德医学院蚕业研究所,河北承德067000

出　　处：《果树学报》2024年第4期690-702,共13页Journal of Fruit Science

基　　金：河北省“三三三人才工程”资助项目(A202101051);河北省教育厅一般项目(QN2020236);河北省自然科学基金项目(C2019406113,H2023406059);承德市基础研究专项(202205B069);承德医学院校级课题(202214);大学生创新创业训练计划项目(2022040,2023034,2023141)。

摘　　要：【目的】探究桑葚发育过程中可溶性糖和有机酸代谢及转录表达水平,揭示桑葚品质形成的分子机制。【方法】以青果期(W1)、转色期(W2)、成熟期(W3)白色桑葚为试验材料,分别测定3个阶段可溶性糖和有机酸含量及转录组变化,并基于转录组与代谢组联合分析揭示调控可溶性糖和有机酸代谢的分子机制。【结果】共检测到64种代谢物,其中有机酸52种、可溶性糖12种。分析发现,蔗糖、葡萄糖和D-果糖为桑葚中主要可溶性糖类物质,苹果酸、柠檬酸和琥珀酸为桑葚中主要有机酸类物质。转录组测序共获得58.65 Gb Clean Data,差异基因分析发现W3 vs W1组获得的差异基因数量最多为9098个。而KEGG富集分析表明,W2 vs W1和W3 vs W2组中差异基因富集到与糖酸代谢相关的通路,主要为淀粉和蔗糖代谢及三羧酸循环通路,在W2 vs W1组中有52个上调的差异基因富集到淀粉和蔗糖代谢,27个上调的差异基因富集到柠檬酸循环,在W3 vs W2组中有27个上调的差异基因富集到淀粉和蔗糖代谢。代谢组和转录组关联分析表明,NINV、HK、CS、ACO、MDH和ICDH是桑葚糖酸积累的关键调控基因。荧光定量分析(qRTPCR)表明,关键调控基因在不同发育时期表达上调,与转录组中表达趋势一致。【结论】基因NINV、HK、CS、ACO、MDH和ICDH在桑葚成熟中可溶性糖和有机酸的合成与代谢中具有重要调控作用,初步揭示了桑葚口感变化的生物学基础。【Objective】Through the systematic study of the metabolism and molecular mechanism of sugar and organic acids,the mechanism of fruit taste formation was well revealed.In this study,we in-vestigated the metabolism of soluble sugar and organic acid and transcriptome expression levels during the development of mulberry(Morus alba)in order to reveal the molecular mechanism of fruit quality formation of mulberry.【Methods】White mulberry fruits were used as experimental materials at green-ing stage(W1),transforming stage(W2)and ripening stage(W3).The content and transcriptome of the soluble sugar and organic acid at three stages were determined separately,and the molecular mechanism of regulation of the soluble sugar and organic acid metabolism were analyzed based on the combination of transcriptome and metabolome.By exploring the key differential genes regulating the synthesis and metabolism of the soluble sugar and organic acid during mulberry ripening,the metabolic network was proposed to elucidate the influence of fruit development on the metabolism of the soluble sugar and or-ganic acid.The UPLC-MS/MS targeted metabolomics method was used to detect the changes of metab-olites at 3 developmental stages,and the Cluster analysis was performed on the obtained different sub-stances.The expression of the differential genes was analyzed by qRT-PCR and the obtained differential genes were further analyzed through KEGG(Kyoto encyclopedia of genes and genomes)pathway en-richment analysis.【Results】A total of 64 metabolites were detected,including 52 organic acids and 12 soluble sugars.According to the cluster analysis of different substances,the results showed that there were obvious changes in sugar and acid metabolism during the mulberry fruit development.Through the data analysis,it was found that the sucrose,glucose and D-fructose were the main soluble sugars in mulberry fruits,and their contents continued to increase during the development of mulberry fruits,and reached a peak at W3.The malic acid,citric acid

关 键 词：桑葚 代谢组 转录组 可溶性糖 有机酸 

分 类 号：S663.2[农业科学—果树学]