基于靶向代谢组学分析嫁接对茶树代谢物及其黄酮类代谢途径的影响

Effects of grafting on metabolites and flavonoid metabolic pathways of tea plant based on widely targeted metabolomics analysis

  • 摘要: 目的 分析茶树嫁接前后茶叶中代谢物变化及其黄酮类代谢途径,为凌云白毫茶老茶园换种改造以及茶产业高质量发展提供理论参考。方法 以茶树品种桂热2号为接穗,凌云白毫茶为砧木的嫁接茶树为研究对象,采集嫁接前和嫁接后的桂热2号1芽2叶样本,分别标记为C和B样本,利用广泛靶向代谢组学技术检测嫁接前后茶叶中代谢物含量,对样本进行主成分分析(PCA)及正交偏最小二乘判别分析(OPLS-DA),采用极差法归一化处理代谢物含量,对差异代谢物进行聚类分析,依据log2 Fold Change≥2或log2 Fold Change≤0.5且变量重要性投影(VIP)≥1的标准筛选差异代谢物,并通过KEGG数据库对黄酮类差异代谢物的相关通路进行分析。结果 通过PCA分析和OPLS-DA分析均可将B和C样本完全分离,第一主成分(PC1)的贡献率为89.80%,第二主成分(PC2)的贡献率为3.04%。聚类分析结果显示,2个样本的代谢物存在显著差异,并且嫁接后黄酮类物质变化最大。茶叶中代谢物表现出显著的差异。在桂热2号茶树嫁接前后2个样本中共检测出3495个差异代谢物,其中有2919个具有显著差异的代谢物;与C样本相比,B样本中有2042个显著差异代谢物VIP上调和877个显著差异代谢物VIP下调。对桂热2号茶树嫁接前后排名前20的显著差异代谢物进行分析,结果显示,B样本中共有17个差异代谢物差异倍数明显高于C样本,差异代谢物主要集中在黄酮苷元Ⅲ的生物合成、不饱和脂肪酸的生物合成、花青素合成、淀粉和蔗糖代谢、咖啡因代谢等KEGG代谢通路中。此外,嫁接前后2个样本中有665个黄酮类物质具有显著差异,与黄酮类显著差异代谢物相关的主要通路主要集中在类黄酮生物合成通路、花青素生物合成通路、异黄酮生物合成通路、黄酮和黄酮醇生物合成通路,上述4个通路的显著差异代谢物分别有30、10、10、23个,主要为黄酮类物质,且大部分含量表现为上调。结论 茶树桂热2号通过嫁接会使茶叶中的代谢物发生显著变化,促进黄酮类物质在茶叶中积累,进而提高茶叶品质。

     

    Abstract: Objective This study aimed to analyze changes in metabolites and flavonoid metabolic pathways in leaves of Lingyun Baihao tea before and after grafting, so as to provide theoretical references for the replanting and transformation and industrial high quality development of Lingyun Baihao tea in old tea gardens in the future.Method Taking Guire 2 as the scion and Lingyun Baihao tea as the rootstock, the grafted tea plants were used as research subjects. One bud and two leaves samples of Guire 2 were collected before and after grafting and labeled as C and B samples, respectively. Widely targeted metabolomics technology was used to detect the metabolite content in tea before and after grafting to perform principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA). The metabolite content was normalized using the range method. Cluster analysis was performed on differential metabolites, which were screened based on the criteria of log2 Fold Change≥2 or log2 Fold Change≤0.5 and variable importance in projection (VIP)≥1. Pathways related to differential metabolites of flavonoids were analyzed based on the KEGG database.Result Both PCA and OPLS-DA could completely separate sample B and sample C. The contribution rate of the first principal component (PC1) was 89.80%, and the contribution rate of the second principal component (PC2) was 3.04%. Cluster analysis results showed significant differences in metabolites between the two samples, with the greatest changes observed in flavonoids after grafting. The metabolites in tea showed significant differences. A total of 3495 metabolites were detected in two tea plant samples of Guire 2 before and after grafting, of which 2919 metabolites showed significant differences. Compared with sample C, sample B had 2042 metabolites with significantly up-regulated VIPs and 877 metabolites with significantly down-regulated VIPs. The analysis of the top 20 significantly differential metabolites in tea plants of Guire 2 before and after grafting showed that sample B had 17 differential metabolites whose fold changes were significantly higher than those in sample C; the differential metabolites were mainly concentrated in KEGG metabolic pathways such as flavonoid glycoside Ⅲ biosynthesis, unsaturated fatty acid biosynthesis, anthocyanin synthesis, starch and sucrose metabolism, and caffeine metabolism. Moreover, 665 flavonoids showed significant differences between the two samples before and after grafting. The main pathways related to significantly differential metabolites of flavonoids were primarily concentrated in the flavonoid biosynthesis pathway, anthocyanin biosynthesis pathway, isoflavone biosynthesis pathway, and flavonoid and flavonol biosynthesis pathway, and these four pathways had 30, 10, 10, and 23 significantly differential metabolites (mainly flavonoids) whose contents were mostly up-regulated.Conclusion Grafting of tea plant Guire 2 can significantly alter the metabolites in tea leaves, promote the accumulation of flavonoids in tea leaves, and thereby improve tea quality.

     

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