紫化茶树花青素与儿茶素时空变化特征及花青素代谢关键基因表达分析

Spatiotemporal variation characteristics of anthocyanins and catechins and key genes expression analysis of purple-leaf tea cultivars

  • 摘要: 目的 探究紫化茶树花青素与儿茶素时空变化特征并筛选花青素代谢关键基因,为解析紫化茶树品质成分时空分布规律及云南特色茶树资源的高效利用提供科学依据。方法 以紫娟(紫色芽叶)和云抗10号(绿色芽叶,对照品种)2个茶树品种为研究对象,分别于2023年3、4、5、7、9和10月采集2个茶树品种的1芽2叶新梢,于5月8日采摘紫娟1芽6叶新梢,按照叶位分离第1~6节位叶片(ZJ-1L~ZJ-6L)及对应茎段(ZJ-1S~ZJ-6S)。测定样品中总花青素含量及儿茶素单体组分含量,分析其动态变化规律,并进行主成分分析(PCA)、聚类分析和相关分析。使用实时荧光定量PCR检测紫娟茶树中花青素代谢关键基因的相对表达量。结果 不同采样时间紫娟茶树新梢总花青素含量均显著高于云抗10号(P<0.05,下同)。不同采样时间紫娟茶树新梢中,总花青素含量9月10日为峰值(5.81 μmol/g),同一采样时间,样品ZJ-2L中总花青素含量最高,为2.86 μmol/g,显著高于其他叶位和茎组织。云抗10号茶树新梢总儿茶素含量均值(210.94 mg/g)为紫娟均值(153.08 mg/g)的1.38倍,2个茶树品种儿茶素单体均以表没食子儿茶素没食子酸酯(EGCG)为主。相关分析结果显示,2个茶树品种总花青素含量与总儿茶素含量呈负相关,而同一时间采样的紫娟茶树新梢总花青素含量与总儿茶素含量呈极显著正相关(P<0.01,下同)。聚类分析结果显示,紫娟和云抗10号2个茶树品种的样品各自聚在一起,且采样时间相近的紫娟样品间距离较近;紫娟茶树新梢叶片和茎组织样品明显分开,样品ZJ-1L和ZJ-2L聚在同一分支,二者再与样品ZJ-1S聚在一起,这3个样品中的总花青素和儿茶素单体组分的丰度普遍较高。紫娟茶树新梢成熟茎(样品ZJ-4S、ZJ-5S、ZJ-6S)中的儿茶素单体组分的丰度普遍较低。实时荧光定量PCR检测结果显示,紫娟茶树3个样品MYB转录因子基因(CsMYB75)相对表达量均显著高于云抗10号茶树。结论 夏季后期至秋季初期紫娟茶树1芽2叶总花青素含量较高,紫娟茶树新梢的总儿茶素含量低于绿色芽叶茶云抗10号。CsMYB75基因持续高表达是紫娟茶树维持高花青素含量的核心调控因子。

     

    Abstract: Objective This study aimed to investigate spatiotemporal variation characteristics of anthocyanin and ca-techin in purple leaf tea plants and screen key genes of anthocyanin metabolism, so as to provide a scientific basis for elucidating the spatiotemporal distribution patterns of quality components in purple-leaf tea and utilizing of tea tree resources in Yunnan.Method Two tea plant cultivars, Zijuan (with purple buds and leaves) and Yunkang 10 (with green buds and leaves), were used as research objects in this study. Tender shoots consisting of one bud and two leaves were collected from both cultivars respectively in March, April, May, July, September, and October in 2023. On May 8, shoots of Zijuan with one bud and six leaves were collected and separated into leaves according to their positions (labeled as ZJ-1L to ZJ-6L) corresponding to stem segments (ZJ-1S to ZJ-6S). The contents of total anthocyanin and catechin monomers in collected samples were determined to analyze their dynamic variation patterns, followed by principal component analysis (PCA), cluster analysis, and correlation analysis. The relative expression of key genes of anthocyanin metabolism in Zijuan were detected using real-time fluorescence quantitative PCR.Result The total anthocyanin content in new shoots of Zijuan tea plant was significantly higher than that of Yunkang 10 (P<0.05, the same below). Across different sampling times, the total anthocyanin content in new shoots of Zijuan tea plant peaked on September 10th (5.81 μmol/g). At the same sampling time, sample ZJ-2L exhibited the highest total anthocyanin content (2.86 μmol/g), which was significantly higher than that of other leaf positions and stem tissues. The average total catechin content in new shoots of Yun-kang 10 (210.94 mg/g) was 1.38 times that of Zijuan (153.08 mg/g). Epigallocatechin gallate (EGCG) was the predominant catechin monomer in both tea cultivars. Correlation analysis revealed that total anthocyanin content and total catechin content in both cultivars were negatively correlated; however, the total anthocyanin content and total catechin content in Zijuan new shoots at the same sampling time had a highly significant positive correlation (P<0.01, the same below). Cluster analysis results showed that, samples from the two tea cultivars, Zijuan and Yunkang 10, clustered respectively, and Zijuan samples collected at similar time points were clustered closer to each other. The leaf and stem tissue samples of Zijuan new shoots were clearly separated; ZJ-1L and ZJ-2L were clustered on the same branch, and they were further clustered with ZJ-1S; these three samples generally exhibited higher abundances in total anthocyanin and catechin monomers. When Zijuan new shoots grew maturer, their sample stems (ZJ-4S, ZJ-5S, and ZJ-6S) generally exhibited low abundances of these catechin monomers. Real-time fluorescence quantitative PCR analysis revealed that the relative expression of the MYB transcription factor gene (CsMYB75) in three Zijuan samples were significantly higher than those in Yunkang 10.Conclusion During the late summer to early autumn, a high content of total anthocyanin in one-bud-two-leaf shoots of the Zijuan is observed, and the catechin content in Zijuan tea plant is significantly lower than that in Yun-kang 10 (with green buds and leaves). The sustained high expression of the CsMYB75 gene is identified as the key regulatory mechanism for the consistently high anthocyanin accumulation in Zijuan tea plant.

     

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