高茶氨酸茶树新品系‘福黄1号’黄化变异机理

doi:10.3864/j.issn.0578-1752.2022.09.012

摘要/Abstract

摘要：

【目的】分析茶树黄化变异相关的代谢和转录机制,探究高茶氨酸茶树新品系‘福黄1号’的黄化变异和高茶氨酸形成机理。【方法】以茶树‘福安大白茶’及其黄化突变种质‘福黄1号’为试验材料,利用超微电镜、广泛靶向代谢组学、靶向代谢组学及转录组学联合分析,确定茶树黄化变异相关的色素、代谢物及转录组数据。【结果】超微结构显示,‘福黄1号’的叶绿体类囊体呈现丝状,基粒片层排列散乱不规则,片层间疏松,存在许多异常的囊泡。色素含量测定表明,叶绿素a和叶绿素b含量显著下降,叶绿素a/b比率下降,相关基因SGR表达显著下调,黄化叶片中光捕获叶绿素a/b蛋白（LHC）表达显著下调。类胡萝卜素总含量虽然差异不大,但各组分含量显著变化,玉米黄质为唯一显著增加的组分,其调控基因VDE表达显著上调,其余组分含量均下降。与‘福安大白茶’相比,‘福黄1号’中共鉴定到680个差异表达基因（DEGs）和57个显著变化的代谢物（SCMs）。KEGG富集分析表明,SCMs和DEGs显著富集到氨基酸生物合成、谷胱甘肽代谢以及TCA循环等途径。此外,与碳和氮代谢相关的通路也被激活。通过靶向测定,共鉴定到19种游离氨基酸,新品系‘福黄1号’游离氨基酸含量显著高于‘福安大白茶’,达到97.13 mg∙g^-1,其中茶氨酸为66.90 mg∙g^-1,占氨基酸含量的68.89%,而精氨酸含量达到8.46 mg∙g^-1,是‘福安大白茶’的56.4倍。调控氨基酸合成的GOGAT和GLU的表达量上调1.17倍和3.17倍。【结论】‘福黄1号’的芽叶色泽主要受叶绿素、类胡萝卜素和类黄酮等色素代谢的影响,SGR和4种LHCs的共同作用也可能影响叶绿体的生物合成来调节叶片色泽。‘福黄1号’茶氨酸含量显著高于‘福安大白茶’的原因主要是泛素化相关的蛋白质水解酶表达上调蛋白降解能力加强,叶绿素和其他含氮分子生物合成的减少,以及黄化叶中碳骨架的缺乏,氨基和氮资源被更有效地储存,使得与氨基酸合成相关的氮代谢激活,茶氨酸合成前体物质之一的谷氨酸积累,这可能促使茶氨酸成为黄化叶中显著积累的含氮化合物。

关键词: 茶树, 福黄1号, 黄化, 代谢, 转录, 组学技术

Abstract:

【Objective】 The aim of this study was to analyze metabolic and transcriptional mechanism of etiolated variation in tea plant, and to explore the albino mutation of a new homotheanine tea cultivar Fuhuang 1 and its formation mechanism of homotheanine. 【Method】 The experimental materials were Fuan Dabaicha and Fuhuang 1. The combined analysis of ultramicroelectron microscopy, widely targeted metabolism, targeted metabolomics and transcriptomics was used to clarify the pigments, metabolism and transcriptome data related to the albino tea.【Result】The results of ultrastructure showed that the chloroplast thylakoids of Fuhuang 1 was filamentous, with irregular arrangement of basal granule lamellae and many abnormal vesicles. Chlorophyll a and chlorophyll b content, chlorophyll a/b ratio, SGR gene expression and light-harvesting chlorophyll a/b protein (LHC) expression were significantly reduced in yellow leaves. Although the total content of carotenoids did not change significantly, the content of each component changed significantly. Zeaxanthin was the only component with significant increase, and the expression of its regulatory gene VDE was significantly up-regulated, and the content of other components decreased. Compared with Fuan Dabaicha, 680 differentially expressed genes (DEGs) and 57 significantly changed metabolites (SCMs) were identified in Fuhuang 1. KEGG enrichment analysis showed that SCMs and DEGs were significantly enriched in pathways related to amino acid biosynthesis, glutathione metabolism and TCA cycle. In addition, the pathways related to carbon and nitrogen metabolism were also activated. A total of 19 free amino acids were identified by targeted determination. The free amino acid content of Fuhuang 1 was 97.13 mg∙g^-1, which was significantly higher than that of Fuan Dabaicha. Theanine content was 66.90 mg∙g^-1, accounting for 68.89% of amino acids, while the content of arginine was 8.46 mg∙g^-1, which was 56.4 times as more as that of Fuan Dabaicha. The expression levels of GOGAT and GLU, which regulated amino acid synthesis, were up-regulated in Fuhuang 1 by 1.17 and 3.17 times higher than that in Fuan Dabaicha. 【Conclusion】 The leaf color of Fuhuang 1 was mainly influenced by chlorophyll, carotenoids and flavonoids. The combined action of SGR and the four LHCs might also affect chloroplast biogenesis to regulate leaf color. The content of theanine in Fuhuang 1 was significantly higher than that in Fuan Dabaicha. The main reason was that the up-regulation of ubiquitination-related protein hydrolase activity and the enhanced protein degradation ability. The biosynthesis of chlorophyll and other nitrogen-containing molecules was reduced, the carbon skeleton in yellow leaves was lacking, amino and nitrogen resources were stored more effectively. The activation of nitrogen metabolism associated with amino acid synthesis led to the accumulation of glutamate, one of the precursors of theanine synthesis. Theanine accumulated significantly in yellow leaves.

Key words: tea plant, Fuhuang 1（Camellia sinensis）, albino, metabolomics, transcriptomics, omics technology

林馨颖,王鹏杰,杨如兴,郑玉成,陈潇敏,张磊,邵淑贤,叶乃兴. 高茶氨酸茶树新品系‘福黄1号’黄化变异机理[J]. 中国农业科学, 2022, 55(9): 1831-1845.

LIN XinYing,WANG PengJie,YANG RuXing,ZHENG YuCheng,CHEN XiaoMin,ZHANG Lei,SHAO ShuXian,YE NaiXing. The Albino Mechanism of a New High Theanine Tea Cultivar Fuhuang 1[J]. Scientia Agricultura Sinica, 2022, 55(9): 1831-1845.

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样本名称 Sample	原始序列 Raw read	过滤序列 Clean read	过滤碱基 Clean base (G)	整体测序错误率 Error rate (%)	Q20 (%)	Q30 (%)	GC含量 GC content (%)
GF-1	47303628	46072126	6.91	0.03	96.65	90.46	44.83
GF-2	47291032	46122320	6.92	0.03	96.37	89.87	44.58
GF-3	57641310	56256308	8.44	0.03	96.27	89.63	44.70
YF-1	46752324	45637674	6.85	0.03	96.47	90.07	44.41
YF-2	45390834	44391162	6.66	0.03	95.62	88.54	44.39
YF-3	46546464	45474928	6.82	0.03	96.54	90.23	44.64
合计 Total	290925592	283954518	42.6

组分 Content	福安大白茶 Fuan Dabaicha	福黄1号 Fuhuang 1	组分 Content	福安大白茶 Fuan Dabaicha	福黄1号 Fuhuang 1
r-氨基丁酸 GABA	0.14±0.00a	0.39±0.37a	缬氨酸 Valine	0.14±0.00b	0.26±0.00a
丙氨酸 Alanine	0.47±0.00a	0.50±0.02a	异亮氨酸 Isoleucine	0.16±0.00b	0.44±0.14a
茶氨酸 Theanine	17.42±0.64b	66.90±2.69a	谷氨酸 Glutamate	2.47±0.15a	4.27±0.35a
脯氨酸 Proline	0.44±0.00b	1.36±0.00a	谷氨酰胺 Glutamine	0.91±0.89b	4.12±0.26a
赖氨酸 Lysine	0.20±0.01b	0.74±0.10a	精氨酸 Arginine	0.15±0.00b	8.46±1.48a
亮氨酸 Leucine	0.15±0.00a	0.46±0.28a	丝氨酸 Serine	1.06±0.06a	1.94±0.13a
天冬氨酸 Aspartate	0.21±0.41b	4.35±0.44a	天冬酰胺 Asparagine	0.15±0.00b	1.32±0.12a
酪氨酸 Tyrosine	0.22±0.00a	0.25±0.02a	组氨酸 Histidine	0.07±0.01b	0.26±0.02a
色氨酸 Tryptophan	0.22±0.01a	0.26±0.01a	甘氨酸 Glycine	0.06±0.01a	0.09±0.02a
苏氨酸 Threonine	0.19±0.01b	0.60±0.04a	总量 Total content	24.88±0.44b	97.13±5.42a