紫芽茶树品种&#x02018;紫娟'花青素合成酶基因CsANS1的克隆与功能分析

范延艮; 萧越; 孟凡月; 刘文杰; 张颖; 孙平; 张丽霞; 任丽军

doi:10.13305/j.cnki.jts.2025.05.004

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茶叶科学 ›› 2025, Vol. 45 ›› Issue (5) : 757-769. DOI: 10.13305/j.cnki.jts.2025.05.004

研究报告

紫芽茶树品种‘紫娟'花青素合成酶基因CsANS1的克隆与功能分析

范延艮¹, 萧越¹, 孟凡月², 刘文杰¹, 张颖¹, 孙平¹, 张丽霞^1,*, 任丽军^1,*

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Cloning and Functional Analysis of the Anthocyanin Synthase Gene CsANS1 in the Purple Bud Tea Variety ‘Zijuan'

FAN Yangen¹, XIAO Yue¹, MENG Fanyue², LIU Wenjie¹, ZHANG Ying¹, SUN Ping¹, ZHANG Lixia^1,*, REN Lijun^1,*

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摘要

花青素具有抗氧化、抗癌等多种健康功效,是功能型作物育种的重要靶标。花青素合成酶（ANS）是花青素生物合成关键酶,其在多种作物中已被证实可调控花青素积累。然而,茶树ANS基因（CsANS）的功能研究仍较滞后。基于‘紫娟'等4种叶色茶树比较转录组数据,鉴定出‘紫娟'紫叶中高表达的CsANS1;基因表达分析显示,CsANS1在‘紫娟'中的表达量显著高于其他品种及CsANS家族其他成员（CsANS2和CsANS3）。此外,组织表达分析表明CsANS1在‘紫娟'叶片中表达量最高,其表达水平随芽叶发育呈现先升高后降低的变化趋势。系统进化分析表明,ANS基因家族成员均具有高度保守的Fe(Ⅱ)/2OG dioxygenase结构域,且家族成员间的序列相似性高于种间同源基因。基因比较测序和蛋白三维结构预测分析发现,‘紫娟'CsANS1编码区存在2个特异非同义变异,其中165位氨基酸变异可能影响局部蛋白构象。进一步通过VIGS沉默CsANS1表达,‘紫娟'新梢花青素含量显著降低,叶片颜色变浅。本研究证实CsANS1是‘紫娟'芽叶花青素高水平积累的关键基因,可为茶树高花青素品种的分子育种提供重要靶点。

Abstract

Anthocyanins exhibit diverse health benefits in humans, including antioxidant and anticancer effects, making them a key target for functional crop breeding. Anthocyanin synthase (ANS), a pivotal enzyme in anthocyanin biosynthesis, has been demonstrated to regulate anthocyanin accumulation in various crops. However, functional studies on ANS genes in tea plants (Camellia sinensis) remain limited. In this study, we identified a specifically highly expressed CsANS1 gene in the purple leaves of tea cultivar ‘Zijuan' through comparative transcriptome analysis of four tea cultivars with distinct leaf colors. Gene expression analysis reveals that CsANS1 expression in ‘Zijuan' was significantly higher than in other cultivars and other ANS family members (CsANS2 and CsANS3) in tea plants. Tissue-specific expression analysis further demonstrats that CsANS1 exhibited the highest expression level in ‘Zijuan' leaves, showing an expression trend of first increasing and then decreasing with the development of buds and leaves. Phylogenetic analysis indicats that sequence similarity among ANS gene copies within species is higher than that among interspecies homologs, and all members of the tea ANS family possess the highly conserved Fe(Ⅱ)/2OG dioxygenase domain. Comparative gene sequencing and protein tertiary structure prediction analyses reveal two non-synonymous mutations in the coding region of CsANS1 in ‘Zijuan', with the amino acid variation at position 165 potentially affecting local protein conformation. Following VIGS-mediated silencing of CsANS1, the anthocyanin content in ‘Zijuan' shoots significantly decreased, accompanied by a lightening of leaf color. In conclusion, this study confirms that CsANS1 is a key gene regulating high-level anthocyanin accumulation in ‘Zijuan' buds and leaves, providing an important molecular target for breeding high-anthocyanin tea cultivars.

导出引用

范延艮, 萧越, 孟凡月, 刘文杰, 张颖, 孙平, 张丽霞, 任丽军. 紫芽茶树品种‘紫娟'花青素合成酶基因CsANS1的克隆与功能分析[J]. 茶叶科学. 2025, 45(5): 757-769 https://doi.org/10.13305/j.cnki.jts.2025.05.004

FAN Yangen, XIAO Yue, MENG Fanyue, LIU Wenjie, ZHANG Ying, SUN Ping, ZHANG Lixia, REN Lijun. Cloning and Functional Analysis of the Anthocyanin Synthase Gene CsANS1 in the Purple Bud Tea Variety ‘Zijuan'[J]. Journal of Tea Science. 2025, 45(5): 757-769 https://doi.org/10.13305/j.cnki.jts.2025.05.004

中图分类号： S571.1

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