梅花肉桂醇脱氢酶基因的鉴定和功能分析
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北京林业大学园林学院/花卉种质资源创新与分子育种北京市重点实验室/国家花卉工程技术研究中心/ 城乡生态环境北京实验室,北京100083

作者简介:

研究方向为花卉种质资源与遗传育种,E-mail: 1309707639@qq.com;

通讯作者:

包菲,研究方向为花卉种质资源与遗传育种,E-mail : baofei@bjfu.edu.cn

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基金项目:

国家自然科学基金 (32271910);北京市大学生科学研究与创业行动计划 (S202310022093)


Identification and Functional Analysis of Cinnamyl Alcohol Dehydrogenase PmCAD) Genes in Prunus mume
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Affiliation:

School of Landscape Architecture, Beijing Forestry University/ Beijing Key Laboratory of Ornamental Plants Germplasm Innovation and Molecular Breeding/ National Engineering Research Center for Floriculture/ Beijing Laboratory of Urban and Rural Ecological Environment, Beijing 100083

Fund Project:

Foundation projects: National Natural Science Foundation of China (32271910); College Student Research and Career-creation Program of Beijing (S202310022093)

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    摘要:

    梅花(Prunus mume Siebold & Zucc.)品种丰富,花香香型多样,其中肉桂醇和乙酸肉桂酯是梅花粉色花品种特有的花香成分。在植物体内,肉桂醛在肉桂醇脱氢酶 (CAD) 的催化下还原生成肉桂醇,肉桂醇又在乙酰转移酶的催化下生成乙酸肉桂酯。因此,肉桂醇不仅是梅花重要的特征花香成分,还是合成乙酸肉桂酯的重要前体。本研究基于梅花二代基因组数据鉴定了56个PmCAD同源基因,发现梅花PmCAD家族成员可分为5个亚组,第I、II和III亚组的蛋白保守基序组成较相似,与第IV和V亚组的差别较大,推测它们之间可能存在功能分化。染色体片段复制和串联复制是PmCAD在基因组中复制进化的两种形式。梅花PmCAD基因启动子区域广泛存在脱落酸、茉莉酸甲酯、水杨酸响应元件和分生组织表达相关元件,推测PmCAD基因主要参与梅花的生长发育和抗逆响应。分析与多物种来源CAD蛋白共建树,结果表明位于第III亚组的3个PmCAD可能参与梅花花香合成过程。不同组织器官热图分析表明这3个PmCAD基因都在花器官中表达。基因相对表达量分析发现它们在花开放不同阶段的表达规律不同,在花开放后表达水平较高。体外酶活试验验证了这3个PmCAD都具有肉桂醇脱氢酶活性,因此认为它们共同参与梅花重要花香成分肉桂醇的合成。亚细胞定位分析发现它们在细胞内的分布存在差异,推测它们可能在细胞的不同部位发挥作用。本研究揭示了梅花PmCAD基因的进化和功能分化,为梅花重要花香成分的合成研究奠定了理论基础和为花香分子育种提供了理论依据。

    Abstract:

    Prunus mume Siebold & Zucc., known for its numerous varieties and diverse floral scents, includes cinnamyl alcohol and cinnamyl acetate as unique floral scent components in pink varieties. In plants, cinnamaldehyde is reduced to cinnamyl alcohol under the catalytic action of cinnamyl alcohol dehydrogenase (CAD), which in turn forms cinnamyl acetate under the catalysis of acetyltransferase. Therefore, cinnamyl alcohol is not only a significant floral scent component in P. mume, but also a crucial precursor for the synthesis of cinnamyl acetate. This study identified 56 PmCAD homologous genes based on the updated genome of P. mume. These PmCAD family members can be categorized into five subgroups. The protein conserved motif composition of subgroups I, II, and III is relatively similar, with significant differences from subgroups IV and V, suggesting potential functional divergence. Chromosome segmental duplication and tandem duplication were identified as two forms of PmCAD replication in genome evolution. The promoters of PmCAD genes were found to extensively contain abscisic acid, methyl jasmonate, salicylic acid responsive elements, and meristem related elements, suggesting that PmCAD genes are mainly involved in the growth, development, and stress response. Phylogenetic analysis of CAD from P. mume and other species indicated that the three PmCAD genes located in subgroup III may be involved in floral scent synthesis. Heat map analysis in different tissues and organs indicated that the three PmCAD were all expressed in flowers. Furthermore, analysis of relative gene expression revealed that the expression patterns of the three candidate PmCAD genes varied at different stages of flowering, with higher expression levels after the flowers bloom. The in vitro enzyme activity analysis confirmed that all three PmCAD possess cinnamyl alcohol dehydrogenase activity, indicating that they jointly participate in the synthesis of cinnamyl alcohol. Subcellular localization analysis revealed differences in their intracellular distribution, suggesting distinct cellular roles. This study sheds light on the evolution and functional differentiation of the PmCAD genes in P. mume, laying a theoretical foundation for the synthesis of key floral scent components and providing theoretical bases for floral fragrance molecular breeding in P. mume.

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刘巳塬,罗瑜杭,邓思航,等.梅花肉桂醇脱氢酶基因的鉴定和功能分析[J].植物遗传资源学报,2025,26(2):356-368.

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  • 收稿日期:2024-05-16
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  • 在线发布日期: 2025-01-23
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