梅花花香合成相关肉桂醇脱氢酶基因(PmCAD)的鉴定和功能分析
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花卉种质资源创新与分子育种北京市重点实验室

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国家自然科学基金面上项目(32271910)


Floral Scent Synthesis Related Cinnamyl Alcohol Dehydrogenase (PmCAD) Genes Identification and Functional Analysis in Prunus mume
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Beijing Key Laboratory of Ornamental Plants Germplasm Innovation and Molecular Breeding,National Engineering Research Center for Floriculture,School of Landscape Architecture,Beijing Forestry University

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The National Natural Science Foundation of China (32271910)

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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PmCADs同源基因,发现梅花PmCADs家族可分为五个亚组,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PmCADs在基因组中复制进化的两种形式;梅花PmCADs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PmCADs基因主要参与梅花的生长发育和抗逆响应;多物种来源CAD共建树分析,表明位于第III亚组的3个PmCADs基因可能参与梅花花香合成过程,不同组织器官热图分析表明它们都在花器官中表达。基因相对表达量分析发现梅花3个候选PmCADs基因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PmCADs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PmCADs基因的进化和功能分化,为梅花重要花香成分的合成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. has many varieties and diverse floral scent types, among which cinnamyl alcohol and cinnamyl acetate are unique floral scent components in pink varieties of P. mume. In plants, cinnamaldehyde is reduced to cinnamyl alcohol under the catalysis of cinnamyl alcohol dehydrogenase (CAD), which in turn generates cinnamyl acetate under the catalysis of acetyltransferase. Therefore, cinnamyl alcohol is not only an important floral scent component in P. mume, but also an important precursor for the synthesis of cinnamyl acetate. This study identified 56 PmCADs homologous genes based on the second-generation genome of P. mume. It was found that the PmCADs family can be divided into five subgroups, and 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 differentiation between them. Chromosome fragment replication and tandem replication were identified as two forms of PmCADs replication in genome evolution. It was found that the promoter regions of PmCADs genes extensively contained abscisic acid, methyl jasmonate, salicylic acid responsive elements, and meristem related elements suggesting that the PmCADs genes are mainly involved in the growth, development, and stress response of P. mume. Phylogenetic analysis of CAD from P. mume and other species suggested that the three PmCADs genes located in subgroup III may be involved in the floral scent synthesis in P. mume. Heat map analysis of PmCADs genes in different tissues and organs indicated that the three PmCADs were all expressed in flowers. Furthermore, analysis of relative gene expression revealed that the expression patterns of the 3 candidate PmCADs 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 PmCADs have cinnamyl alcohol dehydrogenase activity, indicating that they jointly participate in the synthesis of cinnamyl alcohol, an important floral scent component of P. mume. Subcellular localization analysis revealed differences in their distribution within the cell, suggesting that they may play roles in different parts of the cell. This study revealed the evolution and functional differentiation of the PmCADs genes in P. mume, laying theoretical foundations for the synthesis of important floral scent components and providing theoretical bases for floral fragrance molecular breeding in P. mume.

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  • 收稿日期:2024-05-16
  • 最后修改日期:2024-06-21
  • 录用日期:2024-09-09
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