辣椒MATE基因家族鉴定及其与类黄酮积累的关联分析

Identification of MATE gene family and its association with flavonoid accumulation in pepper (Capsicum annuum L.)

  • 摘要: 【目的】 从辣椒全基因组中鉴定多药和毒性化合物排出蛋白(CaMATE)家族成员,分析其在辣椒不同组织中的表达模式及其与类黄酮积累的关联,为进一步研究CaMATE基因家族在辣椒类黄酮转运中的生物学功能,以及选育和改良高类黄酮含量辣椒品种提供理论依据。【方法】 以4个不同基因型辣椒品种(HJ10-1、HJ11-3-1、CJ12-17-1和0622-1-3-2-1-3-1)的果皮为试验材料,利用生物信息学方法鉴定CaMATE基因家族成员,并系统分析该家族成员的基本理化性质、基因结构、系统发育关系。通过分析4个不同基因型辣椒品种在不同组织中的表达模式,筛选调控辣椒类黄酮积累的CaMATE家族基因。通过实时荧光定量PCR检测8个CaMATE家族基因的表达情况。对CaMATE家族基因与类黄酮组分在4个辣椒品种中高表达的转录因子进行相关分析。【结果】 从辣椒基因组中鉴定出61个CaMATE基因家族成员,命名为CaMATE1~CaMATE61,这些成员随机且不均匀分布于13条染色体上;其编码蛋白包含2~12个跨膜结构、3种类型的MATE保守结构域,主要定位于细胞膜。CaMATE家族基因启动子区域的顺式作用元件主要涉及激素响应、生长发育和防御或胁迫响应。系统发育分析将MATE成员分为6个亚家族(Ⅰ~Ⅵ),辣椒和番茄MATE家族成员间的相似性较高。CaMATE家族成员在不同组织中的表达模式存在较大差异,CaMATE3CaMATE6CaMATE23基因的相对表达量在4个辣椒品种中均较高。CaMATE25基因的相对表达量仅在CJ12-17-1辣椒花后30 d的果皮样品中呈较高水平。实时荧光定量PCR检测结果与转录组测序结果一致。相关分析结果显示,CaMATE3CaMATE25CaMATE38CaMATE51基因均与超过15个类黄酮组分呈较强相关,并且这4个基因与bHLH96和HD-ZIP3高表达转录因子也呈较强相关。【结论】 共鉴定出61个CaMATE基因家族成员。在辣椒果实中高表达的基因(如CaMATE3CaMATE25等)与多个类黄酮组分存在较强相关性,推测这些基因参与调控辣椒类黄酮的转运过程。筛选出MYB113、bHLH96和HD-ZIP3为潜在类黄酮转运CaMATE家族基因的上游调控因子。

     

    Abstract: 【Objective】 Multidrug and toxic extrusion compound transporter family members (CaMATE) were identified from the whole genome of pepper,and their expression patterns in different pepper tissues and their association with flavonoid accumulation were analyzed,which provided theoretical basis for the subsequent research on the biological function of CaMATE gene family in pepper flavonoid transport and the breeding and improvement of new pepper varieties with high flavonoid content.【Method】 Peel of four pepper varieties with different genotypes(HJ10-1, HJ11-3-1, CJ12-17-1, and 0622-1-3-2-1-3-1) were used as the research materials for this study. CaMATE gene family members were identified by bioinformatics methods, and the basic physicochemical properties, gene structure, phylogenetic relationship of the family members were systematically analyzed. The CaMATE genes regulating the accumulation of flavonoids in pe-pper were screened by the expression patterns in different tissues of four pepper varieties with different genotypes, and the expression of eight CaMATE family genes were detected by real-time fluorescence quantitative PCR. Furthermore, the correlation analysis on CaMATE family genes with highly expressed transcription factors in flavonoid components in four pepper varieties was performed.【Result】 A total of 61 CaMATE gene family members were identified from the pepper genome,named CaMATE1-CaMATE61, which were unevenly distributed on chromosome 13. Their encoded proteins contained 2-12 transmembrane structures and 3 types of MATE conserved domains, which were mainly located in the cell membrane. The cis-acting elements in the promoter region of CaMATE family genes were mainly involved in hormone response, growth and development, and defense or stress response. Phylogenetic analysis showed that the MATE members were divided into six subfamilies (I-VI), and the similarity of MATE members between pepper and tomato was high. The expression patterns of CaMATE family members in different tissues were quite different, among which CaMATE3CaMATE6 and CaMATE23 were highly expressed in four pepper varieties. The relative expression level of CaMATE25 gene was only high in the peel of CJ12-17-1 pepper at 30 d after flowering. The results of real-time fluorescence quantitative PCR were consistent with those of transcriptome sequencing. Correlation analysis showed that CaMATE3CaMATE25CaMATE38 and CaMATE51 genes were strongly correlated with more than 15 flavonoid components, and these four genes were also strongly correlated with the highly expressed transcription factors, bHLH96, and HD-ZIP3.【Conclusion】 The 61 members of CaMATE gene family are identified from the pepper genome. The highly expressed genes (Such as CaMATE3 and CaMATE25) in pepper fruits have strong correlation with multiple flavonoid components, suggesting that they are involved in regulating the transport process of pepper flavonoids. Moreover, MYB113, bHLH96, and HD-ZIP3 are screened to be upstream regulators of potential flavonoid transport CaMATE family genes.

     

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