中国辣椒AAT基因家族的鉴定及表达分析

    Identification and Expression Analysis of the AAT Gene Family in Chinese Pepper

    • 摘要:
      目的 中国辣椒(Capsicum chinense)是辣椒主要栽培种之一,种内多数品种的果实呈现由支链酯类物质形成的浓郁果香。醇酰基转移酶(Alcohol acyl-CoA transferase,AAT)催化支链酯类合成的最后一步反应,对支链酯类含量有重要影响。鉴定中国辣椒中AAT基因家族成员并分析其组织表达模式,可为其功能研究提供参考。
      方法 通过生物信息学分析和实时荧光定量PCR对中国辣椒的AAT基因家族进行鉴定及表达特征分析。
      结果 从中国辣椒基因组中鉴定到10个AAT基因,分布于6条染色体上,分别命名为CcAAT1-10。理化性质预测结果显示,AAT基因家族编码的氨基酸序列长度在256~683 aa之间,分子质量范围为29~77 kDa,等电点范围为5.34~8.79,平均亲水系数均为负值,不稳定指数在23.76~51.02之间。蛋白质结构预测显示,CcAAT的二级结构以α-螺旋和无规则卷曲为主,三级结构差异较大。亚细胞定位预测发现CcAAT均定位于细胞质中,其基因启动子上存在16种顺式调控元件。时空表达分析显示,CcAAT5CcAAT6在辣椒各器官中均未检测到表达,CcAAT8在花和果实中特异表达,CcAAT1CcAAT3CcAAT7在叶片中高表达,CcAAT4在根系中高表达。
      结论 明确了中国辣椒中AAT基因家族成员和表达模式,推测CcAAT8可能是影响果实支链酯类物质含量的关键基因。

       

      Abstract:
      Objective Capsicum chinense is one of the main cultivated species of pepper, which generally has a strong fruit aroma formed by branched chain esters. AAT (alcohol acyl-CoA transferase) catalyzes the last step in the synthesis of branched chain esters and has an important impact on its content. Identification of the AAT gene family in C. chinense and analysis on its expression patterns will provide references for its functional studies.
      Method Bioinformatics and real-time quantitative PCR analysis were applied to identify the AAT gene family of C. chinense and analyze its expression patterns.
      Result Ten members of the AAT gene family were identified and named CcAAT1-CcAAT10 according to their distribution order on six chromosomes. Physicochemical properties prediction revealed that the length of amino acid sequence encoded by AAT gene family range from 256 to 683 aa, with their molecular mass ranging from 29 to 77 kDa, and isoelectric point ranging from 5.34 to 8.79. Average hydrophobic coefficients of them were all negative values. Their instability indexes ranged 23.76 to 51.02. Protein structure prediction showed that their secondary structure was dominated by α-helices and irregular convolutions, and the tertiary structure varied widely. Subcellular localization prediction revealed that all CcAAT were located in the cytoplasm and 16 cis-regulatory elements were found on their gene promoter. Spatiotemporal expression analysis showed that CcAAT5 and CcAAT6 had no detectable expression, CcAAT8 was specifically expressed in flowers and fruits, CcAAT1, CcAAT3 and CcAAT7 were highly expressed in leaves, and CcAAT4 was highly expressed in roots.
      Conclusion It clarified the expression pattern of the AAT gene family in C. chinense and it was hypothesized that CcAAT8 might be a key gene affecting the content of branched chain esters.

       

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