蒲瓜 JMJ 组蛋白去甲基化酶家族基因鉴定及表达分析

    Identification and Expression Analysis of the JMJ Histone Demethylase Family Genes in Lagenaria siceraria

    • 摘要:
      目的 由含JMJ组蛋白去甲基化酶(Histone demethylases,HDMs)介导的去甲基化对染色质“构象”及其引起的复制、转录等生理活动具有重要作用。鉴定蒲瓜(Lagenaria siceraria)JMJ家族基因并解析其表达模式,为了解其生长发育及逆境响应机制提供依据。
      方法 利用HMM在蒲瓜全基因组中鉴定JMJ组蛋白去甲基化酶家族基因,通过生物信息学方法、RNA-Seq数据及RT-qPCR分析JMJ组蛋白去甲基化酶家族基因在蒲瓜根、子叶、真叶、下胚轴、顶芽、雌花、雄花及卷须的表达模式。
      结果 从蒲瓜基因组鉴定到17个JMJ家族成员,分为5个亚家族,按染色体分布命名为LsiJMJ1~LsiJMJ17。理化性质分析和亚细胞定位预测结果显示,蒲瓜JMJ蛋白长度为412~1 852 aa,多数为弱酸性核定位蛋白。染色体定位和共线性分析发现,蒲瓜JMJ基因不均匀分布于9条染色体,含1对基因高度同源基因,且与拟南芥、甜瓜等物种存在多对同源基因。保守基序分析表明,同一亚组的蛋白具有相似的motif。基因结构和编码蛋白结构域分析显示,多数同源基因具有相似的外显子- 内含子结构,编码序列含6~33个外显子。顺式作用元件分析发现,JMJ家族基因含有胁迫反应、激素响应、生长发育调节、光响应以及其他类元件。蛋白网络互作分析表明,LsiJMJ3与LsiJMJ9、LsiJMJ2、LsiJMJ12间可能存在蛋白共表达,LsiJMJ12、LsiJMJ5和LsiJMJ7或LsiJMJ11两两之间可能存在蛋白共表达。表达分析表明,LsiJMJ4LsiJMJ9在所有组织中显著高表达,尤其在雌花和顶芽中表达量最高,LsiJMJ11在雄花中特异高表达,LsiJMJ8在5种组织中低表达。
      结论 成功鉴定出17个JMJ家族成员,其保守结构域可能在它们去甲基化酶活方面发挥重要作用。蒲瓜JMJ家族基因的表达可能受发育及逆境胁迫影响,推测LsiJMJ4LsiJMJ7LsiJMJ9可能参与基础生长发育过程,如雌花发育或顶芽分化调控,LsiJMJ11与雄花发育相关,LsiJMJ8可能受特定条件诱导表达。

       

      Abstract:
      Objective Demethylation mediated by histone demethylases (HDMs) containing the JMJ domain plays a crucial role in chromatin conformation and the physiological activities it governs, such as replication and transcription. Identifying members of the JMJ histone demethylase gene family in bottle gourd (Lagenaria siceraria) and analysing their expression patterns could provide a reference for studying its growth and development.
      Method The genome-wide identification of the JMJ histone demethylase family genes in bottle gourd was performed by HMM, and the expression patterns of the JMJ histone demethylase family genes were analysed by bioinformatics, RNA-Seq data and RT-qPCR in the roots, cotyledons, real leaves, hypocotyls, terminal buds, pistillate flowers, staminate flowers and tendrils of bottle gourd.
      Result 17 JMJ family members was identified from bottle gourd genome, dividing into 5 subfamilies, named LsiJMJ1-LsiJMJ17 according to chromosome distribution. Physical and chemical property analysis and subcellular localisation prediction results showed that L. siceraria JMJ proteins ranged in length from 412 to 1852 aa, and most were weakly acidic nuclear localisation proteins. Chromosome localisation and collinearity analysis found that LsiJMJ genes were unevenly distributed across nine chromosomes, containing one pair of highly homologous genes, and multiple pairs of homologous genes with species such as Arabidopsis thaliana and Cucumis melo. Conserved motif analysis indicated that proteins within the same subgroup share similar motifs. Additionally, gene structure and encoded protein domain analysis revealed that most homologous genes exhibit similar exon-intron structures, and encoded sequences contain approximately 6 to 33 exons. Cis-acting element analysis identified stress response, hormone response, growth and development regulation, light response, and other types of elements in JMJ family genes. Protein network interaction analysis suggests that LsiJMJ3 co-express with LsiJMJ9, LsiJMJ2, and LsiJMJ12, while LsiJMJ12, LsiJMJ5, and LsiJMJ7 or LsiJMJ11 co-express in pairs. Expression analysis indicates that LsiJMJ4 and LsiJMJ9 are significantly highly expressed in all tissues, particularly in pistillate flowers and terminal buds, LsiJMJ11 is specifically highly expressed in staminate flowers, and LsiJMJ8 is lowly expressed in five tissue types.
      Conclusion Seventeen members of the JMJ family have been successfully identified. Their conserved domains may play an important role in influencing the demethylase activity. The expression of LsiJMJs may be affected by development and abiotic stress. For instance, LsiJMJ4, LsiJMJ7 and LsiJMJ9 were involved in basic growth and development processes such as pistillate flower development or terminal bud differentiation regulation, LsiJMJ11 is related to staminate flower development, and LsiJMJ8 may be induced to express under specific conditions.