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Abstract：
As an important genus in Orchidaceae, Cymbidium has rich ecological diversity and significant economic value. DNA binding with one zinc finger (Dof) proteins are pivotal plant-specific transcription factors that play crucial roles in the growth, development, and stress response of plants. Although the Dof genes have been identified and functionally analyzed in numerous plants, exploration in Orchidaceae remains limited. We conducted a thorough analysis of the Dof gene family in Cymbidium goeringii, C. ensifolium, and C. sinensis. In total, 91 Dof genes (27 CgDofs, 34 CeDofs, 30 CsDofs) were identified, and Dof genes were divided into five groups (I-V) based on phylogenetic analysis. All Dof proteins have motif 1 and motif 2 conserved domains and over half of the genes contained introns. Chromosomal localization and collinearity analysis of Dof genes revealed their evolutionary relationships and potential gene duplication events. Analysis of cis-elements in CgDofs, CeDofs, and CsDofs promoters showed that light-responsive cis-elements were the most common, followed by hormone-responsive elements, plant growth-related elements, and abiotic stress response elements. Dof proteins in three Cymbidium species primarily exhibit a random coil structure, while homology modeling exhibited significant similarity. In addition, RT-qPCR analysis showed that the expression levels of nine CgDofs changed greatly under heat stress. CgDof03, CgDof22, CgDof27, CgDof08, and CgDof23 showed varying degrees of upregulation. Most upregulated genes under heat stress belong to group I, indicating that the Dof genes in group I have great potential for high-temperature resistance. In conclusion, our study systematically demonstrated the molecular characteristics of Dof genes in different Cymbidium species, preliminarily revealed the patterns of heat stress, and provided a reference for further exploration of stress breeding in orchids.
摘要：
作为兰科的重要属，兰具有丰硕的生态多样性和显著的经济价值。DNA与一个锌指（Dof）蛋白的结合是关键的植物特异性转录因子，在生长中起着至关重要的作用，发展,和植物的应激反应。尽管Dof基因已经在许多植物中被鉴定和功能分析,兰科的探索仍然有限。我们对春兰的Dof基因家族进行了彻底的分析，C.敏叶，还有C.sinensis.总的来说,91个Dof基因（27个CgDofs，34CeDofs,确定了30CsDofs），根据系统发育分析，将Dof基因分为五组(I-V)。所有Dof蛋白都具有基序1和基序2保守结构域,并且超过一半的基因含有内含子。Dof基因的染色体定位和共线性分析揭示了它们的进化关系和潜在的基因复制事件。CgDofs中的顺式元素分析，CeDofs,和CsDofs启动子显示光响应的顺式元件是最常见的，其次是激素反应性元素，植物生长相关元素，和非生物应激反应元件。三种大花兰属植物中的Dof蛋白主要表现出无规卷曲结构，而同源建模表现出显著的相似性。此外,RT-qPCR分析表明，在热胁迫下，9种CgDofs的表达水平发生了很大变化。CgDof03、CgDof22、CgDof27、CgDof08和CgDof23显示不同程度的上调。大多数在热应激下上调的基因属于I组，表明I组中的Dof基因具有巨大的耐高温潜力。总之,我们的研究系统地证明了多夫基因在不同大花兰物种中的分子特征，初步揭示了热应激的模式，为进一步探索兰花逆境育种提供参考。
