目的 对半夏生长素响应因子（auxin response factors，ARF）基因（PtARF）进行全基因组鉴定及不同温度和脱落酸（abscisic acid，ABA）调控下的表达模式解析，以期为解析PtARF的生物学功能提供理论依据。方法 基于已发表的半夏全基因组数据，利用生物信息学方法对PtARF基因家族成员进行鉴定，并系统分析其理化性质、染色体定位、系统进化关系、保守结构域、基因结构及启动子顺式作用元件；采用实时荧光定量PCR（quantitative real-time PCR，qRT-PCR）技术，分析半夏在高温（35 ℃）、低温（20 ℃）及ABA处理下的PtARF表达特征。结果 共鉴定出22个PtARF基因（PtARF1～PtARF22），分为3个亚家族；编码氨基酸长度为202～1 205 aa，相对分子质量为23 177.42～135 132.80，等电点（pI）为5.38～9.59。基因结构分析显示，所有成员基因均含有B3保守结构域；启动子顺式作用元件预测显示，PtARF基因启动子区分布温度响应元件和ABA响应元件。基因组共线性分析显示，片段复制事件在PtARF基因家族进化中起关键作用。qRT-PCR结果显示，所有PtARFs基因在高温（35 ℃）胁迫下表达量显著上调，低温（20 ℃）胁迫对大部分基因无显著影响；ABA处理可诱导PtARF基因表达水平升高。结论 从全基因组水平系统鉴定了半夏ARF基因家族，揭示了不同PtARF成员对温度胁迫及ABA处理的表达模式差异，为进一步阐释PtARF在半夏“高温倒苗”生理过程中的潜在功能奠定基础。

Objective To perform genome-wide identification of auxin response factor (ARF) genes (PtARF) in Pinellia ternata and analyze their expression patterns under different temperature stress and abscisic acid (ABA) treatments, aiming to provide a theoretical basis for deciphering the biological functions of PtARF. Methods Based on the published whole-genome data of Pinellia ternata, members of the PtARF gene family were identified using bioinformatic methods. Systematic analyses were conducted on their physicochemical properties, chromosomal locations, phylogenetic relationships, conserved domains, gene structures, and promoter cis-regulatory elements. Quantitative real-time PCR (qRT-PCR) was employed to analyze the expression profiles of PtARF genes in P. ternata under high-temperature (35 ℃), low-temperature (20 ℃), and abscisic acid (ABA) treatments. Results A total of 22 PtARF genes (PtARF1 to PtARF22) were identified and classified into three subfamilies. The encoded proteins ranged from 202 to 1 205 amino acids (aa) in length, with relative molecular masses between 23 177.42 and 135 132.80 and isoelectric points (pI) ranging from 5.38 to 9.59. Gene structure analysis revealed that all members contain the conserved B3 DNA-binding domain. Prediction of promoter cis-regulatory elements indicated the spread presence of temperature-responsive and abscisic acid (ABA)-responsive elements in the promoter regions of PtARF genes. Genomic collinearity analysis demonstrated that segmental duplication events played a crucial role in the evolution of the PtARF gene family. qRT-PCR results showed that the expression levels of all PtARF genes were significantly up-regulated under high-temperature (35 ℃) stress. In contrast, low-temperature (20 ℃) stress had no significant impact on most genes. ABA treatment induced elevated expression levels of PtARF genes. Conclusion This study comprehensively identified the ARF gene family in P. ternata at the whole-genome level, revealing distinct expression patterns among different PtARF members in response to temperature stress and ABA treatment. These findings provide molecular insights for elucidating the potential functions of PtARFs in Pinellia ternata’s high-temperature-induced seedling wilt process.

R286.12

国家自然科学基金资助项目（82104329，32401226）；河南省重点研发专项（251111310500，231111312700，241111310200）；中国博士后科学基金特别资助项目（2014T170252）；河南省国际科技合作项目（242102521056）；河南省科技攻关（242102310549）；河南中医药大学科研苗圃工程资助项目（MP2024-56）