目的 研究西红花Crocus sativus AP2/ERF转录因子家族以及响应激素信号诱导柱头药效成分生物合成，为西红花柱头次生代谢调控网络研究奠定基础。方法 基于西红花基因组数据，通过隐马尔可夫模型搜索鉴定西红花中的AP2/ERF转录因子，并利用生物信息学分析软件和在线网站分析基因的理化性质、染色体定位、系统发育进化树、保守结构域和基序以及启动子的顺式作用元件；利用高效液相色谱（high performance liquid chromatography，HPLC）和实时荧光定量PCR（quantitative real-time PCR，qRT-PCR）分别测定赤霉素（gibberellin A3，GA3）和茉莉酸甲酯（methyl jasmonate，MeJA）处理后柱头中西红花苦苷、西红花苷Ⅰ和西红花苷Ⅱ的含量及其生物合成相关功能基因和部分CsAP2/ERFs基因的表达量。结果 西红花中含有88个AP2/ERF转录因子，分布在8条染色体上；系统发育和保守基序显示CsAP2/ERFs分类于脱水响应元件结合蛋白（DREB）、乙烯响应因子（ERF）和soloist 3个亚家族，其中DREB和ERF共计81个CsAP2/ERFs；GA3和MeJA处理对西红花苷Ⅰ和西红花苷Ⅱ的合成均有促进作用，但对西红花苦苷合成的作用相反；CsAP2/ERF1、9、2、33和34受激素信号诱导在柱头发育过程中表达量发生显著变化，且GA3和MeJA诱导变化趋势相反;基因表达相关性分析发现该5个CsAP2/ERFs在激素作用下分别与不同的功能基因具有显著的相关性，表明CsAP2/ERFs响应激素信号调控柱头中脱辅基类胡萝卜素化合物生成的关键基因进而影响西红花药效成分的生物合成。结论 鉴定了西红花中的CsAP2/ERFs转录因子家族，阐明了相关功能基因和部分CsAP2/ERFs受GA3和MeJA处理的诱导表达模式，为探索西红花响应激素信号诱导调控西红花脱辅基类胡萝卜素类成分生物合成调控研究奠定了基础。

Objective This study aims to characterize the AP2/ERF transcription factor family in Crocus sativus and examine the biosynthesis of active components in the stigma induced by responsive hormone signals, laying the foundation for the research on the secondary metabolic regulatory network of C. sativus stigmas. Methods AP2/ERF transcription factors were searched and identified using hidden Markov model (HMM) based on C. sativus genome data. Bioinformatics analysis software and online websites were performed to characterize their physicochemical properties, chromosomal locations, phylogenetic tree, conserved domains/motifs, and cis-acting elements of promoters. The contents of picrocrocin, crocin I and crocin II in the stigma after treatment with gibberellin A3 (GA3) and methyl jasmonate (MeJA) were determined by high performance liquid chromatography (HPLC) and quantitative real-time PCR (qRT-PCR), respectively. Meanwhile, the expression levels of the related biosynthesis functional genes and some CsAP2/ERFs genes were also measured. Results A total of 88 AP2/ERFs transcription factors were identified in C. sativus, which distributed across eight chromosomes. Phylogenetic analysis combined with the examination of conserved motifs indicated that CsAP2/ERFs can be categorized into three subfamilies: DREB, ERF, and soloist. Among these, a total of 81 CsAP2/ERFs were classified within the DREB and ERF subfamilies. Both GA3 and MeJA promoted the synthesis of crocin I and crocin II, but had the opposite effect on the synthesis of picrocrocin. CsAP2/ERF1, 9, 2, 33 and 34 were induced by hormone signals to undergo significant changes in their expression levels during the development of the stigma. Moreover, the changes induced by GA3 and MeJA were in opposite trends. Correlation analysis of gene expression revealed that five CsAP2/ERFs were significantly correlated with different functional genes under the influence of hormones, indicating that CsAP2/ERFs responsive hormone signals regulated the key genes for the generation of de-prothetic group carotenoid compounds in the stigms, thereby affecting the biosynthesis of the active ingredients of C. sativus. Conclusion This study identifies CsAP2/ERFs transcription factor family in C. sativus and clarifies the expression patterns of related functional genes and some CsAP2/ERFs after treatment with GA3 and MeJA. These findings provide a foundational basis for investigating the regulatory mechanisms of de-prothetic group apocarotenoid biosynthesis in C. sativus induced by responsive hormonal signaling.

R286.12

国家自然科学基金项目(81703667); 嘉兴市公益性研究计划项目(2023AY11048,2025CGZ056); 嘉兴大学校级SRT项目(8517241041)