目的 深入研究西红花苷生物合成途径，基于西红花Crocus sativus柱头进行全长转录组测序，挖掘可能参与西红花苷生物合成调控的bZIP转录因子基因。方法 通过PacBio Sequel高通量测序技术对西红花柱头进行全长转录组测序及生物信息学分析，从中鉴定具有完整结构域的全长CsbZIPs，分析其编码蛋白的理化性质、保守基序、系统进化及互作网络，并筛选可能参与西红花苷生物合成调控的候选基因。结果 全长转录组测序共获得37 732条Isoforms序列，平均长度为2 358 bp，N50长度为3 594 bp；CsbZIPs蛋白主要为定位到细胞核中的不稳定亲水蛋白，氨基酸数目57～607 aa，相对分子质量6 696.55～65 982.04，等电点5.01～12.16；64个CsbZIPs蛋白都包含典型碱性区域和亮氨酸拉链区域，根据其与拟南芥AtbZIPs系统发育关系被分为11个亚族，其中A亚族含CsbZIPs最多；基因表达模式分析及转录组与代谢组联合分析结果表明，E、S及A亚族的CsbZIPs基因，尤其是E亚族的CsbZIP44、CsbZIP54、CsbZIP55、CsbZIP62、CsbZIP64等基因可能参与西红花苷生物合成的转录调控。结论 较为全面地揭示了可能参与西红花苷生物合成的bZIP转录因子基因，为深入研究西红花苷生物合成调控提供了丰富的数据基础。

Objective To investigate the biosynthesis pathway of crocin, identify bZIP transcription factor genes that may be involved in the biosynthesis regulation of crocin, based on the full-length transcriptome sequencing of saffron stigma. Method Sequel high-throughput sequencing technology was used to perform full-length transcriptome sequencing and bioinformatics analysis on the stigma of saffron. Identifying full-length CsbZIPs with complete domains, and analyzing its physicochemical properties, conserved motifs, phylogenetic evolution, and interaction networks of their encoded proteins. Screening candidate genes that may participate in the regulation of saffron biosynthesis. Results A total of 37 732 Isoforms were obtained, with an average length of 2 358 bp and an N50 length of 3 594 bp; The CsbZIP proteins are mainly unstable hydrophilic proteins located in the nucleus, with amino acid number of 57—607 aa, the relative molecular weight of 6 696.55—65 982.04, and isoelectric point of 5.01—12.16; All of the 64 CsbZIP proteins contain typical alkaline regions and leucine zipper regions. Based on the phylogenetic relationship with AtbZIPs, they are divided into 11 subfamilies, in which Group A containing the most CsbZIPs; The results of gene expression pattern analysis and transcriptome metabolome association analysis indicate that CsbZIP genes in E, S, and A subgroups, especially CsbZIP44, CsbZIP54, CsbZIP55, CsbZIP62, CsbZIP64 genes in E subgroups, may be involved in the transcriptional regulation of crocin biosynthesis. Conclusion The data mining comprehensively reveals the bZIP transcription factor genes that may be involved in the biosynthesis regulation of crocin, provides a rich data foundation for in-depth research on the biosynthesis regulation of crocin.

R286.12

国家自然科学基金项目（81703667）;浙江省基础公益研究计划项目（LGN21C150004）;嘉兴市公益性研究计划项目（2023AY11048）;嘉兴市科技特派员专项（2021K117）