生物技术通报2025，Vol.41Issue(9)：195-206,12.DOI:10.13560/j.cnki.biotech.bull.1985.2025-0231
百合LoAPS1克隆及其在休眠解除过程的功能分析
Cloning of the LoAPS1 and Its Function Analysis during the Process of Dormancy Release in Lilium
摘要
Abstract
[Objective]ATP sulfurylase(APS1)is a key enzyme in the first step of the sulfate assimilation process.Studying the function of APS1 during dormancy release in Oriental Lilium'Siberia'provides important theoretical guidance for understanding the function of sulphur metabolic pathways in dormancy release in lily bulbs.[Method]The full-length sequence of LoAPS1 gene was obtained from different stages of dormancy relase by TA cloning,and its bioinformatics analysis was performed.Combined with the Lanzhou lily genome,the cis-acting elements of APS1 promoter were analyzed.Using the transcriptome database of different phases of dormancy release in lily and the riboflavin promotion of dormancy release in lily,the expression patterns of APS1 genes and related genes of sulphur metabolism pathway were analyzed.The subcellular localization of LoAPS1 protein was verified by tobacco subcellular localization transient transformation technique.The effect of silencing LoAPS1 on the process of dormancy release in lily bulbs was verified by virus induced gene silencing(VIGS)technique.[Result]The LoAPS1 gene had an ORF length of 1 413 bp,GenBank accession number was WMQ58782.1,and encoded 470 amino acids.The LoAPS1 protein contained two conserved structural domains,ATP-sulfurylase and PUA-like,with subcellular localization in chloroplasts,and might interact with sulphur metabolism pathways,such as APR,APK and SIR.The amino acid sequence of LoAPS1 had a high affinity with Musa acuminata and Zingiber officinale.The lily APS1 promoter contained four transcriptional start sites,including light-responsive elements,MYB-binding sites,and signal-responsive elements for methyl jasmonate(MeJA),gibberellin(GA3),and abscisic acid(ABA).The expression of LoAPS1 and related genes of sulfur metabolism pathway were down-regulated during the process of dormancy release of lily.The results of the VIGS and RT-qPCR showed that inhibition of LoAPS1 expression promoted the dormancy release in lily bulbs.[Conclusion]LoAPS1 is localized in chloroplasts and may respond to hormone signaling such as MeJA,GA3 and ABA,and inhibition of LoAPS1 expression promotes the dormancy release in lily bulbs.关键词
LoAPS1/基因克隆/病毒介导的基因沉默技术/生物信息学分析/亚细胞定位/休眠解除/百合Key words
LoAPS1/gene cloning/VIGS/bioinformatics analysis/subcellular localization/dormancy release/Lilium引用本文复制引用
徐小萍,杨成龙,和兴,郭文杰,吴健,方少忠..百合LoAPS1克隆及其在休眠解除过程的功能分析[J].生物技术通报,2025,41(9):195-206,12.基金项目
国家自然科学基金项目延伸研究项目(GJYS202407),福建省自然科学基金青年创新基金项目(2023J05064),国家自然科学基金青年项目(32302595) （GJYS202407）
