作物学报2026，Vol.52Issue(4)：1268-1276,9.DOI:10.3724/SP.J.1006.2026.54079
甘薯IbOPR2基因启动子克隆及调控因子的筛选
Cloning of the IbOPR2 gene promoter and identification of regulatory factors in sweetpotato
摘要
Abstract
Jasmonic acids(JAs),as endogenous plant hormones,play a crucial role in the plant response to salt stress.12-oxophytodienoate reductase(OPR),a key enzyme in JA biosynthesis,remains poorly understood in terms of its expression regulation.To identify transcription factors involved in regulating IbOPR2 expression,this study used the salt-tolerant sweetpo-tato cultivar'Haida 7791'to clone the promoter sequence of the IbOPR2 gene.Bioinformatics analysis revealed various cis-acting elements,and transcriptional activation assays confirmed promoter activity.Yeast one-hybrid(Y1H)screening was conducted to identify transcription factors interacting with the promoter,and point-to-point Y1H assays were used to validate specific binding.A 1964 bp promoter sequence upstream of IbOPR2 was successfully cloned,containing recognition motifs such as MYBHv1,MYB,and WRKY,along with stress-responsive elements(MYB,MYC)and MeJA-responsive elements,indicating strong tran-scriptional activation potential.Five candidate transcription factors—PHL7,ZFP16,SKIP11,TGA2,and ERF2—were identified using MYC and MeJA response elements as bait in Y1H assays.Follow-up point-to-point verification confirmed that IbPHL7,IbZFP16,and IbSKIP11 specifically bind to the IbOPR2 promoter.Dual-luciferase reporter assays further demonstrated that these three transcription factors could activate the IbOPR2 promoter.Moreover,qRT-PCR analysis showed that the expression of IbPHL7,IbZFP16,and IbSKIP11 is responsive to salt stress in sweetpotato.Taken together,these results suggest that these tran-scription factors may function as upstream regulators of IbOPR2,modulating its expression under salt stress conditions.This study represents the first identification of transcriptional regulators of sweetpotato IbOPR2,providing a foundation for further investigation into its regulatory mechanisms in response to salt stress.关键词
甘薯/IbOPR2/酵母单杂/转录因子/启动子Key words
sweetpotato/IbOPR2/yeast one-hybrid/transcription factor/promoter引用本文复制引用
王懿涵,李富昌,刘意,朱国鹏..甘薯IbOPR2基因启动子克隆及调控因子的筛选[J].作物学报,2026,52(4):1268-1276,9.基金项目
本研究由"崖州湾"菁英人才基金项目(SKJC-JYRC-2024-22)和财政部和农业农村部国家现代农业产业技术体系建设专项(CARS-10-Sweetpotato)资助.This study was supported by the"Yazhou Bay"Elite Talent Project Fund(SKJC-JYRC-2024-22)and the China Agriculture Research System of MOF and MARA(CARS-10-Sweetpotato). （SKJC-JYRC-2024-22）
