南方农业学报2025，Vol.56Issue(8)：2480-2489,10.DOI:10.3969/j.issn.2095-1191.2025.08.009
水稻PHD17基因启动子克隆及冷胁迫应答分析
Promoter cloning and analysis of the cold-stress response of PHD17 gene in rice
摘要
Abstract
[Objective]This study aimed to clone promoter sequence of gene OsPHD17 in PHD(plant homeodomain)of rice and analyze its response to cold stress,so as to provide a theoretical reference for elucidating the cold-stress re-sponse mechanism of gene OsPHD17.[Method]Quantitative real-time PCR(qPCR)was employed to analyze expression pattern of gene OsPHD17 under cold stress treatment.Recognition elements of transcriptional factors of the promoter were predicted based on PlantRegMap database.The promoter sequence(length of 1160 bp upstream of the transcription start site)of gene OsPHD17 was cloned and inserted to vector pCAMBIA3301 to obtain plant expression vector POsPHD17-GUS.Transgenic Arabidopsis thaliana lines were generated via Agrobacterium tumefaciens-mediated transformation.Ex-pression of GUS reporter genes under cold stress were detected by RT-PCR and qPCR to analyze the change in promoter transcriptional activity under cold stress conditions.[Result]The promoter of gene OsPHD17 included recognition ele-ments of the transcriptional factors regulating growth and development(TALE,LBD,MIKC_MADS,G2-like,BBR-BPC)and recognition elements of the transcriptional factors regulating cold-stress response(ERF,C2H2,bHLH,bZIP,and NAC).Expression of the gene OsPHD17 was significantly up-regulated at 3.0 hours of cold stress(P<0.05,the same below);the expression peaked at 12.0 h with approximately 11.7-fold increase compared to the expression at 0 h,indi-cating a rapid cold-stress response of the gene OsPHD17.The cloned promoter sequence was 1160 bp in length,and trans-genic Arabidopsis thaliana lines were successfully established.RT-PCR detection showed that T3 generation were homo-zygous with GUS genes expression of POsPHD17-GUS transgenic Arabidopsis thaliana lines,indicating that the promoter of gene OsPHD17 had transcriptional activity.Under cold stress of 4℃,the two homozygous T3 lines of Arabidopsis thali-ana showed a highly significant(P<0.01)or significantly up-regulated relative expression of the GUS gene in the two lines compared to the CK group(untreated),demonstrating an increase of transcriptional activity of promoter of gene Os-PHD17.[Conclusion]The promoter of gene OsPHD17 that exhibits transcriptional activity is capable of modulating downstream gene expression.Transcriptional activity is markedly upregulated under cold stress conditions,and expres-sion of gene OsPHD17 is significantly upregulated,thereby confirming the important regulatory role of gene OsPHD17 in cold-stress response of rice.关键词
水稻/OsPHD17/启动子克隆/冷胁迫/表达分析Key words
rice/OsPHD17/promoter cloning/cold stress/expression analysis分类
农业科技引用本文复制引用
董伟峰,孙晓丽,贺子天,杨珺凯,李艳肖,吴浩,GEORGE Bawa,谢辉,孙明哲,沈阳..水稻PHD17基因启动子克隆及冷胁迫应答分析[J].南方农业学报,2025,56(8):2480-2489,10.基金项目
中央引导地方科技发展专项(ZY24QY0201) （ZY24QY0201）
黑龙江省"双一流"学科协同创新成果项目(LJGXCG2023-072) （LJGXCG2023-072）
黑龙江八一农垦大学研究生创新科研项目(YJSCX2024-Y04) （YJSCX2024-Y04）
黑龙江八一农垦大学大学生创新创业项目(202210223055) Central Guidance for Special Topics on Local Scientific and Technological Development(ZY24QY0201) （202210223055）
Heilongjiang Province"Double First-Class"Discipline Collaborative Innovation Achievement Project(LJGXCG2023-072) （LJGXCG2023-072）
Heilongjiang Bayi Agricultural University Graduate Innovative Research Project(YJSCX2024-Y04) （YJSCX2024-Y04）
Heilongjiang Bayi Agricultural University College Students'Innovation and Entrepreneurship Project(202210 223055) （202210 223055）
