DU Shuangjiang,LIU Shiliang,JIA Yunhe,et al.Cloning and Functional Analysis of Watermelon ClMADS18 Transcription Factor[J].Northern Horticulture,2026,(17):10-18.[doi:10.11937/bfyy.20260271]
西瓜ClMADS18基因克隆与响应低温胁迫功能解析
- Title:
- Cloning and Functional Analysis of Watermelon ClMADS18 Transcription Factor
- 文章编号:
- 1001-0009(2026)17-0010-09
- Keywords:
- watermelon; MADS-box; cold stress; gene cloning; expression analysis
- 分类号:
- S651
- 文献标志码:
- A
- 摘要:
- 以西瓜品种“1338”、哥伦比亚拟南芥和本氏烟草为试材，采用生物信息学分析、基因克隆、亚细胞定位、实时荧光定量PCR及转基因等方法，对ClMADS18基因进行了系统研究，以期探究西瓜MADS-box基因家族成员在低温胁迫响应中的功能。结果表明：ClMADS18基因全长771 bp，编码256个氨基酸，蛋白分子量28 665.32 kDa，等电点6.16，属定位于细胞核的酸性疏水性稳定蛋白。该基因在西瓜叶片中表达量最高，其表达受低温、ABA、GA3、SA及MeJA等多种信号诱导。过表达ClMADS18的拟南芥在4 ℃下发芽能力显著高于野生型。综上所述，ClMADS18基因在西瓜响应低温胁迫过程中发挥重要作用，可作为西瓜耐低温育种的潜在候选基因。
- Abstract:
- Taking the watermelon cultivar ‘1338’,Columbia Arabidopsis thaliana,and Nicotiana benthamiana as test materials,a systematic study was conducted on the ClMADS18 gene through bioinformatics analysis,gene cloning,subcellular localization,real-time quantitative PCR,and transgenic methods,in order to explore the function of MADS-box gene family members in watermelon under low-temperature stress.The results showed that the full-length sequence of ClMADS18 was 771 bp,encoding a protein of 256 amino acids with a molecular weight of 28 665.32 kDa and an isoelectric point of 6.16.The encoded protein was identified as a stable,acidic,and hydrophobic protein localized in the nucleus.ClMADS18 exhibited the highest expression level in watermelon leaves,and its transcription was induced by multiple signals including low temperature,abscisic acid (ABA),gibberellic acid 3 (GA3),salicylic acid (SA),and methyl jasmonate (MeJA).The germination rate of Arabidopsis thaliana overexpressing ClMADS18 was significantly higher than that of the wild-type under 4 ℃ treatment.In conclusion,ClMADS18 played an important role in watermelon’s response to low-temperature stress,and could be used as a potential candidate gene for low-temperature tolerance breeding in watermelon.
参考文献/References:
［1］GRAMZOW L,THEISSEN G.A hitchhiker′s guide to the MADS world of plants［J］.Genome Biology,2010,11(6):214.［2］DAZ-RIQUELME J,LIJAVETZKY D,MARTNEZ-ZAPATER J M,et al.Genome-wide analysis of MIKCC-type MADS box genes in grapevine［J］.Plant Physiology,2009,149(1):354-369.［3］PA〖AKRˇD〗ENICOV L,de FOLTER S,KIEFFER M,et al.Molecular and phylogenetic analyses of the complete MADS-box transcription factor family in Arabidopsis:New openings to the MADS world［J］.The Plant Cell,2003,15(7):1538-1551.［4］ARORA R,AGARWAL P,RAY S,et al.MADS-box gene family in rice:Genome-wide identification,organization and expression profiling during reproductive development and stress［J］.BMC Genomics,2007,8(1):242.［5］WANG Y,ZHANG J,HU Z,et al.Genome-wide analysis of the MADS-box transcription factor family in Solanum lycopersicum［J］.International Journal of Molecular Sciences,2019,20(12):2961.［6］WANG Z,CHANG J,HAN J,et al.Genome-wide reidentification and expression analysis of MADS-box gene family in cucumber［J］.International Journal of Molecular Sciences,2025,26(8):3800.［7］WANG P,WANG S,CHEN Y,et al.Genome-wide analysis of the MADS-box gene family in watermelon［J］.Computational Biology and Chemistry,2019,80:341-350.［8］IMMINK R G H,GADELLA T W J Jr,FERRARIO S,et al.Analysis of MADS box protein-protein interactions in living plant cells［J］.PNAS,2002,99(4):2416-2421.［9］KAUFMANN K,MELZER R,THEIBEN G.MIKC-type MADS-domain proteins:Structural modularity,protein interactions and network evolution in land plants［J］.Gene,2005,347(2):183-198.［10］XU N,CHU Y,CHEN H,et al.Rice transcription factor OsMADS25 modulates root growth and confers salinity tolerance via the ABA-mediated regulatory pathway and ROS scavenging［J］.PLoS Genetics,2018,14(10):e1007662.［11］HANNAH M A,HEYER A G,HINCHA D K.A global survey of gene regulation during cold acclimation in Arabidopsis thaliana［J］.PLoS Genetics,2005,1(2):e26.［12］张慧娜,齐秀玲,申晓萍,等.西瓜起源与演化研究进展［J］.中国农学通报,2016,32(35):232-236.［13］陈哲,梁瀚元.我国西瓜产业发展与贸易趋势研究［J］.经营与管理,2019(9):114-122.［14］KORKMAZ A,DUFAULT R J.Developmental consequences of cold temperature stress at transplanting on seedling and field growth and yield.I.watermelon［J］.Journal of the American Society for Horticultural Science,2001,126(4):404-409.［15］侯伟,孙爱花,杨福孙,等.低温胁迫对西瓜幼苗光合作用与叶绿素荧光特性的影响［J］.广东农业科学,2014,41(13):35-39.［16］彭金光,孙玉宏,师瑞红,等.10℃低温对西瓜幼苗耐性生理指标的影响［J］.安徽农学通报,2006,12(10):42-45.［17］李亮,刘杭,张桂池,等.不同低温需求量砂梨休眠相关基因PpMADS13的特征及表达模式［J］.亚热带植物科学,2016,45(1):42-47.［18］李元元.低温诱导烟草早花研究与烟草MADS-box基因的同源克隆［D］.北京:中国农业科学院,2011.［19］叶红红,尹明安,朱敏,等.番茄果实特异表达载体pBI121-2A11-HB-1的构建及其对农杆菌的转化［J］.食品科学,2014,35(19):174-179.［20］ZHANG X,PEI H,LV Z,et al.Genome-wide characterization of MADS genes in sweet cherry and the role of PavMADS14 in response to cold stress［J］.Scientia Horticulturae,2025,352:114453.［21］CHEN R,MA J,LUO D,et al.CaMADS,a MADS-box transcription factor from pepper,plays an important role in the response to cold,salt,and osmotic stress［J］.Plant Science,2019,280:164-174.［22］沈威,滕瑞敏,李辉,等.茶树MADS-box转录因子基因的克隆 与非生物胁迫响应分析［J］.茶叶科学,2017,37(6):575-585.［23］GUO F X,CHEN Y,LI X R,et al.Enhancement of low-temperature tolerance in watermelon (Citrullus lanatus) seedlings by cool-hardening germination［J］.Australian Journal of Experimental Agriculture,2007,47(6):749.［24］LU J,SHIREEN F,CHENG F,et al.High relative humidity improve chilling tolerance by maintaining leaf water potential in watermelon seedlings［J］.Plant Physiology and Biochemistry,2021,166:818-826.［25］FATIMA M,MA X,ZHANG J,et al.Genome-wide analysis of MADS-box genes and their expression patterns in unisexual flower development in dioecious spinach［J］.Scientific Reports,2024(14):18635.［26］WAN S,LIANG B,YANG L,et al.The MADS-box family gene PtrANR1 encodes a transcription activator promoting root growth and enhancing plant tolerance to drought stress［J］.Plant Cell Reports,2023,43(1):16.［27］马鑫磊.谷子MADS-box和Ⅲ型PRX基因家族的生物信息学分析及干旱胁迫下的表达分析［D］.秦皇岛:河北科技师范学院,2022.［28］LIU Q,ZHANG G Y,CHEN S Y.Structure and regulation of plant transcription factors［J］.Chinese Science Bulletin,2000,45(14):1465-1474.［29］刘仕梁.西瓜ClMADS18转录因子克隆及功能分析［D］.哈尔滨:东北农业大学,2023.
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备注/Memo
第一作者简介：杜双江（2001-），男，硕士研究生，研究方向为蔬菜遗传育种。E-mail：245522491@qq.com.责任作者：王学征（1978-），女，博士，教授，博士生导师，现主要从事西瓜甜瓜种质资源研究利用及遗传育种等研究工作。E-mail：xz6206815@163.com.基金项目：国家西甜瓜产业技术体系资助项目（CARS-025）。收稿日期：2026-01-21