生物技术通报2023，Vol.39Issue(11)：261-269,9.DOI:10.13560/j.cnki.biotech.bull.1985.2023-0133
三个甘蓝WRKY基因的克隆及其对非生物胁迫的表达
Cloning of Three Cabbage WRKY Genes and Their Expressions in Response to Abiotic Stress
摘要
Abstract
WRKY proteins are a class of transcription factors involved in plant growth and development,biotic and abiotic stress,and other biological processes.To study the response mechanism of cabbage WRKY genes under stress of adversity may lay the foundation for further research on the stress tolerance function of BoWRKYs.BoWRKY40(BolC02g035760.2J),BoWRKY46(BolC04g031460.2J),and BoWRKY70(BolC04g035730.2J)from cabbage(Brassica oleracea var.capitata L.)seedlings were cloned by PCR approach.The pSuper1300:BoWRKYs-GFP expression vector was constructed by homologous recombination method for subcellular localization.Response patterns of BoWRKYs genes to abscisic acid(ABA),PEG8000 and salt stress were analyzed by RT-qPCR.As results,the length of CDS regions of BoWRKY40,BoWRKY46 and BoWRKY70 were 873,831 and 864 bp,which encoded proteins of 290,276 and 287 amino acids,respectively.The predicted molecular weights of BoWRKY proteins were 32.41,32.08 and 32.52 kD with theoretical isoelectric points(pI)of 6.77,6.18 and 5.62.The conserved structural domain of the WRKY superfamily'WRKYGQK'was present in the proteins,and multiple alignment analysis revealed that BoWRKYs had high conservation with WRKYs in Arabidopsis(Arabidopsis thaliana),tomato(Solanum lycopersicum L.),maize(Zea mays L.),and cotton(Gossypium hirsutum L.).Subcellular localization analysis showed that BoWRKYs were located in the nucleus.Fluorescence quantitative PCR results showed that WRKY family genes were induced in different degrees under ABA,PEG8000 and salt stress.Among them,BoWRKY40 responded positively to three kinds of stress,BoWRKY46 and BoWRKY70 were up-regulated under ABA and salt stress,and their expressions gradually decreased after PEG8000 treatment.These results indicated that the expressions of cabbage BoWRKY40,BoWRKY46,and BoWRKY70 were closely related to stress responses.关键词
甘蓝/基因克隆/非生物胁迫/亚细胞定位/表达分析Key words
cabbage/gene cloning/abiotic stress/subcellular localization/expression analysis引用本文复制引用
杨旭妍,赵爽,马天意,白玉,王玉书..三个甘蓝WRKY基因的克隆及其对非生物胁迫的表达[J].生物技术通报,2023,39(11):261-269,9.基金项目
黑龙江省省属高等学校基本科研业务费科研项目(145209513),齐齐哈尔大学研究生创新项目(YJSCX2021031),黑龙江省普通高等学校青年创新人才培养计划(UNPYSCT-2017155) （145209513）
