生物技术通报2024，Vol.40Issue(6)：152-160,9.DOI:10.13560/j.cnki.biotech.bull.1985.2023-1127
大白菜BrCYP83B1基因的克隆及表达分析
Cloning and Expression Analysis of BrCYP83B1 Gene in Chinese Cabbage
摘要
Abstract
[Objective]The cytochrome P450 family is an important enzyme system involved in glucosinolate synthesis in cruciferous plants,where in the CYP83 subfamily plays a predominant role in core structure formation,this work aims to investigate the Chinese cabbage(Brassica rapa ssp.Pekinensis)CYP83B1 gene functional role.[Method]RT-PCR was used to clone the CYP83B1 gene.Bioinformatics analysis software was utilized to predict the encoded protein's physicochemical properties,homology of the encoded protein,and promoter cis-acting elements.The expression pattern of BrCYP83B1 was analyzed by RT-qPCR,and a plant overexpression vector was constructed for further experimentation.[Result]The cDNA length of BrCYP83B1 was 1 500 bp,encoding a total of 499 amino acids.The protein belonged to cytochrome P450 superfamily and predominantly located in the cytoplasm.Its secondary structure primarily comprised of α-helixes and irregular coil.Homologous comparison illustrated that BrCYP83B1 had close relationship with Brassica napus L and Brassica oleracea L.var.italica.The BrCYP83B1 promoter contained cis-acting elements that were involved in the response to salicylic acid(SA),abscisic acid(ABA),and methyl jasmonate(MeJA),suggesting that the expression of BrCYP83B1 gene may be regulated by hormones.The expression of BrCYP83B1 was detected in various plant organs,including the roots,stems,leaves,flowers and fruits from RT-qPCR results.Notably,the highest expression was observed in the leaves.Moreover,BrCYP83B1 significantly presented induction upon treatment with MeJA,while its expression was repressed by SA.Additionally,ABA treatment initially up-regulated and subsequently down-regulated the gene.[Conclusion]BrCYP83B1 may be involved in the response regulation of Chinese cabbage to hormones.关键词
大白菜/BrCYP83B1/基因克隆/植物激素/表达分析/超表达载体Key words
Brassica rapa ssp.pekinensis/BrCYP83B1/gene cloning/plant hormones/expression analysis/overexpression vector引用本文复制引用
王玉书,赵琳琳,赵爽,胡琦,白慧霞,王欢,曹业萍,范震宇..大白菜BrCYP83B1基因的克隆及表达分析[J].生物技术通报,2024,40(6):152-160,9.基金项目
国家自然科学基金项目(31401908),黑龙江省自然科学基金项目(C2016056),黑龙江省省属高等学校基本科研业务费科研项目(145209513,145209319,135509702),黑龙江省普通高等学校青年创新人才培养计划(UNPYSCT-2017155),黑龙江省高等教育教学改革研究与实践项目(SJGY20220406) （31401908）
