农业生物技术学报2024，Vol.32Issue(1)：60-69,10.DOI:10.3969/j.issn.1674-7968.2024.01.006
甘蓝型油菜BnaNAC14.1基因的克隆与表达模式分析
Cloning and Expression Pattern Analysis of BnaNAC14.1 Gene in Brassica napus
摘要
Abstract
NAC transcription factors are widely involved in plant growth and development and abiotic stress response.In this study,the BnaNAC14.1 gene was cloned from the'Zhongshuang 11'cDNA of Brassica napus by RT-PCR.Bioinformatics analysis showed that the coding sequence of BnaNAC14.1 gene was 1 908 bp,and encoded 635 amino acids,including the NAM conserved domain.The pCAMBIA1305.1-35S-sGFP-BnaNAC14.1 fusion expression vector was constructed and transformed into tobacco(Nicotiana tabacum)leaf cells by Agrobacterium tumefaciens-mediated transformation.The results of subcellular localization showed that BnaNAC14.1 localized in the nucleus;The recombinant vector pGBKT7-BD-BnaNAC14.1 was transformed into yeast(Saccharomyces cerevisiae)strain Y2HGold,and blue monoclonal colonies were observed on SD/-Trp+X-α-gal medium which indicated that the BnaNAC14.1 had transcriptional activation activity.The expression pattern demonstrated that the BnaNAC14.1 had relatively higher expression levels in flowers and development seeds than that in roots,stems and leaves.The results of qRT-PCR showed that BnaNAC14.1 gene was significantly up-regulated by salt,drought and exogenous hormones abscisic acid(ABA),and inhibited by indoleacetic acid(IAA)and methyl jasmonate(MeJA).The above results preliminarily identified that BnaNAC14.1 gene might be involved in abiotic stress and hormone response,and play an important role in seed development.This study provides theoretical support for further study on the BnaNAC14.1 biological function and rapeseed breeding with high quality and stress resistance.关键词
甘蓝型油菜/NAC转录因子/基因克隆/亚细胞定位/表达模式Key words
Brassica napus/NAC transcription factor/Gene cloning/Subcellular localization/Expression pattern分类
农业科技引用本文复制引用
章冰馨,祝金博,刘翠,李青洋,崔鹏,刘宏波..甘蓝型油菜BnaNAC14.1基因的克隆与表达模式分析[J].农业生物技术学报,2024,32(1):60-69,10.基金项目
国家自然科学基金(32071929) （32071929）
浙江省自然科学基金(LY21C130001) （LY21C130001）
浙江省"十四五"农业新品种选育重大科技专项子课题(2021C02064-2-1) （2021C02064-2-1）
