甘蓝型油菜BnaNAC14.1基因的克隆与表达模式分析OACSTPCD
Cloning and Expression Pattern Analysis of BnaNAC14.1 Gene in Brassica napus
NAC转录因子广泛参与植物生长发育及非生物逆境胁迫反应.本研究采用RT-PCR方法,从甘蓝型油菜(Brassica napus)'中双11'cDNA中克隆BnaNAC14.1基因(GenBank No.XM_022712555.2).生物信息学分析表明,BnaNAC14.1基因编码区全长1 908 bp,编码635个氨基酸,包含NAM保守结构域;构建pCAMBIA1305.1-35S-sGFP-BnaNAC14.1融合表达载体,利用根癌农杆菌(Agrobacterium tumefaciens)介导法转化烟草(Nicotiana tabacum)叶片细胞,亚细胞定位结果显示,该转录因子定位于细胞核;以酵母(Saccharomyces cerevisiae)表达载体pGBKT7-BD-BnaNAC14.1转化酵母菌株Y2HGold,在SD/-Trp+X-α-gal培养基中显示蓝色,表明该转录因子具有转录激活活性.表达模式分析表明,BnaNAC14.1基因在甘蓝型油菜花和种子中表达量显著高于根、茎和叶;qRT-PCR结果显示,BnaNAC14.1基因受盐、干旱及外源激素脱落酸(abscisic acid,ABA)的诱导,在吲哚乙酸(indoleacetic acid,IAA)和茉莉酸甲酯(methyl jasmonate,MeJA)处理下表达受到抑制.上述结果初步明确BnaNAC14.1基因参与逆境胁迫和激素应答,可能在种子发育中发挥重要作用.本研究为后续深入探讨BnaNAC14.1生物学功能和优质抗逆油菜育种提供基础资料.
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.
章冰馨;祝金博;刘翠;李青洋;崔鹏;刘宏波
浙江农林大学现代农学院,杭州 311300
农业科学
甘蓝型油菜NAC转录因子基因克隆亚细胞定位表达模式
Brassica napusNAC transcription factorGene cloningSubcellular localizationExpression pattern
《农业生物技术学报》 2024 (001)
BnGPAT9调控甘蓝型油菜甘油脂合成及低磷胁迫介导脂质重塑的分子机制研究
60-69 / 10
国家自然科学基金(32071929);浙江省自然科学基金(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)
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