甘蓝型油菜BnNAC61基因的克隆及表达特征分析  

Cloning and expression characteristics of BnNAC61 gene in Brassica napus

作　　者：郝丽芬[1,2] 燕孟娇[2] 皇甫海燕 宋培玲[2] 房永雨[2] 贾晓清[2] 李子钦[2] 韩冰[1] HAO Li-fen;YAN Meng-jiao;HUANGFU Hai-yan;SONG Pei-ling;FANG Yong-yu;JIA Xiao-qing;LI Zi-qin;HAN Bing(College of Life Sciences,Inner Mongolia Agricultural University,Hohhot 010011,China;Inner Mongolia Academy of Agricultural and Animal Husbandry Sciences,Hohhot 010031,China)

机构地区：[1]内蒙古农业大学生命科学学院,内蒙古呼和浩特010011 [2]内蒙古农牧业科学院,内蒙古呼和浩特010031

出　　处：《中国油料作物学报》2021年第3期452-461,共10页Chinese Journal of Oil Crop Sciences

基　　金：内蒙古科技计划项目(2019GG343);内蒙古科技成果转化专项(2019CG014)。

摘　　要：NAC(即NAM、ATAF和CUC)转录因子参与植物生长、发育、衰老和多种逆境胁迫反应的调控。为阐释甘蓝型油菜BnNAC61的表达特征,本研究采用RT-PCR方法,从甘蓝型油菜总cDNA中克隆BnNAC61基因。生物信息学分析表明,其CDS全长846 bp,编码281个氨基酸,N端含有NAM保守结构域;其启动子区存在W-box、响应抗性和胁迫、ABA、MeJA的顺式作用元件。利用烟草瞬时表达系统进行亚细胞定位,结果发现BnNAC61定位在细胞核。酵母试验表明,BnNAC61属于转录激活子,转录激活区位于136~208 aa区段。利用qRT-PCR研究BnNAC61的表达模式,结果表明,黑胫病菌(Leptosphaeria biglobosa)接种处理后,BnNAC61被显著诱导上调表达;逆境胁迫PEG、NaCl、4℃处理后BnNAC61被不同程度地诱导表达,表现先升高后降低的趋势;激素SA、MeJA和ACC处理后Bn⁃NAC61表达量显著高于对照(P<0.05),而激素ABA处理后,BnNAC61表达被抑制。上述结果表明BnNAC61是参与多种逆境胁迫的转录因子,尤其可能在茉莉酸和乙烯信号途径中发挥重要作用。NAC(NAM,ATAF and CUC)are involved in the regulation of plant growth,development,senescence and and various stress responses.To elucidate expression characteristics of BnNAC61 in Brassica napus,Bn⁃NAC61 gene was cloned from total cDNA of B.napus by RT-PCR.Bioinformatics analysis showed that the total length of CDS was 846 bp,encoding 281 amino acids,and N-terminal contained the conserved domain of NAM.Its cis-acting elements of W-box,response to resistance and stress,ABA and MeJA(jasmonic acid methyl ester)were found in promoter region.Subcellular localization using the transient expression system of tobacco showed that BnNAC61 was localized in nucleus.Yeast assay showed that BnNAC61 belonged to the transcriptional activator,and the transcriptional activation region was located at the 136-208 aa.By qRT-PCR,it was found that BnNAC61 expression was significantly up-regulated after inoculation with Leptosphaeria biglobosa.Moreover,BnNaC61 expression was induced by PEG,NaCl and 4℃stress,and showed a first increasing trend.BnNAC61 expressions after salicylic acid,MeJA and ACC(aminocyclopropane carboxylic acid)treatments were significantly higher than that of the control(P<0.05),although it was inhibited after ABA treatment.These results indicated that BnNAC61 is a transcription factor involved in various stress,especially in signaling pathways of MeJA and ethylene.

关 键 词：甘蓝型油菜 BnNAC61基因 亚细胞定位 转录激活 表达分析 

分 类 号：S565.4[农业科学—作物学]