【摘要】 WRKY是植物中特有的具有锌指结构的转录因子家族之一,在植物非生物逆境胁迫应答中起着重要的调控作用。为研究金沙柚WRKY在铝胁迫条件下的作用,研究在前期构建的金沙柚铝胁迫转录组数据基础上,利用生物信息学方法从金沙柚根中得到了能显著响应铝胁迫的3个WRKY基因(Cg4g001470、Cg7g003890和Cg6g010690),开放阅读框长度分别为966、933和993 bp,各编码321、310和330个氨基酸,分别命名为CgWRKY40、CgWRKY70-1和CgWRKY70-2,氨基酸序列和结构分析显示,这3个基因编码的蛋白分别属于WRKY蛋白的IIa、III a和III a类型,PSORT亚细胞定位预测均定位于细胞核。实时荧光定量PCR分析表明,3个WRKY基因在金沙柚的嫩根、老根、茎和叶的表达量存在较大差异,在老根的表达量均最高;在铝胁迫处理后,均能出现不同程度的上调表达,相对表达量分别为处理前的4.58、1.52和12.06倍。在铝胁迫条件下,水杨酸能较大幅度的调控3个WRKY基因的上调表达,分别为对照的133.80、33.39和114.23倍;过氧化氢调控3个WRKY基因出现显著上调表达,分别为对照的86.40倍、7.93倍和102.32倍;水杨酸与过氧化氢共同作用下能调控3个WRKY基因出现不同程度的上调表达,分别为对照的51.81倍、1.51倍、37.72倍。上述研究表明,3个WRKY基因均能参与铝胁迫响应,且均可以被水杨酸、过氧化氢、水杨酸与过氧化氢调控上调表达。更多还原

【Abstract】 WRKY is a large family of plant?specific transcription factors with zinc finger motif,which plays an important role in regulating plantresponse to abiotic stress.In this study,3 WRKY genes(Cg4 g001470,Cg7 g003890 and Cg6 g010690 were identified,named CgWRKY40,CgWRKY70-1 and CgWRKY70-2),which could significantly respond to aluminum stress,using bioinformatics method from transcriptome database of Jinsha pomelo.The open reading frame lengths of the three WRKY genes were 966,933 and 993 bp,and en?coded 321,310 and 330 amino acids,which belonged to WRKY protein type IIa,III,and III a,respectively.PSORT predicted that these three proteins were all localized in the nucleus.Real-time fluorescent quantitative PCR analysis showed that the expression levels of the three WRKY genes varied in the tender roots,old roots,stems and leaves of Jinsha pomelo tissues,and the highest expression level was in the old roots.The expression pattern of the three genes were various after aluminum stress treatment,and up-regulated to 4.58,1.52 and4.58 times that of the control.The three genes could be up-regulated by salicylic acid under aluminum stress,and the relative expression increased up to 133.80,33.39 and 114.23 times that of the control,respectively.All the three genes could be up-regulated by hydrogen peroxide under aluminum stress,and their relative expres?sion increased up to 86.40,7.93,and 102.32 times that of the control.The three genes could be up-regulated by salicylic acid and hydrogen peroxide under aluminum stress,and their relative expression increased up to51.81,1.51,37.72 times that of the control.The research results showed that the three WRKY genes were in?volved in aluminum stress response,and could be up-regulated by salicylic acid,hydrogen peroxide,salicylic acid and hydrogen peroxide.更多还原