互花米草SaNAC36转录因子克隆及表达分析

Cloning and expression analysis of SaNAC36 transcription factor in Spartina alterniflora

以互花米草(Spartina Alterniflora)为材料,利用三代全长转录组测序数据以及生物信息学手段获取SaNAC36的转录本序列.序列分析显示,SaNAC36基因全长CDS为942 bp,编码313个氨基酸.序列比对结果表明,SaNAC36蛋白具有保守的结构域且与水稻(Oryza sativa)、高粱(Sorghum bicolor)以及玉米(Zea mays)等植物具有高度同源性.qRT-PCR结果显示,SaNAC36基因具有显著的组织表达差异性,在互花米草叶片中表达量最高,且在盐、ABA以及干旱处理下表达量均呈下降趋势.亚细胞定位和自激活试验证明,SaNAC36蛋白在细胞核内表达且具有转录激活活性.同时在拟南芥(Arabidopsis thaliana)中过表达SaNAC36产生植株生长矮小的表型,进一步表明SaNAC36蛋白是互花米草生长发育及非生物胁迫应答调控的重要转录因子.

In order to explore the function of NAC36 transcription factor in Spartina alterniflora,the sequence of SaNAC36 transcript was first obtained by the third generation of full-length transcriptome sequencing data of S.alterniflora and then analyzed via bioinformatics methods.Sequence analysis indicated the CDS length of SaNAC36 gene was 942 bp,which encoded 313 amino acids.Sequence alignment results showed that SaNAC36 protein had conserved domains and high homology with proteins in Oryza sativa,Sorghum bicolor and Zea mays.The results of qRT-PCR showed that the expressions of SaNAC36 gene varied significantly among different tissues,with the maximum expression level occurring in the leaves of S.alterniflora,and also presented down-regulated trend under salt,ABA and drought stress.Subcellular localization and self-activation experiments showed that SaNAC36 protein expressed in the nucleus and had transcriptional activation activity.At the same time,it was confirmed that the overexpression of the SaNAC36 gene in Arabidopsis thaliana produced dwarf related phenotypes,further suggesting that SaNAC36 protein as an important transcriptional factor invovled in the regulation of growth and development and abiotic stress response in S.alterniflora.