Cloning and Expressional Analysis Under Abiotic Stress of the Protein Kinase Gene AsSnRK2.10 in Oat (Avena sativa)
XIANG Dian-Jun1, MAN Li-Li2*, WANG Qing-Xiang1, WANG Xiao-Dong1, ZHANG Wei-Wei1, LIU Peng1, LI Zhi-Gang1
1. College of Agriculture,Inner Mongolia University for Nationalities,Tongliao 028042,China; 2. College of Life Sciences and Food Engineering,Inner Mongolia University for Nationalities,Tongliao 028042,China
Abstract Sucrose non-fermented related protein kinase 2 (SnRK2) family members are plant-specific serine/threonine kinases involved in response to abiotic stresses. In this study,to explore the biological roles of SnRK2 gene in growth,development,and abiotic stress tolerance in oat (Avena sativa),one of the SnRK2 family members,designated AsSnRK2.10 (GenBank No. MN729578),was isolated from oat variety 'Monida' by reverse transcription-PCR (RT-PCR). The molecular characteristics of AsSnRK2.10 encoded protein were predicted by bioinformatics analysis. Then subcellular localization of target protein was performed using a transient expression system in tobacco (Nicotiana benthamiana). And lastly,the expression profiles of AsSnRK2.10 in different tissues and under abiotic stress conditions were detected by qRT-PCR,respectively. The results showed that AsSnRK2.10 gene contained a 1 086 bp of open reading frame (ORF),which encoded a polypeptide of 361 amino acid polypeptide with a predicted molecular weight of 40.66 kD and a theoretical isoelectric point of 4.75. The AsSnRK2.10 protein contained the SnRK2-specific functional domains,including a protein kinase adenosine triphosphate (ATP)-binding signaling region,a N-myristoylation site,a serine/threonine protein kinase active site,an activation loop,a transmembrane helix site,a domain 1 required for the osmotic stress response,and a domain 2 required for the abscisic acid (ABA) response. The smallest secondary structure element of the AsSnRK2.10 protein was beta turn,and the largest was random coil. Phylogenetic analysis showed that the AsSnRK2.10 protein had the closest evolutionary relationship with SnRK2 members from seven species,and was clustered into one branch. The subcellular localization analysis revealed that the AsSnRK2.7 protein was localized primarily in the nucleus. Tissue-specific expression showed that AsSnRK2.10 was expressed in the roots,stems,leaves,and panicles of oat during different development stages. However,the highest transcript levels of the AsSnRK2.10 gene in the roots,stems,leaves,and panicles occurred at the heading stage,filling stage,tillering stage,and heading stage,respectively. Furthermore,AsSnRK2.10 gene was involved in ABA signal transduction and positively responded to dehydration,high salinity,and low-temperature stresses. The results provide a theoretical basis for further study on the biological function of AsSnRK2.10 gene and its application in oat stress resistance breeding.
XIANG Dian-Jun,MAN Li-Li,WANG Qing-Xiang, et al. Cloning and Expressional Analysis Under Abiotic Stress of the Protein Kinase Gene AsSnRK2.10 in Oat (Avena sativa)[J]. 农业生物技术学报, 2020, 28(11): 1923-1935.
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