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Cloning and expression analysis of two WRKY transcription factors from the rare recretohalophyte Reaumuria trigyna

WANG Jia;ZHENG Lin-lin;GU Tian-pei;WANG Xue-feng;WANG Ying-chun;College of Life Science,Inner Mongolia University;Inner Mongolia and Monsod Drought Resistance Greening Corporation Limited;  
Reaumuria trigynais an endangered small shrub thriving in vast arid areas of Inner Mongolia.This species has developed a distinct morphology and physiology to adapt to the semi-desert environment.The WRKY gene family are plant-specific transcription factors which play important roles in the regulation of plant development,metabolism,and diverse biotic and abiotic stresses.Based on R.trigynatranscriptome data,two cDNA fragments(annotated as WRKY transcription factors)which were significantly upregulated after salt treatment were selected.Using rapid amplification of cDNA ends(RACE),these two WRKY transcription factors were cloned from R.trigyna.Blasted in the NCBI database,the homology to Arabidopsis AtWRKY33 were 79% and 87%,so they were named RtWRKY33-1and RtWRKY33-2(GenBank accession number KF421158 and KF421159),respectively.The full-length of the RtWRKY33-1was 2163 bp and included a 1681 bp open reading frame(ORF)which encoded 573 amino acids.The full-length of RtWRKY33-2was 2155 bp and included a 1776 bp ORF encoding 591 amino acids.Amino acid sequence analysis indicated that they were typical of class I WRKY transcription factors.Protein structure prediction analysis found that the primary structure and secondary structure in the WRKY domain of the amino acid sequence had structures characteristic of high similarity.However,in the non-conserved domains,especially the N-terminus(1to 80 aa range),before the first WRKY domain(190to 240 aa range)and between two WRKY domains(430to 450 aa range),there were significant differences that may influence the function of these two genes.Semi-quantitative RTPCR analysis showed that four abiotic stresses can induce the expression of these two genes.However,RtWRKY33-2was mainly inducted by salt,cold and ABA,while RtWRKY33-2responded faster to drought stress.These two genes in R.trigyna may play a different rolein abiotic stress responses.This research is a basis for study on the regulation mechanism of WRKY33 transcription factors in R.trigyna.
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