Isolation and Expression Analysis of a Novel Abiotic Stress-Induced Gene W89 from Wheat

来源 :Agricultural Sciences in China | 被引量 : 0次 | 上传用户:ggf9988998
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Water stress and cold stress are important factors restricting plant growth.However,there is little knowledge on the function of stress-responsive genes in plants.Therefore,it is necessary to clone some important genes to study the mechanism of plant adaptation to abiotic stress for improvement of plant resistance.A putative water stress-induced gene,W89,was cloned from the cDNA library of drought-treated wheat seedlings by phage hybridization in situ,and its entire length was obtained using 5′-rapid amplification of cDNA ends(RACE)and reverse transcription-polymerase chain reaction(RT-PCR).The full-length cDNA of W89 consists of 2 392 bp and contains a 1 896 bp open reading frame(ORF) encoding a 631 amino acid protein.Southern blot analysis indicated that W89 was a single-copy gene.RT-PCR analysis revealed that the expression of W89 was upregulated by drought,cold,and abscisic acid(ABA).Amino acid sequence analysis discovered that W89 had a conserved region of DUF248(pfam03141),which contained a methyltransferase domain with a sterile alpha motif(SAM)-binding motif.Phylogenetic analysis showed that W89 was 66%identical to Oryza sativa dehydration-responsive protein(BAD67956).It was supposed that W89 was a novel dehydration-responsive protein encoding gene.On the basis of the functions of methyltransferase and the SAM-binding motif,the SAM-binding motif of W89 was supposed to be connected with other proteins or transcription factors to transduce stress signals and finally regulate the expression of stress-responsive genes on the early stage of drought stress. Water stress and cold stress are important factors restricting plant growth. However, there is little knowledge on the function of stress-responsive genes in plants. Wherefore, it is necessary to clone some important genes to study the mechanism of plant adaptation to abiotic stress for improvement of plant resistance. A putative water stress-induced gene, W89, was cloned from the cDNA library of drought-treated wheat seedlings by phage hybridization in situ, and its entire length was obtained using 5’-rapid amplification of cDNA ends (RACE ) and reverse transcription-polymerase chain reaction (RT-PCR). The full-length cDNA of W89 consists of 2 392 bp and contains a 1 896 bp open reading frame (ORF) encoding a 631 amino acid protein. Southern blot analysis that W89 was a single-copy gene. RT-PCR analysis revealed that the expression of W89 was upregulated by drought, cold, and abscisic acid (ABA). Amino acid sequence analysis discovered that W89 had a conserved region of DUF248 (pfam03141), whic h contained a methyltransferase domain with a sterile alpha motif (SAM) -binding motif. Phylogenetic analysis showed that W89 was 66% identical to Oryza sativa dehydration-responsive protein (BAD67956). It was supposed that W89 was a novel dehydration-responsive protein encoding gene.On the basis of the functions of methyltransferase and the SAM-binding motif, the SAM-binding motif of W89 was supposed to be connected with other proteins or transcription factors to transduce stress signals and finally regulate the expression of stress-responsive genes on the early stage of drought stress.
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