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Function Analysis Of Two WRKYs From Caragana Intermedia

Posted on:2018-05-25Degree:MasterType:Thesis
Country:ChinaCandidate:M Z MaoFull Text:PDF
GTID:2310330518955926Subject:Biochemistry and Molecular Biology
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WRKY proteins are very vital transcription factors in plants.They have been reported to be involved in the regulation of plant development,biotic and abiotic stress responses,signalling pathways,etc.In this study,two genes encoding WRKY transcription factors from Caragana intermedia were cloned,and their functions were preliminarily identified,the detailed results are as following:1.The two WRKYs were named CiWRKY6 and CiWRKY45 respectively.The ORF of CiWRKY6 is 570 bp which encodes 189 amino acids.The ORF of CiWRKY45 is 900 bp which encodes 299 amino acids.The gDNA sequence of CiWRKY45 is 1554 bp which contains four exons and three introns.According to structure and evolution analysis,CiWRKY6 and CiWRKY45 belong to group ?a and group ?c of the WRKY transcription factor family respectively.2.CiWRKY6 and CiWRKY45 were both localized in the nuclei by transforming the GFP fusion constructs into Arabidopsis mesophyll protoplast.3.The expression patter analyzed by qRT-PCR found that CiWRKY6 and CiWRKY45 were induced via various abiotic stresses and ABA.Transcript of CiRKY6 and CiWRKY45 was dramatically induced by drought,salt and high pH,whereas it was lightly regulated by ABA,cold and heat.The expression of CiWRKY45 in leaf tissue is higher than that in stem and root.4.CiWRKY6 overexpression led to reduce drought tolerance compared with WT.The MDA content of OE was more than WT under drought stress,and the transgenic plants had a lower survival rate than WT.5.Transgenic Arabidopsis overexpressing CiWRKY45 delayed senescence compared with WT,and the senescence-related genes was down-regulated.
Keywords/Search Tags:Caragana intermedia, WRKY transcription factor, Drought, Senescence, Arabidopsis thaliana
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