Cloning and functional analysis of universal stress protein gene EuUSP1 in Eucommia ulmoides
Universal stress proteins(USPs)play a crucial role in protecting plants against various stressors.In this study,we aimed to investigate the molecular structure and function of Eucommia ulmoides univer-sal stress protein gene,EuUSP1.Tissue culture seedlings of E.ulmoides were used as materials,and Eu-USP1 cDNA was cloned through RT-PCR and RACE techniques.Bioinformatics analysis was conducted to gain insights into the gene.An overexpression vector was constructed and genetically transformed into Samsung tobacco for functional analysis.Spatiotemporal expression patterns were analyzed using qPCR technology.The results revealed that the transcriptional cDNA of EuUSP1 has a total length of 782 bp,with an open reading frame(ORF)length of 495 bp,a 5'untranslated region(UTR)of 149 bp,and a 3'UTR of 138 bp.It encodes a protein consisting of 164 amino acid residues.The expression level of EuUSP1 in young stems of E.ulmoides was higher than those in young leaves and roots.Furthermore,its expression showed a significant increase in July compared to other months.The transgenic overexpression of EuUSP1 did not significantly affect the morphology of tobacco plants.However,under drought stress conditions,the transgenic plants expressing EuUSP1 gene exhibited reduced wilting.After 10 days of drought stress,the activities of catalase(CAT),superoxide dismutase(SOD),and peroxidase(POD)enzymes in the leaves of the transgenic tobacco plants were significantly higher than those in the wild-type plants,and the respective enzyme activity values were 237.3,533.65,and 8 800 U·g-1.Additionally,the malondialdehyde(MDA)content before drought stress was significantly lower than that in the transgenic plants,with a value of 6.99 nmol·g-1.Overall,our findings suggest that EuUSP1 enhances drought resistance in E.ulmoides.
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