Studies On The Expression Pattern Of PsnPR10 Gene To Response Alternaria Alternate And Salt Stress In Populus Simonii X P.nigra

In this study,the PsnPR10 gene from Populus simonii×P.nigra was cloned,and the expression pattern of PsnPR10 gene in response to Alternaria alternata,salt and SA was analyzed,which laid a solid foundation for studying the mechanism of PR10 protein family involved in plant growth and development and defense response regulation.The results are as follows.A 477bp cDNA fragment of the PsnPR10 gene from Poplus simonii × P.nigra was cloned and sequenced.Its open reading frame encoded 158 amino acids,the molecular weight of the putative protein was 17.6 kDa,the isoelectric point was 5.20,and the instability coefficient was 34.32.The total average hydrophobic index of was-0.186,the hydrophilic regions were evenly distributed,and the number of hydrophilic amino acids was large,belong to hydrophilic protien.The expression vector of pBI121-PsnPR10-GFP fusion protein was constructed,and subcellular localization indicated PsnPR10is located in the nucleus.The expression pattern of PsnPR10 gene in Poplus simonii × P.nigra under SA,MeJA,NaCl and leaf blight stress for 0,3,6,12,24 and 48 hours showed a trend of "induction to recovery".The promoter of PsnPR10 gene in Populus simonii was cloned and analyzed by on-line analysis software.It was found that it contains G-box,W-box,MYB,methyl jasmonate response element and other important cis acting elements.The transgenic Arabidopsis thaliana with GUS gene expression driven by PsnPR10 promoter was cultivated by Agrobacterium tumefaciens Infection.GUS staining showed that the promoter had an activity to driven GUS gene expression,and GUS gene expression had tissue characteristics,expressed in leaves and roots,but not in stems.The recombinant PsnPR10 protein was induced to express under 37℃ and 0.1 M IPTG.A large amount of PsnPR10 protein was obtained in the supernatant by ultrasonic lysis.The purified PsnPR10 recombinant protein was obtained by using amlose resin.The RNase activity of PsnPR10 recombinant protein was detected and it was found that PsnPR10 has RNase activity in vitro.