Subcellular Localization And Functional Study Of Suaeda Salicin Protein (SsDHN)

In this study,sub-cellular localization technology and pull-down technology were used to study the dehydrin gene of Suaeda salsa from the cell structure and protein level.At the same time,by observing the salt-stress condition,the tobacco and wild-type tobacco were transgenic with the dehydrin gene.Changes in subcellular ultrastructure reveal the salt tolerance mechanism of the dehydrin gene in plants.The main conclusions are as follows:1.Firstly,the amino acid sequence of SsDHN gene was clustered by bioinformatics technology.The results showed that the dehydrin protein SsDHN was a mixed protein,which was closely related to the dehydrated protein of clover,and had a close relationship with the dehydrin protein of Momordica charantia.Far,this is consistent with the sequence homology alignment.2.In this experiment,the subcellular localization of Suaeda dehydrin protein was studied by transient expression method.By constructing the Super1300-SsDHN-GFP fusion expression vector and using transient expression method,the results were observed by confocal microscopy.The results showed that the dehydrin gene was localized on the chloroplast.3.The experiment studied the changes of chloroplast ultrastructure of wild type tobacco and trans dehydrin gene tobacco under different salt concentrations.Under salt stress,the chloroplast of transgenic dehydrin gene was damaged but the damage was not significant.Tobacco transgenic to the sulphate gene,there were many starch granules produced under high salt stress,which made the concentration of the solution in the cell.Higher than before,ensuring the absorption of water from plant cells can alleviate the decrease of water in plant cells caused by salt stress.The shape of chloroplasts in wild-type tobacco leaves expands,and a small part of chloroplasts agglomerates and is directed to the center of cells.The trend of movement,the phenomenon of small wall chloroplast body separation is obvious.In a few chloroplasts,the thylakoid membrane dissolves and ruptures,and small vacuoles appear.The matrix sheets are condensed into bundles,and some positions are expanded and arranged disorderly.The base layer is partially blurred.No starch granules appear,the number of hunger particles is small,and the volume is small.The test results show that the transhydrin gene tobacco is more salt tolerant than the wild type tobacco.4.A prokaryotic expression vector DE3-Pet32a-SsDHN was constructed to induce the expression of Suaeda salsa dehydratin gene.The target protein was detected by SDS-PAGE electrophoresis,and the results were in accordance with the expected size.Then,the SsDHN protein labeled with His was purified by Ni-NTA resin method,pull-down analysis was carried out,and the uniform bands were analyzed by mass spectrometry.Immunoglobulin,four Phytophthora tobacco proteins,Arabidopsis thaliana proteins,fructose diphosphate aldolase I and endoglucanase were obtained,which may interact with dehydration proteins,laying a foundation for further research.