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The Functional Analysis Of Thellungiella Halophila TRXh And Comparative Proteomic Study Of Thellungiella Ecotypes

Posted on:2007-11-04Degree:MasterType:Thesis
Country:ChinaCandidate:Z B SunFull Text:PDF
GTID:2120360182497223Subject:Developmental Biology
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The plant was subjected to various stresses in its lifecycle, such as the drought, not suitabletemperature, ozone and the ultraviolet ray irradiation, high salt and the virus etc., the occurrences ofthese stresses were often accompanied with ROS and the secondary oxidize stress. The livingcreature can regulate the redox state through a variety of paths or ways, and the regulation withthioredoxin is a way of these kinds.The first part of this thesis studied the expression pattern,enzyme activity of Thellungiellahalophila TRXh, effection of overexpressing a Thellungiella halophila TRXh on the stress toleranceof Arabidopsis. We also created the Th TRXh silence vector, and got some gene silence transgenicThellungiella halophila plants.The stress tolerance of the ThTRXh-overexpression Arabidopsis plants was studied.Thegermination ratio of transgenic plants was higher than that of the wild type in medium containingMV or in that containing NaCl. The seedling growth of the transgenic plants was more severelyinhibited, comparaed with that of the wild type. Under the stresses of MV or NaCl, the root growthsof the transgenic and the wild type were both restrained. The taproot growth of the transgenic plantwas better than that of the wild type under the both stresses, while the lateral root number of thetransgenic plants was smaller than that of the wild type. The Th TRXh gene silence lines of theThellungiella halophila were screened. Under the normal growth condition, there is not any obviousdifference between the gene silence lines and the wild type Thellungiella halophila. Some furtherstudies should be taken to know the functions of the Th TRXh. The proteomics can be implied in many fields such as the protein expression level,the sequenceof amino acids, the post-translational modification, the protein location and the protein-proteininteraction, which cannot be solved by genomics. The proteome of many plants, such asArabidopsis, rice and corn, has been studied, but there was no paper concerning the proteomics ofThellungiella reported so far. Thellungiella, the model system to study plant salt-tolerance, hasmany EST data, and is very close to Arabidopsis. Due to these advantages, the genomic data andprotein database of Arabidopsis can be used in the proteimics study of Thellungiella halophila. Thesecond part of this thesis established a platform of two-dimension gel electrophoresis combining theuse of Proteomic Protein Picker to study the protein expression difference of Thellungiellahalophila, and optimized the procedure of protein extraction and isoelectric focusing. We took aprimary study of the proteomic differences among the Thellungiella ecotypes, some different spotswere found, and a few of them were identified by MS. After the identification of the differentprotein dots from Thellungiella, the results were checked in the protein database of Arabidopsis.Unfortunately, the results are not very well-pleasing. To improve the result, we performed the in-geldigest using BSA and directly loaded the enzyme-digested sample. In this way, well MSidentification results were obtained. The reasons of these phenomena might concern two aspects.On the one hand,perhaps the peptide MS map of BSA is detailed while searching proteins fromThellungiella in the protein database of Arabidopsis covers a pretty wide span. On the other hand,maybe this method causes a great loss in protein amount and therefore leads to the amount ofloading protein lower than that acquired for the assurance of the accuracy of MS identification. Onthe whole, this part lays a foundation for the proteomics study of Thellungiella.
Keywords/Search Tags:Thellungiella halophila, oxidize stress, salt tolerance, Th TRXh, proteomics, ecotype
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