Analysis Of Different Proteins Of Fuzz Mutant In Gossypium.arboreum

The differentiation and initiation of the cotton fiber decide the final quality and the yield of the fiber. The relative research on protein can help us regulate the development of cotton fiber and get more benefits by genetic engineering and has important significance in application and theories. Protein is the direct executor for expressing the biological function with a lot of bio-information. It is so complex that we can not predict the structure and kinetics by analysis of the genome. Therefore the research of the structure, function and the location of protein in cell growing periods is urgently needed.The whole proteins were extracted by TCA-Acetone and phenol saturated ways. And then the two methods were evaluated. TCA-Acetone way which was easy to make the proteinases be denatured maybe get more kinds of proteins potentially, but the crude powder of proteins included many impurity components with lower concentration solution and yield of the protein compared with that by phenol saturated way. On the other hand, the phenol saturated way give us more kinds of proteins and the PI of these proteins are in a wide distribution with a simple manipulation technique. Finally we choose the phenol saturated method as optimal way in the later analysis for cotton protein.We got the whole proteins of diploid and tetraploid cotton species by phenol saturated way. we found that both of diploid and tetraploid cotton species almost had the similar content of proteins and alterative tendency after comparing the produce ratios of the protein powders. Excepting some abundant constitutive proteins the profiles of SDS-PAGE between the two species are different which also show their different genetic background. Thereby we can use the profiles as a powerful tool to distinguish the cotton species. The valuable different proteins between the wild type and mutant were not be found, and can not provide enough information for research of proteomics due to the low resolving power of the protein gels by SDS-PAGE.Through the observation of the phenotype and agricultural treats, we draw conclusions as followed, the phenotype between the wild type and mutant are very similar excepting the plant pigment expression and the existence of fuzz. Subsequently we confirm that the exactly time of fuzz initiation of G. arboretum (DPL971) was 5 days post anthesis by observing the epidermis cells of different growing period ovules with the microscope, SEM and paraffin section techniques.The proteomics of 5 DPA ovules of DPL971 and DPL972 were compared after extracting the whole proteins, 2-DE, matching the spots, constructing the proteins groups, evaluating the quality of the gels. Screening by statistic means and artificial observation, we gained 59 different protein spots including 5 particular spots in the gel of wild type and 14 spots in the gel of mutant, and 14 proteins of predominant expression in wild type and 26 proteins in mutant. These protein spots could be received repeatedly and stably after being screened scientifically and express the difference between wild type and mutant. We hypothesize that there would exist some reversal system to control the initiation of the fuzz.Our result can help people to identify and clone the functional genes and provide the information to understand the complex mechanism of plant cell differentiation.