| In recent years, the structure and function of CENPA Has Drawn Wide Attention, Among them the relationship between the expression or abnormal structure of CENPA in the process of cell division and the abnormal chromosomes separation has become a hot issue,by which people study the function of CENPA. In the study, we make’linyou2018’wheat as a material and extract CENPA Respectively with direct acid extraction and acid extraction-ethanol precipitation and acid extraction-TCA precipitation methods from it, then for CENPA,we do quantitative,testing and identification Respectively with Bradford method,sds-page and indirect immunofluorescene technology, compare the three different extraction methods, to find a suitable method of protein extraction wheat. On this basis, we use direct acid extraction method extract CENPA of wheat which were divided into CK(not treated), B(UV-B), BL(UV-B+He-Ne laser),L(He-Ne laser) four different treatments.The content of CENPA of the four treatments in different treat days were measured In order to study the effects of UV-B and He-Ne laser on the CENPA, as well as to further investigate relationships between CENPA and abnormal chromosomes separation. Moreover,in the study, Nitric oxide synthase (NOS) and nitrate reductase (NR) were extracted, Quantitative and compared in order to study the effects of UV-B and He-Ne laser on the enzymes related nitric oxide synthesis, as well as to Preliminary discuss the mechanism which enzymes related nitric oxide synthesis response to UV-B stress. The results were showed as follows:(1) it could make better to utilize the direct acid extraction methods on CENPA extraction,Using this method,the protein was got simply and efficiently, the direct acid extraction method produce the purer sample,much more clearer bands and highest resolution. This method can form a good foundation for the future, purification and quantitative immune imprinting hybrid.So we decide to select the direct acid extraction methods as the extract method of CENPA in the subsequent experiment.(2) Five days of experimental treatment of wheat, Enhance UV-B radiation and He-Ne laser irradiation has a significant effect on the content of CENPA in wheat seeding root and leaves,as well as both the effect are roughly the same, after enhanced UV-B irradiation, content of CENPA in wheat seeding roots and leaves dropped significantly (P<0.01) compared with the control group showing a certain inhibiting effect. after a single He-Ne laser treatment, the content of CENPA has no significant difference with the control group, which indicates the effect of laser on CENPA content is not adverse and in combination treatment of He-Ne laser and enhanced UV-B radiation, the wheat CENPA content shows Obviously higher than the UV-B treatment group, and close to CK group showing a certain inhibiting effect,which indicate a certain dose of He-Ne laser has some repair to the CENPA of wheat which has been damaged by enhanced UV-B radiation.(3) after enhanced UV-B irradiation, the content of nitric oxide synthase (NOS) of roots in Wheat seedling obviously increased; after a single He-Ne laser treatment, the content of NOS Significantly increasedhas no significant difference with the control group; While in combination treatment of He-Ne laser and enhanced UV-B radiation, the content of NOS shows higher than those in the control group and UV-B treatment group,which showed that UV-B and He-Ne laser can resistance to stress joint togther and promote each other. after enhanced UV-B irradiation, the activity of Nitrate reductase (NR) in roots of Wheat seedling obviously droped; after He-Ne laser treatment, the activity of Nitrate reductase (NR) rised; in combination treatment of He-Ne laser and enhanced UV-B radiation, the lower activity of NR caused by the UV radiation-B has been significantly improved,that is to say He-Ne laser can enhance the resistance ability to damage caused by UV-B radiation. |
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