The Drought Resistant Function Research Of TaCDPK1 In Wheat(Triticum Aestivum L.)

Under natural conditions,the growth and development of wheat will be affected by a variety of stress environments,resulting in a serious decline in production and quality.Therefore,it is important to explore how to improve the resistance and quality of wheat through modern biotechnology.Now,people have found protein kinase could response under some stresses induced,like drought,high-salt,low temperature and so on.It is necessary to explore the function of protein kinase to enhance the resistance of plant.The previous experimental work of our laboratory had cloned a wheat calcium dependent protein from the wheat,and was named as TaCDPK1.To study the function of TaCDPK1 in wheat,This paper mainly studies TaCDPK1 from two aspects.On the one hand,through the study of the bioinformatics analysis,subcellular localization,GUS staining analysis of promoter,expression pattern under abiotic stress of TaCDPK1,there preliminarily summarized the structure and function of TaCDPK1.On the other hand,it is mainly to explore the phenotypic of TaCDPK1 transgenic Arabidopsis under drought stress.In addition,the ABA signaling pathway and GA signaling pathway involved in TaCDPK1 were explored.The main results are as follows:1.Through some bioinformatics analysis websites,It can be inferred that TaCDPK1 has kinase region and four EF-Hand domains.The protein content of TaCDPK1 protein was 59.3 kDa.The protein expression of TaCDPK1 gene is expressed mainly in cell membrane and nucleus.By tissue specific quantitative analysis,it was found that TaCDPK1 was mainly expressed in stem and leaf.GUS staining analysis of promoter indicated that TaCDPK1 gene was mainly expressed in stem,leaf and stamen.Analysis of expression patterns under different stress treatments indicated that TaCDPK1 was induced by drought and ABA.In addition,the interaction between TaCDPK1 and the selected protein TaRan1 was verified by pull-down and BiFC and the interaction place was located in the nucleus and cell membrane.2.The TaCDPK1 was transferred to Arabidopsis thaliana by Agrobacterium-mediated transformation and we found that TaCDPK1 could promote the germination and the increase of root surface of transgenic Arabidopsis thaliana under drought stress.The expression of drought stress related genes(RD29A,COR47,DREB2 A,RAB18,KIN1,KIN2)was promoted at the same time.It can be inferred that TaCDPK1 improves the drought resistance of transgenic Arabidopsis by promoting the expression of stress related genes.3.Expression pattern analysis shows that TaCDPK1 induced by ABA expression.The stomatal aperture of transgenic Arabidopsis was smaller than wild type under different ABA concentration.The NCED3,a key kinase involve in the synthesis pathway of ABA,was obviously increased after drought stress in transgenic Arabidopsis.then,we can surmise that TaCDPK1 might involved in the signaling pathway of ABA response.4.Under PAC treament,the germination rate of overexpressed strains was significantly greater than that of WT.And in the hypocotyl experiment,the hypocotyl length of over-expression lines was longer than WT in GA and PAC.That indicated that TaCDPK1 overexpression strains are not sensitive to PAC.The experiment also found that some gene expressions related to GA synthesis in overexpression strains decreased under GA treatment,for example,KS,KAO1,KAO2,etc.It illustrated that TaCDPK1 may be involved in GA pathway.The above results provide a theoretical basis for further study on the mechanism of TaCDPK1 gene to improve plant resistance.It also provides theoretical reference and practical guidance for the improvement of wheat varieties of anti-stress transgenic engineering.