Cloning,expression And Identification Of CtOleosin Family Genes In Safflower

Safflower,1-2 yeas old herbaceous plant of the compositae,is widely used as an oil crop with an oil content as high as 35-47,and its linoleic acid content is as high as 73-80,which has been known as the "king of linoleic acid".In this study,safflower was used as the object of research.The full-length cDNA fragment of CtOleosin family genes were cloned by RT-PCR,and its physicochemical properties and structural characteristics were analyzed by bioinformatics.The genes of this family were detected in different parts,different developmental stages and different substancesby qRT-PCR method.The CtOleosin family genes were cloned,and plant superexpression vector was constructed to transform into Arabidopsis thaliana.The transgenic Arabidopsis thaliana was obtained and then to study its function.The specific findings are as follows:1.The best internal reference was EF1?.The qRT-PCR results showed that the CtOleosin family genes had similar expression patterns.The expression of CtOleosin genes was the most significant in seeds and flowers,and hardly expressed in roots,stems,leaves,cotyledons and cotyledons.The expression levels of this family genes were gradually increased in the different developmental stages of the seed,and reached the highest level at 28 d,and then decreased;After treatment with ABA,SA and NaCl,family genes were found to be significantly affected.2.The full-length cDNA fragment of the CtOleosin genes were cloned by RT-PCR,and the sequence were analyed by bioinformatics.The results showed that the CtOleosin1-8 family genes encoded 161,137,160,185,211,224,148,161 amino acids,respectively.CtOleosin genes exist transmembrane regions,and have no signal peptide.All family genes contain proline motifs,which are members of the Oleosin family.3.The CtOleosin family gene was successfully cloned,and the plant overexpression vector pOBT-CtOleosin was constructed and transferred into wild type Arabidopsis thaliana.The T3 generation of CtOleosin1-8 was 4 strains,1 strain,5 strain,4 strain,3 strain,respectively.4.Phenotypic analysis and functional identification of transgenic Arabidopsis thaliana showed that the 100-grain weight of transgenic Arabidopsis thaliana seeds in generation T3 was slightly higher than that of transgenic leaves in generation T3 of wild type seeds,and the pods and grain size of transgenic Arabidopsis thaliana were slightly larger than those of wild type,and the oil content of transgenic Arabidopsis thaliana is higher than that of wild type.Compared with wild-type and transformed into CtOleosin gene,seed germination rate and protein content have different degrees.