Cloning And Functional Analysis Of PP2-A1 From Cucumber Phloem Protein

We used cucumber phloem protein PP2-A1 as the research objective and Arabidopsis thaliana as the main experiment material.The target gene was cloned and transformed into wild type Arabidopsis thaliana by Agrobacterium mediated method,and then we screened and cultivated the T₂ positive transgenic plants.We verified the defense function of the target gene by measuring and analyzing the growth indexes and physiological indexes of the same generation of wild-type and transgenic Arabidopsis.It will provide theoretical support for the cultivation and production of cucumber in the future.The main results are as follows:?1?According to the specific primers of phloem protein 2-A1 gene and cucumber cDNA by RT-PCR program,we got the objective liquid and approved that the target gene is successfully cloned,through the transformation of Escherichia coli,DNA sequencing and protein homology analysis;By using Agrobacterium mediated transformation of Arabidopsis thaliana,we successfully screened homozygous transgenic Arabidopsis plants.And we continue to culture it until geting the T₂ generation which as the next step experimental object.?2?Abiotic stress is an example in the aspect of gene function verification.In the same period,the germination potential,germination rate and germination index of transgenic Arabidopsis is different from that of wild-type Arabidopsis,and higher than that of wild-type Arabidopsis.The results showed that the germination ability of transgenic Arabidopsis seed is stronger than that in the saline environment.The POD and CAT activity of transgenic Arabidopsis thaliana is higher than that of wild type Arabidopsis thaliana under different salt concentration.It is proved that the transgenic Arabidopsis have stronger defense ability.?3?The results of study on the biological stress by using Botrytis cinerea and Phytophthora show that the transgenic Arabidopsis growth better than the wild type;Physiological indicators show POD and CAT activity of transgenic Arabidopsis are higher than that of wild type Arabidopsis after treatment.These results show that the ability of transgenic Arabidopsis is stronger than that of the wild type Arabidopsis in the process of preventing fungal infection.In summary,based on successful cloning of the target gene and study on the defense function of transgenic Arabidopsis thaliana.The above results showed that the performance of transgenic Arabidopsis thaliana is more positive resistance response which has strong defense functionin by using transgenic Arabidopsis through salt stress and fungal infection treatment.