Research on Improving Plant Persistence Drought Resistance by Inducible Pathways.
|Course||Biochemistry and Molecular Biology|
|Keywords||CBF3 gene WIN1 gene RD29A promotor LEA14 promotor Drought-resistance Transcription factor Inducible promoter Phenotypic markers Plant expression vector construction|
Using transcription factors to enhance drought resistance of crops is currently a hot research. In order to overcome the disadvantages that constitutively expressing transcription factor genes can affect the traits of transgenic plants, in this study we have constructed a kind of bivalent plant expression vector with functions which are cascade amplification and phenotypic marker. This expression system, such as in drought stress conditions, can be induced to express through the cascade Amplification way. It might be able to increase the resistance of plants while increasing the leaf surface wax accumulation, then the phenotype is easy to be identified.We first carried out stress tests with wild-type Arabidopsis thaliana, through the PR-PCR semi-quantitative methods to test the expression of CBF3 gene, WIN1 gene, RD29A gene, LEA14 gene under the conditions of stress, to provide technical support for constructing plant expression vector.Cold-induced transcription factor gene CBF3, synthetic wax-related genes WIN1, drought-induced gene RD29A’s promoter and cold-induced gene LEA14’s promoter were cloned from the Arabidopsis by using the PCR method then. And we use CBF3 transcription factor which can regulate downstream genes RD29A and LEA14’s promote respectively to drive the expression of gene WIN1 and CBF3.We funally successfully constructed the bivalent plant expression vector RD29AP-CBF3/LEA14P-WIN1/pCAMBIA2201.The recombinant plasmids RD29AP-CBF3/pBI121, LEA14P-WINl/pBI121, RD29AP-CBF3/LEA14P-WINl/pCAMBIA2201 were transferred into Agrobacterium tumefaciens GV3101. Using leaf discs method, the gene were transferred into tobacco cell. Transferred leafs were selected on solid medium containing Kanamycin. Transgenic tobacco plants were found containing purpose gene by PCR.Owing to time constraints, this study just has preliminary results. This experiment provides a practical new ideas for the future breeding of transgenic drought-resisitance plants for using pollen-tube pathway to transform cotton, and has laid a foundation of enhancing the screening efficiency to select transgenic cotton which were transformed through pollen-tube pathway in cotton fields.