The Expression Of CFPS-M And CFPS-2M Gene And Resistance In Arabidopsis

Deinococcus radiodurans is one of the strongest radiation tolerance bacterium, CFPS is a gene which located in Deinococcus R1 genome.By analysis of the NCBI data found that CFPS and LEA76 belongs to the homologous gene family, belong to the third group of LEA genes.The third race LEA proteins contain multiple copies of11 amino acid sequence of primitive(TAQAAKEKAGE).In this study,CFPS gene is divided into five structure, containing 11 amino acid repeat sections from 104 to 160 amino acid structure domain, named CFPS-M; Contains twice as much as CFPS-M repeat segment structure domain, named CFPS-2M.We build the recombinant vector of the PCAMBIA2300-M and PCAMBIA2300-2M,Using the method of agrobacterium mediated the two genes into arabidopsis, obtain transgenic positive seedlings, and under a variety of abiotic stress resistance research analysis. The main results are as follows:(1)Successfully build the plant expression vector of PCAMBIA2300-M and PCAMBIA2300-2M, Using the method of freezing and thawing will the restructure carrier of PCAMBIA2300-M and PCAMBIA2300-2M into agrobacterium GV3103, via Kan resistance screening and colony PCR appraisal successfully imported into agrobacterium.(2)Choose the Arabidopsis thaliana as receptors,by agrobacterium infect the CFPS-M and CFPS-2M gene into the Arabidopsis thaliana, by means of Kan resistance selection and PCR identification of the genomic DNA, and real-time fluorescent quantitative PCR identification, selected 18 positive strains seedlings of CFPS-M and21 positive strains seedlings of CFPS-2M, and breeding selection to T3 generation homozygous positive seedlings.(3)Under a variety of abiotic stress, the relative arabidopsis seed germination rate and relative root length of transgenic plants is greater than the wild type plants.and the effect of CFPS-2M gene which contains double copy of CFPS-M is more significant in Arabidopsis, such as under the abiotic stress of drought, salinity, high temperature, lowtemperature are more prominent.(4)Under a variety of abiotic stress, the physiological and biochemical indexes of the transgenic Arabidopsis showed that the content of malondialdehyde of transgenic plants is significantly reduced than the wild type controls(P < 0.05).the content of osmotic regulation substances(soluble sugar, soluble protein and proline) is significantly higher than nontransgenic plants,and the activity of antioxidant enzyme system(SOD,POD, CAT) also greatly increased significantly higher than the control.These results suggest that the transgenic plants by reducing the accumulation of harmful substances in plants of MDA, and improve the content of osmotic regulation substances and increse the activity of antioxidant enzyme system to improve the ability of plants to resist the abiotic stress.(5)The resistance of CFPS-2M transgenic plant is more stronger than the CFPS-M transgenic plant.indicate that increased the copies of CFPS- M gene is enhance the effects of the gene expression.