Physiological And Biochemical Differences And Evaluation Of FKBP12 Gene Of Sugarcane With Different Combinations

Sugarcane is one of the best raw materials for producing fuel ethanol. The energy cane cultivars required high biomass, strong resisitance to botic and abotic stress as well as the good adaptability. The research was focused on the relationship of seven physiological and biochemical indexes to the biomass at the last growth stage of seeding, and on the application of FKBP12 gene to increase the drought-resisitance by genetic engineering. The results were showed as follows:1. Photosynthetic rate and chlorophyll content had positive effects on biomass in the cross combinations, but carotenoid content arose weak negative effect. The photosynthetic rate and chlorophyll content showed no significant positive correlation, while the positive correlation between chlorophyll content and carotenoid content assumed to be significant. IAA oxidase activity, protein content and soluble sugar content in leaf had negative effects while leaf sucrose content produced weak-positive effect on biomass of F1 seedlings.2. Using the designed PCR primers and the expression analysis through Real-time fluorescence quantitative PCR, the results showed the expression of FKBP12 gene was up-regulated at 45-folds after PEG stress for 36h, indicated the role of the FKBP12 gene in the resisitance to sugarcane drought.3. Two plant expression vectors: environmental inducible expression vector Rd29A-FKBP12-Hyg and the RNAi vector PTCK-FKBP12 were successfully constructed.4. Using gene gun bombardment method, FKBP12 gene super-expression vector PMI-FKBP12 was transformed into callus of sugarcane variety FN95-1702. Fifteen regeneration plants had PMI higher activity stained by chlorophenol red; Two from nineteen regeneration plants expressed at higher level of mRNA of FKBP12 gene exposed to water stress using Real-time fluorescence quantitative PCR, but they were only putative and still need further validation.