Maize Transformation With Trehalose-6-Phosphate Synthase Gene From Selaginella Pulvinata

Drought is the main limiting factor for maize production. The use of transgenic technology modified tolerance to drought of maize varieties has become a major technical means now. Transgenic technology has to break the reproductive isolation between species compared with traditional hybridzation, and then transferred a useful genes of one species to other specise. It’s expanded the useful genes in a wider range of species to spread. In study that how to improve drought tolerance in maize, At home and abroad researchers have used SOD, the DREB and other drought-resistant gene transformation of maize to improve drought tolerance, however, these genes can not make corns showed good tolerance to drought. So explore the use of higher drought tolerance, suitable for corn physiological metabolism of the drought-resistant gene is the key to effective improve the drought-resistant of mazie. Trehalose in stress tolerance of plants has a significant role in promoting. So, improve the content of trehalose in the corn and to improve the drought tolerance of corn, and it has great significance to plant genetic improvement and corn production.This study by cloning Selaginella pulvinata trehalose-6-phosphate synthase SpTPSl gene and its modified SpTPSI△genes and construct the plant expression vector and used to transform maize embryonic calli of’18-599’by Agrobacterium-mediated transformation. Regenerated plants were obtained after culture callus and selection and differentiation. Transgenic positive plants were obtained after detected by PCR and provide a reference for the cultivation of genetically modified drought-resistant varieties.1. The Plant expression vector was constructed to target to trehalose-6-phosphate synthase gene and its modified genesAccording to the tutor group has cloned trehalose-6-phosphate synthase gene and its modified genes sequence from the Selaginella pulvinata and restriction sites were added to the fragments by PCR amplification. Then inserted into the plant expression vector and the new vectors were named as PTU-SpTPS1and PTU-SpTP·S1△. 2. Transformation of maize embryonic calliThe immature embryos of maize inbred line "18-599" were used to induce embryonic calli, Agrobacterium-mediated transformation of maize. Selection medium with different concentration of herbicide were utilized to select the resistant embryonic calli, which were transferred to differentiation and regeneration medium, differentiation of the90regenerated plants and transplanted to the field of72survived.3. Detection of regenerated plants40and32regenerated plants were obtained from the resistant calli transformed with SpTPS1and SpTPS1△genes.2and0TO plants were certified to be positive by PCR detection of target genes,15and9TO plants were certified to be positive by PCR detection of marker genes.Based on positive result of molecular level detection and field observations of the plant phenotype on this study, the transgenic plants is expected to provide material for further cultivate drought-tolerant maize varieties.