Regeneration Potential of Five Cultivars of Canola (Brassica Napus L.) And Agrobacterium-mediated Transformation with the Salt Tolerance Gene mtlD

In order to improve the salt tolerance of some canola genotypes to be suitable for cultivation in Kingdom of Saudi Arabia (KSA), the salt tolerance gene (mt1D) encoding for mannitol-1-phosphate dehydrogenase was introduced to the canola plants using the agrobacterium-mediated transformation. To achieve that, five canola genotypes were selected as a relatively salt tolerance among 15 canola genotypes tested. The regeneration potential of the selected five genotypes was tested. A high percentage of callus induction ranged from 95 to 99% among the 5 cultivars with no significant differences were obtained within two weeks in the presence of 1 mg/ l of 2, 4-D. However, the cultivars showed a varied response to shoot regeneration. The cultivars Paketol and Sarow-8 showed higher regeneration frequency (66% and 61.5%, respectively) compared with the other cultivars using 4.5 mg/l BA. Hence the cultivars Paketol and Sarow-8 were genetically transformed with the salt tolerance gene (mt1D) encoding for mannitol-1-phosphate dehydrogenase. The mt1D gene was isolated from Escherichia coli using PCR cloning, sequenced, sub-cloned in the plant expression vector pBI121 and transferred to the hypocotyls explants exceeded from 6 days seedlings of canola plants. After infection with Agrobacterium, calli were transferred to the regeneration medium supplemented with 100 mg/L kanamycine. Transformation of the regenerated shoots was confirmed by PCR assay. The transformation efficiency calculated as 18 and 11% in cultivars Paketol and Sarwo-8, respectively.