Studies On Genetic Transformation Of AtTIP5;1 Gene Into Oncidium And Saintpaulia Ionantha

In this study, Oncidium PLBs and Saintpaulia ionantha leaves were used as material to establish a performance regeneration and regeneration system. Based on the established system, AtTIP5;1 was transformed into Oncidium and Saintpaulia ionantha through Agrobacterium and transgenic plants were obtained and analyzed. The study for the further study of AtTIP5;1 gene about resistance and shorten the vegetative growth stage, and Oncidium, Saintpaulia ionantha gene engineering breeding laid the experimental foundation. The results described as following.1. The effects of auxin NAA, IBA and cytokinin 6-BA on the regeneration of tissue culture of Oncidium PLBs were studied. The results showed that MS medium supplemented with 1.0mg·L-1 6-BA and 0.5mg·L-1 NAA was the optimal combination for PLBs proliferation, giving the highest proliferation coefficient 3.87. During adventitious bud differentiation, MS medium supplemented with 2.0mg·L-16-BA and 0.5 mg·L-1 NAA produced the best bud induction rate of 86.67%. At the stage of roots inducing, added 0.5mg·L-1 IBA only could promote the best roots induction rate (86.67%).2. Established Agrobacterium-mediated gene transformation system for Oncidium PLBs and successfully transformed AtTIP5;1 into Oncidium. The optimal transformation conditions were:pre-trained PLBs for 2d, infected for 15min, co-cultured for 2d with 100umol·L-1 AS additive. For selecting transformed Oncidium PLBs, the most appropriate hygromycin and cefotaxime concentration were 20mg·L-1 and 300-400 mg·L-1, respectively. Our data demonstrated that Oncidium transgenic plants could resist boron toxicity. The expression of AtTIP5;1 in transgenic plants could significantly abate PLBs browning, abate the decrease of dry matter percentage, soluble sugar and soluble protein content and the increase of POD, SOD content which all caused by high boron stress. Our data suggests that AtTIP5;1 can help Oncidium transgenic plants to resist boron toxicity.3. The effects of auxin NAA, IB A and cytokinin 6-BA on the regeneration of Saintpaulia ionantha leaves in vitro were studied. The results showed that during adventitious bud differentiation, MS medium supplemented with 0.5 mg·L-1 6-BA plus 0.1 mg-L’1 NAA induced most of buds, resulting in a 100% bud induction rate. Meanwhile, the buds grew healthily and strongly. At the stage of roots inducing, 0.1mg·L-1 NAA could promote roots grew strikingly, generating the highest average root number of 16.72.4. Established Agrobacterium-mediated gene transformation system for Saintpaulia ionantha leaves and successfully transform AtTIP5;1 into Saintpaulia ionantha. The optimal transformation conditions were:pre-trained leaves for 2d, infected for 15min, co-cultured for 2d with 100umol·L-1 AS additive. For selecting transformed Saintpaulia ionantha leaves, the most appropriate hygromycin and cefotaxirne concentration were 40mg·L-1 and 300 mg·L-1, respectively. Our data showed that Saintpaulia ionantha transgenic plants could resist boron toxicity. The expression of AtTIP5;1 in transgenic plants could significantly abate tissue browning and leaf morphing, abate the decrease of dry matter percentage, soluble sugar and soluble protein content and the increase of POD, SOD content which all caused by high boron stress. Our results indicate that AtTIP5; 1 can help Saintpaulia ionantha transgenic plants to resist boron toxicity.