Optimization Of Regeneration System Of Sea Island Cotton And Study On The Genetic Transformation Of NAC Transcripts Family Gene

This study is further to optimize plant regeneration system of the Sea Island cotton and obtain regenerated plants by inducing somatic embryogenesis of Sea Island cotton hypocotyl of different genotypes; And with the embryogenic callus as receptor, transform NAC transcripts family with agrobacterium mediation and obtain transgenic plant, in order to lay the foundation for molecular breeding of drought-resistance. The major results are as follows:1. By exploring effects of the ratio of hormones, different IBA contents and different carbohydrate sources to differentiatial ability of Sea Island cotton embryogenic callus, activated carbon contents on the rooting of regenerated plants, further to optimize plant regeneration system of the Sea Island cotton. The results showed that the best hormones ratio was 0.1 mg/L 2,4-D+0.1 mg/L KT, the best media was 0.05 mg/L IBA,30 g/L glucose and 0.1% activated carbon.2. To further screen optimized agrobacterium-mediated gene transfer system for raising transformation efficiency of exogenous gene in Sea Island cotton, appropriate time of infection and co-cultivation had been confirmed by treating embryogenic callus of Xinhai 30 with the same physiological state and GFP gene respectively as receptor materials and reporter gene, and the time of transformation and co-cultivate were treated as experimental factors. The results indicated that the most suitable time of infection and co-culivate for agrobacterium-mediated gene transfer system of Sea Island cotton was 15 min and 24 h, respectively.3. Transgenic plants of NAC transcripts family were obtained by embryogenic callus of Sea Island cotton Xinhai 30 using the agrobacterium-mediated method. Embryogenic callus of Xinhai 30 were pretreated 10 d, co-culture 24 h after soaking 15 min in the bacilli of OD value was 0.5. After that, transferring the embryogenic callus to media contained 50 mg/L kanamycin and 400 mg/L cephamycin for inducing differentiation of somatic embryos, which was able to obtain 15% transgenic somatic embryos, and 5% of which enabled to obtain normal regenerated plants. PCR tests showed that NAC2,NAC3,NAC6 of NAC Transcripts Family had been transferred into regenerated cotton plants.