Studies On Transformation Of Soybean Cotyledonary Nodes By Agrobacterium Tumefaciens

Agrobacterium twnefacien-mediated soybean cotyledonary node transformation system was optimizated, according to the characteristic of A.. twnefacien -mediated transformation and soybean cotyledonary node regeneration system. Bacillus thuringiensis crylA gene was introduced into soybean successfully. Transformed plants were examined by PCR and PCRSouthern reaction, and 4 of the plants give positive reaction. These results show that the Bt gene has integated into receiver?s genorne.Optimization of soybean cotyledonary node regeneration:?MMS medium is obviously better than B5BA and MS medium in increasing adventitious buds. The results indicated that BA greatly influence the production of adventitious buds by germinating experiment of three geminating media,B5BA B5 and Agar. The suitable concentrations of BA~ NAA and proline in MMS medium are 2mg/I ~. O.02mgIl~ 2g/I respectively?The number of cotyledon influences production of adventitious buds ,and the suitable number of cotyledon is halfThe factors influencing A.tumefacien-mediated soybean transformation were studied with the section of soybean hypocotyl from 7-day-old seedlings as explants and resistant callus as target ,The results indicate:?Preculture of explants on hypertonic medium before infection of A.tumefacien can raise the transformation frequecy of the section of soybean hypocotyl. The optimium duration of pre culture is 1 day.?Both the concentration of acetosyingone(AS) and PH value at stages of infection and coculture influence the transformation frequency of soybean, The suitable PH value is between 5.4and 5.6,when it exceeds 5.8,the transformation frequency decreases obviously ,The suitable AS concentration l00J.LM.?Sonication of explants can greatly increase the frequency of gus gene expression and transformation after it infected by A.twnefaciens, and the suitable treatment duration is 60 seconds.?The age of explant influences the transformation frequency greatly, The best age of explant is 7 days.Postgr&luate:Liu Zhaojun Major: Crop Genetics and BreedingSuperviser:Prof. Yang Qingkai...