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作 者:韩继成[1] 陈宁[1] 陈霜莹[1] 章德明[1] 蔡文启[2]
机构地区:[1]河北省农林科学院昌黎果树研究所,昌黎066600 [2]中国科学院微生物研究所,北京100080
出 处:《分子植物育种》2006年第5期670-674,共5页Molecular Plant Breeding
基 金:河北省自然科学基金(399488)资助.
摘 要:本研究利用无核白、红地球、黑王和红宝石这4个无病毒葡萄试管苗的叶片为外植体,在NN69培养基上获得了稳定高频率的再生植株,并对抗生素浓度、根癌农杆菌不同活化方法、葡萄叶片不同叶位、切割叶片时的不同液体、聚乙烯吡咯烷酮(酚类吸附剂)和维生素C(抗氧化剂)对葡萄叶片遗传转化的影响进行了研究。结果表明,葡萄叶片对卡那霉素敏感,并因不同品种而稍有差异;叶片不同叶位中以取顶端第一幼叶时转化效率最高;根癌农杆菌活化方法中以活化18h后,添加等体积新鲜LB培养基,再培养5h时转化效率最高;聚乙烯吡咯烷酮和维生素C对遗传转化不但没有明显的促进作用,而且明显抑制叶片的再生率。The effects of the concentration of kanamycin and carbenicillin, leaf at different leaf order, cultured methods for Agrobacterium tumefaciens, PVP and vitamin C were studied on the ratio of genetic transformation of the leaves of virus-free grape based on high ratio and stable plant regeneration acquired on NN69 medium. The result showed that the grape leaves were very sensitive to kanamycin and the sensitivity varied with different varieties. The highest transformation rate of the adventitious bud was obtained from the first top leaf, the A. tumefaciens cultured for 18 hours, added equal volume of LB medium and cultured for another 5 hours. The PVP and vitamin C not only have no clear effects on the transformation but also restrain the regeneration rate of the leaves.
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