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以生产上优良旱稻(OryzasativaL.)新品种旱稻297、旱稻10号等的幼胚愈伤组织为转化受体材料,用基因枪法把抗Basta除草剂的bar基因导入了这些品种的愈伤组织,经两轮Basta抗性筛选和分化获得了再生植株,对再生植株进行PCR扩增和Southern杂交检测,T0和T1代Basta抗性实验表明,bar基因已整合到旱稻的基因组DNA中,并在T1代继续表达。对各品种幼胚培养的诱导、分化培养基实验表明,MB和MS培养基可作为这5个品种的愈伤组织诱导培养基,改良的RMB2、RMS2培养基可显著地提高愈伤组织的分化频率。实验所获得的转基因植株和建立的遗传转化系统,为旱稻的抗除草剂分子育种和其它基因转化奠定了初步的基础。
The immature embryos calli derived from Oryza sativa L. (Oryza sativa L.) cultivars Upland 297 and Upland 10 were transformed into recipient materials. The bar gene of anti-Basta herbicide was introduced into the callus of these cultivars by particle bombardment. After two rounds of Basta resistance screening and differentiation, regenerated plants were obtained, and PCR amplification and Southern hybridization were carried out on the regenerated plants. The Basta resistance test in T0 and T1 generation showed that the bar gene was integrated into the genomic DNA of upland rice, Continue to express on behalf of. Induction and differentiation medium experiments of immature embryos of each variety showed that MB and MS medium could be used as callus induction medium of these five varieties. Improved RMB2 and RMS2 medium could significantly improve the differentiation of callus frequency. The experimentally obtained transgenic plants and the established genetic transformation system laid the preliminary foundation for the resistance breeding of herbicide and other gene transformation of upland rice.