To Transfer Anti-herbicide Gene into Upland Rice (Oryza sativa)Cultivars
La Honggui1 Wang Huaqi1** Huang Danian2 Hua Zhihua2 Yan Meixian2 Gao Zhenyu2 Feng Xiujing1
(1.Center of Upland Rice, College of Agronomy and Biotechnology, China Agricultural University,Beijing Key Laboratory of Crops Genetic Improvement, Beijing 100094, China; 2.China National Rice Research Institute, Hangzhou 310006, China)
摘要摘要:以生产上优良旱稻(Oryza sativa L. )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。
Abstract:Abstract: The anti-herbicide gene bar was transferred into immature embryos calli from elite cultivars of upland rice(Oryza sativa L. )297, 10 and so on with help of particle bombardment. Some regeneration plantlets were acquired after two selections on media containing herbicide Basta and differentiation. Further molecular test by both PCR amplification and Southern blotting and experiments in fields demonstrated that extrogenous gene bar was integrated into T0 genomes and inherited to T1. The tests of induction and differentiation media indicated that MB and MS were better for induction of calli and the modified media of RMB2 and RMS2 promoted differentiation frequency of calli remarkably. The obtained transgenic plants and established genetic transformation system of the experiment were available for further anti-herbicide molecular breeding and other genetic transformation.