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以水稻愈伤组织和悬浮细胞系作为基因枪转化的外植体,把水稻花粉特异性基因PS1启动子与barnase构成的嵌合基因导入籼稻,获得籼稻五个品种Basmati 1、青油占、胜优2号,明恢63,新山占29的转基因植株.试验以两个质粒PS1- barnase和pILTAB227共转化的方法,以潮霉素B作为筛选因子,选择抗性愈伤及再生植株.获得的barnase转基因植株的育性比未转化的对照明显降低,表现为部分不育和完全不育.除barnase阳性转化株平均株高比对照有所降低外,营养器官发育正常,雌性可育.完全不育的植株花粉粒畸形,不能被I-KI溶液染色,但它们与正常植株杂交能够获得杂交种子.转基因植株的Sonthern杂交分析表明,bamase基因普遍为多拷贝整合.对转基因植株白交及与非转化株测交后代以点杂交分析bamase转基因的遗传行为,发现转基因的遗传符合1～3个插入位点的孟德尔遗传模式.
Rice callus and suspension cell lines were used as explants for gene gun transformation. The chimeric gene consisting of the rice pollen-specific gene PS1 promoter and barnase was introduced into indica rice to obtain five varieties of indica, Basmati 1 You 2, Minghui 63, Johor Bahru (JB) accounted for 29. The experiment selects the resistant calli and the regenerated plants with the method of co-transformation of two plasmids PS1-barnase and pILTAB227, using hygromycin B as screening factor. The sterility of barnase transgenic plants was significantly lower than that of the untransformed control plants, which showed partial sterility and complete sterility. The vegetative organs developed normally and the female fertile except the average plant height of barnase-positive transformants was lower than the control The fertile plants pollen grains were deformable and could not be stained with I-KI solution, but they could obtain hybrid seeds by crossing with normal plants.The Sonthern hybridization analysis of transgenic plants showed that the bamase gene was generally multi-copy integrated.The transgenic plants Transgenic plants were tested for genetic crosses by spot hybridization and found that the genetic inheritance of the transgene met the Mendelian inheritance pattern of 1 to 3 insertion sites.