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以ps1-barnase(brn)为目的基因,pHcintG(PG)为选择/标记基因进行共转化,以PDS-1000-氦气基因枪介导,将brn及PG基因转化到水稻台北309及秋光的核DNA中,得到了转ps1-barnase基因的工程雄性不育植株。以悬浮细胞作为基因枪轰击的靶材料,转化植株再生频率较初级愈伤组织的为高。转brn基因植株的其他主要性状与供体亲本无显著差异,但却表现不育。其不育的程度在不同的植株之间表现不同。在转brn基因植株中观察到全不育(占全部brn阳性植株的40.6％)、高不育(占15.6％)及半不育的个体(占43.7％)。全不育的转基因植株自交完全不能结实(结实率为零),除个别植株外,花粉完全不被I-KI染色;而人工授以正常的花粉则可以获得杂交种子。而brn基因的阴性植株及未进行转化的对照植株则完全可育,表明转基因植株之雄性不育乃brn基因所致。结果表明,brn基因在水稻中是完全可以正常表达的,其表达的时期推测在花粉母细胞减数分裂前至花粉形成之间的整个时期。
The target gene ps1-barnase (brn) was selected and pHcintG (PG) was co-transformed with the selection / marker gene. Based on the PDS-1000-Helium gene gun, the brn and PG genes were transformed into rice Taipei 309 and the kernel of autumn light DNA, the engineered male sterile plants transformed with the ps1-barnase gene were obtained. Using suspension cells as target material for bombardment, the frequency of plant regeneration was higher than that of primary callus. The other major traits of transgenic plants were not significantly different from those of the donor but showed infertility. The level of infertility varies between plants. Infertility (accounting for 40.6% of all brn-positive plants), high infertility (15.6%) and semi-infertile individuals (43.7%) were observed in transgenic brn plants. All inbred transgenic plants were completely self-fertile (seed setting rate was zero), pollens were not completely stained by I-KI except for individual plants, while hybrid seeds could be obtained by artificially handing normal pollen. However, the negative plants of brn gene and the untransformed control plants were completely fertile, indicating that the male sterility of the transgenic plants is caused by the brn gene. The results showed that the brn gene was completely expressed in rice and its expression period was presumed to be in the entire period between pre-meiosis of pollen mother cells and pollen formation.