高效的水稻基因枪转化和可育转基因植株再生

从水稻品种台北309成熟胚诱导生长3~4周的愈伤组织，经过1~3次继代培养后，可以形成大量颗粒状胚性愈伤组织，适用于基因枪转化实验。将分别含有雪花莲凝集素(GNA)基因、h pt基因的质粒pIP860和p35H混合包裹在金粉上轰击转化上述胚性愈伤组织，在含30~50mg/L潮霉素的培养基上进行筛选、预再生、再生及长根培养。使用干燥处理

Efficient Production of Fertile Transgenic Rice Following Particle Bombardment

Many globular embryogenic calli suitable for particle bombardment were produced, after 3-4 week old calli derived from mature embryos of rice variety Taipei 309 were subcultured for 1-3 times. The embryogenic calli were bombarded with gold particles coated with the plasmid pIP860 containing snowdrop lectin (GNA) gene and the plasmid p35H containing hpt gene. Selection, pre-regeneration, regeneration and rooting were carried out on media with 30-50 mg/L hygromycin. Partial desiccation treatment was used to increase the efficiency of regeneration. The effective method of transplanting regenerated plantlets was established. Transformed plantlets were recovered within 13-21 weeks after mature seeds were plated. From 536 bombarded embryogenic calli, 2783 hygromycin resistant plants of 199 lines were obtained. DNA dot blot analysis showed that 73. 4% of resistant plants contained GNA and hpt genes. Southern blot analysis confirmed the integration of GNA gene in the genome of transgenic rice. 89. 3% of the transgenic plants were fertile. Mendelian segregation of the foreign genes was demonstrated by molecular analysis of R1 plants.