基因枪法将GAI基因导入巴西橡胶的研究

为了探讨利用基因工程技术进行橡胶树品质改良的可行性,用基因枪轰击巴西橡胶(Hevea brasiliensis)愈伤组织,将GAI矮化基因导入橡胶受体中,通过50mg L-1卡那霉素筛选鉴定,优化了基因枪法转化橡胶的各种参数。结果表明,DNA金弹与靶细胞的距离为9cm,每皿轰击一次时,胚状体诱导率可以达到1.87%。经过GUS组织染色和PCR扩增鉴定,初步确定GAI基因已经整合到橡胶基因组中。

Transfer of GAI Gene into Hevea brasiliensis by Particle Bombardment

GAI gene was transferred to calli from anther culture of Hevea brasiliensis by particle bombardment to develop dwarf rubber trees.Plant material was bombarded with plasmid PBI121 containing kanamycin resistant,GUS and GAI genes.Kanamycin concentration at 50 mg L-1 was used as selection pressure after concentration gradient experiments.The experiments showed that each plate of Hevea calli,which was bombarded once by gold particles coated with the plasmid and the distance between particles and target cells at 9 cm,had good results,the embryoid induction rate reaching to 1.87%.It was confirmed that GAI gene had been inserted into Hevea genome as detected by histochemical staining using X-gluc and PCR analysis.