基因枪法介导GNA基因遗传转化甘蔗的研究

GNA Gene Genetic Transformation of Sugarcane by Particle Bombardment

目的:将含有雪花莲外源凝集素(GNA)基因的植物表达载体用基因枪法分别导入一个果蔗和一个糖蔗品种中,以期获得转基因植株。方法:将GNA基因插入到植物表达载体上,构建出不同选择标记、不同启动子的表达载体,并用基因枪法将之导入甘蔗胚性愈伤组织,分别在G418、PPT和Hyg的选择压力下,筛选抗性植株,并进行分子杂交鉴定。结果:通过斑点杂交和PCR-Southern杂交证明GNA基因已整合到甘蔗基因组中。结论:用基因枪法成功获得了含有GNA基因的甘蔗转化株,为培育抗甘蔗绵蚜(Ceratovacuna lanigeraZehnther)的新品种提供了基础。

Objectives： Plant expression vectors containing snowdrop lectin gene （ Galanthus niralis aggluthinin, GNA） were tranformed by particle bombardment into one fruit- useful and one sugar- useful sugarcane culivars respectively, in order to obtain transgenic sugarcane plants, Methods： Plant expression vectors were constructed by inserting GNA gene with different promoter and selection markers. Sugarcane＇s embryonic calli was transformed by particle bombardment. The resistant calli and shoots were sereened under the selective pressure of PPT and G418, which would be identified by molecular hybridization. Results： The transformed plants were detected by PCR and PCR- Southern blot and proved GNA gene had been integrated into sugarcane genome. Conclusion： Transformed sugareane plants with GNA gene were obtained by particle bombardment successfully. The result would he the basis for breeding new sugarcane cultivars resistant to woolly aphids （ Ceratovacuna lanigera Zehnther）.