基因枪法介导的魔芋遗传转化研究

Genetic Transformation of Amorphophallus Mediated by Gene-Gun

利用基因枪轰击法建立了魔芋的遗传转化体系。试验结果表明 ,氦气压力 /轰击距离以 7.5 8GPa/9cm最适。在轰击前 4h和轰击后 16h用附加有 0 .4mol/L甘露醇的MS培养基进行高渗处理、轰击后恢复培养6～ 7d ,有利于减少逆境伤害 ,增加转化成功的几率。早期选用 1mg/LBasta作为选择压 ,而后期的筛选使用 2mg/L较高浓度的Basta可以获得真正的转化子。对获得的 91株抗性植株进行了PCR检测 ,结果显示 2 1株呈阳性 ,表明PAT基因和AGPS1反义基因均已整合进魔芋基因组。

Genetic transformation system of Amorphophallus wa s established via gene-gun .Results showed that the optimum helium gas pressure/b ombardment distance was 7.58 Gpa/9 cm. MS medium with addition of 0.4 mol/L man nit was used to do hypertonic treatment 4 hours before bombardment and 16 hours after bombardment . This could decrease adversity damage and increase the probab ility of transformation success. To obtain transgenic plants during screening,th e selected pressure was 1 mg/L Basta at the early stage and 2 mg/L Basta at the late stage.21 plants of 91 resistant plants tested by PCR were positive. It pro ved that PAT gene and AGPS1 antigene were integrated into the Amophoph allus genome.