基因枪转化甘蔗愈伤组织体系的优化

Optimizing of genetic transformation of sugarcane calli via PDS-1000/He

以生产上广泛应用的甘蔗商业品种新台糖16号的愈伤组织为受体材料,以含有GUS基因的植物表达载体pBI121为外源DNA,利用基因枪PDS 1000/He,设定不同的参数,将pBI121轰击新台糖16号愈伤组织.经GUS染色,结果发现在发射距离1/4in、可裂膜压力1100psi、轰击距离6cm时转化效率最高.

Calli of sugarcane cultivar 'ROC'16 were selected as the targets for bombardment. The plant expression vector pBI121 with GUS gene was selected as the extrinsic DNA to coat microcarriers. The bombardment parameters of PDS-1000/He were adjusted and the calli were bombed. The calli were assayed with a colorimetric GUS assay. The results showed that the efficiency of transformation was high when distance between rupture disk and macrocarrier, the rupture pressure, distance between stopping screen and target shelf were 1/4 inch, 1 100 psi, 6 cm, respectively.