辐照对结缕草农杆菌介导法遗传转化的影响

EFFECT OF GAMMA IRRADIATION ON Agrobacterium-MEDIATED GENETIC TRANSFORMATION OF JAPANESE LAWNGRASS(Zoysia japonica Steud.)

以结缕草成熟种子诱导产生的胚性愈伤组织为材料 ,研究了钴 60伽玛射线辐照对农杆菌介导法遗传转化的影响。结果表明 ,在辐照剂量为 1～ 4Gy时 ,GUS瞬间表达率随剂量的增加逐渐提高 ,但在辐射剂量高于 4Gy时 ,GUS瞬间表达率随剂量的增加而下降。综合考虑辐照后愈伤组织的存活率和GUS瞬间表达率 ,初步认为 2Gy为最佳辐照剂量。进一步观察表明 ,辐照处理后以继代培养 36h最适合农杆菌介导感染。

The effects of gamma irradiation on Agrobacterium-mediated genetic transformation of were investigated in the current paper, using embryonic calli derived from the mature seeds of Japanese lawngrass(Zoysia japonica Steud.). The result indicated that the GUS transient expression rates were enhanced with the increasing doses when treated by doses lower than 4Gy, however it would be decreased when treated by doses higher than 4Gy. Based on the survival rate and GUS transient expression rate, 2Gy is the optimal dose for Agrobacterium-mediated genetic transformation. Further observation found that 36 hours reculture after gamma irradiation is the most appropriate for Agrobacterium infection.