小麦基因枪高效转化体系的建立

Establishment of an Efficient Bombardment Transformation System in Wheat

摘要:

以小麦幼胚为基因枪转化受体,用含GUS和bar基因的质粒pDM80 3对小麦幼胚进行转化,并对幼胚大小、轰击距离、质粒和金粉用量等参数对GUS瞬时表达率及表达量的影响进行了研究。幼胚以1 0~1 5mm最为适宜,6cm和9cm各轰击一次、质粒0.5μg/枪及金粉300μg/枪,GUS瞬时表达率及表达量最高。通过对以上参数的优化,建立了一个高效的小麦基因枪转化体系。

Abstract:

An efficient transformation system for wheat was established by improving transformation conditions for the particle bombardment of immature embryos.Particle bombardment was carried out using pDM803 containing the reporter gene β-glucuronidase(GUS) and bar gene.Transformation parameters,such as the size of immature embryos,the distance of bombardment,the amount of gold particles and plasmid used per bombardment were analyzed.The immature embryo of 1.0~1.5 mm was optimal.The transient expression of GUS reached the highest when the immature embryos were bombarded twice at the distance of 6 cm and 9 cm,using 300 μg gold particles and 0.5 μg plasmid per bombardment.