Abdollahi M R, Moieni A, Mousavi A, Salmanian A H, Jalali Javaran M, Majdi M
Department of Plant Breeding, Faculty of Agriculture, Tarbiat Modarres University, P.O. Box 14115-336, Tehran, Iran.
Pak J Biol Sci. 2007 Sep 15;10(18):3141-5. doi: 10.3923/pjbs.2007.3141.3145.
A new method for transformation of rapeseed microspore-derived embryos (MDEs), based on microwounding of MDEs by particle bombardment prior to inoculation with an Agrobacterium suspension was reported. In this study, effects of two transformation systems (integrated bombardment and Agrobacterium transformation system and the singular bombardment) on transient GUS expression in hypocotyls of rapeseed (Brassica napus L. cv. PF704) MDEs were studied. Bombardment parameters were: helium pressure, 1350 psi; distance between stopping screen and target tissue, 9 cm, gold particles size of 1.0 microm and chamber vacuum pressure, 24 in Hg. In integrated system A. tumefaciens strain AGL1 carrying the binary vector pCAMBIA3301 was used. Bombarded hypocotyls of MDEs were inoculated with Agrobacteria at OD600 = 1.0 for 10 min or OD600 = 0.25 for 24 h. Integrated transformation system increased the mean number of blue spots about 2-2.5 fold compared to singular bombardment and inoculated hypocotyls with OD600 = 1.0 for 10 min produced the highest number of blue spots per bombardment (743 +/- 23.67).
报道了一种基于在用农杆菌悬浮液接种之前通过粒子轰击对油菜小孢子衍生胚（MDEs）进行微创伤的油菜籽MDEs转化新方法。在本研究中，研究了两种转化系统（联合轰击和农杆菌转化系统以及单一轰击）对油菜（甘蓝型油菜L. cv. PF704）MDEs下胚轴中瞬时GUS表达的影响。轰击参数为：氦气压力1350磅力/平方英寸；阻挡屏与靶组织之间的距离9厘米；金颗粒大小1.0微米；腔室真空压力24英寸汞柱。在联合系统中，使用携带二元载体pCAMBIA3301的根癌农杆菌菌株AGL1。用OD600 = 1.0的农杆菌接种轰击后的MDEs下胚轴10分钟，或用OD600 = 0.25的农杆菌接种24小时。与单一轰击相比，联合转化系统使蓝色斑点的平均数增加了约2至2.5倍，并且用OD600 = 1.0的农杆菌接种轰击后的下胚轴10分钟时，每次轰击产生的蓝色斑点数量最多（743±23.67）。
