Petolino J F, Hopkins N L, Kosegi B D, Skokut M
Plant Transformation and Gene Expression, Dow AgroSciences LLC, 9330 Zionsville Rd., Indianapolis, IN 46268, USA e-mail:
Plant Cell Rep. 2000 Jul;19(8):781-786. doi: 10.1007/s002999900180.
The present study was designed to establish embryogenic callus as a target tissue for whisker-mediated transformation of maize (Zea mays L.). Silicon carbide whiskers were used to deliver the bar and uidA (GUS) genes into embryogenic maize callus. Samples of osmotically-treated Type II callus were vigorously agitated in the presence of whiskers and plasmid DNA using a standard laboratory vortex or a modified dental amalgamator. On average, three transgenic callus lines were obtained per 100 samples treated. Plants were regenerated from several GUS-expressing callus lines and DNA analyses confirmed stable integration and inheritance. As with other direct DNA delivery methods involving embryogenic maize callus, integration patterns of the inserted DNA appeared to be complex. Although currently less efficient than microparticle bombardment on a per target basis, whisker-mediated transformation of embryogenic callus represents a viable method for transgenic maize production.
本研究旨在建立胚性愈伤组织作为玉米(Zea mays L.)晶须介导转化的靶组织。碳化硅晶须用于将bar和uidA(GUS)基因导入胚性玉米愈伤组织。使用标准实验室涡旋仪或改良的牙科汞齐搅拌机,在晶须和质粒DNA存在的情况下,对经渗透处理的II型愈伤组织样品进行剧烈搅拌。平均而言,每处理100个样品可获得三个转基因愈伤组织系。从几个表达GUS的愈伤组织系再生出植株,DNA分析证实了稳定整合和遗传。与其他涉及胚性玉米愈伤组织的直接DNA递送方法一样,插入DNA的整合模式似乎很复杂。虽然目前在每个靶标的基础上比微粒轰击效率低,但晶须介导的胚性愈伤组织转化是生产转基因玉米的一种可行方法。