Zhang L, Rybczynski J J, Langenberg W G, Mitra A, French R
Department of Plant Pathology and Center for Biotechnology, University of Nebraska, Lincoln, NE 68583-0722, USA, , , , , , US.
The Botanical Garden-Center for Biological Diversity and Conservation of the Polish Academy of Sciences, PL-02-773 Warsaw, Poland, , , , , , PL.
Plant Cell Rep. 2000 Jan;19(3):241-250. doi: 10.1007/s002990050006.
A method for producing large numbers of transgenic wheat plants has been developed. With this approach, an average of 9.7% of immature embryo explants were transformed and generated multiple self-fertile, independently transformed plants. No untransformed plants, or escapes, were regenerated. This transformation procedure uses morphogenic calli derived from scutellum tissue of immature embryos of Triticum aestivum cv. Bobwhite co-bombarded with separate plasmids carrying a selectable marker gene (bar) and a gene of interest, respectively. Transformed wheat calli with a vigorous growth phenotype were obtained by extended culture on media containing 5.0 mg/l bialaphos. These calli retained morphogenic potential and were competent for plant regeneration for as long as 11 months. The bar gene and the gene of interest were co-expressed in T0 progeny plants. This wheat transformation protocol may facilitate quantitative production of multiple transgenic plants and significantly reduce the cost and labor otherwise required for screening out untransformed escapes.
一种生产大量转基因小麦植株的方法已经开发出来。通过这种方法，平均9.7%的未成熟胚外植体被转化，并产生了多个可自交结实、独立转化的植株。没有再生出未转化的植株或逃逸植株。这种转化程序使用源自普通小麦品种“博比特”未成熟胚盾片组织的形态发生愈伤组织，分别与携带选择标记基因（bar）和目的基因的单独质粒共轰击。通过在含有5.0 mg/l双丙氨膦的培养基上延长培养，获得了具有旺盛生长表型的转化小麦愈伤组织。这些愈伤组织保持了形态发生潜力，并且在长达11个月的时间内都有能力再生植株。bar基因和目的基因在T0代植株中共同表达。这种小麦转化方案可能有助于定量生产多个转基因植株，并显著降低筛选未转化逃逸植株所需的成本和劳动力。
