Gordon-Kamm W. J., Spencer T. M., Mangano M. L., Adams T. R., Daines R. J., Start W. G., O'Brien J. V., Chambers S. A., Adams W. R., Willetts N. G., Rice T. B., Mackey C. J., Krueger R. W., Kausch A. P., Lemaux P. G.
Discovery Research, DEKALB Plant Genetics, Eastern Point Road, Groton, Connecticut 06340.
Plant Cell. 1990 Jul;2(7):603-618. doi: 10.1105/tpc.2.7.603.
A reproducible system for the generation of fertile, transgenic maize plants has been developed. Cells from embryogenic maize suspension cultures were transformed with the bacterial gene bar using microprojectile bombardment. Transformed calli were selected from the suspension cultures using the herbicide bialaphos. Integration of bar and activity of the enzyme phosphinothricin acetyltransferase (PAT) encoded by bar were confirmed in all bialaphos-resistant callus lines. Fertile transformed maize plants (R0) were regenerated, and of 53 progeny (R1) tested, 29 had PAT activity. All PAT-positive progeny analyzed contained bar. Localized application of herbicide to leaves of bar-transformed R0 and R1 plants resulted in no necrosis, confirming functional activity of PAT in the transgenic plants. Cotransformation experiments were performed using a mixture of two plasmids, one encoding PAT and one containing the nonselected gene encoding [beta]-glucuronidase. R0 plants regenerated from co-transformed callus expressed both genes. These results describe and confirm the development of a system for introduction of DNA into maize.
已开发出一种用于培育可育转基因玉米植株的可再生系统。利用微粒轰击法，用细菌基因bar转化来自玉米胚性悬浮培养物的细胞。使用除草剂双丙氨膦从悬浮培养物中筛选转化愈伤组织。在所有抗双丙氨膦的愈伤组织系中均证实了bar的整合以及由bar编码的膦丝菌素乙酰转移酶（PAT）的活性。再生出可育的转基因玉米植株（R0），在检测的53个后代（R1）中，有29个具有PAT活性。分析的所有PAT阳性后代均含有bar。对bar转化的R0和R1植株的叶片进行除草剂局部施用，未出现坏死现象，证实了PAT在转基因植株中的功能活性。使用两种质粒的混合物进行共转化实验，一种编码PAT，另一种含有编码β-葡萄糖醛酸酶的非选择基因。从共转化愈伤组织再生的R0植株表达了这两个基因。这些结果描述并证实了一种将DNA导入玉米的系统的开发。
