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Transformation of Maize Cells and Regeneration of Fertile Transgenic Plants.

机译:玉米细胞的转化和可育转基因植物的再生。

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摘要

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.
机译:已经开发了用于产生可育的转基因玉米植物的可再现系统。使用微粒轰击,用细菌基因棒转化来自胚发生玉米悬浮培养物的细胞。使用除草剂双丙氨磷从悬浮培养物中选择转化的愈伤组织。 bar的整合和由bar编码的草丁膦丝氨酸乙酰转移酶(PAT)的酶活性在所有对双丙氨磷耐药的愈伤组织中得到证实。再生了能育的转化玉米植物(R0),测试的53个子代(R1)中29个具有PAT活性。分析的所有PAT阳性子代均包含bar。将除草剂局部施用到经bar转化的R0和R1植物的叶子上不会导致坏死,从而证实了PAT在转基因植物中的功能活性。使用两种质粒的混合物进行共转化实验,一种质粒编码PAT,一种含有未选择的编码β-葡糖醛酸糖苷酸酶基因。从共转化的愈伤组织再生的R0植物表达了两个基因。这些结果描述并证实了将DNA引入玉米的系统的开发。

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