Efficient transformation and regeneration of rice small cell groups.
Efficient transformation and regeneration of rice small cell groups.
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水稻小细胞群的高效转化与再生。
DOI：
10.1073/pnas.88.15.6389
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发表时间：
1991
影响因子：
11.1
通讯作者：
Douglas Testa
中科院分区：
文献类型：
--
作者：
Nancy Y. Lee;Yunzhu Wang;Jinshui Yang;K. Ge;Shu Huang;Jiazhen Tan;Douglas Testa
已经使用比以前使用的更简单、更快和更有效的方法（例如，原生质体转化和再生方法）。从已建立的水稻细胞系悬浮培养物制备50-100个细胞的小细胞群。用含有β-葡萄糖醛酸酶基因的质粒转化具有完整细胞壁的小细胞群，并由聚乙二醇介导。β-葡萄糖醛酸酶活性测定表明，转化频率约为7%。从转化的愈伤组织再生的植物的根和叶中检测到β-葡萄糖醛酸酶活性。确定每个细胞的一个或两个拷贝的β-葡萄糖醛酸酶基因整合到从转化的愈伤组织分离的水稻染色体DNA中。这种转化和再生方法广泛适用于双子叶和单子叶植物，特别是那些对原生质体再生有抗性的品种。
Transgenic plants have been regenerated from small cell groups of rice using a simpler, faster, and more efficient method than used previously (e.g., protoplast transformation and regeneration methods). Small cell groups of 50-100 cells were prepared from established suspension cultures of rice cell lines. Transformation of small cell groups with intact cell walls was carried out with a plasmid harboring the beta-glucuronidase gene and was mediated by polyethylene glycol. Assay of beta-glucuronidase activity indicated that the frequency of transformation was about 7%. beta-Glucuronidase activity was detected in the roots and leaves of plants regenerated from transformed calli. One or two copies of the beta-glucuronidase gene per cell were determined to be integrated into the rice chromosomal DNA isolated from transformed calli. This method of transformation and regeneration is widely applicable to both dicotyledonous and monocotyledonous plants, especially those varieties that are resistant to regeneration from protoplasts.