Transformation of Indica rice (Oryza sativa L.) mediated by Agrobacterium tumefaciens
Transformation of Indica rice (Oryza sativa L.) mediated by Agrobacterium tumefaciens
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根癌农杆菌介导的籼稻（Oryza sativa L.）转化
DOI：
10.1093/oxfordjournals.pcp.a078292
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发表时间：
1992
影响因子：
4.9
通讯作者：
Hsin
中科院分区：
文献类型：
--
作者：
M. Chan;Tse‐Min Lee;Hsin
建立了农杆菌介导的籼稻基因转移体系。CV. Taichung Native 1），并确定了农杆菌介导转化的最适条件。从3至4天龄的幼苗切下的茎、叶和根用携带有β-葡萄糖醛酸酶（GUS）和新霉素磷酸转移酶（NPTII）基因嵌合衍生物的质粒的农杆菌菌株感染。用抗生素G418筛选后，只有根存活并形成愈伤组织。Southern杂交分析和酶活性测定证实了嵌合基因在水稻基因组中的整合。获得了37.5%的转基因愈伤组织转化率。此外，与virprobes杂交的缺失表明转化子未被农杆菌污染。马铃薯悬浮培养物（PSC）在无培养基存在下的转化完全失败表明PSC对于Ti介导的籼稻转化是必需的。在用共整合载体pGV 2260：：NG 1感染后观察到高转化效率。
A system has been developed for the transfer of genes byAgrobacterium tumefaciensto indica rice (Oryza sativaL. cv. Taichung Native 1), and optimal conditions for suchAgrobacterium-mediated transformation have been determined. Excised stems, leaves, and roots from 3- to 4-day-old seedlings were infected with strains ofAgrobacteriumthat carried plasmids with chimeric derivatives of genes for β-glucuronidase (GUS) and neomycin phosphotransferase (NPTII). After selection with the antibiotic G418, only roots survived and formed calli. The integration of chimeric genes in the rice genome was demonstrated by Southern analysis and assays of enzymatic activity. We obtained a transformation frequency of 37.5% for transgenic callus. In addition, the absence of hybridization withvirprobes indicated that transformants were not contaminated withAgrobacterium. The complete failure of transformation in the absence of medium from potato suspension cultures (PSC) indicates that PSC is essential for Ti-mediated transformation in indica rice. A high efficiency of transformation was observed upon infection with the cointegrate vector pGV2260::NG1.