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农杆菌介导的水稻(Oryza sativa L.)高效转化及T-DNA边界序列分析

Efficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA.

作者信息

Hiei Y, Ohta S, Komari T, Kumashiro T

机构信息

Plant Breeding and Genetics Research Laboratory, Japan Tobacco Inc., Shizuoka.

出版信息

Plant J. 1994 Aug;6(2):271-82. doi: 10.1046/j.1365-313x.1994.6020271.x.

Abstract

A large number of morphologically normal, fertile, transgenic rice plants were obtained by co-cultivation of rice tissues with Agrobacterium tumefaciens. The efficiency of transformation was similar to that obtained by the methods used routinely for transformation of dicotyledons with the bacterium. Stable integration, expression and inheritance of transgenes were demonstrated by molecular and genetic analysis of transformants in the R0, R1 and R2 generations. Sequence analysis revealed that the boundaries of the T-DNA in transgenic rice plants were essentially identical to those in transgenic dicotyledons. Calli induced from scutella were very good starting materials. A strain of A. tumefaciens that carried a so-called 'super-binary' vector gave especially high frequencies of transformation of various cultivars of japonica rice that included Koshihikari, which normally shows poor responses in tissue culture.

摘要

通过水稻组织与根癌农杆菌共培养,获得了大量形态正常、可育的转基因水稻植株。转化效率与常规用于双子叶植物细菌转化的方法所获得的效率相似。通过对R0、R1和R2代转化体的分子和遗传分析,证明了转基因的稳定整合、表达和遗传。序列分析表明,转基因水稻植株中T-DNA的边界与转基因双子叶植物中的基本相同。由盾片诱导的愈伤组织是非常好的起始材料。携带所谓“超级双元”载体的根癌农杆菌菌株对包括越光在内的各种粳稻品种具有特别高的转化频率,而越光在组织培养中通常反应较差。

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