Microprojectile-mediated transient and integrative transformation of rice embryogenic suspension cells: effects of osmotic cell conditioning and of the physical configuration of plasmid DNA

被引：25

作者：

Nandadeva, YL ^{[1
]}

Lupi, CG ^{[1
]}

Meyer, CS ^{[1
]}

Devi, PS ^{[1
]}

Potrykus, I ^{[1
]}

Bilang, R ^{[1
]}

机构：

[1] ETH Zurich, Inst Plant Sci, CH-8092 Zurich, Switzerland

来源：

PLANT CELL REPORTS | 1999年 / 18卷 / 06期

关键词：

Oryza sativa; embryogenic cell suspension; particle bombardment; single-stranded DNA; plasmolysis;

D O I：

10.1007/s002990050611

中图分类号：

Q94 [植物学];

学科分类号：

071001 ;

摘要：

Microprojectile-mediated transient and integrative transformation Frequencies in rice (Oryza sativa cv. Taipei 309) embryogenic suspension cells were studied as a function of various parameters. Mannitol at concentrations of 0.5 and 0.6 M was best for osmotic preconditioning of the cells for transient, but not For integrative transformation, for which sucrose yielded the best and most reliable results. Denaturation of the transforming plasmid DNA Frier to bombardment improved transient and: integrative transformation frequencies two to three fold. Delivery of double-stranded plasmids in linear form had no effect on transient transformation when compared to supercoiled plasmid DNA, but led to an overall two fold increase in integrative transformation frequency. This shows that optimized protocols for generating transgenic plants should not be based, exclusively on transient gene expression assays.

引用

页码：500 / 504

页数：5

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