Laser‐mediated gene transfer in rice
作者:
Yande Guo,
Hong Liang,
Michael W. Berns,
期刊:
Physiologia Plantarum
(WILEY Available online 1995)
卷期:
Volume 93,
issue 1
页码: 19-24
ISSN:0031-9317
年代: 1995
DOI:10.1034/j.1399-3054.1995.930104.x
出版商: Munksgaard International Publishers
关键词: Cell osmotic pressure;laser microbeam;Oryza sativa;rice;transformation
数据来源: WILEY
摘要:
We report an effective system for introducing exogenous DNA into cells of embryonic calli ofOryza sativaL. cv. Japonica. Plant cells were pretreated in hypertonic buffer to draw some of the water from the cells and were then put into medium of less negative osmotic potential containing exogenous DNA and treated immediately with a laser microbeam (λ=355 nm) to puncture holes in the cell wall and membrane. The pretreatment of the cells generated a gradient of osmotic pressure between the inside and outside of the cells, which facilitated the uptake of material into cells through the laser perforations. Bright yellow‐green fluorescence could be detected inside cells that had been bathed in a solution containing the fluorescent molecule calcein. β‐Glucuronidase (GUS) genes were successfully introduced into rice cells as indicated by gene expression both in post‐treated cells and in plantlets derived from kanamycin‐resistant calli that had been treated by th
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