| 摘要 |
Process for the transformation of a plant genotype, wherein plant protoplasts are isolated from any plant tissue and cultured in one of the nutrient media customarily used for culturing plant protoplasts; before the actual transformation, the said protoplasts are pre-incubated at a temperature of from 4.degree. to 10.degree.C for from 20 minutes to 6 hours in a pre-incubation medium containing Ca2+, K+ and Na+ ions and also a suitable carbon source; the protoplasts are then separated from the pre-incubation medium and resuspended in the actual transformation medium which contains as an essential component from 0.1 to 60 mM Mg2+ ions in the presence or absence of Ca2+ ions; directly thereafter a DNA sample, containing one or more genes under the control of expression signals active in plants and also a supporting DNA, is added to the transformation solution; from 0.1 to 10 minutes later, a plasma membrane-modifying agent in a concentration of from 10 % to 30 % is added, and the protoplasts and DNA sample are incubated in the said transformation solution for a period that is sufficient for the incorporation of the DNA into the protoplasts. The transfer of the genetic material into the plant cell is carried out directly, without the use of a natural plant-infective system such as a plant bacterium or plant virus, and without transfer by insects or phytopathogenic fungi, by joint incubation in a suitable incubation medium of the DNA to be transformed and plant protoplasts. In this manner, desired genes can very simply and efficiently beg transferred to plant material, resulting in plants with improved properties.
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