Method for transforming monocotyledons
AssigneeJapan Tobacco Inc.
InventorsYokoh Hiei, Toshihiko Komari
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Abstract
A method for transforming a monocotyledon by which the time required from transformation to regeneration of a plant is shorter so that the frequency of emergence of mutants is smaller than the conventional methods, which may be generally applied even to the plants for which the regeneration method from a protoplast to a plant has not been established, and with which the preparation of the material to be subjected to the method is easy. That is, the present invention provides a method for transforming a monocotyledon comprising transforming a cultured tissue during dedifferentiation process or a dedifferentiated cultured tissue of said monocotyledon with a bacterium belonging to genus Agrobacterium containing a desired gene.
Claims25
A method for transforming a monocotyledon callus, comprising contacting a cultured tissue of a monocotyledon during dedifferentiation wherein said dedifferentiation is obtained by culturing an explant on a dedifferentiation-inducing medium for not less than 7 days or a dedifferentiated cultured tissue of a monocotyledon, with a bacterium belonging to the genus Agrobacterium containing a desired gene.
A method for transforming a monocotyledon with a desired gene, comprising:
- contacting a cultured tissue of said monocotyledon during dedifferentiation thereof, or a dedifferentiated cultured tissue of said monocotyledon, with a suspension of Agrobacterium tumefaciens having a cell population of 10.sup.6 to 10.sup.11 cells/ml for 3-10 minutes and then culturing said cultured tissue of said monocotyledon during dedifferentiation thereof, or said dedifferentiated cultured tissue of said monocotyledon, on a solid medium for several days together with said Agrobacterium tumefaciens, or
- adding said Agrobacterium tumefaciens to culture medium in which said cultured tissue of said monocotyledon during dedifferentiation thereof or said dedifferentiated cultured tissue of said monocotyledon is cultured, and continuously culturing said cultured tissue of said monocotyledon during dedifferentiation or said dedifferentiated cultured tissue of said monocotyledon together with said Agrobacterium tumefaciens,
- wherein said dedifferentiated cultured tissue of said monocotyledon is selected from the group consisting of a tissue cultured during the process of callus formation which is cultured for not less than 7 days after an explant is placed on a dedifferentiation-inducing medium and a callus, and
- wherein said Agrobacterium tumefaciens contains plasmid pTOK162, and said desired gene is present between border sequences of the T region of said plasmid pTOK162, or wherein said desired gene is present in another plasmid contained in said Agrobacterium tumefaciens.
Dependencies are inferred from the claim text.