Method for preparing transformed cucumis melo

20060168699

10/501087

August 01, 2002

The present invention relates to a method of preparing a transformed Cucumis melo using Acrobacterium tumefaciens, more particularly, to a method for preparing a transformed Cucumis melo, which comprises the steps of: (a) inoculating a cotyledon from Cucumis melo with Agrobacterium tumefaciens harboring a suitable vector; (b) regenerating the inoculated cotyledon in a regeneration medium containing 3.0-8.0 mg/l of kinetin as growth regulator and 0.5-3.0 mg/l of IAA (Indole-3-acetic acid) and culturing the inoculated cotyledon to obtain regenerated shoots; and (c) culturing the regenerated shoots on a rooting medium to obtain the transformed Cucumis melo.

Lee, Sun (Daejon City, KR); Yoo, Je-Geun (Daejon City, KR)

1. method for preparing a transformed Cucumis melo, which comprises the steps of: (a) inoculating a cotyledon from Cucumis melo with Agrobacterium tumefaciens harboring a vector, in which the vector is capable of inserting into a genome of a cell from Cucumis melo and contains the following sequences: (i) a replication origin operable in the cell from Cucumis melo; (ii) a promoter capable of promoting a transcription in the cell from Cucumis melo; (iii) a structural gene operably linked to the promoter; and (iv) a polyadenylation signal sequence, (b) regenerating the inoculated cotyledon in a regeneration medium containing 3.0-8.0 mg/1 of kinetin as growth regulator and 0.5-3.0 mg/1 of IAA (Indole-3-acetic acid) and culturing the inoculated cotyledon to obtain regenerated shoots; and (c) culturing the regenerated shoots on a rooting medium to obtain the transformed Cucumis melo.

2. The method according to claim 1, wherein an amount of kinetin in the regeneration medium of step (b) is 5.0-7.0 mg/l.

3. The method according to claim 1, wherein an amount of IAA in the regeneration medium of step (b) is 1.0-2.0 mg/l.

4. The method according to claim 1, wherein an amount of NAA in the rooting medium of step (c) is 0.08-0.2 mg/l.

5. The method according to claim 1, wherein the step (a) is executed in an inoculating medium containing 3.0-8.0 mg/l of kinetin, 0.5-3.0 mg/1 of IAA and 50-200 μM of acetosyringone.

6. The method according to claim 1, wherein the regeneration medium further comprises 0.5-2.0 mg/l of CuSO4.

7. A method for preparing a transformed Cucumis melo, which comprises the steps of: (a) inoculating an cotyledon from Cucumis melo with Agrobacterium tumefaciens harboring a vector in an inoculating medium containing 3.0-8.0 mg/l of kinetin, 0.5-3.0 mg/l of IAA and 50-200 μM of acetosyringone, in which the vector is capable of inserting into a genome of a cell from Cucumis melo and contains the following sequences: (i) a replication origin operable in the cell from Cucumis melo; (ii) a promoter capable of promoting a transcription in the cell from Cucumis melo; (iii) a structural gene operably linked to the promoter; and (iv) a polyadenylation signal sequence, (b) regenerating the inoculated cotyledon in a regeneration medium containing 5.0-7.0 mg/l of kinetin and 1.0-2.0 mg/l of IAA as growth regulators and CuSO4 as a regenerating accelerating agent to obtain regenerated shoots; and (c) culturing the regenerated shoots to obtain the transformed Cucumis melo on a rooting medium containing 0.08-0.2 mg/l of NAA.

8. A transformed Cucumis melo prepared by the method according to claim 1.

800294/000

12; 01

edspap.20060168699