Melon (Cucumis melo) - Patents granted to Biosem

Process for the production of transgenic plantlets having diploid phenotype from genetically transformed explants, said plantlets belonging to the species Cucumis melo and containing at least one gene introduced by the intermediary of Agrobacterium tumefaciens , comprising the following steps: A) inducing genetically transformed shoot buds from cotyledons of Cucumis melo in a shoot bud induction medium without forming calli, wherein the cotyledons are obtained from embryos which have germinated from 0 to 4 days before being contacted with A. tumefaciens, wherein the induction medium comprises about 440 to about 2,200 mg/L of calcium chloride calculated as CaCl₂R₂H₂ O, and about 0.3 to about 1.13 mg/L 6-benzyl aminopurine (BAP); and
B) forming genetically transformed plantlets from genetically transformed shoot buds, wherein the step of forming comprises: (i) culturing the genetically transformed shoot buds in a medium having 6-benzyl aminopurine (BAP) until the shoot buds have reached a height of at least 3 mm; and
(ii) transferring and incubating the shoot buds in a suitable macro-element plant cell culture medium sufficiently to form the genetically transformed plantlets.

Process for production of transgenic plantlets with a diploid phenotype, from genetically transformed explants, said plantlets belonging to the species Cucumis melo and containing at least one gene introduced by the intermediary of A. tumefaciens comprising the following steps: A) inducing genetically transformed shoot buds from cotyledons of Cucumis melo in a shoot bud induction medium without forming calli, wherein the cotyledons are obtained from embryos which have germinated from 0 to 4 days before being contacted with A. tumefaciens, and wherein the induction medium comprises about 440 to about 2,200 mg/L of calcium chloride calculated as CaCl₂R₂H₂ O, about 0.3 to about 1.13 mg/L of 6-benzyl aminopurine (BAP), and about 0 to about 1.3 mg/L indole-3-acetic acid (IAA); and
B) forming genetically transformed plantlets from the genetically transformed shoot buds, wherein the step of forming comprises: (i) culturing the shoot buds in a medium having 6-benzyl aminopurine (BAP) until the shoot buds have reached a height of at least 3 mm; and
(ii) transferring and incubating the shoot buds in a suitable macro-elements plant cell culture medium comprising:

• KH₂PO₄ from about 50 to about 100 mg/L;
• MgSO₄from about 75 to about 300 mg/L;
• CaCl₂R₂H₂O from about 500 to about 2500 mg/L;
• KNO₃ from about 750 to about 1200 mg/L; and
• NH₄NO₃ from about 150 to about 200 mg/L sufficiently to form the genetically transformed plantlets.

Transgenic plants having a diploid phenotype belonging to the species Cucumis melo comprising at least one DNA sequence introduced by the intermediary of Agrobacterium tumefaciens.

Transgenic plant tissue having a diploid phenotype belonging to the species Cucumis melo comprising at least one DNA sequence that confers resistance to cucumber mosaic virus.

Transgenic plants having a diploid phenotype and belonging to the species Cucumis melo comprising at least one DNA sequence that confers resistance to Cucumber mosaic virus.

Transgenic plants having diploid phenotype, belonging to the species Cucumis melo, and containing at least one gene introduced by the intermediary of Agrobacterium tumefaciens, wherein the plant tissue is produced from genetically transformed explants by a process comprising the following step: A) inducing genetically transformed shoot buds from cotyledons of Cucumis melo in a shoot bud induction medium without forming calli, wherein the cotyledons are obtained from embryos which have germinated from 0 to 4 days before being contacted with A. tumefaciens, wherein the induction medium comprises about 440 to about 2,200 mg/L of calcium chloride calculated as CaCl₂R₂H₂ O, and about 0.3 to about 1.3 mg/L 6-benzyl aminopurine (BAP); and
B) forming genetically transformed plantlets from genetically transformed shoot buds, wherein the step of forming comprises: (i) culturing the genetically transformed shoot buds in a medium having 6-benzyl aminopurine (BAP) until the shoot buds have reached a height of at least 3 mm; and
(ii) transferring and incubating the shoot buds in a suitable macro-element plant cell culture medium sufficiently to form the genetically transformed plantlets.

Process for production of transgenic, phenotypically normal plantlets from genetically transformed explants, said plantlets belonging to the species Cucumis melo and containing at least one gene, which has been introduced through Agrobacterium tumefaciens, characterized by the following steps: A) induction of genetically transformed shoot buds from cotyledons of Cucumis melo which have germinated for 0 to 4 days and, after this period, have been brought into contact with A. tumefaciens, the induction being carried out on an induction medium for genetically transformed shoot buds which comprises all of the minerals, salts and vitamins normally required for the induction of shoot buds from genetically non-transformed explants and containing, amongst the mineral salts, approximately 440 to approximately 2,200 mg/L of calcium chloride calculated as CaCl₂R₂H₂O, and approximately 0.8 to approximately 1.2% of bacto-agar or agar-agar, said induction medium being supplemented with approximately 0.3 to about 1.13 mg/L of 6-benzyl aminopurine (BAP); and approximately 0 to approximately 1.3 mg/L indole-3-acetic acid (IAA);
B) culturing the resulting genetically transformed shoot buds in two successive stages, the first of these culture stages taking place on a plant cell culture medium containing a cytokinin and, more particularly, 6-benzyl aminopurine (BAP), and the second stage, which is carried out when the shoot buds have reached a length of at least 3 mm, taking place on a plant cell culture medium containing, as macroelements:

• KH₂ PO₄ approximately 50 to approximately 100 mg/L
• MgSO₄ approximately 75 to approximately 300 mg/L
• CaCl₂R₂H₂O approximately 500 to approximately 2500 mg/L
• KNO₃ approximately 750 to approximately 1200 mg/L
• NH₄NO₃ approximately 150 to approximately 200 mg/L.

Cell culture medium suitable for the development of shoot buds into plantlets in the course of the regeneration of a plant, characterized in that it contains, as macro-elements:

• KH₂ PO₄ approximately 50 to approximately 100 mg/L
• MgSO₄R2H₂O approximately 75 to approximately 300 mg/L
• CaCl₂R₂H₂O approximately 1000 to approximately 2500 mg/L
• KNO₃ approximately 750 to approximately 1200 mg/L
• NH₄NO₃ approximately 150 to approximately 200 mg/L.

Shoot bud induction medium composed of a plant cell culture medium, which comprises all the minerals, salts and vitamins normally required for inducing shoot buds from non-genetically transformed explants and containing, amongst its mineral salts, calcium chloride, and bacto-agar or agar-agar, characterized in that the CaCl₂ content of this medium is 1000 to 2200 mg/L calculated as CaCl₂ R₂H₂O, and the bacto-agar or agar-agar content is 0.8 to 1.2%, said medium being supplemented with 0.3 to about 2.0 mg/L of BAP and 0 to 1.3 mg/L of IAA.