Claims
- 1. A method for producing transgenic Impatiens plants, comprising the steps of:
- (a) introducing an expression vector into a plant tissue explant via Agrobacterium to produce a transformed explant, wherein said expression vector comprises a selectable marker gene and a second foreign gene, or
- (a') introducing two expression vectors into said plant tissue explant via Agrobacterium to produce a transformed explant, wherein one of said expression vectors comprises a selectable marker gene, and wherein the second of said expression vectors comprises a second foreign gene;
- (b) culturing said transformed explant on a selection medium;
- (c) culturing said transformed explant on regeneration medium; and
- (d) recovering fertile transgenic plants from said transgenic explants capable of transmitting said foreign gene to progeny.
- 2. The method of claim 1, wherein prior to introducing said explant, said explant is pre-cultured in (1) in MS medium comprising from approximately 0.5 mg/L to 2 mg/L TDZ followed by (2) MS medium comprising auxin and cytokinin.
- 3. The method of claim 2, wherein said auxin is approximately 0.05 to 0.2 mg/L NAA and said cytokinin is approximately 1 mg to 6 mg/L Zeatin.
- 4. The method of claim 2, wherein said first pre-culturing occurs for approximately 5 days and said second pre-culturing occurs for approximately 48 hours.
- 5. The method of claim 1, wherein said regeneration medium is MS medium with approximately 0.05 to 0.2 mg/L NAA and approximately 1 to 6 mg/L Zeatin.
- 6. The method of claim 1, wherein said marker is the nptII gene.
- 7. The method of claim 6, wherein said selection medium is MS medium comprising auxin, and cytokinin with 500 mg/L carbenicillin, 100 mg/L. kanamycin, and 100 mg/L cefotaxmine.
- 8. The method of claim 7, wherein said auxin is approximately 0.05 to 0.2 mg/L NAA and said cytokinin is approximately 1 to 6 mg/L Zeatin.
- 9. The method according to claim 7, wherein said regeneration medium is the same as said selection medium.
- 10. The method of claim 1, wherein said Impatiens plant tissue explant is selected from the group consisting of shoot tips, hypocotyl tips, and node regions.
- 11. The method of claim 10, wherein said Impatiens plant tissue explant is obtained from Impatiens shoot tip.
- 12. The method of claim 1, wherein said expression vector that comprises said second foreign gene further comprises a promoter, wherein said promoter is selected from the group consisting of Cauliflower Mosaic Virus (CaMV) 35S promoter, the enhanced 35S promoter, the UBQ3 promoter, the UBQ10 promoter, the UBQ11 promoter, the UBQ14 promoter, the TEFA 1 promoter, the rolC promoter, and the Commelina Yellow Mottle Virus promoter, wherein the expression of said second foreign gene is under the control of said promoter.
- 13. The method of claim 11, wherein said promoter is a CaMV 35S promoter.
- 14. The method of claim 1, wherein the expression of said second foreign gene confers resistance to disease caused by an organism selected from the group consisting of virus, bacterium, fungus, and insect.
- 15. The method of claim 1, wherein said second foreign gene confers resistance to a virus, and wherein said virus resistance gene disrupts the function of said virus, and wherein said virus-disrupting gene is selected from the group consisting of viral coat protein, 2'-5' oligonucleotide synthetase, viral genome antisense RNA, and pokeweed antiviral protein.
- 16. The method of claim 1, wherein said second foreign gene confers resistance to an insect, and wherein said insect resistance gene is selected from the group consisting of tryptophan decarboxylase, lectin, and Bacillus thuringiensis toxin.
- 17. The method of claim 16, wherein said lectin is Galanthus nivalis lectin.
- 18. The method of claim 1, wherein said second foreign gene confers resistance to a bacterium or a fungus, and wherein said second foreign gene encodes a polypeptide selected from the group consisting of chitinase, a .beta.-1,3-glucanase, ribosome-inactivating protein, lytic peptide, and plant defensin.
- 19. The method of claim 18, wherein said plant defensin is radish seed Rs-AFP2.
- 20. The method of claim 1, wherein said second foreign gene is operatively linked with a DNA molecule encoding pea vicilin signal peptide.
- 21. The method of claim 1, wherein said transgenic Impatiens comprises an expression vector that further comprises a third foreign gene.
- 22. The method of claim 1, wherein said second foreign gene encodes chitinase, and wherein said third foreign gene encodes .beta.-1,3-glucanase.
- 23. The method of claim 1, wherein the expression of said second foreign gene confers insensitivity to ethylene, and wherein said second foreign gene encodes a mutated ethylene receptor.
- 24. The method of claim 23, wherein said mutated ethylene receptor gene is the Arabidopsis etr-1 gene or a tomato NR gene.
- 25. The method of claim 1, wherein said second foreign gene is the Vitreoscilla hemoglobin gene.
- 26. The method of claim 1, wherein said second foreign gene is an isopentenyl transferase gene, wherein the expression of said isopentenyl transferase gene is under the control of a promoter of a senescence-associated gene.
- 27. The method of claim 26, wherein said promoter is the Arabidopsis SAG12 gene promoter.
- 28. The method of claim 1, wherein said second foreign gene encodes a polypeptide having a MADS box domain.
- 29. The method of claim 28, wherein said second foreign gene is selected from the group consisting of the PLENA gene, the SQUAMOSA gene, the DEFICIENS A gene, the GLOBOSA gene, the APTELA1 gene, the APETALA3 gene, the AGAMOUS gene, the OsMADS24 gene, the OsMADS45 gene, and the OsMADS1 gene.
- 30. The method of claim 1, wherein said second foreign gene encodes a protein that modifies plant habit.
- 31. The method of claim 1, wherein said second foreign gene confers resistance to INSV, wherein said second foreign gene encodes INSV S, M, and L RNAs.
- 32. The method of claim 1, wherein said second foreign gene confers resistance to TSWV, wherein said second foreign gene encodes TSWV viral movement protein.
- 33. The method of claim 1, wherein said second foreign gene confers fragrance, wherein said second foreign gene is selected from the group consisting of linalool synthase gene and limonene synthase gene.
- 34. The method of claim 1, wherein said second foreign gene confers resistance to drought, salinity or a cold, wherein said second foreign gene is selected from the group consisting of E. coli MnSOD gene, asparagine synthetase gene or promoter, Delta.sup.1 -Pyrroline-5-Carboxylate Synthetase gene, bacterial fructan gene, CAP85 and CAP160 genes, and trg-31.
- 35. A fertile transgenic Impatiens plant produced by the method of claim 1.
- 36. A fertile transgenic Impatiens plant having stably integrated in the plant genome a foreign gene, wherein said transgenic Impatiens plant is capable of transmitting said foreign gene to progeny.
- 37. Seeds and progeny of said transgenic Impatiens plant of claim 36 which are transformed with said foreign gene.
Parent Case Info
This Appln claims the benefit of U.S. Provisional 60/058,902 filed Sep. 12, 1997.
Foreign Referenced Citations (1)
Number |
Date |
Country |
0 354 687 |
Feb 1990 |
EPX |