Claims
- 1. A process for culturing plant cell aggregates to form calli or plantlets which comprises:
- (a) uniformly plating isolated protoplasts in a solidified culture medium, the solidifying component of which is agarose in the absence of agaropectin,
- (b) incubating segments of the solidified culture medium, which segments contain the protoplasts, in a liquid nutrient solution until the protoplasts form plant cell aggregates, and
- (c) culturing the plant cell aggregates under conditions sufficient for the formation of calli or plantlets until the plant cell aggregates have reached a callus or plantlet stage.
- 2. A process according to claim 1, which comprises transferring the segments into 1 to 10,000 times their volume of a liquid nutrient solution suitable for culturing plant cells, and continuation of culturing in the temperature range from 0.degree. to 40.degree. C.
- 3. A process according to claim 1, wherein the incubation step of the process occurs at an incubation temperature in the range of from about 12.degree. to about 30.degree. C.
- 4. A process according to claim 2, wherein the segments are put into 5 to 10,000 times their own volume of a nutrient solution suitable for culturing plant cells.
- 5. A process according to claim 4, wherein the segments are put into 20 to 100 times their own volume of a nutrient solution suitable for culturing plant cells.
- 6. A process according to claim 1, wherein the solidified culture medium is incubated in the liquid nutrient solution not later than 7 days after plating with protoplasts.
- 7. A process according to claim 6, wherein segmentation is effected 3 to 4 days after plating.
- 8. A process according to claim 1, which comprises the use of segments of equal size and regular shape and having an average diameter of 1 to about 100 mm.
- 9. A process according to claim 8, which comprises the use of segments having an average diameter of 2 to 60 mm.
- 10. A process according to claim 9, wherein the segments are discs, spheres, cubes, prisms or cones having an average diameter of 2 to 10 mm.
- 11. A process according to claim 1, which comprises the use of protoplasts which are isolated from a plant of a species selected from the group consisting of Nicotiana tabacum, Hyoscyamus muticus, Lycopersicon esculentum, Crepis capillaris, Brassica rapa, and Petunia hybrida.
- 12. A process according to claim 1, wherein the solidifying agarose is selected from the group consisting of SEA PLAQUE LMT.RTM., LMP.RTM., TYPE VIII.RTM., HGT.RTM., HGTP.RTM., LE.RTM., STANDARD LMT.RTM. and SEA PREP.RTM. agarose.
- 13. A process for culturing plant cell aggregates from plant cells formed from protoplasts to form calli or plantlets, which comprises:
- (a) culturing protoplasts under conditions in which the protoplasts regenerate cell walls to form plant cells;
- (b) plating the plant cells formed from protoplasts in a solidified culture medium, the solidifying component of which is agarose in the absence of agaropectin;
- (c) incubating segments of the solidified culture medium, which segments contain the plant cells, in a liquid nutrient solution until the plant cells form plant cell aggregates; and
- (d) culturing the plant cell aggregates under conditions sufficient for the formation of calli or plantlets until the plant cell aggregates have reached a callus or plantlet stage.
- 14. A process according to claim 13, wherein the solidified culture medium is incubated in the liquid nutrient solution not later than 7 days after the plant cells are cultured to form protoplasts.
Priority Claims (1)
Number |
Date |
Country |
Kind |
2317/83 |
Apr 1983 |
CHX |
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Parent Case Info
This application is a continuation of application Ser. No. 053,582, filed 5/18/87 which is a continuation of application Ser. No. 801,375, filed 11/22/85, which is a continuation of application Ser. No. 601,876, filed 4/19/84, all now abandoned.
US Referenced Citations (4)
Non-Patent Literature Citations (4)
Entry |
Maramorosch et al; Practical Tissue Culture Applications, Academic Press, N.Y., 1979, pp. 31-34. |
Reinert et al., Plant Cell and Tissue Culture; Springer-Verlag, N.Y., 1982, pp. 40-45. |
Singleton et al., Dictionary of Microbiology; John Wiley & Sons, New York; .COPYRGT. 1978; p. 7. |
Staba, E. John; Plant Tissue Culture as a Source of Biochemicals; CRC Press Inc., Boca Raton, Fla., .COPYRGT. 1980; pp. 100-102. |
Continuations (3)
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Number |
Date |
Country |
Parent |
53582 |
May 1987 |
|
Parent |
801375 |
Nov 1985 |
|
Parent |
601876 |
Apr 1984 |
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