Abstract
Synchronization of somatic embryogenesis at high frequency is a useful system for the mass production of embryos. Many attempts have been carried out, however, it was difficult to obtain the system in which most of the initial embryogenic cells or cell clusters synchronously differentiate to embryos. In carrot suspension cultures, high frequency, synchronous embryogenesis systems (following three systems) have been established.
(1) Small spherical single cells from suspension cultures obtained by sieving and density gradient centrifugation in Percoll solutions differentiated to embryogenic cell clusters at high frequency when they were cultured in a medium containing 2,4-dichlorophenoxyacetic acid (0.05 micromolar), zeatin (1 micromolar) and mannitol (0.2 molar). (2) Embryogenic cell clusters from suspension cultures obtained by sieving, density gradient centrifugation in Ficoll solutions, and subsequent centrifugation at a low speed for a short time synchronously differentiated to embryos, especially globular embryos at high frequency, when they were cultured in a medium containing zeatin (0.1 micromolar) but no auxin. (3) Embryogenic cell clusters obtained by above method are cultured at cell densities of 2×103 cell clusters ml-1. Globular embryos which were sieved from embryos induced synchronously differentiated to torpedo-shaped embryos at high frequency when they were cultured at densities below 150 globular embryos ml-1.
Using these systems, the whole process of embryogenesis from single cells to whole plants could be synchronously induced at high frequency.
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Abbreviations
- ABA:
-
abscissic acid
- 2,4-d :
-
2,4-dichlorophenoxyacetic acid
- GA3 :
-
gibberellin A3
- IAA:
-
indoleacetic acid
- NAA:
-
naphthylacetic acid
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Osuga, K., Komamine, A. Synchronization of somatic embryogenesis from carrot cells at high frequency as a basis for the mass production of embryos. Plant Cell Tiss Organ Cult 39, 125–135 (1994). https://doi.org/10.1007/BF00033920
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DOI: https://doi.org/10.1007/BF00033920