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Recovery of somatic embryos and plantlets from protoplast cultures of Larix × eurolepis

Summary

Somatic embryos and plantlets were regenerated from protoplasts of hybrid larch (Larix × eurolepis) isolated from two embryogenic callus and cell suspension culture lines (L1 and L2). L2, which was highly embryogenic, consistently yielded protoplasts that gave rise to somatic embryos. Centrifugation on a discontinuous medium/Percoll density gradient resulted in accumulation of embryogenic protoplasts in one of the Percoll interfaces. First division frequencies were in the range of 28–39% in line 1 and 18–20% in line 2 in both liquid and agarose-solidified culture media. The critical factor in maintaining high viability of cultures was lowering of osmotic pressure by dilution of the initial medium. The first somatic embryos were detected in 23- to 28-day-old cultures. Some of these developed into plants that were transferred to soil.

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References

  1. von Aderkas P, Bonga JM (1988) Formation of haploid embryoids of Larix decidua: early embryogenesis. Amer. J. Bot. 75:690–700

  2. von Aderkas P, Klimaszewska K, Bonga JM J. Diploid and haploid embryogenesis in Larix leptolepis, L. decidua, and their reciprocal hybrids. Can. J. For. Res. (in press).

  3. Attree SM, Bekkaoui F, Dunstan DI, Fowke LC (1987) Regeneration of somatic embryos from protoplasts isolated from an embryogenic suspension culture of white spruce (Picea glauca). Plant Cell Reports 6:480–483

  4. Bekkaoui F, Saxena PK, Attree SM, Fowke LC, Dunstan DI (1987) The isolation of culture of protoplasts from an embyrogenic cell suspension culture of Picea glauca (Moench) Voss. Plant Cell Reports 6:476–479

  5. Bekkaoui F, Pilon E, Raju DSS, Crosby WL, Dunstan DI (1988) Transient gene expression in electroporated Picea glauca protoplasts. Plant Cell Reports 7:481–484

  6. Durzan DJ, Gupta PK (1987) Somatic embryogenesis and polyembryogenesis in Douglas-fir cell suspension cultures. Plant Science 52:229–235

  7. Gupta PK, Durzan DJ (1987) Somatic embryos from protoplasts of loblolly pine proembryonal cells. Bio/Technology 5:710–712

  8. Gupta PK, Dandekar AM, Durzan DJ (1988) Somatic proembryo formation and transient expression of a luciferase gene in Douglas-fir and loblolly pine protoplasts. Plant Science 58:85–92

  9. Gupta PM, Durzan J (1987) Biotechnology of somatic polyembryogenesis and plantlet regeneration in loblolly pine. Bio/Technology 5:147–151

  10. Hakman I, Fowke LC (1987) An embryogenic cell suspension culture of Picea glauca (White spruce). Plant Cell Reports 6:20–22

  11. Kirby EG (1988) Recent advances in protoplast culture of horticultural crops: conifers. Scientia Horticulture 37:267–276

  12. Klimaszewska K (1989) Plantlet development from immature zygotic embryos of hybrid larch through somatic embryogenesis. Plant Science 63: 95–103

  13. Laine E, David H, David A (1988) Callus formation from cotyledon protoplasts of Pinus oocarpa and Pinus patula. Physiol. Plant. 72:374–378

  14. Nagmani R, Becwar MR, Wann SR (1987) Single-cell origin and development of somatic embryos in Picea abies (L.) Karst. (Norway spruce) and P. glauca (Moench) Voss (white spruce). Plant Cell Reports 6:157–159

  15. Widholm, JM (1972) The use of fluorescin diacetate and phenosafranine for determining viability of cultured plant cells. Stain Technology 47:189–194

  16. Wilson SM, Thorpe TA, Moloney MM (1989) PEG-mediated expression of GUS and CAT genes in protoplasts from embryogenic suspension cultures of Picea glauca. Plant Cell Reports 7:704–707

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Communicated by F. Constabel

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Klimaszewska, K. Recovery of somatic embryos and plantlets from protoplast cultures of Larix × eurolepis . Plant Cell Reports 8, 440–444 (1989). https://doi.org/10.1007/BF00269044

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Keywords

  • Cell Suspension
  • Somatic Embryo
  • Suspension Culture
  • Density Gradient
  • Critical Factor