Regeneration of diploid and tetraploid plants from callus-derived protoplasts of Agapanthus praecox ssp. orientalis (Leighton) Leighton

Abstract

protoplasts was developed for the Liliaceous ornamental plant, Agapanthus praecox ssp. orientalis (Leighton) Leighton `Royal Purple Select' (2n=2x=32).Viable protoplasts were routinely isolated from leaf-derived embryogenic calluses with yields of 0.8 to 1.5x10 protoplasts per g FW of calluses. Protoplasts started to divide 5 to 7 days after isolation, and protoplast-derived colonies consisting of 50 to 100 cells were obtained after 1 month. A plating efficiency of 0.8% was obtained after 2 months of culture using a gellan gum-solidified medium containing 1 mg 1-1 each of PIC and BA under continuous illumination. Protoplastderived calluses produced somatic embryos at a frequency of 46.7 % on PGR-free medium, whereas 68.3 % of the calluses regenerated adventitious shoots on a medium containing 1 mg 1-1 BA. Somatic embryos and adventitious shoots developed into plantlets, which were successfully transplanted to pots. Flow cytometric analysis and chromosome observation revealed that both diploid and tetraploid plants were regenerated from protoplasts.

This is a preview of subscription content, access via your institution.

References

  1. Buiteveld J & Creemers-Molenaar J (1994) Plant regeneration from protoplasts isolated from suspension cultures of leek (Allium ampeloprasum L.). Plant Sci. 100: 203-210

    Google Scholar 

  2. Johnson AAT & Veilleux RE (2001) Somatic hybridization and applications in plant breeding. Plant Breed. Rev. 20: 167-225

    Google Scholar 

  3. Kunitake H & Mii M (1990) Somatic embryogenesis and plant regeneration from protoplasts of asparagus (Asparagus officinalis L.). Plant Cell Rep. 8: 706-710

    Google Scholar 

  4. Larkin PJ & Scowcroft RW(1981) Somaclonal variation - a novel source of variability from cell cultures for plant improvements. Theor. Appl. Genet. 60: 197-214

    Google Scholar 

  5. Mii M, Yuzawa Y, Suetomi H, Motegi T & Godo T (1994) Fertile plant regeneration from protoplasts of a seed-propagated cultivar of LiliumΧformolongi by utilizing meristematic nodular cell clumps. Plant Sci. 100: 221-226

    Google Scholar 

  6. Mukhopadhyay S & Desjardins Y (1994) A comparative study on mode of culture and plant regeneration from protoplast-derived somatic embryos of two genotypes of Asparagus officinalis. Plant Sci. 100: 97-104

    Google Scholar 

  7. Murashige T & Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15: 473-497

    Google Scholar 

  8. Mor Y, Halevy AH, Kofranek AM & Reid MS (1984) Postharvest handling of Lily of the Nile flowers. J. Amer. Soc. Hort. Sci. 109: 494-497

    Google Scholar 

  9. Nakano M, Hosokawa K, Oomiya T & Yamamura S (1995) Plant regeneration from protoplasts of Gentiana by embedding protoplasts in gellan gum. Plant Cell Tiss. Org. Cult. 41: 221-227

    Google Scholar 

  10. Nakano M, Sakakibara T, Suzuki S & Saito H (2000) Decrease in the regeneration potential of long-term cell suspension cultures of Lilium formosanum Wallace and its restoration by the auxin transport inhibitor, 2,3,5-triiodobenzoic acid. Plant Sci. 158: 129-137

    Google Scholar 

  11. Saito H & Nakano M (2001) Flow cytometric analysis of partially-synchronized suspension cultures of Hemerocallis hybrida. Plant Biotechnol. 18: 229-231

    Google Scholar 

  12. Singh RJ (1993) Plant Cytogenetics. CRC Press, Boca Raton, Florida

    Google Scholar 

  13. Supaibulwatana K & Mii M (1997) Organogenesis and somatic embryogenesis from young flower buds of Agapanthus africanus Hoffmanns. Plant Biotechnol. 14: 23-28

    Google Scholar 

  14. Suzuki S & Nakano M (2001) Organogenesis and somatic embryoregeneration genesis from callus cultures in Muscari armeniacum Leichtl. ex Bak. In Vitro Cell. Dev. Biol. Plant 37: 382-387

    Google Scholar 

  15. Suzuki S, Supaibulwatana K, Mii M & Nakano M (2001a) Prosource duction of transgenic plants of the Liliaceous ornamental plant Agapanthus praecox ssp. orientalis (Leighton) Leighton via Agrobacterium-mediated transformation of embryogenic calli. Plant Sci. 161: 89-97

    Google Scholar 

  16. Suzuki S, Oota M & Nakano M (2001b) Embryogenic callus induction from leaf explants of the Liliaceous ornamental plant, Agapanthus praecox ssp. orientalis (Leighton) Leighton. Hismode tological study and response to selective agents. Sci. Hort. 95: 123-132.

    Google Scholar 

  17. Widholm JM (1972) The use of fluorescein diacetate and phenosafranine for determining viability of cultured plant cells. Stain Tech. 47: 189-194

    Google Scholar 

Download references

Author information

Affiliations

Authors

Corresponding author

Correspondence to Masaru Nakano.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Nakano, M., Tanaka, S., Oota, M. et al. Regeneration of diploid and tetraploid plants from callus-derived protoplasts of Agapanthus praecox ssp. orientalis (Leighton) Leighton. Plant Cell, Tissue and Organ Culture 72, 63–69 (2003). https://doi.org/10.1023/A:1021290106221

Download citation

  • agapanthus
  • embryogenic callus
  • plant regeneration
  • ploidy level
  • protoplast