Skip to main content
SpringerLink
Log in

Protoplast regeneration from gametophytes and sporophytes of some species in the order Laminariales (Phaeophyceae)

  • Original Papers
  • Published:
Protoplasma Aims and scope Submit manuscript

Summary

Protoplasts were isolated from sporophytes and from gametophyte cultures of several species in the order Laminariales. For each example, the isolation and culture procedures were investigated systematically, to identify conditions leading to plant regeneration. After dedifferentiation through a filamentous stage, protoplasts isolated from adultLaminaria saccharina sporophytes regenerated polystichous bladelets. In contrast, cells isolated fromLaminaria digitata sporophytes proved recalcitrant in culture, except when the donor plants were undifferentiated sporelings. The most critical factors for protoplast development were the origin of explants, the osmoticum used for cell isolation, cultivation in plain seawater, and the absence of stress during the first two weeks of culture. We also found that protoplast isolation from the sporophytes of members of the Laminariales results in the release of hydrogen peroxide, up to 5–120 μM final concentration in the macerating medium, a characteristic which may be related to protoplast recalcitrance. Protoplasts isolated from the gametophytic phase readily regenerated into normal gametophytes, capable of gametogenesis and producing sporophytes by fertilization.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

F.W.:

fresh weight

MES:

(2-N-morpholino)ethane-sulfonic acid

References

  • Ar Gall E, Chiang YM, Kloareg B (1993) Isolation and regeneration of protoplasts fromPorphyra dentata andPorphyra crispata. Eur J Phycol 28: 277–283

    Article  Google Scholar 

  • —, Asensi A, Marie D, Kloareg B (1996) Parthenogenesis and apospory in the Laminariales: a flow cytometry analysis. Eur J Phycol 31: 369–380

    Article  Google Scholar 

  • Benet H, Bruss H, Duval J-C, Kloareg B (1994) Photosynthesis and photoinhibition in protoplasts of the marine brown algaLaminaria saccharina. J Exp Bot 271: 211–220

    Google Scholar 

  • Björk M, Haglund K, Ramazanov Z, Garcia-Reina G, Perdersén M. (1992) Inorganic-carbon assimilation in the green seaweedUlva rigida C. Ag. (Chlorophyta). Planta 187: 152–156

    Article  Google Scholar 

  • Boyen C, Bertheau Y, Barbeyron T, Kloareg B (1990a) Preparation of guluronate lyase fromPseudomonas alginovora for protoplast isolation inLaminaria. Enzyme Microb Technol 12: 885–890

    Article  CAS  Google Scholar 

  • —, Kloareg B, Polne-Fuller M, Gibor A (1990b) Preparation of alginate lyase from marine molluscs for protoplasts isolation in brown algae. Phycologia 29: 173–181

    Google Scholar 

  • Butler DM, Ostgaard K, Boyen C, Evans LV, Jensen A, Kloareg B (1989) Isolation conditions for high yields of protoplasts fromLaminaria saccharina andLaminaria digitata (Phaeophyceae). J Exp Bot 40: 1237–1246

    Article  CAS  Google Scholar 

  • Chen LCM (1986) Cell development ofPorphyra miniata (Rhodophyceae) under axenic culture. Bot Mar 29: 435–439

    Google Scholar 

  • Collén J, Pedersén M (1994) Stress induced oxidative burst byEuchema platycladum (Rhodophyta). Physiol Plant 92: 417–422

    Article  Google Scholar 

  • Criqui MC, Jamet E, Parmentier Y, Marbach J, Durr A, Fleck J (1992a) Isolation and characterization of a plant cDNA showing homology to animal glutathione peroxidases. Plant Mol Biol 18: 623–627

    Article  PubMed  CAS  Google Scholar 

  • Criqui MC, Plesse B, Durr A, Marbach J, Parmentier Y, Jamet E, Fleck J (1992b) Characterization of genes expressed in mesophyll protoplasts ofNicotiana sylvestris before the re-initiation of the DNA replicational activity. Mech Dev 38: 121–132

    Article  PubMed  CAS  Google Scholar 

  • Cutler AJ, Saleem M, Coffey MA, Loewen MK (1989) Role of oxidative stress in cereal protoplast recalcitrance. Plant Cell Tissue Organ Culture 18: 113–127

    Article  CAS  Google Scholar 

  • De Marco A, Roubelakis-Angelakis KA (1996) The complexity of enzymic control of hydrogen peroxide concentration may affect the regeneration potential of plant protoplasts. Plant Physiol 110: 137–145

    PubMed  Google Scholar 

  • Ducreux G, Kloareg B (1988) Plant regeneration from protoplasts ofSphacelaria (Phaeophyceae). Planta 174: 25–29

    Article  Google Scholar 

  • Fleck J, Durr A, Lett M, Hirth L (1979) Changes in protein synthesis during the initial stage of life of tobacco protoplasts. Planta 145: 279–285

    Article  CAS  Google Scholar 

  • — — — — (1980) Comparison of proteins synthesized in vivo and in vitro by mRNA from isolated protoplasts. Planta 148: 453–454

    CAS  Google Scholar 

  • Grosset J, Meyer Y, Chartier Y, Kauffmann S, Legrand M, Frittig B (1990) Tobacco mesophyll protoplasts synthesize 1,3-β-glucanase, chitinases and “osmotins” during in vitro culture. Plant Physiol 92: 520–527

    PubMed  CAS  Google Scholar 

  • Hahne G, Lörz H (1988) Release of phytotoxic factors from plant cell walls during protoplast isolation. J Plant Physiol 132: 345–350

    CAS  Google Scholar 

  • Ishii S (1987) Generation of active oxygen species during enzymic isolation of protoplasts from oat leaves. In Vitro Cell Dev Biol 239: 653–658

    Google Scholar 

  • — (1988) Factors influencing protoplast viability of suspension-cultured rice cells during isolation process. Plant Physiol 88: 26–29

    PubMed  CAS  Google Scholar 

  • Kloareg B, Quatrano R (1987) Enzymatic removal of the cell walls from zygotes ofFucus distichus (L.) Powell (Phaeophyta). Hydrobiologia 151/152: 123–129

    Article  Google Scholar 

  • —, Polne-Fuller M, Gibor A (1989) Mass production of viable protoplasts fromMacrocystis pyrifera (L.) C. Ag. (Phaeophyta). Plant Sci 62: 105–112

    Article  Google Scholar 

  • Kuhlenkamp R, Muller DG (1994) Isolation and regeneration of protoplasts from healthy and virus-infected gametophytes ofEctocarpus siliculosus (Phaeophyceae). Bot Mar 37: 525–530

    Article  Google Scholar 

  • Mejjad M, Loiseaux de Goer S, Ducreux G (1992) Protoplast isolation, development and regeneration in different strains ofPilayella littoralis (L.) Kjellm (Phaeophyceae). Protoplasma 169: 42–48

    Article  Google Scholar 

  • Meyer Y, Chartier Y (1981) Hormonal control of mitotic development in tobacco protoplasts: two-dimensional distribution of newly-synthetised proteins. Plant Physiol 68: 1273–1278

    Article  PubMed  CAS  Google Scholar 

  • — —, Grosset J, Marty I, Brugidou C, Marinho P, Rivera R (1993) Gene expression in mesophyll protoplasts. In: Roubelakis-Angelakis KA, Tranh Tan Van K (eds) Morphogenesis in plants: molecular approaches. Plenum, New York, pp 221–236

    Google Scholar 

  • Mieth H, Speth V, Ebel J (1986) Phytoalexin production by isolated soybean protoplasts. Z Naturforsch 41: 193–201

    CAS  Google Scholar 

  • Polne-Fuller M, Gibor A (1984) Developmental studies inPorphyra. I Blade differentiation inPorphyra perforata as expressed by morphology, enzymatic digestion and protoplast regeneration. J Phycol 20: 609–616

    Article  Google Scholar 

  • — — (1986) Calluses, cells, and protoplasts in studies toward genetic improvement of seaweeds. Aquaculture 57: 117–123

    Article  Google Scholar 

  • Provasoli L (1986) Media and prospects for cultivation of marine algae. In: Watanabe A, Hattori A (eds) Cultures and collections of algae. Proceedings U.S.-Japan Conference, Hakone. Japanese Society for Plant Physiologists, Kyoto, pp 63–75

    Google Scholar 

  • Reddy CRK, Fujita Y (1991) Regeneration of plantlets fromEnteromorpha (Ulvales, Chlorophyta) protoplasts in axenic culture. J Appl Phycol 3: 265–275

    Article  Google Scholar 

  • —, Migita S, Fujita Y (1989) Protoplasts isolation and regeneration of three species ofUlva in axenic culture. Bot Mar 32: 483–490

    Article  Google Scholar 

  • Roubelakis-Angelakis KA (1993) An assessment of possible factors contributing to recalcitrance of plant protoplasts. In: Roubelakis-Angelakis KA, Tranh Tan Van K (eds) Morphogenesis in plants: molecular approaches. Plenum, New York, pp 201–219

    Google Scholar 

  • Rusig AM, Le Guyader H, Ducreux G (1994) Dedifferentiation and microtubule reorganization in the apical cell protoplast ofSphacelaria (Phaeophyceae). Protoplasma 179: 83–94

    Article  Google Scholar 

  • Saga N, Kudo T (1989) Isolation and culture of protoplasts from the green algaMonostroma angicava. J Appl Phycol 1: 25–30

    Article  Google Scholar 

  • Sawabe T, Ezura Y (1996) Regeneration fromLaminaria japonica Areschoug (Laminariales, Phaeophyceae) protoplasts isolated with bacterial alginase. Plant Cell Rep 15: 892–895

    Article  CAS  Google Scholar 

  • Siminis CI, Kanellis AK, Roubelakis-Angelakis K (1994) Catalase is differentially expressed in dividing and non-dividing protoplasts. Plant Physiol 105: 1375–1383

    PubMed  CAS  Google Scholar 

  • Wolff SP (1994) Ferrous ion oxidation in presence of ferric ion indicator xylenol orange for measurement of hydroperoxides. Methods Enzymol 233: 182–189

    Article  CAS  Google Scholar 

  • Wu CY, Lin GH (1987) Progress in the genetics and breeding of economic seaweeds in China. Hydrobiologia 151/152: 57–61

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Centre d’Etudes d’Océanologie et de Biologie Marine, CNRS, F-29682, Roscoff Cedex, France

    H. Benet, E. Ar Gall, A. Asensi & B. Kloareg

Authors
  1. H. Benet
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. E. Ar Gall
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. A. Asensi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. B. Kloareg
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Benet, H., Ar Gall, E., Asensi, A. et al. Protoplast regeneration from gametophytes and sporophytes of some species in the order Laminariales (Phaeophyceae). Protoplasma 199, 39–48 (1997). https://doi.org/10.1007/BF02539804

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02539804

Keywords

Search

Navigation