Journal of Applied Phycology

, Volume 21, Issue 2, pp 255–264 | Cite as

Morphogenetic responses from protoplasts and tissue culture of Laminaria digitata (Linnaeus) J. V. Lamouroux (Laminariales, Phaeophyta): callus and thalloid-like structures regeneration



The regeneration of meristematic tissues from sporophytes of Laminaria digitata was studied by protoplast and tissue culture. Sequential treatment of explants in sterile seawater with 1% Betadine for 5 min, 1% commercial bleach for 1–2 min and 2% antibiotic treatment supplemented with 1 μM GeO2 overnight enabled viable explants as high as 55%. Different morphogenetic responses were observed from tissue culture on media supplemented with plant growth regulators alone or in combination, mainly filamentous calluses up to 50% according to the media. Dark green compact calluses were observed on two combinations: 4 μM Pi + 2 μM N-(2-chloro-4-pyridyl)-N’-phenylurea (CPPU) and 0.04 μM Pi + 0.44 μM 6-benzylaminopurine. Thalloid-like structures comparable to adventitious buds were regenerated on medium supplemented with 4 μM Pi + 0.45 μM zeatin but at low frequency suggesting a strong genotypic effect. Friable calluses were developed from protoplasts in enriched medium with polyamines and containing 0.40 μM CPPU + 0.45 μM 2,4-dichlorophenoxyacetic acid. In order to produce protoplasts, a one-step enzymatic protocol was developed and yields reached 22 × 106 protoplasts per gram of fresh weight.


Adventitious bud Callus Laminaria digitata Plant growth regulators Protoplast Meristematic explant 



This research was supported by a grant of Conseil Régional de Basse-Normandie. The authors would like to thank Dr Gérard Baux and Dr Philippe Fossier (CNRS, Neurobiologie Cellulaire et Moléculaire) for providing crude extracts of Aplysia vaccaria.


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© Springer Science+Business Media B.V. 2008

Authors and Affiliations

  1. 1.UMR IFREMER 100 - Physiologie et Ecophysiologie des Mollusques Marins, IFR 146 ICOREUniversité de Caen Basse-NormandieCaen CedexFrance

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