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Evaluation of the in vitro methods for micropropagation of Chondracanthus acicularis (Roth) Fredericq (Gigartinales, Rhodophyta): tissue culture and production of protoplasts

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Abstract

Tissue was cultured and protoplasts isolated from the carrageenophyte Chondracanthus acicularis with the aim of developing micropropagation as an alternative to harvesting raw material from natural beds. Both adventitious shoots and filamentous calluses were induced by tissue culture on medium solidified with 0.4–1 % (w/v) agar. Adventitious shoots were mainly produced from discoid bases while filamentous calluses were mainly induced from basal zones and sub-apical explants. A gradient of the regeneration ability was observed from the top to the bottom of the thallus. The discoid base was the most reactive explant and produced the highest number of adventitious shoots compared to basal zones and sub-apical explants, irrespective of the concentration of agar. Protoplasts were isolated enzymatically from the whole thallus using a combination of cellulase R-10 Onozuka, macerozyme R-10, and crude extract of the gland gut of algivorous molluscs. The highest mean yield of protoplasts (1.2 × 106 protoplasts g−1 fresh weight) was obtained after 16 h of digestion with an enzyme mixture containing 2 % (w/v) cellulase R-10, 1 % (w/v) macerozyme R-10 Onozuka, 4 % (v/v) crude extract of gut gland of Haliotis, 0.8 M mannitol, 50 mM sodium citrate, 0.3 % (w/v) bovine serum albumin. Depending on the conditions, mean protoplast yields ranged from 3.14 × 105 to 1.2 × 106 protoplasts g−1 fresh weight. Different factors (storage duration, mannitol, sodium citrate, crude extract of the gland gut of algivorous molluscs) were tested to improve the yield of protoplasts but none has a significantly effect.

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Acknowledgments

Moussa Yagame Bodian was financially supported by the “Ministère de l’Enseignement Supérieur des Universités des Centres Universitaires Régionaux et de la Recherche Scientifique” of Senegal via the foundation FIRST (Fonds d’Impulsion de la Recherche Scientifique et Technique).

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Authors and Affiliations

  1. Université de Caen Basse-Normandie, Esplanade de la Paix, CS 14032, 14032, CAEN, cedex 5, France

    Moussa Yagame Bodian, Nadège Lafontaine, Maria Matard, Isabelle Mussio & Anne-Marie Rusig

  2. UNICAEN, BioMEA, 14032, Caen, France

    Moussa Yagame Bodian, Nadège Lafontaine, Maria Matard, Isabelle Mussio & Anne-Marie Rusig

  3. CNRS INEE, FRE3484, 14032, Caen, France

    Moussa Yagame Bodian, Nadège Lafontaine, Maria Matard, Isabelle Mussio & Anne-Marie Rusig

  4. Faculté des Sciences et Techniques, Laboratoire de Botanique et de Biodiversité, Département de Biologie Végétale, Université Cheikh Anta Diop, BP 5005, Dakar-Fann, Dakar, Sénégal

    Moussa Yagame Bodian

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  1. Moussa Yagame Bodian
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  2. Nadège Lafontaine
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Correspondence to Isabelle Mussio.

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Bodian, M.Y., Lafontaine, N., Matard, M. et al. Evaluation of the in vitro methods for micropropagation of Chondracanthus acicularis (Roth) Fredericq (Gigartinales, Rhodophyta): tissue culture and production of protoplasts. J Appl Phycol 25, 1835–1845 (2013). https://doi.org/10.1007/s10811-013-0042-3

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