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Embryogenic cell suspension cultures of garlic (Allium sativum L.) as method for mass propagation and potential material for genetic improvement

Féréol L., Chovelon V., Causse S., Kalumvueziko M.L., Kahane R.. 2005. In : Guangshu Liu (ed.), Dewei Zhu (ed.). Proceedings of the Fourth International Symposium on Edible Alliaceae, Beijing, China, April 21-26, 2004. Louvain : ISHS [Belgique], p. 65-74. (Acta Horticulturae, 688). International Symposium on Edible Alliaceae. 4, 2004-04-21/2004-04-26, Pékin (Chine).

Embryogenic calluses were induced from young leaf explants of garlic (Allium sativum L). Four cultivars, "Rouge de la Réunion", "Messidrome", "Morasol" and "Printanor" have been successfully tested. These calluses expressed up to 90% of embryogenic calluses differentiating globular somatic embryos after 2 months on N6 modified medium supplemented with 2,4-D (0.1 mg/l) and kinetin (0.5 mg/l). Embryogenic calluses were used to establish cell suspension cultures of the above-mentioned cultivars. Friable calluses were induced from compact ones, and could give rise to the production of cell suspension cultures composed of small aggregates of embryogenic cells. These suspension cultures were maintained in liquid medium based on N6 modified macro-nutrients and supplemented with 2,4-D/benzyladenine (0.3 mg/l/0.1 mg/l). The packed cell volume (PCV) of the suspension cultures increased 2-fold in a 2-week period. These cell suspension cultures led to successful regeneration of mature embryos and their conversion into plantlets. Optimal embryo regeneration efficiency was obtained after plating on semi-solid medium base on N6 macro-nutrients and a balance in 2,4-D/Kinetin (0.1 mg l/0.5 mg/l). A large number of somatic embryos (potentially 8 x 10x9 to 10x11) could be produced per year for each cultivar. The conversion into plantlet was approximately 50%. Plants were successfully acclimatised in greenhouse. Histological analyses were performed along the suspension cultures and regeneration process, and helped for establishing the sequence of culture media. The somatic embryogenic nature was confirmed by single cell origin and polar development of the regenerants. This protocol was used in a goal of mass propagation of garlic plants true to the original type. It would be a key tool for biotechnologies in genetic improvement of garlic.

Mots-clés : allium sativum; embryogénèse somatique; tissu végétal; régénération; multiplication des plantes; technique de culture; amélioration des plantes

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