New explant for somatic embryogenesis induction and plant regeneration from diploid banana (‘Calcutta 4´, Musa AA) Jorge López Torres 1* , Rafael G, Kosky 2 , Nery Montano Perez 1 , Damicela Reinaldo Alvarez 1 , Ayme Rayas Cabrera 1 , Manuel Cabrera Jova 1 , Arletys Santos Pino 1 Victor Medero Vega 1 , Milagros Basail Pérez 1 , Nicolas Roux 3 *Autor para correspondencia 1 Instituto de Investigaciones de Viandas Tropicales (INIVIT). Apdo 6, Sto Dgo, Villa Clara, CP 53 000, Cuba. e-mails: jlopez@inivit.co.cu; jlopezbiotec@yahoo.es) 2 Instituto de Biotecnología de las Plantas (IBP), Universidad Central Marta Abreu de Las Villas, Carretera Camajuaní km 5.5, Santa Clara, Villa Clara, CP 54 830, Cuba. 3 Parc Scientifique Agropolis II 34 397 Montpellier, Cedex 5 – France ABSTRACT A method has been developed for plant regeneration via embryogenic cell suspensions from diploid cultivar ‘Calcutta 4´. For callus induction with embryogenic structures, different plant tissues such as scalps from cauliflower-like meristems and meristematic domes of axillary sprouted buds in combination with several culture media were evaluated as explants. The best embryogenic response (8%) was noticed with meristematic domes of axillary sprouted buds in culture medium Murashige and Skoog salts at 50%, MS vitamins, 30 g l -1 sucrose, 10 mg l -1 ascorbic acid, 1 mg l –1 2,4-D, 0.22 mg l –1 Zeatine and supplemented with 100 mg l –1 malt extract, 100 mg l –1 glutamine, 1 mg l –1 biotin, 200 mg l –1 casein hydrolysate, 4.0 mg l –1 proline and solidified with 2.0 g l –1 Gelrite. Embryogenic cell suspensions were established and the highest increase of cellular biomass with 0.50 ml settled cell volume (SCV) was obtained 18 days after culture initiation. In a RD1 culture medium, embryogenic masses from 1362 to 2480 embryos were formed. An average of 54.5% germinated embryos in Temporal Immersion System (TIS) was obtained. Results were significantly higher in comparison with the use of semisolid culture media (28%). Regenerated plants are in field conditions to value their genetic stability. Keywords: axillary buds, somatic embryogenesis, Temporal Immersion System Abbreviations: gFW - grams of fresh weight; SCV – Settled Cell Volume; MS - Murashige and Skoog (1962) medium; BA- 6-benzyaldenine; 2,4-D - 2,4-dichlorophenoxgace acid; IAA - indole-3-acetic acid. INTRODUCTION Bananas and plantains are important fruits in tropical and subtropical countries and provide employment, nutrition and food security. They are grown in more than 100 countries (Escalant and Jain, 2004), however, due to the appearance of ‘Black Sigatoka’ disease caused by Mycosfaerella figiensis, the yields losses are estimated at 33-76% being necessary the development of biotechnological techniques that can supplement the genetic improvement programs. The integration of genetic engineering into breeding programs may provide powerful tools to overcome these limitations by introducing specific genetic changes that can be utilized for banana improvement within a short period of time. However, these applications require reliable plant regeneration protocols for banana (Khalil et al., 2002; Pérez and Rosell, 2008). On the other hand, the Global Musa Genomics Consortium, uses ‘Calcutta 4’, Musa (AA) as the standard genotype for genomic works and in this respect, it is recommended to develop diploid embryogenic cell suspensions for mutagenesis (INIBA and IPGRI, 2002) but embryogenic cell suspensions from this genotipe are not available (Strosse et al., 2003; Strosse et al., 2004). For that reason is necessary to develop a protocol that allows the regeneration of plants starting from somatic Artículo Científico Biotecnología Vegetal Vol. 12, No. 1: 25 - 31, enero - marzo, 2012 ISSN 1609-1841 (Versión impresa) ISSN 2074-8647 (Versión electrónica)