Indirect organogenesis in Medicago sativa L. var. Gilboa
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Abstract
The alfalfa is considered one of the most important forage plants in livestock feeding all over the world. The introduction in Cuba began about 40 years ago, but difficulties of seed reproductions and varieties well adapted to climate conditions have been the main factors to limit its exploitation. Plant biotechnology offers many techniques that can achieve new varieties with stress tolerance, and adapted to our climate conditions, but all of these have to begin creating the cognitive bases of tissue culture to permit the in vitro plant manipulation. This projection can be developed using an in vitro regeneration via organogenesis protocol. The objective of this work was to obtain shoots formation from morphogenetic callus in Medicago sativa L. var. Gilboa. Different explants were studied for callus formation using three doses (1, 3, 5 mg.L-1) of 2,4-dichlorophenoxyacetic acid (2,4-D) and a control treatment. Three concentration of 2,4- (2, 3 y 4 mg.L-1) combined with 6-benzylaminopurine (2, 3 y 4 mg.L-1) and a MS medium as control treatment were evaluated to achieve proliferation and regeneration of callus. The highest percentage of callus induction was observed for immature leaf explants. Auxin presence was a necessary requirement for a better growth of cellular mass during callus multiplication. Highest percentage of callus formation (85 %) was observed with 3 mg.L-1 of 2,4-D. Callus proliferation was efficient, but shoot formation was onlysuperiorstatistically when 6-BAP (4 mg.L-1)and 2,4-D (2 mg.L-1) were combined into the culture medium.
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References
Abdel Latef, A.A. y Ahmad, P. (2015): Chapter 1. Legumes and breeding under abiotic stress: an overview. En: Legumes under environmental stress yield. Improvement and adaptations. Azooz, M.M. and Ahmad, P. (Eds.) Oxford, UK. p. 2.
Akitha Devi, M.; Sakthivelu, G.; Giridar, P. y Ravishankar (2012): Protocol for augmented shoot organogenesis in selected variety of soybean (Glycine max L. Merr.) Indian J. Exp. Biol., 50: 729-734.
Babar, M.M.; Zaidi, N.S.; Azooz, M.M. and Kazi, A. G. (2015): Chapter 15. Biotechnology approaches to overcome biotic and abiotic stress constraints in legumes. En: Legumes under environmental stress yield. Improvement and adaptations. Azooz, M.M. and Ahmad, P. (Eds.) Oxford, UK, p. 255.
Bingham, E.T.; Hurley, L.V.; Kaatz, D.M. y Saunders, J.W. (1975): Breeding alfalfa which regenerates from callus tissue in culture. Crop Sci., 15: 719-721.
Borém, A. y Fritsche-Neto, R. (2014): Biotechnology and Plant Breeding Applications and Approaches for Developing Improved Cultivars. First edition. Oxford, UK., 8-9.
Brown, D.C.W. y Atanassov, A. (1985): Role of genetic background in somatic embryogenesis in Medicago. Plant Cell Tiss. Org. Cult., 4: 111-122.
Brown, J.; Caligari, P.D.S. y Campos, H.A. (2014) Plant breeding. 2nd edition. Oxford, UK.,185-188.
Chen, T.; Marowitch, J. y Thompson, B. (1987): Genotypic effects on somatic embryogenesis and plant regeneration from callus cultures of alfalfa. Plant Cell Tiss. Org. Cult., 8: 73–81.
Cobb, J.N.; DeClerck, G.; Greenberg, A.; Clark, R.; McCouch, S. (2013): Next-generation phenotyping: requirements and strategies for enhancing our understanding of genotype–phenotype relationships and its relevance to crop improvement. Theoretical Applied Genetics, 126: 867–887.
Collado, R.; García, L.; Angenon, G.; Torres, D.; Romero, C.; Bermúdez, I.; Veitía, N. y Ramírez, M. (2008): Organogénesis indirecta en Phaseolus vulgaris L. cv. CIAP 7247. Biotecnología Vegetal, 8(2): 81-86.
García, L.; Bermúdez, I.; Veitía, N.; Collado, R.; Torres, D.; Romero, C. y Martirena, A. (2012): Regeneración de plantas de cinco variedades de Phaseolus vulgaris L. vía organogénesis directa. Biotecnología Vegetal, 12 (1): 49-52.
García, L.; Pérez, J.; Kosky, R.G.; Bermúdez- Caraballoso, I.; Veitía, N.; Collado, R.; Padrón, Y.; Torres, D.; Romero, C. y Reyes, M. (2010): Regeneración de plantas vía embriogénesis somática en Phaseolus acutifolius A. Gray. Biotecnología Vegetal, 10 (3): 143-149.
Heisey, P.W. y Fuglie, K.O. (2014): Private research and development for crop genetic improvement. En: Genetically engineered crops in America analyses, adoption, trends. F.G. Lawrence (Ed.), Nova Science Publishers, Inc. New York, EEUU. p. 130.
Li, J.J.; Wu, Y.M.; Wang, T. y Liu, J.X. (2009): In vitro direct organogenesis and regeneration of Medicago sativa. Biologia Plantarum, 53 (2): 325-328.
Menéndez, J.; Tang, M. y Aira, A. (1987): Comportamiento de cultivares de alfalfa (Medicago sativa, L.) en Guantánamo. Pastos y Forrajes, 10 (3): 203-211.
Mitten, D.H.; Sato, S.J. y Skokut, T.A. (1984): In vitro regeneration potential of alfalfa germplasm sources. Crop Sci, 24: 943-945.
Murashige, T. y Skoog, F. (1962): A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plantarum, 15: 473-497.
Noaman, M.M. y Ahmad, E. (2004): Development of Alfalfa Tolerant to Salinity Stress Using Organogenesis Technique. Biotechnology, 3: 136-139.
Orcen, N. (2013): In vitro organogenesis and regeneration of alfalfa (Medicago sativa L.) domestic cultivar cv. Kayseri. Fresenius Environmental Bulletin, 22 (9a): 2770- 2774.
Quintana, M.; Carbonell, A. y Nápoles, J.A. (2014): Capítulo III. Algunas técnicas biotecnológicas aplicadas al estudio de las leguminosas. En: Leguminosas en la producción agropecuaria. ACPA (Ed.), Cuba, 34-52.
Rodríguez, M.; Latsague, Mirtha; Chacón, M. y Astorga, Pamela (2014): Inducción in vitro de callogénesis y organogénesis indirecta a partir de explantes de cotiledón, hipocótilo y hoja en Ugni molinae. Rev. Electrónica UACh Bosque (Valdivia) 35 (1).
Srivastava, D.K.; Gambhir, G. and Sharma P. (2014): Chapter 3. Plant cell and tissue culture techniques in crop improvement. En: Biotechnology in agriculture and food processing. Opportunities and challenges. Ed. Panesar, P.S. Marwaha, S.S. CRC Press, EEUU.,75-119.
Volenec, J.J.; Cunningham, S.M.; Haagenson, D.M.; Berg, W.K. y Joern, B.C. (2002): Physiological genetics of alfalfa improvement: past failure, future prospects. Field Crop Res., 75 (2): 97-110.
Zare, N., Valizadeh, M., Tohidfar, M., Mohammadi, S.A., Mal boobi, M.A. y Habashi, A.K. (2009): Selection of regenerative genotypes from Iranian alfalfa cultivars. Journal of Food, Agriculture and Environment, 7 (3 & 4): 567 -572.
Zhang, S. and Lemaux, P.G. (2004): Molecular analysis of in vitro shoot organogenesis. Rev. Crit. Rev. Plant. Sci., 23 (4): 325-335.
Zimmerman, J. (1993): Somatic embryogenesis: a model for early development in higher plants. Plant Cell, 5: 1411–1423.