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Callogenesis De Heliconia Collinsiana GRIGGS in Vitro

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Callogenesis De Heliconia Collinsiana GRIGGS in Vitro Revista Mexicana de Ciencias Agrícolas Vol.4 Núm.8 12 de noviembre - 31 de diciembre, 2013 p. 1175-1186 Callogenesis de Heliconia collinsiana GRIGGS in vitro: establecimiento, inducción y proliferación* Callogenesis of Heliconia collinsiana GRIGGS in vitro: establishment, induction and proliferation Eleodoro Hernández-Meneses1, María Cristina Guadalupe López-Peralta2§ y Andrés Adolfo Estrada-Luna3 1Postgrado en Recursos Genéticos y Productividad (PREGEP)-Fisiología Vegetal. (doromeneses@colpos .mx). 2PREGEP-Genética. Colegio de Postgraduados. 56230, Montecillo, Texcoco, Estado de México. Tel. (595) 9520200 Ext. 1540. [email protected]. 3CINVESTAV-Unidad Irapuato. Departamento de Ingeniería Genética. ([email protected]). §Autora para correspondencia: [email protected]. Resumen Abstract Las heliconias son plantas ornamentales cultivadas como Heliconias are grown as ornamental plants and cut flower flor de corte y maceta. En México se cultivan cada vez pot. In Mexico is increasingly grown in States with tropical más en Estados que disponen de condiciones climáticas climatic conditions. While it is possible in vitro propagation of tropicales. Si bien ya es posible la propagación in vitro de Heliconia by direct organogenesis, it has not yet been able to heliconias mediante organogénesis directa, aún no se han establish protocols for both indirect organogenesis and somatic podido establecer protocolos tanto de organogénesis como embryogenesis for mass propagation of elite genotypes. The embriogénesis somática indirecta para la propagación purpose of this research was to develop a protocol for callus masiva de genotipos élite. La finalidad de ésta investigación induction and proliferation in vitro in Heliconia collinsiana. fue desarrollar un protocolo para la inducción y proliferación Explants of root tips, leaf, petiole and basal cross sections of de callos in vitro en Heliconia collinsiana. Explantes de pseudostem were cultivated in the culture medium of Murashige ápices de raíz, hoja, pecíolo y secciones transversales and Skoog (1962) supplemented with 13.6, 27.1, 54.3, 81.4, basales de pseudotallo se cultivaron en el medio de cultivo 108.6 and 135.7 µm of 2,4-D, dicamba and picloram combined de Murashige y Skoog (1962) suplementado con 13.6, with activated carbon (0 to 0.5 g L-1). The cultures were kept in 27.1, 54.3, 81.4, 108.6 y 135.7 µM de 2,4-D, dicamba y the dark and four, eight and 12 weeks were evaluated percentage picloram combinadas con carbón activado (0 y 0.5 g L-1). of callus induction. Only cross sections calluses induced Los cultivos se mantuvieron en oscuridad y a las cuatro, pseudostem at the baseline at 12 weeks with 81.4 (100%) and ocho y 12 semanas se evaluó el porcentaje de inducción 135.7 (90%) µM of 2,4-D and picloram, respectively, combined de callos. Únicamente las secciones transversales basales with 0.5 g L-1 of activated carbon. This callus by sub-cultivating de pseudotallo indujeron callos a las 12 semanas con them in 0, 4.5 and 9 µM of 2, 4-D in the proliferation stage 81.4 (100%) y 135.7 (90%) µM de 2,4-D y picloram, responded differently. After 16 weeks, only the calluses induced respectivamente, combinados con 0.5 g L-1 de carbón continued to grow with 81.4 µM of 2, 4-D in photoperiod activado. Éstos callos al subcultivarlos en 0, 4.5 y 9 µM de of 16 h. The callus generated with 135.7 µM of picloram 2, 4-D en la etapa de proliferación respondieron diferente. got blackened at four weeks of culture in all the treatments. Después de 16 semanas, sólo continuaron creciendo los callos inducidos con 81.4 µM de 2, 4-D en fotoperiodo de Key words: activated carbon, dicamba, picloram, platanillo. * Recibido: marzo de 2013 Aceptado: octubre de 2013 1176 Rev. Mex. Cienc. Agríc. Vol.4 Núm.8 12 de noviembre - 31 de diciembre, 2013 Eleodoro Hernández-Meneses et al. 16 h. Los callos generados con 135.7 µM de picloram se Introduction ennegrecieron a las cuatro semanas de cultivo en todos los tratamientos evaluados. Heliconias are tropical ornamental plants grown commercially Palabras clave: carbón activado, dicamba, picloram, for the production of cut flowers or as potted plants for their platanillo. bright colors and exotic shapes (Criley, 2000; Loges et al., 2007). The cultivation of these ornamentals in Mexico has increased mainly in the South-East States that have tropical Introducción conditions (Murguia et al., 2007). The main problem held by heliconias for obtaining clonal plantations that can only be propagated by rhizome division, since sexually (seeds) genetic Las heliconias son plantas ornamentales tropicales cultivadas variability is generated, especially when it comes to varieties. comercialmente para la producción de flores para corte, o bien, como plantas de maceta, por sus colores brillantes y Helicona collinsiana is a species cultivated in Mexico as formas exóticas (Criley, 2000; Loges et al., 2007). El cultivo a cut flower and like other species and varieties have high de estas plantas ornamentales en México se ha incrementado degree of natural hybridization limiting its spread by seeds. principalmente en los estados del Sur-Sureste que cuentan Its inflorescence is of pendant type and flowering occurs in con las condiciones tropicales propicias para ello (Murguía et the months of July to November. It reaches a height of 3 to 4.5 al., 2007). El problema principal que presentan las heliconias m and is well suited for cultivation in areas with 50% shade. para obtener plantaciones clonales es que sólo se pueden The inflorescence consists of 12-18 bracts of bright red and propagar mediante división de rizomas ya que sexualmente covered with white wax, the rachis is reddish in color and (semillas) se genera variabilidad genética, especialmente is growth habit is of musoide kind (Berry and Kress, 1991). cuando se trata de variedades. Using the techniques of plant tissue culture in vitro is Helicona collinsiana es una especie cultivada en México an efficient tool to propagate elite genotypes by direct como flor de corte y al igual que otras especies y variedades organogenesis; also the obtained plants are free of disease presentan alto grado de hibridación natural que limita su (Marulanda-Angel et al., 2011). Early studies with this propagación por semillas. Su inflorescencia es tipo colgante technique in heliconias were used by Nathan et al. (1992) y la floración ocurre en los meses de julio a noviembre. who through lateral bud regeneration achieved indirectly Alcanza una altura de 3 a 4.5 m y se adapta muy bien para su in vitro H. psittacorum and found that the plants obtained cultivo en áreas con 50% de sombreado. La inflorescencia produced more than conventionally propagated. This same consta de 12 a 18 brácteas de color rojo brillante y cubiertas type of explant was also efficient for the propagation of H. de cera blanca; el raquis es de color rojizo y su hábito de standleyi (Sosa et al., 2008). In addition to these studies, crecimiento es tipo musoide (Berry y Kress, 1991). which have focused on mass propagation of commercial varieties of heliconias highly available for cultivation El uso de las técnicas del cultivo de tejidos vegetales in in field, we has also explored the possibility of inducing vitro representa una herramienta eficiente para propagar somatic embryos, but up to this date it has not been achieved genotipos élite mediante la organogénesis directa; by establishing an efficient protocol to propagate these además, las plantas obtenidas están libres de enfermedades morphogenic ornamental by this route (Ulysses et al., 2007). (Marulanda-Ángel et al., 2011). Los primeros estudios con esta técnica en heliconias fueron Nathan et al. (1992) This ornamental species and other phylogenetically related, quienes a través de yemas laterales lograron la regeneración failed to obtain somatic embryogenesis directly, which is why, de manera indirecta in vitro de H. psittacorum y encontraron in this investigation we sought callus induction indirectly que las plantas obtenidas produjeron más vástagos que las because this phase is the dedifferentiation of somatic tissue propagadas de forma convencional. Este mismo tipo de and the subsequent acquisition of embryogenic competence. explante también fue eficiente para la propagación de H. Based on these backgrounds, this study aimed to develop a standleyi (Sosa et al., 2008). Además de estos estudios, protocol for callus induction and proliferation in Heliconia que se han enfocado a la propagación masiva de variedades collinsiana by culturing in vitro from different explants in the de heliconias altamente comerciales disponibles para su presence of various auxins and activated carbon. Callogenesis de Heliconia collinsiana GRIGGS in vitro: establecimiento, inducción y proliferación 1177 cultivo en campo, también se ha explorado la posibilidad de Materials and methods inducir embriones somáticos; sin embargo, hasta la fecha no se ha logrado el establecimiento de un protocolo eficiente que permita propagar estas ornamentales mediante esta ruta Plant material and explant types morfogénica (Ulisses et al., 2007). From aseptic seedlings developed in vitro explants were En esta especie ornamental, así como en otras relacionadas obtained of leaf, petiole, and root tips and cross sections filogenéticamente, no se ha podido obtener la embriogénesis (thin layer of cells) in the base of the pseudostem. For the somática de manera directa; es por ello, que en esta leaf explants were cut 1 cm segments with the central leaf of investigación se buscó la inducción de callos de forma main rib while the petioles were used longitudinal sections indirecta porque esta fase constituye la dediferenciación del of 1 cm. In the root explants were obtained 1 cm longitudinal tejido somático y la adquisición subsecuente de competencia sections including the apex. For transverse sections were cut embriogénica. Con base en éstos antecedentes, el presente pseudostem seedling of basal segments of 10 mm in diameter trabajo tuvo como finalidad desarrollar un protocolo para la with 1.2 mm thick (Figure 1).
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