Regeneración in vitro de plantas de piña ( Ananas comosus ) ecotipo amazónico Gobernadora / In vitro regeneration of Amazonian pineapple ( Ananas comosus ) plants ecotype Gobernadora

Resumen En Venezuela existen cultivares y ecotipos de piña (A. comosus) de importancia local, entre ellos los amazónicos, cultivados principalmente por los aborígenes Piaroa. Ellos siembran los propágulos lo cual restringe la disponibilidad de material para el cultivo a gran escala. Se abordó la limitación recurriendo al cultivo de tejidos vegetales para la propagación in vitro de plantas de piña, ecotipo amazónico Gobernadora, mediante embriogénesis somática (ES) y organogénesis adventicia (OA). El material vegetal empleado correspondió a secciones basales e intermedias de hojas. Sólo las secciones de base foliar (SBF) fueron morfogénicamente inducidas. El mayor número de vitroplantas (1,58 plantas/explante) se obtuvo del callo embriogénico inducido en medio MS con Picloram 10 mg.L-1 + Tidiazuron 2 mg.L-1, transferido a MS sin hormonas. En el proceso organogénico, se obtuvo el mayor número de plantas/explante (5) por vía directa en MS con ácido naftalenoacético 5 mg.L-1 + bencilaminopurina 0,25 mg.L-1, transferido a MS. Siendo este último el mejor sistema de cultivo in vitro por su productividad y por ser una ruta que minimiza la variación somaclonal.

Abstract There are a number of pineapple (Ananas comosus) cultivars and ecotypes of local commercial importance in Venezuela, among them the Amazonian ones, cultivated mainly by the aboriginal Piaroa, are of relevance. They sow the propagules, which restricts the availability of material for large-scale cultivation. This limitation was approached by plant tissue culture for in vitro propagation of Amazonian pineapple plants, Gobernadora ecotype, through somatic embryogenesis (ES) and adventitious organogenesis (OA). Basal and intermediate sections of leaves were tested. Only the leaf base sections (FBS) were morphogenically induced. The highest number of vitroplants (1.58 plants / explant) was obtained from the embryogenic callus induced in MS medium with Picloram 10 mg.L-1 + Thidiazuron 2 mg.L-1, transferred to MS medium without hormones. In the organogenic process, the highest number of plants / explants (5) was obtained directly in MS with naphthaleneacetic acid 5 mg.L-1 + benzylaminopurine 0.25 mg.L-1, transferred to MS. The latter being the best in vitro culture system due to its productivity and for being a method that minimizes somaclonal variation.

Análisis de patrones morfológicos y anatómicos en la embriogénesis somática del banano Williams (AAA) / Analysis of the morphological and anatomical patterns in the somatic embryogenesis of Williams banana (AAA)

La embriogénesis somática representa una herramienta esencial en el mejoramiento genético y en la micropropagación clonal masiva de bananos mejorados. En el presente trabajo se analizaron los patrones morfológicos y anatómicos que ocurren durante la embriogénesis somática del banano Williams, dirigidos a conocer y mejorar este proceso. En la investigación se establecieron suspensiones celulares embriogénicas (SCE) a partir de callo embriogénico obtenido de manos florales inmaduras masculinas, las cuales originaron abundantes embriones que regeneraron plantas. Hacia los tres meses de cultivo se detectaron embriones somáticos (ES) primarios color blanco-crema en las manos florales de los nudos nueve a doce, contados a partir del ápice floral. Al cuarto mes estos ES primarios dieron origen al callo embriogénico, de color blanco crema, estructura granular, con abundantes ES torpedo en su periferia y con una organización celular en tres diferentes zonas. De este callo se cultivaron porciones pequeñas con ES torpedo en medio de multiplicación durante dos meses, dando origen a la SCE I. La misma se tamizó (250 µm) para establecer la SCE II. El sedimento de células y los agregados celulares embriogénicos de ambas SCE se trasladó a medio de maduración. Transcurridos dos meses los embriones maduros se transfirieron a medio de conversión de embriones, lográndose regenerar plantas completas a partir de las dos semanas. Las SCE produjeron numerosos embriones somáticos maduros y mostraron una buena conversión de embriones a plantas y regeneración de plantas. Este sistema de embriogénesis somática permitió la obtención de plantas funcionales en nueve meses.

Somatic embryogenesis represents an essential tool for the genetic improvement and for the mass clonal micropropagation of the improved banana plant. In this present work morphological and anatomical patterns were analyzed in the somatic embryogenesis of Williams banana, to know and enhance this process. In the investigation embryogenic cell suspensions (ECS) were established from embryogenic callus obtained from floral immature male hands, which gave rise to many somatic embryos that regenerated plants. Towards the three months of culture white-cream primary somatic embryos (SE) were detected in the floral hands of the nodes nine to twelve, counted from the floral apex. At the fourth month this primary SE gave origin to a creamy-white embryogenic callus, with granular structure and abundant SE torpedo on its periphery. Cell organization with three different zones was observed in callus. Small portions of this callus were cultivated in the multiplication medium for two months, to originate ECS I. This ECS was filtered through a mesh (250 µm pore size) to establish the ECS II. The sediment of embryogenic cells and cell clusters of the ECS were moved to maturation media. After two months the mature embryos were transferred to conversion medium, and two weeks later, whole plants were developed. The ECS produced numerous mature SE, which showed good conversion of embryos into plants and plant regeneration. This system of somatic embryogenesis permitted the mass production of functional plants in nine months.

Somatic embryogenesis in Solanum tuberosum from cell suspension cultures: histological analysis and extracellular protein patterns.

An embryogenic cell suspension, continuously grown in Murashige and Skoog (MS) medium with 0.5 mg/L of 2,4-dichlorophenoxyacetic acid, was established from friable callus of Solanum tuberosum internode sections. The cell suspension was predominantly composed of cell masses and free embryogenic cells. When transferred to an auxin-free medium with zeatin, somatic embryos (SEs) developed and converted to complete plants when cultured on solid MS medium without growth regulators. The system produced approximately 600 SEs per 50 mL of medium. In this investigation, accumulation of extracellular proteins (EPs) of different molecular weights were found associated to different phases of the embryogenic process. At the initiation of the cell suspension, cell clusters and free cells present in the culture (phase "A") secreted a 78kDa EP, unique to this phase. In phase "B", which is related to embryonic cell determination process, proteins (7-14kDa) were secreted mainly by embryogenic cells. In phase "C", SEs in different developmental stages secreted protein of 32 kDa, which appeared as a particular feature of the phase. EPs of phase "D", secreted by torpedo and mature embryos, had molecular weights between 20 and 50 kDa. Further studies will be necessary to identify these proteins and link them to previously identified somatic embryogenesis-related proteins. Histological analysis of the potato embryogenesis in liquid media showed unicellular origin of the SE.

Plant regeneration of Anthurium andreanum cv Rubrun

To establish an efficient regeneration system for Anthurium andreanum cv Rubrun, seeds from plant spadixes were germinated on a medium supplemented with 2.2 muM BA. After 2 weeks, 74 percent of the seeds germinated and four weeks later, micro-cuttings from these plantlets were subcultured on a medium containing 4.4 muM BA and 0.05 muM NAA. On average, 3.6 shoots per explant were obtained. Four weeks old in vitro plants from germinated seeds and the plantlets obtained from micro-cuttings, showed callus proliferation at the stem base. These tissues were subcultured on a medium supplemented with 8.9 muM BA and 2.7 muM NAA. After 6 weeks of culture, about 43.8 plantlets per square cm of callus were obtained. Anatomical studies showed the organogenic nature of these calli. Anthurium andreanum plants regenerated by organogenesis were transferred to pots and a rate of 80 percent of plant acclimatization was obtained.

[Improvement of somatic embryogenesis process in sugarcane Venezuelan cultivars]. / Optimización del proceso de embriogénesis somatica en variedades venezolanas de caña de azúcar.

Efficient embryogenic callus formation (70%) in many sugarcane cultivars, has been established using young leaf explants cultivated on modified Murashige and Skoog medium containing 13 microM 2,4-dichlorophenoxiacetic acid (2,4-D). However, Venezuelan sugarcane cultivars V78-1 and V75-6 produced only 30% of embryogenic callus when were cultured in these conditions. In order to improve somatic embryogenesis in these Venezuelan cultivars, embryogenic calli were induced using different media: C3 (13 microM 2,4-D); C7 (31.5 microM 2,4-D); Cd (30 microM Dicamba) and C5BA (22.5 microM 2,4-D, 22.2 microM N6-Benzyladenine). After 45 days of induction, the highest embryogenic callus production (90%), was observed in both cultivars, when they were cultured on Cd medium. Moreover, plant development from somatic embryos originated in this callus, was evident four days after incubation on regeneration medium (without hormones), while the somatic embryos originated from calli growing in C3, C7 and C5BA media, gave rise to plants eight days after incubation on regeneration medium.