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Título en ingles: New alternative for micropropagation in temporary immersion system of plantain cultivar "INIVITPV-2011" (AAB). Resumen El trabajo fue desarrollado en el Laboratorio de Biotecnología Vegetal del Instituto de Investigaciones en Viandas Tropicales (INIVIT) con el objetivo de incrementar el coeficiente de multiplicación en el cultivar de plátano vianda "INIVITPV-2011" (AAB) en el Sistema de Inmersión Temporal (SIT). Se estudiaron diferentes tiempos de inmersión (10, 20 (control) y 30 minutos) y frecuencias de inmersión (3, 6 (control) y 8 horas) en frascos Nalgene de 10 L de capacidad, se estudió además el volumen de medio de cultivo por explante (20, 40 (control) y 60 ml de medio de cultivo/explante), así como el tiempo de subcultivo (15, 18, 21 (control) y 25 días) y la densidad de explantes por frasco (20, 40 (control), 60 y 80 explantes/frasco de cultivo). Se utilizó el medio de cultivo de multiplicación MS suplementado 2,0 mg.L-1 de 6-BAP; 3,5 mg.L-1de AIA; 30,0 g.L-1 de sacarosa; 10,0 mg.L-1 de ácido ascórbico. Los resultados obtenidos permitieron establecer una metodología para la micropropagación en el Sistema de Inmersión Temporal del cultivar de plátano vianda "INIVITPV-2011" (AAB) la cual consistió en utilizar un tiempo de inmersión de 10 minutos con una frecuencia cada 3 horas, es decir, 8 inmersiones al día, además para cada frasco de 10 L se inocularon 60 explantes y la renovación con 3600 ml de medio de cultivo y un tiempo de cultivo de 18 días permitió alcanzar la mayor productividad del material en fase de multiplicación. Estos resultados, utilizando el Sistema de Inmersión Temporal permitieron establecer una metodología eficiente para la micropropagación del cultivar de plátano vianda "INIVITPV-2011" (AAB). Palabras clave: propagación in vitro; tiempo; frecuencia; inmersión; multiplicación. Abstract This work was developed at the Plant Biotechnology Laboratory from the Research Institute of Tropical Root and Tuber Crops (INIVIT) in order to increase the multiplication coefficient in plantain cultivar "INIVITPV-2011" (AAB) in Temporary Immersion Systems (TIS). Different immersion times (10, 20 (control) and 30 minutes) and immersion frequencies (3, 6 (control) and 8 hours) in 10 L Nalgene flasks, and culture medium volume per explants (20, 40 (control) and 60 ml culture medium / explants) were studied, as well as, subculture time (15, 18, 21(control) and 25 days) and explants density per flask (20, 40 (control), 60 and 80 explants / culture flask). The multiplication culture medium MS supplemented with 2.0 mg.L-1 6-BAP, 3.5 mg.L-1 IAA, 30.0 gL-1 sucrose, 10.0 mg.L-1 ascorbic acid was used. Results obtained allowed to establish a methodology for micro-propagation of plantain cultivar "INIVITPV-2011" (AAB) in temporary immersion system, which consisted of using a 10 minute immersion time with 3 hour frequency (8 immersions per day). Besides, 60 explants were inoculated in each 10 L flask, and the renewal with 3600 ml culture medium and 18 day culture time allowed to obtain the highest productivity in the multiplication stage. These results, using the temporary immersion system allowed to establish a method for micro-propagation of plantain cultivar "INIVITPV-2011" (AAB). Key words: in vitro propagation; time; frequency; immersion; multiplication.

The use of meristems for the in vitro establishment of Colocasia esculenta (L.) Schott cv. 'INIVIT MC-2012' decreases bacterial contamination but is required to increase its growth. The objective of this work was to determine the effect of 6-Benzylaminopurine and indoleacetic acid on its in vitro establishment. Various concentrations of the two growth regulators were included in the culture medium and with the best combination the explants were cultured in shaking and static liquid culture medium. With the use of the culture medium constituted by 80% of the salts and vitamins MS, 30 g l-1 sucrose, 0.1 g l-1 myo-inositol, 0.1 mg l-1 6-BAP, 0.05 mg l-1 of AIA and culture in agitation were obtained meristems with the appropriate morphological characteristics to transfer to the multiplication phase at 28 days of culture. Keywords: orbital agitator, propagation, taro

Título en inglés: Multiplication of banana clone 'FHIA-18' (AAAB) in Temporary Immersion System Resumen Con el propósito de desarrollar un protocolo para la multiplicación del clon de banano ´FHIA-18´ (AAAB) en sistema de inmersión temporal, se definieron como objetivos del trabajo determinar el efecto del tiempo (5, 10 y 15 minutos) y la frecuencia de inmersión (3, 6 y 8 horas por día), así como la influencia de diferentes combinaciones de reguladores del crecimiento (2,0; 3,0 y 4,0 mg.L-1 de 6-BAP y 2,0; 2,5; 3,0; 3,5 y 4,0 mg.L-1 de 6 AIA), el efecto del volumen de medio de cultivo por planta (20, 30, 40 y 50 ml/explante) y la densidad de explantes por frasco de cultivo (30, 50, 70 y 90 explantes/frasco) para incrementar el coeficiente de multiplicación. Con el empleo de un tiempo de 10 minutos y una frecuencia de inmersión cada tres horas, se alcanzaron los mejores resultados en cuanto al número de explantes obtenidos. Con este tiempo y frecuencia de inmersión los explantes presentaron el mayor diámetro del pseudotallo. Para cada frasco de 10,0 L se inocularon 70 explantes y la renovación con 2800 ml de medio de cultivo (40 ml/explante) con un tiempo de cultivo de 21 días permitió alcanzar la mayor productividad del material en fase de multiplicación. Además al utilizar las sales MS suplementadas con 3,0 mg.L-1 de 6-BAP; 2,0 mg.L-1 de AIA; 10,0 mg.L-1 de ácido ascórbico, se logró disminuir el crecimiento innecesario de los tallos y hojas de los brotes en la fase de multiplicación y por lo tanto un mayor número de explantes. Palabras claves: coeficiente, densidad de explantes, tiempo, frecuencia, volumen de medio de cultivo. Abstract In order to develop a protocol for multiplication of Banana clone 'FHIA-18' (AAAB) in temporary immersion systems, the following working objectives were defined: to determine the effect of immersion time (5, 10 and 15 minutes) and frequency (3, 6 and 8 hours per day), as well as, the influence of different combinations of growth regulators (2,0; 3,0 and 4,0 mg.L-1 de 6-BAP and 2,0; 2,5; 3,0; 3,5 and 4,0 mg.L-1 de 6 AIA), the volume effect of culture medium plants (20, 30, 40 and 50 ml/explant) and explants density per culture flask (30, 50, 70 y 90 explants/flask) to increase the multiplication coefficient. With 10 minutes immersion time and an immersion frequency every three hours, the best results were achieved in relation to the number of explants obtained. With this immersion time and frequency, explants showed the highest pseudostem diameter. Seventy explants were inoculated in each 10,0 L culture flask. The highest productivity at the multiplication phase was achieved with a culture medium renewal of 2800 ml (40 ml/explants), and a 21 day culture time. In addition to using MS salts supplemented with 3.5 mg.L-1 of 6-BAP, 1.30 mg.L-1 of IAA, 10.0 mg.L-1 ascorbic acid, the unnecessary growth of stems and leaves of shoots in the multiplication phase was reduced and; therefore, a greater number per explant. Keywords: coefficient, explants density, time, frequency, culture medium volume.

Temporary Immersion Systems (TIS) had been used for in vitro multiplication of different cultivars of Musa spp. achieving high coefficients of multiplication. This investigation was carried out to focussed on the multiplication of the plantain `INIVITPV06-30' (Musa AAB) in TIS. Independent and consecutive experiments were realized to determine the effect of immersion period, immersion frequency, culture medium volume, time of culture and number of explants per flask on the multiplication of this cultivar in TIS. The best results were obtained with 10 minutes immersions of every 6 hours as immersion frequency, 40 ml culture medium per explant, 90 explants per culture flask and 18 days of culture. Plantain cultivar `INIVITPV06-30' (Musa AAB) was multiplied in TIS. The multiplication coefficient and the quality of plant material were high allowing the scaling of them in the tissue culture commercial laboratories. Key words: explants, micropropagation, multiplication coefficient.

Necessity to produce high quality planting material has required searching new alternatives to increase the efficiency of in vitro propagation methods and their automation, as in Temporary Immersion Systems (TIS). This work was aimed to multiply the hybrid ‘FHIA-25’ (Musa spp. AAB) in TIS. The effect of different culture medium volumes per explant and densities of planting materials per culture flask at the same immersion frequency was determined. These two factors showed influence on the evaluated variables. A volume of 40 ml of culture medium per explant and densities of 80 explants per flask were selected to multiply this cultivar in TIS. These results permitted to increase in vitro production of high-quality plants for rooting stage. Key words: multiplication coefficient, culture flasks, liquid culture medium

The need of producing high quality planting material has required the searching of new alternatives to increase the efficiency of in vitro propagation methods and their automation, such as Temporary Immersion Systems. This work has been carried out to increase the multiplication coefficient in mass propagation of hybrid ‘FHIA-21’ (AAAB) in Temporary Immersion Systems. Different culture medium volumes per explant and densities of planting materials per unit were studied at the same immersion frequency. The highest multiplication coefficient rate, (14.99 and 16.02 respectively), was obtained when 40 ml culture medium volume were used at a density of 70 explants/flask. The use of Temporary Immersion System allowed increasing the multiplication coefficient in hybrid ‘FHIA-21’ (AAAB) and the highest quality multiplication coefficient for rooting stage and further acclimatization in field conditions. Key words: hybrid, liquid culture medium, multiplication coefficient