Temporary immersion system in plant micropropagation. Plant Cell Tiss Org Cult

Centre de Coopération Internationale en Recherche Agronomique pour le Développement – Amis (CIRAD-AMIS), CIRAD
Plant Cell Tissue and Organ Culture (Impact Factor: 2.13). 05/2002; 69(3):215-231. DOI: 10.1023/A:1015668610465


Temporary immersion systems for plant micropropagation have been described and grouped into 4 categories according to operation: tilting and rocker machines; complete immersion of plant material and renewal of the nutrient medium; partial immersion and a liquid nutrient renewal mechanism; complete immersion by pneumatic driven transfer of liquid medium and without nutrient medium renewal. The positive effects of temporary immersion on micropropagation are indicated for shoot proliferation and microcuttings, microtuberization and somatic embryogenesis. Immersion time, i.e. duration or frequency, is the most decisive parameter for system efficiency. Optimizing the volume of nutrient medium and the volume of the culture container also substantially improves efficacy, especially for shoot proliferation. Temporary immersion also generally improves plant material quality. It results in increased shoot vigour and in the frequency of morphologically normal somatic embryos. Hyperhydricity, which seriously affects cultures in liquid medium, can be eliminated with these culture systems or controlled by adjusting the immersion times. Plant material propagated by temporary immersion can perform better during the acclimatization phase than material obtained on semi-solid or in liquid media. Successful regeneration of plants, after direct sowing on soil of Solanum tuberosum microtubers and Coffea arabica somatic embryos produced in temporary immersion bioreactors, has been demonstrated. As could be expected when using liquid medium for micropropagation, several estimations confirm large gains in efficacy from temporary immersion. The parameters most involved in reducing production costs include: (1) the drastic reduction in work; (2) reduction in shelving area; (3) reduction in the number of containers used; (4) better biological yields. Scaling-up somatic embryogenesis and shoot proliferation procedures involving temporary immersion systems in order to commercialize this process are now taking place.

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    • "On the other hand in a temporary immersion system, Jiménez et al. (1999) observed that the best growth of potato shoots at an immersion frequency of 5 min for every 3 h. Etienne and Berthouly (2002) reported that 1 h of immersion per 6 h interval was useful for microtuberization of potato. In our NSB system, a nutrient spray frequency of once per hour was optimum for shoot growth and microtuberization. "
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    ABSTRACT: A laboratory scale bioreactor system has been developed using nutrient spray technology for in vitro mass production of potato microtubers. Its effectiveness on the production of microtubers was investigated and compared with conventional liquid and semi-solid culture systems through bioreactor. Optimal culture conditions such as spray intervals, varying concentrations of 6-benzylaminopurine (BAP) and explants density were determined for the NSB. In order to determine optimal spray intervals, liquid medium was sprayed inside the NSB at different intervals (½, 1–4 h) of which the 1 h interval resulted in the highest number of shoots (3.47) and length (8.99 cm). Number of microtubers produced (5.13) was highest with 1 h intervals and fresh weight of microtubers (0.90 g) was highest for ½ h interval. Different concentrations of BAP (0.5, 1.0 and 1.5 mg/L) were used to evaluate its effect on microtuberization. It was observed that number and diameter of microtubers were increased (5.31 and 0.96 cm) when 0.5 mg/L BAP was supplemented in MS medium. We found fresh weight of microtubers (0.97 g) was increased when 1.0 mg/L BAP were added to the medium. In order to determine suitable explants density, single nodes grouped into five categories e.g 30, 45, 60, 75, and 90 and placed in the NSB system. A density of 60 explants resulted in increases in shoot length (17.5 cm) number of internodes (12.5) and with highest amount of chlorophyll (40.2 mg/g) as well as with highest number and fresh weight of microtubers (4.43 and 0.89 g, respectively). Out of the three culture systems, the NSB performed best where 1.5–2.0 fold increases in shoot growth and microtuberization without hyperhydration. The NSB also produced the highest number (4.67), fresh weight (0.86 g) and diameter (0.78 cm) of microtubers. From this study we may conclude that the NSB system has good potential for commercial mass production of potato micro-tuber.
    Scientia Horticulturae 08/2015; 192:369-374. DOI:10.1016/j.scienta.2015.06.014 · 1.37 Impact Factor
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    • "La implementación de metodologías de propagación basadas en el empleo de medios de cultivo líquidos que prescinden del agente gelificante, definitivamente reducen los costos de producción (Preil 2005; Aggarwal y Barna 2004). Además, éstos facilitan la absorción de nutrientes por parte de los tejidos, disminuyen el tiempo requerido para el desarrollo de los cultivos (Alcaraz-Meléndez et al., 2002); simplifican las operaciones de preparación y dispensado del medio, permitiendo establecer métodos de control y muestreo en cada etapa y sustentan el desarrollo propuestas para la automatización de los procesos in vitro (Satyahari 2005; Etienne y Berthouly 2002). "
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    ABSTRACT: Título en imgles: Liquid medium culture: an approach for the commercial micropropagation of aloe (Aloe barbadensis Mill.) Título corto: Un avance para la micropropagación comercial de zábilaResumen: La micropropagación es una alternativa para la producción comercial de plantas de zábila (Aloe barbadensis Mill.) limitada por los altos costos de producción. Con el objetivo de prescindir de los agentes gelificantes, reduciendo costos, se comparó el medio de cultivo líquido con el medio de cultivo gelificado en las diferentes etapas de micropropagación de la zábila. En la etapa de establecimiento se observó mayor porcentaje de explantes contaminados en el medio de cultivo líquido estático (25.55%) que en el medio gelificado (11.11%); y aunque el resto de los explantes se establecieron independientemente de la condición del medio de cultivo, en el medio líquido alcanzaron mayor altura (3.81 cm) que en el medio gelificado (3.03 cm). En la etapa de multiplicación, la altura de los explantes (entre 4.43 y 6.01 cm) fue superior en los recipientes de inmersión temporal automatizado (RITA®) en comparación con el medio gelificado (entre 3.24 y 3.42 cm); sin diferencias significativas entre el número de brotes/explante. Todos los brotes enraizaron a los 30 días independientemente del medio de cultivo empleado (líquido estático y gelificado), sin observar variaciones en la altura del brote y, número y longitud de las raíces. El empleo de los medios de cultivo líquidos y la implementación de los sistemas de inmersión temporal tipo RITA® permiten reducir los costos de producción al prescindir de los agentes gelificantes, lo que representa un avance para la micropropagación comercial de zábila. Palabras clave: Cultivo de tejidos, agentes gelificantes, RITA®, sistemas de inmersión temporal.Abstract: Micropropagation is considered a successful alternative for aloe (Aloe barbadensis Mill.) plant production. However, it has limited use due to the high production cost. Liquid media were compared to agar-gelled medium during all micropropagation stages of aloe to reduce the cost for gelling agent used. In the establishment stage, there was a higher percentage of contaminated explants in static liquid medium (25.55%) than those cultured in agar-gelled medium (11.11%), although all the explants were established independently of the culture medium used, higher height (3.81 cm) was observed in liquid medium than those growing in agar-gelled medium (3.03 cm). In the multiplication stage, explant height was higher in the recipients used for automated temporary immersion system (RITA®) (4.43‑6.01 cm) than those cultured in agar-gelled medium (3.24‑3.42 cm), there was no significant difference for number of shoots/explant. All shoots had roots at 30 days independently of used culture media (static liquid or agar-gelled media). Shoot height, number and root length had similar values in both culture media. The implementation of liquid media and automated temporary Immersion system RITA® may allow to reduce production costs of gelling agent used, it represents an approach for the commercial micropropagation of aloe.Keys words: Tissue culture, gelling agents, RITA®, temporary immersion system.Recibido: junio 15 de 2014 Aprobado: abril 13 de 2015
    05/2015; 17(1):24-31. DOI:10.15446/rev.colomb.biote.v17n1.50669
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    • "The use of temporary immersion systems results in increased multiplication rates and less handling and labor; thus, allowing the reduction of production costs (Feuser et al. 2003, Rech Filho et al. 2005, Steinmacher et al. 2011). Escalona et al. (1999) developed the twin flasks temporary immersion system (TIS-TF) for pineapple micropropagation and, later, other studies reported the efficiency of this system in many other plant species (Etienne and Berthouly 2002, Rech Filho et al. 2005, Niemenak et al. 2008, Steinmacher et al. 2011). Furthermore, the use of TIS resulted in a lower Breeding and Applied Biotechnology 15: 66-71, 2015 rate of somaclonal variation for pineapples, as compared to the permanent immersion system (Feuser et al. 2003). "
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    ABSTRACT: Micropropagation based techniques enable the mass production of pest- and disease-free stock plants. Aiming at optimizing pineapple [Ananas comosus (L.) Merril] micropropagation, a complete randomized experiment with three factors was established: immersion system (2 L twin flasks temporary immersion and permanent immersion); culture medium (supplemented or not with GA3); and shoot height class. The effects on regeneration rate (number of new shoots per 1 g fresh mass (FM) of inoculated microshoots), FM per shoot, and shoot survival were evaluated. Significant interactions (p<0.05) were observed in the regeneration rate, and simple effects (p<0.05) were detected for FM per shoot. All treatments resulted in a survival rate higher than 95%. Altogether, it was found that 2 L twin flasks temporary immersion and MS culture medium free of GA3 allowed the optimization of the protocol used. This treatment was tested with positive response in a biofactory system that uses sterile plastic bottles (5L). © 2015, Brazilian Society of Plant Breeding. All rights reserved.
    01/2015; 15(2):66-71. DOI:10.1590/1984-70332015v15n2a13
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