Temporary immersion systems in plant micropropagation

CIRAD; CATIE; Centre de Coopération Internationale en Recherche Agronomique pour le Développement – Amis (CIRAD-AMIS), CIRAD
Plant Cell Tissue and Organ Culture (Impact Factor: 2.61). 05/2002; 69(3):215-231. DOI: 10.1023/A:1015668610465

ABSTRACT 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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    ABSTRACT: Temporary Immersion Bioreactor (TIB) system has been proven to be an efficient tool for important economic crops micropropagation. This is mainly because the TIB system is capable of improving plant quality and multiplication rates for ease of scaling-up while requiring low production costs. A regular TIB system comprises two separate chambers: one for nutrient medium storage and the other for culturing shoots. Liquid medium is transferred to the culture tank and remains there for minutes, after which it is pumped back to the storage tank for reuse. A control system plays a significant role in manipulating the flow of the liquid medium in the TIB system. A simple control system consists of two digital timers working in parallel, responsible for the operation of electromagnetic valves, which creates pressure difference between a liquid medium and a culturing shoots chamber. Manual programming of the digital timers is a labor-intensive task. The programming task becomes more critical especially when operating multiple TIB systems. Incorrect programming of the digital timers may cause serious damage to the plant production process, and devices and equipment connected to the TIB systems including the operators working at the production site when there is insufficient pressure or overpressure. The objective of this research was to develop the computer software to control the sequence of multiple TIB Systems. Computer software was developed under LABVIEW environment based on parallel computing and state machine architecture. The software consists of three independent modules: 1) graphic user interface (GUI) module, 2) sequence control (SC) module, and 3) hardware interface (HI) module. The GUI module received the information regarding immersion time and immersion duration for each TIB system from the operator. The information was automatically synchronized with the SC module, which initiated the immersion process at the specified time and terminated the process after the immersion duration met the target. The HI module continuously monitored the TIB sequence, assigned by the SC module, and triggered the electromagnetic valves manipulating the air stream in the TIB system to force liquid medium transport. The developed computer software was connected to eight 20-liter TIB systems and tested under sugarcane plant micropropagation. It was observed that the computer software could independently control the sequence of each TIB system in accordance with the specified conditions. It requires less than one-minute changing immersion schedule and immersion time without process interruption. The developed software can provide a convenient and effective way to operate multiple 20-liter TIB systems with safety. It is possible to apply the software to other similar TIB systems.
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    ABSTRACT: The two commercially important apple rootstocks i.e., MM106 and B9 were micropropagated using a liquid culture system. Three different strengths of 0.8% agar solidified PGR free basal MS medium were first tested to optimize the culture media for both the rootstocks. Full strength medium (MS0) supported maximum in vitro growth, multiplication, rooting and survival under field conditions as opposed to quarter and half strength media. When three different volumes of liquid MS0 were tested, highest in vitro growth, multiplication, rooting and also survival under field conditions were achieved in 20 mL liquid MS0. The cost of one litre of liquid medium was also reduced by 8 times to Rs. 6.29 as compared to solid medium. The cost of 20 mL medium was further reduced to Rs. 0.125.
    Indian journal of experimental biology 07/2014; 52(7):748-54. · 1.20 Impact Factor
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    ABSTRACT: The effects of temporary immersion system (TIS) culture on the growth and quality of Dioscorea fordii Prain et Burk and Dioscorea alata plantlets were investigated. Results indicate that TIS promoted the growth and quality of D. fordii and D. alata plantlets. Proliferation rate, shoot length, fresh weight (FW) and dry weight (DW) of shoots, and total biomass production were significantly (P≤0.05) higher in the TIS than in gelled and liquid medium, respectively. The TIS also promoted tuberization of D. fordii, and decreased vitrification of D. alata significantly. The healthy plantlets of D. fordii and D. alata obtained in the TIS would probably have positive effects on transplanting in large-scale commercial production.


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May 15, 2014