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: This study aims at establishing a temporary immersion technique for large-scale propagation of cocoyam (Xanthosoma sagittifolium). Sucrose was experimented with as a determinant factor for shoot multiplication in this culture system. The highest proliferation rate (68 ± 7) occurred with 20 g l−1 sucrose in the culture medium. This concentration appeared to be the optimal amount due to its promoting effect on plantlet development. The acclimatized plantlets showed a continuous effect of sucrose treatment during ex vitro growth, especially in low sucrose concentration. This fact is evidenced by the low survival rate (0.13 ± 0.12) and the poor chlorophyll content (1.180 ± 0.076 mg g−1) recorded on plantlets derived from 15 g l−1 of sucrose. The treatment with 60 g l−1 of sucrose prior to acclimatization was efficient for roots induction and elongation. The analysis of pH revealed a fluctuation from one subculture to another, with an overall pH decrease under all treatments tested and, thus, indicates that plants release proton during growth. This feature had an impact on in vitro plantlets growth. The potentiality of the temporary immersion technique to foster the production of Xanthosoma sagittifolium is discussed.
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    ABSTRACT: Two essays were carried out to evaluate the effect of different types of auxins on root formation and the influence of dark and culture substratum on tetraploid hybrid FHIA-01 proliferation (Musa spp. AAAB). The plant material consisted of tissue culture plantlets of FHIA -01 hybrid tetraploid banana (Musa AAAB). The trial, with a total of 10 replicates per treatment was carried out in each pot containing five explants. For both tests, a combination of two cytokinins was enriched in culture substratum. The results obtained show that regeneration was high in culture substratum with light than substratum without meta-methoxytopolin riboside (M2). The medium M2 to the light induced a higher number of the buds compared to medium dose reduced meta-methoxytopolin riboside (M1). Meanwhile, only explants inoculated on the medium M1 in the dark induced callus. The bud proliferation, induction of root, leaf and the broadcast callus induction are significantly influenced by the different substratum and photoperiod, increasing the explant size, the number of emerged leaves, roots and the number of the weight of explant with buds proliferated. Formulating specific culture media cultivars according to group (ABB or AAA) and the choice of culture conditions (light intensity) would avoid consecutive failures and low proliferation in in vitro culture.
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May 15, 2014