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Annals of Botany 90: 21-29, 2002
© 2002 Annals of Botany Company

Photoautotrophic Culture of Coffea arabusta Somatic Embryos: Development of a Bioreactor for Large-scale Plantlet Conversion from Cotyledonary Embryos

F. AFREEN{dagger},1, S. M. A. ZOBAYED*,{dagger},1 and T. KOZAI1

1 Department of Bioproduction Science, Chiba University, Matsudo, Chiba 271-8510, Japan

* For correspondence. Fax 00 1 519 7670755, e-mail szobayed{at}uoguelph.ca
{dagger} Present address: Department of Plant Agriculture, University of Guelph, Guelph, Ontario, Canada N1G 2W1

Received: 17 December 2001; Returned for revision: 2 February 2002; Accepted: 19 March 2002

Somatic embryos were developed from in vitro-grown leaf discs of Coffea arabusta in modified Murashige and Skoog medium under 30 µmol m–2 s–1 photosynthetic photon flux (PPF). Cotyledonary stage embryos were selected from the 14-week-old cultures and were placed under a high (100 µmol m–2 s–1) PPF for 14 d. These pretreated embryos were grown photoautotrophically in three different types of culture systems: Magenta vessel; RITA-bioreactor (modified to improve air exchange); and a specially designed temporary root zone immersion bioreactor system (TRI-bioreactor) with forced ventilation. The aims of the study were to achieve large-scale embryo-to-plantlet conversion, and to optimize growth of plantlets under photoautotrophic conditions. The plantlet conversion percentage was highest (84 %) in the TRI-bioreactor and lowest in the modified RITA-bioreactor (20 %). Growth and survival of converted plantlets following 45 d of photoautotrophic culture in each of the three culture systems were studied. Fresh and dry masses of leaves and roots of plantlets developed in the TRI-bioreactor were significantly greater than those of plantlets developed in the modified RITA-bioreactor or Magenta vessel. The net photosynthetic rate, chlorophyll fluorescence and chlorophyll contents were also highest in plantlets grown in the TRI-bioreactor. Normal stomata were observed in leaves of plantlets grown in the TRI-bioreactor, whereas they could be abnormal in plantlets from the modified RITA-bioreactor. Survival of the plants after transfer from culture followed a similar pattern and was highest in the group grown in the TRI-bioreactor, followed by plants grown in the modified RITA-bioreactor and Magenta vessel. In addition, ex vitro growth of plants transferred from the TRI-bioreactor was faster than that of plants from the other culture systems.

Key words: CO2 enrichment, embryo-to-plantlet conversion, ex vitro, forced ventilation, in vitro, stomata, survival percentage.


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