אסיף מאגר המחקר החקלאי

Scaled-up proliferation and regeneration of Nerine in liquid cultures Part I. The induction and maintenance of proliferating meristematic clusters by paclobutrazol in bioreactors

Year:

1994

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Volume :

39

Co-Authors:

Ziv, M., Department of Agricultural Botany, The Hebrew University of Jerusalem, PO Box 12, Rehovot, 76100, Israel, The Warburg Center for Biotechnology in Agriculture, The Hebrew University of Jerusalem, PO Box 12, Rehovot, 76100, Israel
Kahany, S., The Warburg Center for Biotechnology in Agriculture, The Hebrew University of Jerusalem, PO Box 12, Rehovot, 76100, Israel
Lilien-Kipnis, H., Department of Ornamental Horticulture, Agricultural Research Organization, PO Box 6, Bet-Dagan, 50250, Israel

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Abstract:

The natural propagation rate of bulb forming Amaryllidaceae including Nerine is low. Conventional micropropagation techniques are labor intensive and therefore expensive. Liquid cultures facilitate: scaling up, automation and cost reduction of micropropagation. Inflorescence-derived explants of Nerine were cultured on 2,4-dichlorophenoxyacetic acid (2,4-d) and 6-benzyladenine (BA) supplemented Murashige & Skoog (MS) medium. Callus-like tissue interspersed with nodular tissue, as well as direct organogenesis developed at the junction between flower pedicel and peduncle. Subculture of nodular tissue to 1-naphthalene acetic acid (NAA), BA and paclobutrazol (PAC) supplemented liquid medium in Erlenmeyer flasks or bubble bioreactors resulted in proliferation of rounded, compact, easily crumbled meristematic clusters. Growth and proliferation in bioreactors were higher than in shaken flasks and were affected differently by the inoculum to medium ratio in the two types of culture vessel. Nerine cultures showed low sensitivity to high aeration rates in bubble bioreactors despite the accumulation of debris. It was therefore possible to increase aeration rates without reducing the proliferation rate or damaging the quality of the meristematic aggregates. The conditions in semi-continuous culture in flasks and bioreactors were more favorable and increased the growth value by 100% and 140%, respectively. The total protein content increased by 180% in flasks and 90% in bioreactors. Although the presence of PAC throughout the culture period decreased growth and proliferation, it was a promotive bioregulator for meristernatic cluster formation. Proembryogenic clusters developed upon the removal of PAC. The use of meristematic clusters for micropropagation in scaled-up liquid cultures is discussed. © 1994 Kluwer Academic Publishers.

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