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GrainGenes Reference Report: PRT-13-188

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Reference	PRT-13-188
Title	Protoplast preparation without centrifugation: plant regeneration of barley (Hordeum vulgare L.
Journal	Plant Cell Reports
Year	1994
Volume	13
Pages	188-192
Author	Golds T Babczinsky J Mordhourst A Koop H
Abstract	Using a modification of the alginate film culture technique we show that it is possible to prepare and culture tobacco mesophyll and barley cell suspension protoplasts without centrifugation Comparable division frequencies and colony development were observed from protoplasts embedded with enzyme and protoplasts purified by centrifugation A 3 x 30 min washing regime was found ti- be the minimum time necessary to remove the enzyme from the gelled alginate matrix The procedure provides a more gentle method for isolating protoplasts It has the additional benefit of recovering all of the cells released from the starting tissue In particular, the smaller protoplasts that are frequently lost during conventional isolation, are maintained In barley, we illustrate the use of the system for recovering plants from embryogenic protoplast-derived calli from the cultivars Dissa and Igri. Finally, using small volumes of enzyme (50 microliter) single cell aggregates were used to isolate and culture protoplasts.
Keyword	[ Hide all but 1 of 12 ] alginate-film-culture-technique alginates cell-suspensions centrifugation cultivar culture-techniques film hordeum vulgare mesophyll nicotiana tabacum protoplast regenerative-ability

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