Barley scutellum protoplasts: isolation, culture and plant regeneration

A - Papers appearing in refereed journals

Nobre, J., Davey, M. R. and Lazzeri, P. A. 1996. Barley scutellum protoplasts: isolation, culture and plant regeneration. Physiologia Plantarum. 98 (4), pp. 868-874. https://doi.org/10.1034/j.1399-3054.1996.980427.x

AuthorsNobre, J., Davey, M. R. and Lazzeri, P. A.
Abstract

Many applications of cereal protoplast culture systems are still limited by the difficulties of regeneration from suspension cells which are the usual protoplast source. The objective of the present study therefore was to investigate the conditions for the development of a culture system for protoplasts capable of plant regeneration isolated directly from immmature scutella of barley The procedure developed involves a two-stage pre-culture of scutellar tissue, followed by vacuum infiltration with cell wall degrading enzymes and the culture of alginate-embedded protoplasts. The pre-culture of the scutella and the co-cultivation of protoplasts with nurse cells were the most important factors for the success of the culture system, but several other parameters affecting protoplast yield, viability and sustained division were identified including the developmental stage of the embryo, the use of cold conditioning periods during preculture, the composition of the pre-culture and protoplast culture medium, and the embedding matrix. Protoplasts isolated from scutellar tissues of barley cvs Dissa, Clipper, Derkado and Puffin were capable of sustained division in culture. Macroscopic protoplast-derived tissues were obtained in an cultivars, except cv. Puffin, and fertile plants were regenerated from cvs Dissa and Clipper 3-4 months after protoplast isolation. The procedure described provides a never approach for the isolation of totipotent protoplasts in barley which avoids the need for suspension cultures.

KeywordsPlant Sciences
Year of Publication1996
JournalPhysiologia Plantarum
Journal citation98 (4), pp. 868-874
Digital Object Identifier (DOI)https://doi.org/10.1034/j.1399-3054.1996.980427.x
Open accessPublished as non-open access
Funder project or code215
415
Project: 011060
ISSN00319317
PublisherWiley

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