A large-scale isolation procedure for cereal mesophyll protoplasts

Chin, J.C. and Scott, K.J. (1979) A large-scale isolation procedure for cereal mesophyll protoplasts. Annals of Botany, 43 (1). pp. 23-32.

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A method suitable for the large-scale isolation of cereal protoplasts from up to 50 g of leaf material is described. Surface-sterilized leaves from cv. of wheat, barley, maize, sorghum and triticale were diced and vacuum infiltrated with enzyme mixture composed of cellulysin (1% w/v), hemicellulase (1% w/v), and macerozyme (0.5% w/v). With this procedure, yields of between 106 and 107 protoplasts/g of leaves can be reproducibly obtained after only 1.5-3 h of enzymatic treatment. These protoplasts were almost 100% viable (as determined by fluorescein diacetate staining) and incorporation of 3H-uridine and 14C-leucine into an acid-insoluble fraction was demonstrated. Almost 33% of the ribosomes of these isolated protoplasts were present as polysomes

Item Type: Article
Author Affiliation: Department of Biochemistry, Queensland University, St. Lucia, Queensland, Australia 4067
Subjects: Plant Production
Divisions: Sorghum
Depositing User: Ms K Syamalamba
Date Deposited: 05 Oct 2012 09:09
Last Modified: 05 Oct 2012 09:09
URI: http://eprints.icrisat.ac.in/id/eprint/8281

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