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: 23-32.

Author Chin J.C.
Scott K.J.
Title A large-scale isolation procedure for cereal mesophyll protoplasts
Journal name Annals of Botany   Check publisher's open access policy
ISSN 0305-7364
Publication date 1979-01-01
Sub-type Article (original research)
Volume 43
Issue 1
Start page 23
End page 32
Total pages 10
Subject 1804 Statistics, Probability and Uncertainty
2604 Applied Mathematics
3207 Social Psychology
3203 Clinical Psychology
2743 Reproductive Medicine
1110 Nursing
1108 Medical Microbiology
Abstract 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 cultivars of wheat, barley, maize, sorghum, and Triticale were diced and vacuum infiltrated with enzyme mixture composed of cellulysin (1 per cent w/v), hemicellulase (1 per cent w/v), and macerozyme (0.5 per cent w/v). With this procedure, yields of between 106 to 107 protoplasts per gram of leaves can be reproducibly obtained after only 1.5-3 h of enzymatic treatment. These protoplasts were almost 100 per cent viable (as determined by fluorescein diacetate staining) and incorporation of 3H-uridine and 14C-leucine into an acid-insoluble fraction was demonstrated. Almost one-third of the ribosomes of these isolated protoplasts were present as polysomes.
Keyword Cereals
Leaf mesophyll
Protoplast isolation
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Unknown

Document type: Journal Article
Sub-type: Article (original research)
Collection: Scopus Import - Archived
 
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Created: Tue, 16 Aug 2016, 14:45:53 EST by System User