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Isolation and culture of gymnosperm root protoplasts (Pinus pinaster)
Author(s) -
Faye M.,
David A.
Publication year - 1983
Publication title -
physiologia plantarum
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.351
H-Index - 146
eISSN - 1399-3054
pISSN - 0031-9317
DOI - 10.1111/j.1399-3054.1983.tb04215.x
Subject(s) - protoplast , axenic , sorbitol , pinus pinaster , botany , gymnosperm , biology , horticulture , biochemistry , genetics , bacteria
Various factors affecting the yield and isolation of axenic protoplast cultures originating from Pinus pinaster root segments (in which most cells are differentiating) were studied. In spite of the use of plant material collected from germinating seeds under aseptic conditions, an additional sterilization with 0.1 % w/v mercuric chloride in 50% ethanol was a prerequisite for obtaining an axenic protoplast culture. A pretreatment with 30 mM cysteine in 0.7 M sorbitol for I h tripled the yield. Cen‐trifugation at lOOg instead of 40 g further increased the yield to 6 × 103 protoplasts per cm of root segment. Viability ranged from 80 to 91%. Cell divisions occurred after a minimum of 7 days of culture.