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The use of colloid gold labelling in the detection of plasma membrane from symbiotic and non‐symbiotic Glycine max root cells
Author(s) -
Ostrowski Elke D.,
Mellor Robert B.,
Werner Dietrich
Publication year - 1986
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.1399-3054.1986.tb02419.x
Subject(s) - protoplast , centrifugation , membrane , biochemistry , glucan , endoplasmic reticulum , sucrose , glycine , biology , differential centrifugation , biophysics , microbiology and biotechnology , amino acid
Glycine max L. Merr. cv. Maple Arrow protoplasts were prepared from both tissue‐cultured root cells and symbiotically‐infected (fix + ) nodule cells. Whilst both cell types showed glucan synthetase II (GS II; EC 2.4.1.29) activity, neither cell type, whole or gently disrupted, showed glucan synthetase I activity. After sucrose density gradient centrifugation one of the several GS II activity peaks co‐sedimented with the single radioactive particulate peak from [ 125 I]‐labelled protoplasts at 1.14 g ml −1 . This peak is presumed to be the plasma membrane peak because labelling of protoplasts with colloidal gold prior to disruption moved the 125 I peak and the corresponding glucan synthetase II activity into denser regions of the gradient, leaving endoplasmic reticulum‐contaminating IDPase (EC 3.1.3.31) and other glucan synthetase II peaks unmoved. Results are discussed in relation to various strategies of plasma membrane isolation.