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Updated: May 5, 2026

A Simple Method for Isolation of Soybean Protoplasts and Application to Transient Gene Expression Analyses
Published on: January 25, 2018
Osmotic stress inhibits thymidine incorporation into soybean protoplast DNA
1Cell Culture and Nitrogen Fixation Laboratory, Plant Physiology Institute, Agricultural Research Service, U.S. Department of Agriculture, 20705, Beltsville, Maryland, USA.
Abstract:
DNA synthesis in protoplasts isolated from soybean cell suspension cultures has been investigated by [(3)H] thymidine uptake and incorporation kinetics. Initial rates of incorporation in exponential and 5-fluorodeoxyuridine synchronized protoplasts are inhibited by increased osmolarities of the medium. The inhibition was not readily reversible during 3 h culture in low osmotic medium. Velocity sedimentation analyses of replicating DNA from such protoplasts shows a complex pattern of inhibition. The inhibition probably effects replicon initiation as well as strand elongation and ligation of replication intermediates.
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