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Subcellular localization of nuclease in barley aleurone
Author(s) -
Holstein Susanne E. H.,
Kobert Bettina,
Hillmer Stefan,
Brown Peter H.,
Ho T.H. David,
Robinson David G.
Publication year - 1991
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.1399-3054.1991.tb02150.x
Subject(s) - aleurone , immunogold labelling , endoplasmic reticulum , golgi apparatus , cell fractionation , biology , vacuole , organelle , biochemistry , nuclease , polyclonal antibodies , immunocytochemistry , subcellular localization , microbiology and biotechnology , cytoplasm , enzyme , antibody , ultrastructure , botany , immunology , endocrinology
The intracellular localization of an endonuclease (nuclease I) in barley aleurone responding to gibberellic acid was investigated by subcellular fractionation and immunocytochemistry with monoclonal and polyclonal antibodies. Organelle separations were performed with aleurone layers and protoplasts; immunefixations were carried out on protoplasts only. Nuclease was detected in fractions from isopycnic sucrose density gradients which were enriched in either endoplasmic reticulum or Golgi apparatus membranes. These two organelles were also labelled by the indirect immunogold method on thin sections. Intensive labelling of protein and developing vacuoles was observed. Therefore, as noted in other plants nuclease in barley is essentially a vacuolar enzyme.