Immunocytological localization of two enzymes involved in berberine biosynthesis.
Using post-embedding immunogold techniques the cytological localization of the two branchpoint enzymes of isoquinoline biosynthesis, berberine bridge enzyme (BBE) and (S)-tetrahydroprotoberberine oxidase (STOX), was demonstrated. Electron-microscopic examination revealed their exclusive compartmentation within vesicles. After these vesicles have fused with the central vacuole, they release their contents, resulting in a characteristic electron-dense precipitate at the tonoplast. Vesicles of similar structure could be identified in young meristematic tissues of roots or shoots from different Berberis species and Papaver somniferum L. The appearance of electron-dense osmiophilic material is strictly correlated with the alkaloid content of the tissue. Immunocytological staining of P. somniferum tissue with antibodies directed against BBE led to a characteristic labeling of electron-dense aggregates in idioblasts that are not connected to the laticifer system. This localization demonstrates the strictly cytological separation of benzophenanthridine and morphine biosyntheses within this plant.
Bock
Armin
A
Department für Biologie I, LMU München, Menzinger Strasse 67, 80638 München, Germany.
Wanner
Gerhard
G
Zenk
Meinhart H
MH
D000470:Alkaloids / Q000378:metabolism; D044182:Benzylisoquinolines; D001599:Berberine / Q000031:analogs & derivatives* / Q000378:metabolism*; D001600:Berberine Alkaloids / Q000378:metabolism; D028142:Berberis / Q000166:cytology / Q000378:metabolism / Q000648:ultrastructure; D002478:Cells, Cultured; D031675:Eschscholzia / Q000166:cytology / Q000378:metabolism / Q000648:ultrastructure; D007150:Immunohistochemistry; D007546:Isoquinolines / Q000378:metabolism; D016253:Microscopy, Immunoelectron; D010088:Oxidoreductases / Q000378:metabolism*; D044925:Oxidoreductases Acting on CH-CH Group Donors; D010089:Oxidoreductases, N-Demethylating / Q000378:metabolism*; D010207:Papaver / Q000166:cytology / Q000378:metabolism / Q000648:ultrastructure; D059828:Plant Cells; D010944:Plants / Q000378:metabolism* / Q000648:ultrastructure; D014617:Vacuoles / Q000378:metabolism / Q000648:ultrastructure
D000470:Alkaloids; D044182:Benzylisoquinolines; D001600:Berberine Alkaloids; D007546:Isoquinolines; D001599:Berberine; C010941:discretamine; D010088:Oxidoreductases; C071690:reticuline oxidase; D044925:Oxidoreductases Acting on CH-CH Group Donors; C056768:tetrahydroprotoberberine oxidase; D010089:Oxidoreductases, N-Demethylating; C039109:isoquinoline; C004645:canadine; C003298:reticuline