Fate of Myrosin Cells: Characterization of Monoclonal Antibodies Against Myrosinase

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Abstract
Immunofluorescence labelling of myrosinase in paraffin sections was used to study the fate, appearance, and distribution of myrosin cells in Brassica napus after seeding. Labelling was achieved using monoclonal antibodies (MAbs) produced against myrosinase. Myrosinase was extracted and purified from seeds of rape (Brassica napus) to homogeneity. Mice were immunized with highly purified myrosinase and after fusion and cloning of the hybridoma cells, specificity was tested with highly purified myrosinase and the antibodies were characterized. One monoclonal antibody inhibited myrosinase activity. Precipitation of myrosinase activity was achieved with antibodies coupled to monosized magnetic polymer particles (immunomagnetic precipitation). Labelled cells were found in all organs and also in vascular tissue. A large proportion of the labelled cells were found in the marginal zone, both in radicles and hypocotyl, supporting the theory that myrosinase and its substrates, the glucosinolates, take part in the defence system of the plants. A description of the morphological development of myrosin cells during seedling growth is presented.

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