Visualisation of transglutaminase-mediated cross-linking activity in germinating pollen by laser confocal microscopy

Autor: Alessia Di Sandro, Donatella Serafini-Fracassini, Alessandra Scarpellini, Elisabetta A.M. Verderio, Stefano Del Duca, Rosa Anna Iorio
Přispěvatelé: Iorio RA, Di Sandro A, Scarpellini A, Del Duca S, Serafini-Fracassini D., Verderio E.
Rok vydání: 2008
Předmět:
Zdroj: Plant Biosystems - An International Journal Dealing with all Aspects of Plant Biology. 142:360-365
ISSN: 1724-5575
1126-3504
DOI: 10.1080/11263500802150886
Popis: Transglutaminases (TGs) are a multigenic family of calcium-dependent protein cross-linking enzymes, which are present in animal and plant cells. We have previously reported the presence of TGs in the cytosol and, more recently, in the cell wall of Malus domestica pollen, where it may be involved in pollen germination and pollen–stylar interactions. In this report we describe a simple method for the in situ visualisation of TG activity in germinating pollen. The method is based on the incorporation, mediated by pollen TG, of a fluorescently labelled exogenous diamine substrate of TG (fluorescein-cadaverine) into endogenous pollen substrates. Following the in situ TG activity reaction, the presence of cross-linked pollen proteins was visualised in fixed specimens of germinated pollen by laser confocal microscopy. Our data indicate the presence of TG cross-linking activity mainly at the apical part of the pollen tube, in the region proximal to the grain, and in the pollen grain itself. In planta, th...
Databáze: OpenAIRE
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