Immunocytochemical localization of pectin and arabinogalactan protein epitopes in regenerating tissues of sugar beet (Beta vulgaris L.)
Magdalena Tomaszewska-Sowa
Magdalena.Tomaszewska-Sowa@pbs.edu.plKatedra Fizjologii i Podstaw Biotechnologii Roślin, Uniwersytet Technologiczno — Przyrodniczy, Bydgoszcz (Poland)
Abstract
Pectin and arabinogalactan proteins, which are components of the cell wall, are significantly associated with the regulation processes of morphogenesis and differentiation of plants. The aim of this study was to characterize the location of the epitopes of pectins and arabinogalactan proteins in tissues of sugar beet (Beta vulgaris L.) regenerating from unfertilized ovules in in vitro cultures. Considering previous results of immunocytochemical analysis of tissues of sugar beet it seemed important to determine the differences in distribution of these compounds in the cells of sugar beet depending on the type of tissue and medium components that determine the regeneration of explants in in vitro cultures. The presence of specific epitopes was determined in callus tissue with visible meristematic regions , as well as in somatic embryos at torpedo stage, growing in the culture of unfertilized ovules. Detection was conducted using specific monoclonal antibodies, and the location of specific oligosaccharide antigens was visualized in situ by reaction with secondary antibodies conjugated with alkaline phosphatase. It has been shown that the four epitopes typical for pectin, which react with antibodies JIM5, JIM7, LM5 and LM6 are common in all examined tissues and cell types. Among the AGP epitopes that typify arabinogalactan proteins there were distinguished those that occur in callus in relatively large quantities (LM2, and JIM14 JIM13), or only in trace amounts, or which in general are not present. In the control reaction, performed without the incubation stage of the primary antibody, the sections were obtained which showed no signs of coloration or minimally colored, which indicates specificity of the immunocytochemistry procedures.
Keywords:
sugar beet, in vitro culture, pectin, proteoglycans AGPs (arabinogalactan proteins), seeds, somatic embryosReferences
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Authors
Magdalena Tomaszewska-SowaMagdalena.Tomaszewska-Sowa@pbs.edu.pl
Katedra Fizjologii i Podstaw Biotechnologii Roślin, Uniwersytet Technologiczno — Przyrodniczy, Bydgoszcz Poland
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