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Schneider, J; Castilho, A; Pabst, M; Altmann, F; Gruber, C; Strasser, R; Gattinger, P; Seifert, GJ; Steinkellner, H; .
(2015): Characterization of plants expressing the human β1,4-galactosyltrasferase gene.
Plant Physiol Biochem. 2015; 92:39-47 FullText FullText_BOKU

Modification of the plant N-glycosylation pathway towards human type structures is an important strategy to implement plants as expression systems for therapeutic proteins. Nevertheless, relatively little is known about the overall impact of non-plant glycosylation enzymes in stable transformed plants. Here, we analyzed transgenic lines (Nicotiana benthamiana and Arabidopsis thaliana) that stably express a modified version of human beta 1,4-galactosyltransferase ((ST)GalT). While some transgenic plants grew normally, other lines exhibited a severe phenotype associated with stunted growth and developmental retardation. The severity of the phenotype correlated with both increased (ST)GalT mRNA and protein levels but no differences Were observed between N-glycosylation profiles of plants with and without the phenotype. In contrast to non-transgenic plants, all (ST)GalT expressing plants synthesized significant amounts of incompletely processed (largely depleted of core fucose) N-glycans with up to 40% terminally galactosylated structures. While transgenic plants showed no differences in nucleotide sugar composition and cell wall monosaccharide content, alterations in the reactivity of cell wall carbohydrate epitopes associated with arabinogalactan-proteins and pectic homogalacturonan were detected in (ST)GalT expressing plants. Notably, plants with phenotypic alterations showed increased levels of hydrogen peroxide, most probably a consequence of hypersensitive reactions. Our data demonstrate that unfavorable phenotypical modifications may occur upon stable in planta expression of non-native glycosyltransferases. Such important issues need to be taken into consideration in respect to stable glycan engineering in plants. (C) 2015 The Authors. Published by Elsevier Masson SAS.
Authors BOKU Wien:
Altmann Friedrich
De Vitoria Pereira Alexandra
Gattinger Pia Simone
Grünwald-Gruber Clemens
Pabst Martin
Seifert Georg
Steinkellner Herta
Strasser Richard
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Find related publications in this database (Keywords)
beta 1,4-Galactosylation
Nicotiana benthamiana
Transgenic plants
Developmental phenotype

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