Oligosaccharides induce changes in protein patterns of regenerating spruce protoplasts

Abstract

AbstractGalactoglucomannan oligosaccharides (GGMOs, d.p. 4–8 and fractions d.p. 3, 4, 5, 6–7), used in culture media for spruce protoplasts derived from callus cells showed a pleiotropic effect. They influenced both, quality and quantity of extracellular proteins in regenerating protoplasts. GGMOs d.p. 4, 5 at pH 6.0 and the mixture of d.p. 4–8 (pH 3.8 and 6.0) after 48 h of culture increased the amount of extra- and intracellular proteins and the viability of protoplasts. The most significant effect on protoplasts viability in the presence of GGMOs d.p. 4–8 without the growth hormone supplementation at pH 3.8, and with lower efficiency in the presence of NAA (1 mg/l) has been observed. The most significant differences were observed in the molecular mass intervals Mr∼17–21 kDa, 25–30 kDa, and 45–60 kDa. After 24 h of culture extracellular acid proteins with Mr∼30; 41; 68; and 90 in media supplemented with GGMOs d.p. 3, 4, 5, 6–7 (pH 6.0) with or without NAA were identified to belong to the group of β-1,3-glucanases. Extracellular proteins p27.5; 30; 41; 50; 52 and 90 exhibited chitinase activity after 24 h of protoplast cultivation. GGMOs probably fulfil a protective role in this process of spruce protoplast regeneration.