Journal Article

Insights into the effects of polygalacturonase FaPG1 gene silencing on pectin matrix disassembly, enhanced tissue integrity, and firmness in ripe strawberry fruits

Paniagua, Candelas, Quesada, Miguel A., Blanco-Portales, Rosario, Cifuentes, Manuel, Posé, Sara, Mercado, José A.

2013

Overview

Antisense-mediated down-regulation of the fruit-specific polygalacturonase (PG) gene FaPG1 in strawberries (Fragaria×ananassa Duch.) has been previously demonstrated to reduce fruit softening and to extend post-harvest shelf life, despite the low PG activity detected in this fruit. The improved fruit traits were suggested to be attributable to a reduced cell wall disassembly due to FaPG1 silencing. This research provides empirical evidence that supports this assumption at the biochemical, cellular, and tissue levels. Cell wall modifications of two independent transgenic antisense lines that demonstrated a >90% reduction in FaPG1 transcript levels were analysed. Sequential extraction of cell wall fractions from control and ripe fruits exhibited a 42% decrease in pectin solubilization in transgenic fruits. A detailed chromatographic analysis of the gel filtration pectin profiles of the different cell wall fractions revealed a diminished depolymerization of the more tightly bound pectins in transgenic fruits, which were solubilized with both a chelating agent and sodium carbonate. The cell wall extracts from antisense FaPG1 fruits also displayed less severe in vitro swelling. A histological analysis revealed more extended cell–cell adhesion areas and an enhanced tissue integrity in transgenic ripe fruits. An immunohistological analysis of fruit sections using the JIM5 antibody against low methyl-esterified pectins demonstrated a higher labelling in transgenic fruit sections, whereas minor differences were observed with JIM7, an antibody that recognizes highly methyl-esterified pectins. These results support that the increased firmness of transgenic antisense FaPG1 strawberry fruits is predominantly due to a decrease in pectin solubilization and depolymerization that correlates with more tightly attached cell wall-bound pectins. This limited disassembly in the transgenic lines indicates that these pectin fractions could play a key role in tissue integrity maintenance that results in firmer ripe fruit.

• Publisher: Oxford University Press
• Subject: adhesion / Biological and medical sciences / Cell Wall / Cell Wall - genetics / Cell Wall - metabolism / Cell Wall - ultrastructure / Cell walls / Chromatography, Gel / Cirques / depolymerization / Down-Regulation / Electrophoresis, Polyacrylamide Gel / firmness / Fragaria / Fragaria - genetics / Fragaria - metabolism / Fragaria - ultrastructure / Fruit / Fruit - genetics / Fruit - metabolism / Fruit - ultrastructure / Fruits / Fundamental and applied biological sciences. Psychology / gel chromatography / Gels / Gene Expression Regulation, Enzymologic / Gene Expression Regulation, Plant / Gene Silencing / genes / genetically modified organisms / genetics / metabolism / Microscopy, Electron, Scanning / pectins / Pectins - metabolism / Plant physiology and development / Plant Proteins / Plant Proteins - genetics / Plant Proteins - metabolism / Plants, Genetically Modified / Plants, Genetically Modified - genetics / Plants, Genetically Modified - metabolism / Plants, Genetically Modified - ultrastructure / polygalacturonase / Polygalacturonase - genetics / Polygalacturonase - metabolism / Polymers / RESEARCH PAPER / Ripening / shelf life / sodium carbonate / Solubilization / Strawberries / Swelling / Transgenic plants / ultrastructure