Abstract
Degradation of pectin, a major component of plant cell wall, is important for fungal necrotrophs to achieve a successful infection. The activities of pectin methylesterases (PMEs) from both plants and pathogens and the degree and pattern of pectin methylesterification are critical for the outcome of plant–pathogen interaction. Partial degradation of pectin by pectin degrading enzymes releases oligogalacturonides (OGs), elicitors of plant defense responses. Few analytical techniques are available to monitor pectin methylesterification-modulating machineries and OGs produced during plant pathogen interaction. In the present study, ruthenium red is presented as useful dye to monitor both Botrytis cinerea mycelium growth and the induction of PME activity in plant tissue during fungal infection. Moreover a simple, inexpensive and sensitive method, named PECTOPLATE, is proposed that allows a simultaneous phenotyping of PME and pectinase activities expressed during pathogen infection and of pectinase potential in generating OGs. The results in the manuscript also indicate that PME inhibitors can be used in PECTOPLATE as a tool to discriminate the activities of plant PMEs from those of pathogen PMEs expressed during pathogenesis.
Highlights
The cell wall (CW) represents a barrier that pathogens need to breach to colonize the plant tissue
Ruthenium Red Allows Staining of Both B. cinerea Mycelium and pectin methylesterases (PMEs) Activity in Arabidopsis Leaf Tissue During Fungal Infection Specific Arabidopsis PME isoforms are induced during B. cinerea infection in Arabidopsis leaves (Raiola et al, 2011; Lionetti et al, 2012)
Progresses have been made in the isolation and characterization of CW and CW degrading enzymes involved in different physiological processes
Summary
The cell wall (CW) represents a barrier that pathogens need to breach to colonize the plant tissue. Emerging evidence points out that dynamic changes in the composition and structure of the CW during infection are exploited by both plant and pathogen to prevail during their interaction (Bellincampi et al, 2014; Lionetti and Metraux, 2014). Pectin is synthesized in the Golgi and secreted in a highly methyl esterified form into the CW. In this compartment, pectin methylesterases (PMEs, E.C. 3.1.1.11; Pfam 01095; CE8, www.cazy.org), catalyze the de-methylesterification of pectin releasing free carboxyl ester groups, protons and methanol
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