PECTOPLATE: the simultaneous phenotyping of pectin methylesterases, pectinases, and oligogalacturonides in plants during biotic stresses

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.

Three Pectin Methylesterase Inhibitors Protect Cell Wall Integrity for Arabidopsis Immunity to Botrytis

“…3, B and F) and the plasma membrane aquaporin fused to mCherry (pm-rk) as a marker of the plasma membrane (Fig. 3, D and H;Nelson et al, 2007;De Caroli et al, 2011, 2015. At 36 h post transformation, the CW was fluorescently labeled by both AtPMEI11-GFP and AtPMEI12-GFP protein fusions (Fig.…”

“…After establishing that AtPMEI10, AtPMEI11, and AtPMEI12 play a role in immunity, we evaluated their possible contribution to the control of PME activity and pectin methylesterification during B. cinerea infection. The level of PME activity was monitored in wild-type, pmei10-1, pmei11-1, and pmei12-1 plants after B. cinerea infection using histochemical and biochemical assays (Lionetti, 2015). A higher increase of pathogen-induced PME activity was observed in mutant plants with respect to the wild type ( Fig.…”

“…Emerging clues indicate that PME activity and the level of pectin methylesterification are precisely and temporally regulated during the course of a plant disease. A steady increase in plant PME activity resulted in a reduced level of pectin methylesterification, which was observed in Arabidopsis during infection by different pathogens (Bethke et al, 2014;Lionetti, 2015). Genomewide transcriptional profiling suggests that specific Arabidopsis PMEs and PMEIs are expressed in response to pathogens and strictly dependent on the pathogen lifestyle (Lionetti et al, 2012;Windram et al, 2012).…”

Efficacy of Ozonized Water for Fungal Decontamination of Fresh Fruit Pieces Decorating Dessert Cakes

“…The filamentous fungi are reportedly involved in the efficient degradation of plant cell wall as they utilize the complex plant cell wall polysaccharides as a sole source of carbon by producing a wide range of hydrolases such as pectinases, cellulases, hemicellulases and amylases. Most fungi isolated from strawberries and other investigated fruits have been proved to be pectolytic and/or cellulolytic producer including Botrytis cinerae (Lionetti, 2015), Rhizopus oryzae (Chowdhury et al, 2017), Mucor spp. (Alves et al, 2002), P. italicum (Alaña et al, 1989).…”

Plant cell wall and the mechanisms of resistance to pathogens

“…Разработан метод мониторинга уровня инфекции по уровню гидролизованного пектина путем фенотипирования пектин-метилтрансфераз, пектиназ и олигогалактуронидов (Lionetti, 2015).…”