Coexpression of Fungal Cell Wall-Modifying Enzymes Reveals Their Additive Impact on Arabidopsis Resistance to the Fungal Pathogen, Botrytis cinerea

SIMPLE SUMMARY: In the present study, we created transgenic Arabidopsis plants overexpressing two fungal acetylesterases and a fungal feruloylesterase that acts on cell wall polysaccharides and studied their possible complementary additive effects on host defense reactions against the fungal pathogen, Botrytis cinerea. Our results showed that the Arabidopsis plants overexpressing two acetylesterases together contributed significantly higher resistance to B. cinerea in comparison with single protein expression. Conversely, coexpression of either of the acetyl esterases together with feruloylesterase compensates the latter’s negative impact on plant resistance. The results also provided evidence that combinatorial coexpression of some cell wall polysaccharide-modifying enzymes might exert an additive effect on plant immune response by constitutively priming plant defense pathways even before pathogen invasion. These findings have potential uses in protecting valuable crops against pathogens. ABSTRACT: The plant cell wall (CW) is an outer cell skeleton that plays an important role in plant growth and protection against both biotic and abiotic stresses. Signals and molecules produced during host–pathogen interactions have been proven to be involved in plant stress responses initiating signal pathways. Based on our previous research findings, the present study explored the possibility of additively or synergistically increasing plant stress resistance by stacking beneficial genes. In order to prove our hypothesis, we generated transgenic Arabidopsis plants constitutively overexpressing three different Aspergillus nidulans CW-modifying enzymes: a xylan acetylesterase, a rhamnogalacturonan acetylesterase and a feruloylesterase. The two acetylesterases were expressed either together or in combination with the feruloylesterase to study the effect of CW polysaccharide deacetylation and deferuloylation on Arabidopsis defense reactions against a fungal pathogen, Botrytis cinerea. The transgenic Arabidopsis plants expressing two acetylesterases together showed higher CW deacetylation and increased resistance to B. cinerea in comparison to wild-type (WT) Col-0 and plants expressing single acetylesterases. While the expression of feruloylesterase alone compromised plant resistance, coexpression of feruloylesterase together with either one of the two acetylesterases restored plant resistance to the pathogen. These CW modifications induced several defense-related genes in uninfected healthy plants, confirming their impact on plant resistance. These results demonstrated that coexpression of complementary CW-modifying enzymes in different combinations have an additive effect on plant stress response by constitutively priming the plant defense pathways. These findings might be useful for generating valuable crops with higher protections against biotic stresses.