Pathenogenesis-related (PR) proteins are extensively used as molecular markers to dissect the signaling cascades leading to plant defense responses. However, studies focusing on the biochemical or biological properties of these proteins remain rare. Here, we identify and characterize a class of apple (Malus domestica) PR proteins, named M. domestica AGGLUTININS (MdAGGs), belonging to the amaranthin-like lectin family. By combining molecular and biochemical approaches, we show that abundant production of MdAGGs in leaf tissues corresponds with enhanced resistance to the bacterium Erwinia amylovora, the causal agent of the disease fire blight. We also show that E. amylovora represses the expression of MdAGG genes by injecting the type 3 effector DspA/E into host cells and by secreting bacterial exopolysaccharides. Using a purified recombinant MdAGG, we show that the protein agglutinates E. amylovora cells in vitro and binds bacterial lipopolysaccharides at low pH, conditions reminiscent of the intercellular pH occurring in planta upon E. amylovora infection. We finally provide evidence that negatively charged polysaccharides, such as the free exopolysaccharide amylovoran progressively released by the bacteria, act as decoys relying on charge–charge interaction with the MdAGG to inhibit agglutination. Overall, our results suggest that the production of this particular class of PR proteins may contribute to apple innate immunity mechanisms active against E. amylovora.
Essential oils (EOs) are actively investigated as an alternative to numerous synthetic biocide products. Due to their large spectra of biological activities, the impact of EOs on non-target organisms should be characterized for biopesticide development purposes. In this study the potential phytotoxicity of Cinnamomum cassia EO (CEO) on apple trees (Malus domestica) were investigated in terms of oxidative burst (glutathione redox state) and damage (malondialdehyde). At 2%, CEO concentration the reduced glutathione leaf content drops from 269.6 ± 45.8 to 143 ± 28.4 nmol g−1FW, after 30 min, illustrating a rapid and strong oxidative burst. Regarding oxidative damage, malondialdehyde increased significantly 24 h post application to 10.7 ± 3.05 nmol g−1FW. Plant defence induction was previously suspected after trans-cinnamaldehyde (CEO main compound) application. Therefore, the elicitor potential was investigated by qRT-PCR, on the expression level of 29 genes related to major defence pathways (PR protein, secondary metabolism, oxidative stress, parietal modification). Multivariate analysis and increased expression levels suggest induction of systemic resistance. Hence, the present research illustrates the dose–dependent phytotoxicity of CEO in terms of lipid peroxidation. Transcriptional data illustrates the elicitor properties of CEO. These findings can help to design pest management strategies considering both their risks (phytotoxicity) and benefits (defence activation combined with direct biocide properties).
Phosphonate-based products have demonstrated diverse abilities to protect crops against pests, with various mode of actions proposed. In this paper, we specifically investigated the potassium phosphonate (KHP) on apple crop. Its performance to control three major apple bioagressors (Venturia inaequalis, Erwinia amylovora and Dysaphis plantaginea) was evaluated under semi-controlled conditions. The product was able to confer significant protection rates (40 to 75% for apple scab, 40% for fire blight and 30% for rosy aphid) which can be explained by its more or less efficient biocidal activity against the three pests, and by its ability to induce apple immunity (PR proteins and secondary metabolites genes). A cumulative effect of treatments as well as the systemic behavior of the product was demonstrated. Fields trials against apple scab and the postharvest disease bull’s eyes rot (Neofabraea vagabunda) were performed on different apple varieties, by applying KHP combined with light pest management programs either reducing (dessert orchards) or suppressing (cider orchards) fungicide applications. KHP was able to reduce apple scab by 70 to 90% on shoots, young and harvested fruits, and bull’s eyes rot by 70 to 90% on harvested fruits. Overall, our results indicate that KHP is useful for the protection of apple trees against its major pests by direct effect and by triggering the host defense system.
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