RESUMEN
Molecular signals released by microbes at the surface of plant roots and leaves largely determine host responses, notably by triggering either immunity or symbiosis. How these signalling pathways cross-talk upon coincident perception of pathogens and symbionts is poorly described in plants forming symbiosis. Nitrogen fixing symbiotic Rhizobia spp. and arbuscular mycorrhizal fungi produce lipo-chitooligosaccharides (LCOs) to initiate host symbiotic programmes. In Medicago truncatula roots, the perception of LCOs leads to reduced efflux of reactive oxygen species (ROS). By contrast, pathogen perception generally triggers a strong ROS burst and activates defence gene expression. Here we show that incubation of M. truncatula seedlings with culture filtrate (CF) of the legume pathogen Aphanomyces euteiches alone or simultaneously with Sinorhizobium meliloti LCOs, resulted in a strong ROS release. However, this response was completely inhibited if CF was added after pre-incubation of seedlings with LCOs. By contrast, expression of immunity-associated genes in response to CF and disease resistance to A. euteiches remained unaffected by LCO treatment of M. truncatula roots. Our findings suggest that symbiotic plants evolved ROS inhibition response to LCOs to facilitate early steps of symbiosis whilst maintaining a parallel defence mechanisms toward pathogens.
Asunto(s)
Aphanomyces/fisiología , Quitina/análogos & derivados , Lípidos/química , Medicago truncatula/inmunología , Medicago truncatula/microbiología , Inmunidad de la Planta , Especies Reactivas de Oxígeno/metabolismo , Transducción de Señal , Quitina/metabolismo , Quitosano , Resistencia a la Enfermedad , Regulación de la Expresión Génica de las Plantas , Medicago truncatula/genética , Oligosacáridos , Enfermedades de las Plantas/inmunología , Enfermedades de las Plantas/microbiología , Inmunidad de la Planta/genética , Raíces de Plantas/genética , Raíces de Plantas/microbiología , Plantones/crecimiento & desarrollo , Plantones/fisiología , Sinorhizobium meliloti/fisiologíaRESUMEN
N-acetylglucosamine-based saccharides (chitosaccharides) are components of microbial cell walls and act as molecular signals during host-microbe interactions. In the legume plant Medicago truncatula, the perception of lipochitooligosaccharide signals produced by symbiotic rhizobia and arbuscular mycorrhizal fungi involves the Nod Factor Perception (NFP) lysin motif receptor-like protein and leads to the activation of the so-called common symbiotic pathway. In rice and Arabidopsis, lysin motif receptors are involved in the perception of chitooligosaccharides released by pathogenic fungi, resulting in the activation of plant immunity. Here we report the structural characterization of atypical chitosaccharides from the oomycete pathogen Aphanomyces euteiches, and their biological activity on the host Medicago truncatula. Using a combination of biochemical and biophysical approaches, we show that these chitosaccharides are linked to ß-1,6-glucans, and contain a ß-(1,3;1,4)-glucan backbone whose ß-1,3-linked glucose units are substituted on their C-6 carbon by either glucose or N-acetylglucosamine residues. This is the first description of this type of structural motif in eukaryotic cell walls. Glucan-chitosaccharide fractions of A. euteiches induced the expression of defense marker genes in Medicago truncatula seedlings independently from the presence of a functional Nod Factor Perception protein. Furthermore, one of the glucan-chitosaccharide fractions elicited calcium oscillations in the nucleus of root cells. In contrast to the asymmetric oscillatory calcium spiking induced by symbiotic lipochitooligosaccharides, this response depends neither on the Nod Factor Perception protein nor on the common symbiotic pathway. These findings open new perspectives in oomycete cell wall biology and elicitor recognition and signaling in legumes.
Asunto(s)
Aphanomyces/citología , Señalización del Calcio/efectos de los fármacos , Pared Celular/química , Quitina/farmacología , Glucanos/farmacología , Medicago truncatula/genética , Medicago truncatula/inmunología , Acetilglucosamina/metabolismo , Señalización del Calcio/genética , Núcleo Celular/efectos de los fármacos , Núcleo Celular/metabolismo , Quitina/química , Cromatografía en Gel , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Genes de Plantas , Glucanos/química , Interacciones Huésped-Patógeno/genética , Interacciones Huésped-Patógeno/inmunología , Espectroscopía de Resonancia Magnética , Espectrometría de Masas , Medicago truncatula/microbiología , Modelos Moleculares , Epidermis de la Planta/citología , Epidermis de la Planta/efectos de los fármacos , Raíces de Plantas/citología , Raíces de Plantas/efectos de los fármacosRESUMEN
Plant LysM proteins control the perception of microbial-derived N-acetylglucosamine compounds for the establishment of symbiosis or activation of plant immunity. This raises questions about how plants, and notably legumes, can differentiate friends and foes using similar molecular actors and whether any receptors can intervene in both symbiosis and resistance. To study this question, nfp and lyk3 LysM-receptor like kinase mutants of Medicago truncatula that are affected in the early steps of nodulation, were analysed following inoculation with Aphanomyces euteiches, a root oomycete. The role of NFP in this interaction was further analysed by overexpression of NFP and by transcriptome analyses. nfp, but not lyk3, mutants were significantly more susceptible than wildtype plants to A. euteiches, whereas NFP overexpression increased resistance. Transcriptome analyses on A. euteiches inoculation showed that mutation in the NFP gene led to significant changes in the expression of c. 500 genes, notably involved in cell dynamic processes previously associated with resistance to pathogen penetration. nfp mutants also showed an increased susceptibility to the fungus Colletotrichum trifolii. These results demonstrate that NFP intervenes in M. truncatula immunity, suggesting an unsuspected role for NFP in the perception of pathogenic signals.