RESUMO
The restriction of plant-symbiont dinitrogen fixation by an insect semiochemical had not been previously described. Here we report on a glycosylated triketide δ-lactone from Nephrotoma cornicina crane flies, cornicinine, that causes chlorosis in the floating-fern symbioses from the genus Azolla. Only the glycosylated trans-A form of chemically synthesized cornicinine was active: 500 nM cornicinine in the growth medium turned all cyanobacterial filaments from Nostoc azollae inside the host leaf-cavities into akinetes typically secreting CTB-bacteriocins. Cornicinine further inhibited akinete germination in Azolla sporelings, precluding re-establishment of the symbiosis during sexual reproduction. It did not impact development of the plant Arabidopsis thaliana or several free-living cyanobacteria from the genera Anabaena or Nostoc but affected the fern host without cyanobiont. Fern-host mRNA sequencing from isolated leaf cavities confirmed high NH4-assimilation and proanthocyanidin biosynthesis in this trichome-rich tissue. After cornicinine treatment, it revealed activation of Cullin-RING ubiquitin-ligase-pathways, known to mediate metabolite signaling and plant elicitation consistent with the chlorosis phenotype, and increased JA-oxidase, sulfate transport and exosome formation. The work begins to uncover molecular mechanisms of cyanobiont differentiation in a seed-free plant symbiosis important for wetland ecology or circular crop-production today, that once caused massive CO2 draw-down during the Eocene geological past.
Assuntos
Gleiquênias , Lactonas , Simbiose , Animais , Lactonas/metabolismo , Gleiquênias/fisiologia , Gleiquênias/microbiologia , Gleiquênias/efeitos dos fármacos , Dípteros/fisiologia , Glicosilação , Cianobactérias/metabolismo , Cianobactérias/fisiologia , Cianobactérias/genética , Nostoc/fisiologia , Nostoc/genética , Nostoc/metabolismo , Folhas de Planta/metabolismo , Folhas de Planta/fisiologiaRESUMO
Alsophila firma is a tree fern that is distributed mainly in tropical montane cloud forest (TMCF) and is considered as a threatened species. Arbuscular mycorrrhizal fungi (AMF) have been proposed as an alternative in rescue programs of endangered species. However, our knowledge about diversity of AMF and mycorrhizal status of the species of TMCF is limited. In Mexico TMCF shows different degrees of conservation because of fragmentation and land use change. In this study, we evaluated the level of colonization, richness and abundances of spores of AMF in three fragments with different conservation status: conserved (100 years), secondary vegetation (17 years) and disturbed. For this, soil samples and roots were collected from five individuals of A. firma per site, with at least 100m away from each other; a total of 100cm of roots were analysed per site. Root samples showed AMF and occasionally dark septate fungi (DSF) colonizations. For the overall study, 19 species of AMF were recorded: Gigaspora (7), Acaulospora (4), Glomus (4), Funneliformis (2), Sclerocystis (1) and Scutellospora (1). The dominant species in the three sites were Funneliformis geosporum and Acaulospora scrobiculata. The highest diversity (H’) and evenness (J’) (p<0.05) were found in the conserved site (H’=1.7, J’=0.66), when compared to the secondary vegetation (H’=1.5, J’=0.61), and the disturbed site (H’=0.74, J’=0.41). Statistical analysis showed that the AMF degree of colonization was significantly higher (p<0.05) in the conserved site; although, the disturbed site showed low richness and abundances of AMF, the degree of root colonization did not differ statistically (p<0.05) with the secondary vegetation site. Chao2 (Richness estimation model) showed that the number of analysed samples were sufficient to represent the structure of the AMF communities with values >90%. The present study confirmed that A. firma is a mycorrhizal species that exhibits high levels of colonization even in disturbed sites. We suggest that F. geosporum and A. scrobiculata may have the potential to inoculate the gametophyte and young sporophyte of A. firma, to support restoration programs, because of their abundances and high tolerance to disturbed sites.
Alsophila firma es un helecho arborescente que crece principalmente en bosque mesófilo de montaña (BMM) y se encuentra en la categoría de riesgo. Los hongos mico- rrízicos arbusculares (HMA) se han propuesto como una alternativa para programas de recuperación de especies nativas en peligro. Sin embargo, sabemos poco sobre la diversidad de HMA y del estatus micorrízico de las espe- cies vegetales del BMM. En México el BMM presenta diferentes grados de conservación debido a la fragmentación y el cambio de uso de suelo. Se evaluó el grado de colonización, riqueza y abundancia de esporas de los HMA presentes en fragmentos de BMM con diferente historia de manejo: conservado (100 años), en recuperación (17 años) y alterado. Se estudiaron muestras de suelo y raíces toma- das de 5 individuos de A. firma por cada sitio, con al menos 100m de distancia entre individuos. Se analizaron en total 100cm de raíz por cada sitio. Las muestras de raíz presentaron colonización principalmente de HMA y ocasionalmente por hongos septados oscuros (HSO). Se encontraron 19 especies: Gigaspora (7), Acaulospora (4), Glomus (4), Funneliformis (2), Sclerocystis (1) y Scutellospora (1). Las especies dominantes en los tres sitios fueron Funneliformis geosporum y Acaulospora scrobiculata. La mayor diversidad (H’) y equitatividad (J’) (p<0.05) se presentaron en el sitio conservado (H’=1.7; J’ 0.66) con respecto al sitio en recuperación (H’=1.5, J’ 0.61) y alterado (H’=0.74, J’=0.41). Los niveles de colonización micorrízica total de raíz fueron estadísticamente mayores (p<0.05) en el sitio conservado. Aunque el sitio alterado presenta una baja riqueza y abundancia de HMA la colonización micorrízica total de raíz no fue estadísticamente diferente (p<0.05) respecto al sitio en recuperación. El estimador Chao2 indica que las muestras analizadas reflejan la estructura general de la comunidad de HMA con valores superiores al 93%. El presente estudio confirma que A. firma es una especie micorrízica con altos niveles de colonización incluso en sitios alterados. Se sugiere que las especies F. geosporum y A. scrobiculata por su alta tolerancia a sitios alterados pueden tener potencial para inocular gametofitos y esporofitos de A. firma para incrementar la supervivencia en programas de restauración.