Host specialization and spatial divergence of bacteria associated with Peltigera lichens promote landscape gamma diversity.

BACKGROUND: Lichens are micro-ecosystems relying on diverse microorganisms for nutrient cycling, environmental adaptation, and structural support. We investigated the spatial-scale dependency of factors shaping the ecological processes that govern lichen-associated bacteria. We hypothesize that lichens function as island-like habitats hosting divergent microbiomes and promoting landscape gamma-diversity. Three microenvironments -thalli, substrates, and neighboring soils- were sampled from four geographically overlapping species of Peltigera cyanolichens, spanning three bioclimatic zones in the Chilean Patagonia, to determine how bacterial diversity, assembly processes, ecological drivers, interaction patterns, and niche breadth vary among Peltigera microenvironments on a broad geographical scale. RESULTS: The hosts' phylogeny, especially that of the cyanobiont, alongside climate as a secondary factor, impose a strong ecological filtering of bacterial communities within Peltigera thalli. This results in deterministically assembled, low diverse, and phylogenetically convergent yet structurally divergent bacterial communities. Host evolutionary and geographic distances accentuate the divergence in bacterial community composition of Peltigera thalli. Compared to soil and substrate, Peltigera thalli harbor specialized and locally adapted bacterial taxa, conforming sparse and weak ecological networks. CONCLUSIONS: The findings suggest that Petigera thalli create fragmented habitats that foster landscape bacterial gamma-diversity. This underscores the importance of preserving lichens for maintaining a potential reservoir of specialized bacteria.

High Andean Steppes of Southern Chile Contain Little-Explored Peltigera Lichen Symbionts.

Peltigera lichens can colonize extreme habitats, such as high-elevation ecosystems, but their biodiversity is still largely unknown in these environments, especially in the southern hemi- sphere. We assessed the genetic diversity of mycobionts and cyanobionts of 60 Peltigera lichens collected in three high Andean steppes of southern Chile using LSU, ß-tubulin, COR3 and ITS loci for mycobionts, and SSU and rbcLX loci for cyanobionts. We obtained 240 sequences for the different mycobiont markers and 118 for the cyanobiont markers, including the first report of ß-tubulin sequences of P. patagonica through modifying a previously designed primer. Phylogenetic analyses, ITS scrutiny and variability of haplotypes were used to compare the sequences with those previously reported. We found seven mycobiont species and eleven cyanobiont haplotypes, including considerable novel symbionts. This was reflected by ~30% of mycobionts and ~20% of cyanobionts haplotypes that yielded less than 99% BLASTn sequence identity, 15 new sequences of the ITS1-HR, and a putative new Peltigera species associated with 3 Nostoc haplotypes not previously reported. Our results suggest that high Andean steppe ecosystems are habitats of unknown or little-explored lichen species and thus valuable environments to enhance our understanding of global Peltigera biodiversity.

Peltigera frigida Lichens and Their Substrates Reduce the Influence of Forest Cover Change on Phosphate Solubilizing Bacteria.

Phosphorus (P) is one of the most critical macronutrients in forest ecosystems. More than 70 years ago, some Chilean Patagonian temperate forests suffered wildfires and the subsequent afforestation with foreign tree species such as pines. Since soil P turnover is interlinked with the tree cover, this could influence soil P content and bioavailability. Next to soil microorganisms, which are key players in P transformation processes, a vital component of Patagonian temperate forest are lichens, which represent microbial hotspots for bacterial diversity. In the present study, we explored the impact of forest cover on the abundance of phosphate solubilizing bacteria (PSB) from three microenvironments of the forest floor: Peltigera frigida lichen thallus, their underlying substrates, and the forest soil without lichen cover. We expected that the abundance of PSB in the forest soil would be strongly affected by the tree cover composition since the aboveground vegetation influences the edaphic properties; but, as P. frigida has a specific bacterial community, lichens would mitigate this impact. Our study includes five sites representing a gradient in tree cover types, from a mature forest dominated by the native species Nothofagus pumilio, to native second-growth forests with a gradual increase in the presence of Pinus contorta in the last sites. In each site, we measured edaphic parameters, P fractions, and the bacterial potential to solubilize phosphate by quantifying five specific marker genes by qPCR. The results show higher soluble P, labile mineral P, and organic matter in the soils of the sites with a higher abundance of P. contorta, while most of the molecular markers were less abundant in the soils of these sites. Contrarily, the abundance of the molecular markers in lichens and substrates was less affected by the tree cover type. Therefore, the bacterial potential to solubilize phosphate is more affected by the edaphic factors and tree cover type in soils than in substrates and thalli of P. frigida lichens. Altogether, these results indicate that the microenvironments of lichens and their substrates could act as an environmental buffer reducing the influence of forest cover composition on bacteria involved in P turnover.

Substrates of Peltigera Lichens as a Potential Source of Cyanobionts.

Photobiont availability is one of the main factors determining the success of the lichenization process. Although multiple sources of photobionts have been proposed, there is no substantial evidence confirming that the substrates on which lichens grow are one of them. In this work, we obtained cyanobacterial 16S ribosomal RNA gene sequences from the substrates underlying 186 terricolous Peltigera cyanolichens from localities in Southern Chile and maritime Antarctica and compared them with the sequences of the cyanobionts of these lichens, in order to determine if cyanobacteria potentially available for lichenization were present in the substrates. A phylogenetic analysis of the sequences showed that Nostoc phylotypes dominated the cyanobacterial communities of the substrates in all sites. Among them, an overlap was observed between the phylotypes of the lichen cyanobionts and those of the cyanobacteria present in their substrates, suggesting that they could be a possible source of lichen photobionts. Also, in most cases, higher Nostoc diversity was observed in the lichens than in the substrates from each site. A better understanding of cyanobacterial diversity in lichen substrates and their relatives in the lichens would bring insights into mycobiont selection and the distribution patterns of lichens, providing a background for hypothesis testing and theory development for future studies of the lichenization process.

Perfil químico y actividad antibacterial de los extractos de Peltigera laciniata (Merrill ex Riddle) Gyeln / Chemical profile and antibacterial activity of extracts from Peltigera laciniata (Merrill ex Riddle) Gyeln

Introducción: los líquenes, al presentar metabolitos secundarios como xantonas, antraquinonas y alcaloides, se han postulado como material con alto potencial biológico (e. g. antibiótico y antiviral), siendo el antibacterial muy promisorio, el cual se determina por medio de antibiogramas por difusión, punto central de esta investigación. Objetivo: evaluar la actividad antibacterial de los extractos de Peltigera laciniata (Merrill ex Riddle) Gyeln. Olmo de hoja cortada. Métodos: el material liquénico se sometió a percolación con etanol 96 por ciento. Al extracto crudo etanólico se le realizó el aislamiento de alcaloides totales y flavonoides totales con adición de HCL 3 por ciento y metanol, respectivamente. Ambas fracciones, fueron monitoreados por cromatografía de capa fina y fraccionados utilizando cromatografía de columna. Los extractos y fracciones se sometieron a bioensayos sobre Escherichia coli y Staphylococcus aureus para la valoración de los halos de inhibición, utilizando como control Sultamicilina. Los ensayos fueron realizados tres veces con 2 réplicas. Resultados: al realizar la separación cromatográfica de los alcaloides, se observó aumento de la inhibición en comparación con la mezcla alcaloidal. La fracción A1 presenta valores de inhibición cercanos al control y presentó los menores valores de inhibición con respecto a los demás tratamientos evaluados. El efecto de la fracción de los flavonoides totales tuvo menor impacto sobre E. coli y S. aureus, sin embargo, es importante destacar la acción antibacterial de los compuestos nitrogenados de tipo alcaloidal sobre microoganismos Gram positivos. Conclusiones: en el perfil químico realizado a partir de los extractos de la especie de estudio se visualizó la presencia de metabolitos secundarios de tipo alcaloide y flavonoide...(AU)

Introduction: Due to their content of secondary metabolites such as xanthones, anthraquinones and alkaloids, lichens have been suggested to be a material of high biological potential (e.g. antibiotic and antiviral). Their very promising antibacterial potential may be determined by diffusion antibiograms, the main concern of the present study. Objective: Evaluate the antibacterial activity of extracts obtained from Peltigera laciniata (Merrill ex Riddle) Gyeln, cutleaf elm. Methods: The lichenic material was percolated with 96 percent ethanol. Total alkaloids and total flavonoids were isolated from the crude ethanolic extract by adding 3 percent HCL and methanol, respectively. Both fractions were monitored by thin-layer chromatography and fractioned by column chromatography. Extracts and fractions were subjected to bioassays against Escherichia coli and Staphylococcus aureus for inhibition haloes, using sultamicillin as control. The assays were conducted 3 times with 2 replications. Results: Upon chromatographic separation of the alkaloids, an increase was observed in inhibition when compared with the alkaloidal mixture. Fraction A1 displayed inhibition values close to the control. Fraction FT showed lower inhibition values than the other treatments evaluated. The fraction of total flavonoids had a lesser impact on E. coli and S. aureus, but alkaloidal nitrogenated compounds had significant antibacterial activity against Gram-positive microorganisms. Conclusions: The chemical profile of extracts from the study species revealed the presence of alkaloidal and flavonoidal secondary metabolites, as well as the antimicrobial effect of the alkaloids contained in the extract and the fraction. This confirms the antibacterial pharmacological potential attributed to the protoberberine core(AU)

Perfil químico y actividad antibacterial de los extractos de Peltigera laciniata (Merrill ex Riddle) Gyeln / Chemical profile and antibacterial activity of extracts from Peltigera laciniata (Merrill ex Riddle) Gyeln

Introducción: los líquenes, al presentar metabolitos secundarios como xantonas, antraquinonas y alcaloides, se han postulado como material con alto potencial biológico (e. g. antibiótico y antiviral), siendo el antibacterial muy promisorio, el cual se determina por medio de antibiogramas por difusión, punto central de esta investigación. Objetivo: evaluar la actividad antibacterial de los extractos de Peltigera laciniata (Merrill ex Riddle) Gyeln. Olmo de hoja cortada. Métodos: el material liquénico se sometió a percolación con etanol 96 por ciento. Al extracto crudo etanólico se le realizó el aislamiento de alcaloides totales y flavonoides totales con adición de HCL 3 por ciento y metanol, respectivamente. Ambas fracciones, fueron monitoreados por cromatografía de capa fina y fraccionados utilizando cromatografía de columna. Los extractos y fracciones se sometieron a bioensayos sobre Escherichia coli y Staphylococcus aureus para la valoración de los halos de inhibición, utilizando como control Sultamicilina. Los ensayos fueron realizados tres veces con 2 réplicas. Resultados: al realizar la separación cromatográfica de los alcaloides, se observó aumento de la inhibición en comparación con la mezcla alcaloidal. La fracción A1 presenta valores de inhibición cercanos al control y presentó los menores valores de inhibición con respecto a los demás tratamientos evaluados. El efecto de la fracción de los flavonoides totales tuvo menor impacto sobre E. coli y S. aureus, sin embargo, es importante destacar la acción antibacterial de los compuestos nitrogenados de tipo alcaloidal sobre microoganismos Gram positivos. Conclusiones: en el perfil químico realizado a partir de los extractos de la especie de estudio se visualizó la presencia de metabolitos secundarios de tipo alcaloide y flavonoide, evidenciando el efecto antimicrobiano de los alcaloides presentes en el extracto y la fracción, lo cual ratifica el potencial farmacológico de tipo antibacterial, atribuido al núcleo Protoberberínico(AU)

Introduction: Due to their content of secondary metabolites such as xanthones, anthraquinones and alkaloids, lichens have been suggested to be a material of high biological potential (e.g. antibiotic and antiviral). Their very promising antibacterial potential may be determined by diffusion antibiograms, the main concern of the present study. Objective: Evaluate the antibacterial activity of extracts obtained from Peltigera laciniata (Merrill ex Riddle) Gyeln, cutleaf elm. Methods: The lichenic material was percolated with 96 percent ethanol. Total alkaloids and total flavonoids were isolated from the crude ethanolic extract by adding 3 percent HCL and methanol, respectively. Both fractions were monitored by thin-layer chromatography and fractioned by column chromatography. Extracts and fractions were subjected to bioassays against Escherichia coli and Staphylococcus aureus for inhibition haloes, using sultamicillin as control. The assays were conducted 3 times with 2 replications. Results: Upon chromatographic separation of the alkaloids, an increase was observed in inhibition when compared with the alkaloidal mixture. Fraction A1 displayed inhibition values close to the control. Fraction FT showed lower inhibition values than the other treatments evaluated. The fraction of total flavonoids had a lesser impact on E. coli and S. aureus, but alkaloidal nitrogenated compounds had significant antibacterial activity against Gram-positive microorganisms. Conclusions: The chemical profile of extracts from the study species revealed the presence of alkaloidal and flavonoidal secondary metabolites, as well as the antimicrobial effect of the alkaloids contained in the extract and the fraction. This confirms the antibacterial pharmacological potential attributed to the protoberberine core(AU)

La relación entre peltigera sp. y rhizophora magle en arroyo de plata (Bolívar), caribe Colombiano / The relationhip between peltigera sp. and rhizophora manglein arroyo de pllata (bolivar), Colombian caribbean region

Objetivo: determinar la distribución y el estado de los rodales de manglar en Arroyo de Plata (Departamento de Bolívar). Metodología: se realizaron labores de campo a través del arbolado, mediante el muestreo biológico de la flora y de los parámetros físicos y químicos del agua y de los suelos a lo largo de transectos dentro del bosque. Resultados: la composición del rodal incluye cinco especies, Rhizophora mangle (mangle rojo, colorao o colorado), Avicennia germinans (mangle salado, prieto, negro o iguanero), Laguncularia racemosa (mangle blanco, amarillo o bobo), Conocarpus erecta (mangle zaragoza) y Pelliciera rhizophorae (mangle piñuelo). Asociado a la especie pionera (R. mangle) se encontró el líquen (Peltigera sp.), cuya relación se lleva a cabo solo cuando el mangle está en estado de degradación y los parámetros fisicoquímicos de las aguas y los suelos, son extremos. Conclusión: Los manglares mostraron características mixtas, tipo maduro ribereño y de cuenca, donde la especie invasora P. rhizophorae presenta el mayor IVI, debido a su alta densidad. El registro de los líquenes Peltigera sp. y su presencia en los manglares del Caribe colombiano no solo es el primero para esta Región, su distribución puede estar restringida a la zona costera, es incidental, y demuestra su gran capacidad de adaptación a los cambios ambientales, y puede ser considerada entre los en bio-indicadores de las características ambientales, especialmente de las alteraciones actuales de los suelos y las aguas.

Objective: To determine the distribution and status of mangrove stands in Arroyo de Plata (Department of Bolívar). Methodology: Field work was conducted through woodland, using biological sampling of the flora and the physical and chemical parameters of water and soil along transects within the forest. Results: The composition of the stand includes five species, Rhizophoramangle (red, colorao or colorado mangrove), Avicenniagerminans (salty, dark color, black or iguanero mangle) Laguncularia racemosa (white, yellow or bobo mangrove), Conocarpus erecta (zaragoza mangle) and Pelliciera rhizophorae (tea mangrove). The lichen (Peltigera sp), was found associated with the pioneer species (R. mangrove) whose relationship is carried out only when the mangrove is in a state of degradation and the physical-chemical parameters of water and soil, are extreme. Conclusion: Mangroves showed mature coastal basin type mixed characteristics, where the invasive species P. rhizophorae has the highest IVI due to its high density. Registration of Peltigera sp lichens and its presence of the Colombian Caribbean mangrove is not only the first for this region, its distribution may be restricted to the coastal zone, it is incidental and demonstrates its ability to adapt to environmental changes, and can be considered between the bio-indicators of environmental characteristics, especially of the the current alterations of soil and water.