Characteristics of spatial and temporal variability in the distribution of biological soil crusts on the Loess Plateau, China.

Biological soil crust (biocrust) is widely distributed on the Loess Plateau and plays multiple roles in regulating ecosystem stability and multifunctionality. Few reports are available on the distribution characteristics of biocrust in this region, which limits the assessment of its ecological functions. Based on 388 sampling points in different precipitation zones on the Loess Plateau from 2009 to 2020, we analyzed the coverage, composition, and influencing factors of biocrust across different durations since land abandonment, precipitation levels, topography (slope aspect and position), and utilization of abandoned slopelands (shrubland, forest, and grassland). On this base, with the assistance of machine learning and spatial modeling methods, we generated a distribution map of biocrust and its composition at a resolution of 250 m × 250 m, and analyzed the spatial distribution of biocrust on the Loess Plateau. The results showed that the average biocrust coverage in the woodlands and grasslands was 47.3%, of which cyanobacterial crust accounted for 25.5%, moss crust 19.7%, and lichen crust 2.1%. There were significant temporal and spatial variations. Temporally, the coverage of biocrust in specific regions fluctuated with the extension of the abandoned durations and coverage of cyanobacterial crust, while moss crust showed a reverse pattern. In addition, the coverage of biocrust in the wet season was slightly higher than that in the dry season within a year. Spatially, the coverage of biocrusts on the sandy lands area on the Loess Plateau was higher and dominated by cyanobacterial crusts, while the coverage was lower in the hilly and gully area. Precipitation and utilization of abandoned land were the major factors driving biocrust coverage and composition, while slope direction and position did not show obvious effect. In addition, soil organic carbon content, pH, and texture were related to the distribution of biocrust. This study uncovered the spatial and temporal variability of biocrust distribution, which might provide important data support for the research and management of biocrust in the Loess Plateau region.

Diversity patterns of vascular and non-vascular epiphytes along tropical dry forest / Patrones de diversidad de epífitas vasculares y no vasculares a lo largo del bosque seco tropical

Abstract Introduction: Epiphytes (vascular and non-vascular) are one of the most diverse groups in the Neotropics, but despite their importance in the functioning of many ecosystems, much of their taxonomy and ecology is still unknown in the dry forest of Colombia. Objective: To compare the diversity patterns and species composition of vascular and non-vascular epiphytes along tropical dry forest remnants of Cauca Valley, Colombia. Methods: Ten permanent plots (50 x 20 m2) were established in tropical dry forest remnants. The epiphytes were sampled in 40-50 trees per plot. Alpha and gamma diversity were calculated using the richness (q0) and Shannon index (q1) (alpha was estimated as the average for phorophytes). Beta diversity was measured using the Whitaker index. To evaluate the relationship between diversities and environmental variables, GLM analysis was used. Results: We found 50 morphospecies of vascular epiphytes, 77 of bryophytes and 290 of lichens. The 𝛼 and 𝛾 diversity of bryophytes from each remnant was significantly explained by temperature. The abundance of lichens per tree was significantly related with the DBH and tree height of each remnant. The 𝛼 diversity of vascular epiphytes shown can be explained by temperature and precipitation. The 𝛾 diversity was strongly influenced by the beta diversity in bryophytes and lichens. This pattern is because the sites with high disturbance present a lower diversity, as a consequence of a homogenizing effect, that is, a low turnover of species between sampling units. Conclusions: Precipitation and temperature affected the diversity of bryophytes and vascular epiphytes, while it did not show a relationship with the lichen's diversity, for which there is not a high congruence between the diversity and composition of these epiphytes.

Resumen Introducción: Los epífitos (vasculares y no vasculares) son uno de los más diversos grupos de plantas en el Neotrópico, pero a pesar de su importancia para el funcionamiento de varios ecosistemas, existen grandes vacíos en su conocimiento taxonómico y ecológico en el bosque seco de Colombia. Objetivo: Comparar los patrones de diversidad y composición de especies de epífitas vasculares y no vasculares a lo largo de remanentes de bosque seco tropical del Valle del Cauca, Colombia. Métodos: Se establecieron diez parcelas permanentes (50 x 20 m2) en remanentes de bosque seco tropical. Las epífitas se muestrearon en 40-50 árboles por parcela. La diversidad alfa y gamma se calculó utilizando la riqueza (q0) y el índice de Shannon (q1) (alfa se estimó como el promedio para los forófitos). La diversidad beta se midió utilizando el índice de Whitaker. Para evaluar la relación entre diversidades y variables ambientales se utilizó el análisis GLM. Resultados: Se encontraron 50 morfoespecies de epífitas vasculares, 77 de briófitas y 290 de líquenes. La diversidad de 𝛼 y 𝛾 de briófitas de cada remanente fue explicada significativamente por la temperatura. La abundancia de líquenes por árbol se relacionó significativamente con el DAP y la altura del árbol de cada remanente. La diversidad 𝛼 de epífitas vasculares que se muestra puede explicarse por la temperatura y la precipitación. La diversidad 𝛾 estuvo fuertemente influenciada por la diversidad beta en briófitas y líquenes. Este patrón se debe a que los sitios con alta perturbación presentan una menor diversidad, como consecuencia de un efecto homogeneizador, es decir, un bajo recambio de especies entre unidades de muestreo. Conclusiones: La precipitación y la temperatura afectaron la diversidad de briófitas y epífitas vasculares, mientras que no mostró relación con la diversidad de líquenes, por lo que no existe una alta congruencia entre la diversidad y composición de estas epífitas.

Depside and Depsidone Synthesis in Lichenized Fungi Comes into Focus through a Genome-Wide Comparison of the Olivetoric Acid and Physodic Acid Chemotypes of Pseudevernia furfuracea.

Primary biosynthetic enzymes involved in the synthesis of lichen polyphenolic compounds depsides and depsidones are non-reducing polyketide synthases (NR-PKSs), and cytochrome P450s. However, for most depsides and depsidones the corresponding PKSs are unknown. Additionally, in non-lichenized fungi specific fatty acid synthases (FASs) provide starters to the PKSs. Yet, the presence of such FASs in lichenized fungi remains to be investigated. Here we implement comparative genomics and metatranscriptomics to identify the most likely PKS and FASs for olivetoric acid and physodic acid biosynthesis, the primary depside and depsidone defining the two chemotypes of the lichen Pseudevernia furfuracea. We propose that the gene cluster PF33-1_006185, found in both chemotypes, is the most likely candidate for the olivetoric acid and physodic acid biosynthesis. This is the first study to identify the gene cluster and the FAS likely responsible for olivetoric acid and physodic acid biosynthesis in a lichenized fungus. Our findings suggest that gene regulation and other epigenetic factors determine whether the mycobiont produces the depside or the depsidone, providing the first direct indication that chemotype diversity in lichens can arise through regulatory and not only through genetic diversity. Combining these results and existing literature, we propose a detailed scheme for depside/depsidone synthesis.

Tree species identity and composition shape the epiphytic lichen community of structurally simple boreal forests over vast areas.

Greatly simplified ecosystems are often neglected for biodiversity studies. However, these simplified systems dominate in many regions of the world, and a lack of understanding of what shapes species occurrence in these systems can have consequences for biodiversity and ecosystem services at a massive scale. In Fennoscandia, ~90% of the boreal forest (~21Mha) is structurally simplified with little knowledge of how forest structural elements shape the occurrence and diversity of for example epiphytic lichens in these managed forests. One form of structural simplification is the reduction of the number and frequency of different tree species. As many lichen species have host tree preferences, it is particularly likely that this simplification has a huge effect on the lichen community in managed forests. In a 40-70 years old boreal forest in Sweden, we therefore related the occurrence and richness of all observed epiphytic lichens to the host tree species and beta and gamma lichen diversity at the forest stand level to the stand's tree species composition and stem diameter. Picea abies hosted the highest lichen richness followed by Pinus sylvestris, Quercus robur, Alnus glutinosa, Betula spp., and Populus tremula. However, P. tremula hosted twice as many uncommon species as any of the other tree species. Stand level beta and gamma diversity was twice as high on stands with four compared to one tree species, and was highest when either coniferous or deciduous trees made up 40-50% of the trees. The stem diameter was positively related to lichen richness at the tree and stand level, but negatively to beta diversity. For biodiversity, these findings imply that leaving a few trees of a different species during forest thinning is unlikely as effective as combining life-boat trees for endangered species with an even tree species mixture.

Melanin from the Lichens Cetraria islandica and Pseudevernia furfuracea: Structural Features and Physicochemical Properties.

Lichens are symbiotic photosynthesizing organisms with thalli formed by fungi and algae/cyanobacteria that possess high stress tolerance. One of the factors that contributes to the ability of a lichen to tolerate harsh environmental conditions is the presence of unique metabolites, including high-molecular-weight dark pigments termed melanins. The chemical composition and structure of lichen melanins remain poorly studied. We analyzed the elemental composition, the main functional groups, and the physicochemical properties of melanin extracted from Cetraria islandica and Pseudevernia furfuracea lichens. Based on the C/N ratio, this pigment is allomelanin. We also identified functional groups that provide photoprotective and antioxidant properties of melanin. Melanin synthesis might be an essential defense mechanism contributing to the survival of lichens under exposure to UV radiation.

Physiological Response and Multi-Elemental Content in Lichens Growing on Agricultural Fences: A Pilot Study Comparing No-Tillage and Organic Cropping.

The objective of this study is to compare the physiological response (content and degradation of photosynthetic pigments, membrane oxidation products and soluble proteins) and multi-element content of Ramalina celastri (lichenized fungi) growing on agricultural fences with no-tillage (associated with transgenic crops and agrochemical application), organic cropping and a non-cultivated area. We found that R. celastri did not differ in its physiological response to agricultural practices, except for the contents of chlorophyll b and phaeophytin a which were high in both cultivated areas. Lichens growing in organic cropping fields have higher arsenic, chromium, uranium and internal transition elements common in the earth's crust, possibly due to the greater resuspension of the material during soil tillage. Lichens that grow on posts close to no-tillage field had higher bromine contents (present in numerous pesticides). We found evidence that R. celastri behaves as a tolerant species to air pollution in agricultural environments.

What Is in Umbilicaria pustulata? A Metagenomic Approach to Reconstruct the Holo-Genome of a Lichen.

Lichens are valuable models in symbiosis research and promising sources of biosynthetic genes for biotechnological applications. Most lichenized fungi grow slowly, resist aposymbiotic cultivation, and are poor candidates for experimentation. Obtaining contiguous, high-quality genomes for such symbiotic communities is technically challenging. Here, we present the first assembly of a lichen holo-genome from metagenomic whole-genome shotgun data comprising both PacBio long reads and Illumina short reads. The nuclear genomes of the two primary components of the lichen symbiosis-the fungus Umbilicaria pustulata (33 Mb) and the green alga Trebouxia sp. (53 Mb)-were assembled at contiguities comparable to single-species assemblies. The analysis of the read coverage pattern revealed a relative abundance of fungal to algal nuclei of ∼20:1. Gap-free, circular sequences for all organellar genomes were obtained. The bacterial community is dominated by Acidobacteriaceae and encompasses strains closely related to bacteria isolated from other lichens. Gene set analyses showed no evidence of horizontal gene transfer from algae or bacteria into the fungal genome. Our data suggest a lineage-specific loss of a putative gibberellin-20-oxidase in the fungus, a gene fusion in the fungal mitochondrion, and a relocation of an algal chloroplast gene to the algal nucleus. Major technical obstacles during reconstruction of the holo-genome were coverage differences among individual genomes surpassing three orders of magnitude. Moreover, we show that GC-rich inverted repeats paired with nonrandom sequencing error in PacBio data can result in missing gene predictions. This likely poses a general problem for genome assemblies based on long reads.

When facilitation meets clonal integration in forest canopies.

Few studies have explored how - within the same system - clonality and positive plant-plant interactions might interact to regulate plant community composition. Canopy-dwelling epiphytes in species-rich forests provide an ideal system for studying this because many epiphytic vascular plants undertake clonal growth and because vascular epiphytes colonize canopy habitats after the formation of nonvascular epiphyte (i.e. bryophyte and lichen) mats. We investigated how clonal integration of seven dominant vascular epiphytes influenced inter-specific interactions between vascular epiphytes and nonvascular epiphytes in a subtropical montane moist forest in southwest China. Both clonal integration and environmental buffering from nonvascular epiphytes increased survival and growth of vascular epiphytes. The benefits of clonal integration for vascular epiphytes were higher when nonvascular epiphytes were removed. Similarly, facilitation from nonvascular epiphytes played a more important role when clonal integration of vascular epiphytes was eliminated. Overall, clonal integration had greater benefits than inter-specific facilitation. This study provides novel evidence for interactive effects of clonality and facilitation between vascular and nonvascular species, and has implications for our understanding of a wide range of ecosystems where both high levels of clonality and facilitation are expected to occur.

Ecophysiology and phylogeny of new terricolous and epiphytic chlorolichens in a fog oasis of the Atacama Desert.

The Atacama Desert is one of the driest and probably oldest deserts on Earth where only a few extremophile organisms are able to survive. This study investigated two terricolous and two epiphytic lichens from the fog oasis "Las Lomitas" within the National Park Pan de Azúcar which represents a refugium for a few vascular desert plants and many lichens that can thrive on fog and dew alone. Ecophysiological measurements and climate records were combined with molecular data of the mycobiont, their green algal photobionts and lichenicolous fungi to gain information about the ecology of lichens within the fog oasis. Phylogenetic and morphological investigations led to the identification and description of the new lichen species Acarospora conafii sp. nov. as well as the lichenicolous fungi that accompanied them and revealed the trebouxioid character of all lichen photobionts. Their photosynthetic responses were compared during natural scenarios such as reactivation by high air humidity and in situ fog events to elucidate the activation strategies of this lichen community. Epiphytic lichens showed photosynthetic activity that was rapidly induced by fog and high relative air humidity whereas terricolous lichens were only activated by fog.

Species diversity patterns in managed Scots pine stands in ancient forest sites.

Continuity in forest habitats is crucial for species diversity and richness. Ancient Scots pine forests are usually under forest management, which disturbs vegetation and causes differentiation in terms of tree stand age. To date, vegetation variability in ancient Scots pine forests has not been examined based on tree stand age classes. In the present study the continuity of a large Scots pine forest complex was investigated, and a system of sampling plots established in five tree stand age classes: initiation stands (4-10 years), young stands (20-35 years), middle-aged stands (45-60 years), pre-mature stands (70-85 years) and mature stands (95-110 years). Species composition, including vascular plants, bryophytes and lichens, on soil, tree trunks, and coarse woody debris, was analyzed. Based on existing classifications systems, forest species and ancient forest species groups were distinguished. In the studied ancient Scots pine forests the species pool and richness were relatively low, and the vegetation consisted mostly of generalist species. Cryptogams, which can grow on diverse substrates, were the most abundant species. Moreover, most species could tolerate both forest and non-forest conditions. Age class forests provided different environmental niches for species. Initiation stands were optimal for terrestrial light-demanding species, and in terms of species composition, initiation stands were most specific. Young stands were most preferred by species on coarse woody debris, and at this stage of stand maturation epiphytic species re-appeared. The oldest stands were not rich in forest specialists, i.e. species of closed forest and ancient forest species. Cryptogams of closed forests inhabited different substrates, and they were not associated only with the oldest stands. The low number of forest specialists in the oldest stands may be a general feature of acidophilus pine forests. However, it may also be a result of the lack of species sources in the vicinity of maturing pine stands. In managed forests a frequent diversity pattern is an increase in a species pool and richness after clear-cut logging. In the present study we obtained higher species pools in initiation and young stands, but richness was similar in all tree stand age classes. This resulted from taking into account species of different substrates (terrestrial, epixylous and epiphytic species) which changed their participation in the vegetation of subsequent stages of tree stand development.

Heavy-metal tolerance of photobiont in pioneer lichens inhabiting heavily polluted sites.

Heavy metals are known for their negative impact on the physiological processes of lichen photobiont. In spite of this, certain lichens are known to be effective pioneers of polluted sites. Cladonia cariosa, C. rei, and Diploschistes muscorum are prominent examples of lichens that spontaneously colonise post-industrial wastes. We examined the effect of total and intracellular Zn, Pb, Cd, As, Cu, and Ni accumulation in the thalli of these species on the physiological parameters of photobiont. Increased accumulation of Zn, Cd, Cu, and Ni in D. muscorum and of Zn and Ni in C. rei negatively affected contents of photosynthetic pigments, whereas concentrations of Pb had a positive effect in all lichen species. Moreover, pigment contents were positively associated with the concentrations of most examined elements in C. cariosa. The results indicate that even if chlorophyll contents reduced, its degradation does not progress. This suggests that metal stress may exert a negative effect on the synthesis rather than on the integrity of chlorophyll. Most importantly, lichen samples of each of the species from polluted sites proved to possess significantly higher FV/FM ratios than those from a reference site; moreover, the contents of elements of lichen thalli positively influenced this parameter. The efficient functioning of the algal component under heavy-metal stress conditions indicates that the examined lichens are well adapted to extremely contaminated substrates.

A new measurement tool to consider for airborne pollutants evaluations using lichens.

An important factor affecting acquisition of pollution elements could be the lichen growth form. The Brunauer-Emmett-Teller theory approach has been used to determinate the specific area surface (BET-area) of solids by gas multilayer adsorption. Taking this standard method as a new tool, we measure the specific thallus area in foliose and fruticose lichens to evaluated area/volume relation for bioaccumulation prospects. Some preliminary results of elemental contents such as REEs (La, Sc, Sr) and pollutants (Cd, Co, Pb) were also measured to support the importance to use for the analysis of these thallus attributes.

Relative humidity, temperature, substrate type, and height of terrestrial lichens in a tropical paramo / Humedad relativa, temperatura, tipo de sustrato y altura de los líquenes del suelo en un páramo tropical

Abstract Lichens are traditionally divided into types such as "crustose", "foliose" and "fruticose", with different shapes and heights. Substrate, temperature and water are thought to affect lichen height, but there are few studies regarding tropical paramo lichens. Along 2015 I measured those variables in the terricolous lichens of the Cerro Buena Vista, paramo (Costa Rica, 9°33' N & 83°45' W). The measurements were taken inside the lichens and in the substrate under them, in 61 randomly located quadrats (50 x 50 cm). Lichens grew taller on (1) warmer ground, (2) wetter ground, (3) the moister Caribbean slope, and (4) the season with heavier rainfall, as expected. Apparently, atmospheric factors are more important than substrate in the determination of temperature, relative humidity and growth of lichens. Physiologically available water seems to be the main determinant of lichen vertical growth in the Buena Vista paramo.(AU)

Resumen Los líquenes se dividen tradicionalmente en morfotipos como "crustoso", "folioso" y "fruticoso", con diferentes formas y alturas. Se cree que el sustrato, la temperatura y el agua afectan la altura de los líquenes, pero existen pocos estudios con respecto a los líquenes tropicales. A lo largo del 2015, medí esas variables en los líquenes del suelo en el páramo de Cerro Buena Vista (Costa Rica, 9 ° 33'N; 83 ° 45'W). Las medidas se tomaron dentro de los líquenes y en el sustrato debajo de ellos, en 61 cuadrantes ubicados al azar (50 x 50 cm). Los líquenes fueron más altos en (1) suelo más cálido, (2) suelo más húmedo, (3) la pendiente del Caribe, que es más húmeda y (4) la temporada con lluvias más intensas, como se esperaba. Aparentemente, los factores atmosféricos son más importantes que el sustrato en la determinación de la temperatura, la humedad relativa y el crecimiento de los líquenes. El agua fisiológicamente disponible parece ser el principal determinante del crecimiento vertical de los líquenes en el páramo del Cerro Buena Vista.(AU)

Use of corticolous lichens for the assessment of ambient air quality along rural-urban ecosystems of tropics: a study in Sri Lanka.

Monitoring of air quality using lichens as bioindicators on the basis of lichen diversity and frequency is limited along rural-urban ecosystems in tropics. This study attempted to assess and correlate the use of corticolous lichens with atmospheric SO2 and NO2 in such an ecosystem in Sabaragamuwa Province in Sri Lanka. Nine sampling locations, each having three subsampling sites with 162 Mangifera indica and Cocos nucifera trees, were selected for the study. The coverage and frequency of lichens found on selected trees were recorded by 400-cm2 grids and identified using taxonomic keys. SO2 and NO2 levels at each site were determined by "Ogawa" passive air samplers. Data of lichen diversity were used to formulate the index of atmospheric purity (IAP). The environmental parameters related to lichen colonization were measured using standard methods. Data were analyzed using MINITAB 17. The mapping of spatial distribution of lichens and air pollutants were done using inverse distance weighting surface interpolation of geographical information system based on IAP values. A negative correlation was observed between IAP and SO2 and NO2 levels. The presence of the genus Pyxine in almost all urban sites indicated that it could be used as a reliable pollutant tolerant indicator in urban ecosystems. In addition, the index-based mapping techniques could be used successfully to see the effect of atmospheric pollution in urban ecosystems. These results conclude that corticolous lichens have the potential to be used as bioindicators of air quality monitoring along rural-urban ecosystems of tropics.

Effects of Biological Soil Crusts on Enzyme Activities and Microbial Community in Soils of an Arid Ecosystem.

Arid ecosystems constitute 41% of land's surface and play an important role in global carbon cycle. In particular, biological soil crusts (BSC) are known to be a hotspot of carbon fixation as well as mineralization in arid ecosystems. However, little information is available on carbon decomposition and microbes in BSC and key controlling variables for microbial activities in arid ecosystems. The current study, carried out in South Mediterranean arid ecosystem, aimed to evaluate the effects of intact and removed cyanobacteria/lichen crusts on soil properties, soil enzyme activities, and microbial abundances (bacteria and fungi). We compared five different treatments (bare soil, soil with intact cyanobacteria, soil with cyanobacteria removed, soil with intact lichens, and soil with lichens removed) in four different soil layers (0-5, 5-10, 10-15, and 15-20 cm). Regardless of soil treatments, activities of hydrolases and water content increased with increasing soil depth. The presence of lichens increased significantly hydrolase activities, which appeared to be associated with greater organic matter, nitrogen, and water contents. However, phenol oxidase was mainly controlled by pH and oxygen availability. Neither fungal nor bacterial abundance exhibited a significant correlation with enzyme activities suggesting that soil enzyme activities are mainly controlled by edaphic and environmental conditions rather than source microbes. Interestingly, the presence of lichens reduced the abundance of bacteria of which mechanism is still to be investigated.

Selecting lichen functional traits as ecological indicators of the effects of urban environment.

Air pollution and the urban heat island effect are known to directly affect ecosystems in urban areas. Lichens, which are widely known as good ecological indicators of air quality and of climatic conditions, can be a valuable tool to monitor environmental changes in urban environments. The objective of this work was to select lichen functional traits and functional groups that can be used as ecological indicators of the effects of urbanization, with emphasis in the Southern subtropics, where this had never been done. For that, we assessed lichen functional composition in urban sites with different population density, which was considered as proxy for grouping sites in two levels of urbanization (low and medium/high). This a priori grouping was based on their significantly differences on air pollutants and land cover. Urbanization and air pollution showed to affect all lichen functional traits, with different responses depending on the functional group. Medium/high density urbanization was associated to an increase on the mean relative abundance of lichens with chlorococcoid green algae, foliose narrow lobes, soredia as the main reproduction strategy, pruinose thallus and containing secondary metabolites for chemical protection. Lower density urbanization showed a higher relative frequency of cyanolichens and lichens with Trentepohlia as the main algae, loosely attached crustose thallus and isidia as the main reproductive structure. The differences found on photobiont and growth form traits in response to the environmental variables used as proxies of microclimatic conditions (forest cover and number of trees around the sampling units), enabled us to detect the urban heat island effect (drier conditions in more urbanized sites).

A simple survey protocol for assessing terrestrial biodiversity in a broad range of ecosystems.

Finding standard cost-effective methods for monitoring biodiversity is challenging due to trade-offs between survey costs (including expertise), specificity, and range of applicability. These trade-offs cause a lack of comparability among datasets collected by ecologists and conservationists, which is most regrettable in taxonomically demanding work on megadiverse inconspicuous taxon groups. We have developed a site-scale survey method for diverse sessile land organisms, which can be analyzed over multiple scales and linked with ecological insights and management. The core idea is that field experts can effectively allocate observation effort when the time, area, and priority sequence of tasks are fixed. We present the protocol, explain its specifications (taxon group; expert qualification; plot size; effort) and applications based on >800 original surveys of four taxon groups; and we analyze its effectiveness using data on polypores in hemiboreal and tropical forests. We demonstrate consistent effort-species richness curves and among-survey variation in contrasting ecosystems, and high effectiveness compared with casual observations both at local and regional scales. Bias related to observer experience appeared negligible compared with typical assemblage variation. Being flexible in terms of sampling design, the method has enabled us to compile data from various projects to assess conservation status and habitat requirements of most species (specifically rarities and including discovery of new species); also, when linked with site descriptions, to complete environmental assessments and select indicator species for management. We conclude that simple rules can significantly improve expert-based biodiversity surveys. Ideally, define (i) a common plot size that addresses multiple taxon groups and management goals; (ii) taxon groups based on field expertise and feasible number of species; (iii) sufficient and practical search time; (iv) a procedure for recording within-plot heterogeneity. Such a framework, combined with freedom to allocate effort on-site, helps utilizing full expertise of observers without losing technical rigor.

Effects of site-specific climatic conditions on the radial growth of the lichen biomonitor Xanthoria parietina.

The protocols commonly applied in surveys with lichens as biomonitors of airborne trace elements require analyses of samples derived from thalli or parts of thalli grown in the last year before sampling, under the postulation that samples of the same size are of the same age. Unfortunately, the influence of ecological site-specific factors on lichen growth is still largely ignored, so that samples of the same size collected in environmentally and climatically diverse sites might actually differ in age. This work aims at quantifying the influence of climatic conditions on the radial growth rates (RaGRs) of Xanthoria parietina, one of the most popular lichen biomonitors. RaGR was monitored in seven populations distributed along an altitudinal transect of 30 km in the Classical Karst (NE Italy), from 20 to 500 m above sea level. For c. 17 months, lobe growth was measured seasonally with a digital calliper, and site-specific climatic variables were monitored by means of thermo-hygrometric sensors and implemented by meteorological data. Finally, the lobe growth of X. parietina was modelled as a function of 18 environmental variables. Results revealed that thalli of relatively dry sites had significantly lower seasonal RaGR with respect to moister ones. Considering that cumulative precipitations were equally distributed along the transect, it was concluded that RaGR of X. parietina is affected negatively by high air temperatures and positively by high relative humidity. The importance of RaGR variation in lichen bioaccumulation studies is critically discussed.

Altered fire regimes cause long-term lichen diversity losses.

Many global ecosystems have undergone shifts in fire regimes in recent decades, such as changes in fire size, frequency, and/or severity. Recent research shows that increases in fire size, frequency, and severity can lead to long-persisting deforestation, but the consequences of shifting fire regimes for biodiversity of other vegetative organisms (such as understory plants, fungi, and lichens) remain poorly understood. Understanding lichen responses to wildfire is particularly important because lichens play crucial roles in nutrient cycling and supporting wildlife in many ecosystems. Lichen responses to fire have been little studied, and most previous research has been limited to small geographic areas (e.g. studies of a single fire), making it difficult to establish generalizable patterns. To investigate long-term effects of fire severity on lichen communities, we sampled epiphytic lichen communities in 104 study plots across California's greater Sierra Nevada region in areas that burned in five wildfires, ranging from 4 to 16 years prior to sampling. The conifer forest ecosystems we studied have undergone a notable increase in fire severity in recent decades, and we sample across the full gradient of fire severity to infer how shifting fire regimes may influence landscape-level biodiversity. We find that low-severity fire has little to no effect on lichen communities. Areas that burned at moderate and high severities, however, have significantly and progressively lower lichen richness and abundance. Importantly, we observe very little postfire lichen recolonization on burned substrates even more than 15 years after fire. Our multivariate model suggests that the hotter, drier microclimates that occur after fire removes forest canopies may prevent lichen reestablishment, meaning that lichens are not likely to recolonize until mature trees regenerate. These findings suggest that altered fire regimes may cause broad and long-persisting landscape-scale biodiversity losses that could ultimately impact multiple trophic levels.

Comparing Sanger sequencing and high-throughput metabarcoding for inferring photobiont diversity in lichens.

The implementation of HTS (high-throughput sequencing) approaches is rapidly changing our understanding of the lichen symbiosis, by uncovering high bacterial and fungal diversity, which is often host-specific. Recently, HTS methods revealed the presence of multiple photobionts inside a single thallus in several lichen species. This differs from Sanger technology, which typically yields a single, unambiguous algal sequence per individual. Here we compared HTS and Sanger methods for estimating the diversity of green algal symbionts within lichen thalli using 240 lichen individuals belonging to two species of lichen-forming fungi. According to HTS data, Sanger technology consistently yielded the most abundant photobiont sequence in the sample. However, if the second most abundant photobiont exceeded 30% of the total HTS reads in a sample, Sanger sequencing generally failed. Our results suggest that most lichen individuals in the two analyzed species, Lasallia hispanica and L. pustulata, indeed contain a single, predominant green algal photobiont. We conclude that Sanger sequencing is a valid approach to detect the dominant photobionts in lichen individuals and populations. We discuss which research areas in lichen ecology and evolution will continue to benefit from Sanger sequencing, and which areas will profit from HTS approaches to assessing symbiont diversity.