Foraging strategies of fungal mycelial networks: responses to quantity and distance of new resources.

Fungal mycelial networks are essential for translocating and storing water, nutrients, and carbon in forest ecosystems. In particular, wood decay fungi form mycelial networks that connect various woody debris on the forest floor. Understanding their foraging strategies is crucial for complehending the role of mycelium in carbon and nutrient cycling in forests. Previous studies have shown that mycelial networks initiate migration from the original woody resource (inoculum) to a new woody resource (bait) if the latter is sufficiently large but not if it is small. However, the impact of energetic costs during foraging, such as the distance to the bait, has not been considered. In the present study, we conducted full-factorial experiments with two factors, bait size (4 and 8 cm3) and distance from the inoculum (1 and 15 cm). An inoculum wood block, colonized by the wood decay fungus Phanerochaete velutina, was placed in one corner of a bioassay dish (24 cm × 24 cm) filled with unsterilized soil. Once the mycelium grew onto the soil to a distance >15 cm from the inoculum, a sterilized new bait wood block (of either size) was placed on the soil at one of the two distances to be colonized by the mycelia from the inoculum. After 50 days of incubation, the baits were harvested, and their dried weight was measured to calculate the absolute weight loss during incubation. The inoculum wood blocks were retrieved and placed on a new soil dish to determine whether the mycelium would grow out onto the soil again. If no growth occurred within 8 days of additional incubation, we concluded that the mycelium had migrated from the inoculum to the bait. The results showed that mycelia in inocula coupled with baits positioned 1 cm away migrated to the baits more frequently than those with baits positioned 15 cm away. A structural equation model revealed that bait weight loss (energy gain) and hyphal coverage on the soil (foraging cost) significantly influenced mycelial migration decisions. These findings suggest that fungal mycelia may employ their own foraging strategies based on energetic benefits.

Local calcium signal transmission in mycelial network exhibits decentralized stress responses.

Many fungi live as mycelia, which are networks of hyphae. Mycelial networks are suited for the widespread distribution of nutrients and water. The logistical capabilities are critical for the extension of fungal survival areas, nutrient cycling in ecosystems, mycorrhizal symbioses, and virulence. In addition, signal transduction in mycelial networks is predicted to be vital for mycelial function and robustness. A lot of cell biological studies have elucidated protein and membrane trafficking and signal transduction in fungal hyphae; however, there are no reports visualizing signal transduction in mycelia. This paper, by using the fluorescent Ca2+ biosensor, visualized for the first time how calcium signaling is conducted inside the mycelial network in response to localized stimuli in the model fungus Aspergillus nidulans. The wavy propagation of the calcium signal inside the mycelium or the signal blinking in the hyphae varies depending on the type of stress and proximity to the stress. The signals, however, only extended around 1,500 µm, suggesting that the mycelium has a localized response. The mycelium showed growth delay only in the stressed areas. Local stress caused arrest and resumption of mycelial growth through reorganization of the actin cytoskeleton and membrane trafficking. To elucidate the downstream of calcium signaling, calmodulin, and calmodulin-dependent protein kinases, the principal intracellular Ca2+ receptors were immunoprecipitated and their downstream targets were identified by mass spectrometry analyses. Our data provide evidence that the mycelial network, which lacks a brain or nervous system, exhibits decentralized response through locally activated calcium signaling in response to local stress.

Surface Area of Wood Influences the Effects of Fungal Interspecific Interaction on Wood Decomposition-A Case Study Based on Pinus densiflora and Selected White Rot Fungi.

Wood decomposer basidiomycetes are the major agents of lignocellulose decomposition in dead wood. As their interspecific interaction affects wood decomposition, difference in interaction area may alter the magnitude of the effects. This study examines the effects of wood surface area on decomposition by interacting basidiomycetes using laboratory incubation experiments with pine sapwood as a model. Two types of pine wood blocks with equal volume but identical surface area were prepared for colonization by one of four white rot basidiomycete species. The colonized wood blocks were then placed on agar media already colonized by the same strain or one of the other species, simulating fungal monoculture and interspecific interactions on wood surface. Results demonstrated that the decay rate of wood was greater in wood with larger surface, and wood decay was accelerated by the interaction of two fungal species in wood with larger surface but not in wood with smaller surface. In contrast, lignin decomposition was influenced by the competitor in wood with smaller surface but not in wood with larger surface. These results suggest that the observed promotion of decay by fungal interspecific interaction might not be attributable to the resource partitioning between fungal species but to the accelerated carbon of competition cost compensation in this case.

Association between corticolous myxomycetes and tree vitality in Cryptomeria japonica.

The bark of live trees provides an important microhabitat for corticolous myxomycetes. However, the association between the presence of myxomycetes and health of host trees has not been studied in detail. In this study, we aimed to investigate the relationship between tree vitality and myxomycetes on the bark of Cryptomeria japonica trees in a montane forest in western Japan. The vitality of trees was categorized into four grades based on the visual assessment of tree shape and leaf density in the upper branches. Myxomycetes on the bark surface were examined using the moist chamber culture method. A decline in tree vitality increased bark pH and decreased electrical conductivity of the bark exudates. Seventeen myxomycete species were recorded in 74 C. japonica trees. The structure of myxomycete communities varied between healthy and unhealthy trees, and species diversity increased as the vitality declined. The relative abundance of Cribraria confusa decreased as the vitality declined, while that of Paradiacheopsis solitaria increased. The results showed that acidophilic myxomycetes grew on healthy C. japonica bark, but changes in bark pH associated with vitality decline led to the weakening of acidity and shifted the community structure; thus, corticolous myxomycete diversity was enhanced as tree vitality decline.

Timing of Resource Addition Affects the Migration Behavior of Wood Decomposer Fungal Mycelia.

Studies of fungal behavior are essential for a better understanding of fungal-driven ecological processes. Here, we evaluated the effects of timing of resource (bait) addition on the behavior of fungal mycelia when it remains in the inoculum and when it migrates from it towards a bait, using cord-forming basidiomycetes. Experiments allowed mycelium to grow from an inoculum wood across the surface of a soil microcosm, where it encountered a new wood bait 14 or 98 d after the start of growth. After the 42-d colonization of the bait, inoculum and bait were individually moved to a dish containing fresh soil to determine whether the mycelia were able to grow out. When the inoculum and bait of mycelia baited after 14 d were transferred to new soil, there was 100% regrowth from both inoculum and bait in Pholiota brunnescens and Phanerochaete velutina, indicating that no migration occurred. However, when mycelium was baited after 98 d, 3 and 4 out of 10 replicates of P. brunnescens and P. velutina, respectively, regrew only from bait and not from inoculum, indicating migration. These results suggest that prolonged periods without new resources alter the behavior of mycelium, probably due to the exhaustion of resources.

Decay stages of wood and associated fungal communities characterise diversity-decomposition relationships.

The biodiversity-ecosystem function relationship is a central topic in ecology. Fungi are the dominant decomposers of organic plant material in terrestrial ecosystems and display tremendous species diversity. However, little is known about the fungal diversity-decomposition relationship. We evaluated fungal community assemblies and substrate quality in different stages of wood decay to assess the relationships between fungal species richness and weight loss of wood substrate under laboratory conditions. Wood-inhabiting fungal communities in the early and late stages of pine log decomposition were used as a model. Colonisation with certain species prior to inoculation with other species resulted in four-fold differences in fungal species richness and up to tenfold differences in the rate of wood substrate decomposition in both early- and late-decaying fungal communities. Differences in wood substrate quality had a significant impact on species richness and weight loss of wood and the relationships between the two, which were negative or neutral. Late communities showed significantly negative species richness-decay relationships in wood at all decay stages, whereas negative relationships in early communities were significant only in the intermediate decay stage. Our results suggest that changes in fungal communities and wood quality during wood decomposition affect the fungal diversity-decomposition relationship.

Effects of wood resource size and decomposition on hyphal outgrowth of a cord-forming basidiomycete, Phanerochaete velutina.

To assess the relationship between resource use and hyphal growth in a cord-forming basidiomycete, Phanerochaete velutina, soil microcosm experiments were conducted using wood blocks of three different sizes in three different soil quantities, thereby simulating the different amounts of available nutrients. The highest percentage weight loss was observed in the smallest wood blocks after a 27-d incubation period in soil microcosms, although the percentage weight loss over the 2-month pure culture colonization prior to inoculation was not significantly different among various block sizes. The greatest hyphal outgrowth was also observed in the smallest wood blocks and was positively associated with wood decay. The slopes of the regression lines between hyphal coverage and percentage wood mass loss were identical among different wood sizes, but the slopes between hyphal coverage and absolute wood mass loss were steeper in the smaller wood blocks than that in largest one. These results suggest that the level of intensity of mycelial foraging for new resources in the soil depends on the percentage of the amount of wood resource utilized, and not on the absolute amount of carbon obtained from the wood.

Ecological memory and relocation decisions in fungal mycelial networks: responses to quantity and location of new resources.

Saprotrophic cord-forming basidiomycetes, with their mycelial networks at the soil/litter interface on the forest floor, play a major role in wood decomposition and nutrient cycling/relocation. Many studies have investigated foraging behaviour of their mycelium, but there is little information on their intelligence. Here, we investigate the effects of relative size of inoculum wood and new wood resource (bait) on the decision of a mycelium to remain in, or migrate from, inoculum to bait using Phanerochaete velutina as a model. Experiments allowed mycelium to grow from an inoculum across the surface of a soil microcosm where it encountered a new wood bait. After colonisation of the bait, the original inoculum was moved to a tray of fresh soil to determine whether the fungus was still able to grow out. This also allowed us to test the mycelium's memory of growth direction. When inocula were transferred to new soil, there was regrowth from 67% of the inocula, and a threshold bait size acted as a cue for the mycelium's decision to migrate for a final time, rather than a threshold of relative size of inoculum: bait. There was greater regrowth from the side that originally faced the new bait, implying memory of growth direction.

The giant mycoheterotrophic orchid Erythrorchis altissima is associated mainly with a divergent set of wood-decaying fungi.

The climbing orchid Erythrorchis altissima is the largest mycoheterotroph in the world. Although previous in vitro work suggests that E. altissima has a unique symbiosis with wood-decaying fungi, little is known about how this giant orchid meets its carbon and nutrient demands exclusively via mycorrhizal fungi. In this study, the mycorrhizal fungi of E. altissima were molecularly identified using root samples from 26 individuals. Furthermore, in vitro symbiotic germination with five fungi and stable isotope compositions in five E. altissima at one site were examined. In total, 37 fungal operational taxonomic units (OTUs) belonging to nine orders in Basidiomycota were identified from the orchid roots. Most of the fungal OTUs were wood-decaying fungi, but underground roots had ectomycorrhizal Russula. Two fungal isolates from mycorrhizal roots induced seed germination and subsequent seedling development in vitro. Measurement of carbon and nitrogen stable isotope abundances revealed that E. altissima is a full mycoheterotroph whose carbon originates mainly from wood-decaying fungi. All of the results show that E. altissima is associated with a wide range of wood- and soil-inhabiting fungi, the majority of which are wood-decaying taxa. This generalist association enables E. altissima to access a large carbon pool in woody debris and has been key to the evolution of such a large mycoheterotroph.

Wood decomposing abilities of diverse lignicolous fungi on nondecayed and decayed beech wood.

We tested the decay abilities of 28 isolates from 28 lignicolous fungal species (Basidiomycota, Ascomycota and Zygomycota) with the pure culture test. We used beech wood powder in varying moisture conditions and decay stages (nondecayed, intermediately decayed and well decayed) as substrates. The weight loss in wood powder was -0.2-17.8%. Five isolates of Basidiomycota (Bjerkandera adusta, Mycena haematopus, Omphalotus guepiniformis, Trametes hirsuta, Trametes versicolor) caused high weight losses in nondecayed wood. We detected significant effects of decay stage on weight loss in wood in most isolates tested, whereas moisture content rarely had an effect on weight loss. Among Basidiomycota and Xylariaceae in Ascomycota weight loss was greater for nondecayed wood than for intermediately and well decayed wood. In contrast four isolates in Ascomycota (Scytalidium lignicola, Trichoderma hamatum, T. harzianum, T. koningii) caused substantial weight loss in intermediately and well decayed wood, although they rarely caused weight loss in nondecayed wood. Zygomycota caused low weight loss in wood. Wood decay stages also affected decomposition of wood chemical components. Acid-unhydrolyzable residue (AUR) decomposition was reduced, whereas holocellulose decomposition was stimulated by some strains of Basidiomycota and Ascomycota in well decayed wood. T. harzianum in particular caused significant weight loss of holocellulose in well decayed wood, although this fungus caused negligible weight loss of both AUR and holocellulose in nondecayed wood. We discuss these changes in the decay patterns of AUR and holocellulose with varying wood decay stages in relation to the role of fungal decomposition of woody debris in forests.

Roles of diverse fungi in larch needle-litter decomposition.

Functional biodiversity of fungi in larch (Larix leptolepis) forests needle-litter decomposition was examined by a pure-culture test. Weight loss of larch-needle litter, utilization pattern of lignocellulose and chemical composition of remaining litter were investigated and compared for 31 isolates in 27 species of basidiomycetes and ascomycetes. Weight loss (% original weight) of litter ranged from -2.0% to 14.2%. Mean weight loss of litter caused by the basidiomycetes was not significantly different from that caused by the ascomycetes. Basidiomycetes caused loss of lignin and carbohydrates in variable proportions, while ascomycetes exclusively attacked carbohydrates without delignification. The content of lignin and nitrogen in remaining litter was not significantly correlated when both basidiomycetes and ascomycetes were included. However, the correlation coefficient was significant when the relationship was examined separately for basidiomycetes, indicating that the degree of selective delignification determined the final nitrogen content in litter. Possible effects of fungal colonization on needle-litter decomposition in larch forests are discussed.