Stratification of the ant species (Hymenoptera,Formicidae) in the urban broadleaf woodlands of the city of Kiev

Investigations were carried out in June–August 2012 and 2013 in broadleaf forests and planted stands of the northern red oak Quercus rubra in Kiev. The ant visitation rates of 3–6 most abundant plant species within each vegetation layer were analyzed. In all, 16 species of ants were found in the broadleaf forest (Quercus robur + Acer spp. + Carpinus betulus), with the dominance of three species: Formica rufa, Lasius fuliginosus, and L. emarginatus. Eight ant species occurred in Q. rubra stands with the undergrowth of Acer campestre and A. platanoides; the dominant species were Lasius fuliginosus and L. emarginatus. The northern red oak stands with conspecific undergrowth had only four ant species with no dominants among them. Plants with the highest ant visitation rates in the herbage layer of the broadleaf forest were the invasive Impatiens parviflora and the native Aegopodium podagraria. Ants were rare or absent on the remaining herbs: Impatiens noli-tangere, Carex sylvatica, Stellaria holostea, and Galium odoratum. The herbage layer was mostly visited by influents, the dominants being represented only by Lasius emarginatus. Within the shrub layer, ants most frequently visited the undergrowth of the maple Acer platanoides, often harboring colonies of the aphid Periphyllus lyropictus; the highest visitation rates were recorded in the dominants L. emarginatus and L. fuliginosus. The arboreal layer had the highest visitation rate and was most often visited by the dominants Lasius fuliginosus, L. emarginatus, and F. rufa. The common oak Q. robur was the most visited tree in broadleaf forests. In addition to providing ants with food (colonies of the aphids Lachnus roboris and Stomaphis quercus, and also phytophagous insects and other invertebrates), these trees were used for nesting by the dominants L. fuliginosus and L. emarginatus. The presence of aphid colonies was also typical of ripe trees and undergrowth of the maple A. platanoides, the second-visited tree in broadleaf forests. The common hornbeam Carpinus betulus was the least frequently visited, though some ant species, usually L. emarginatus, nested in the trunks and branches of old trees. The vertical distribution of ants in broadleaf forests and red oak stands formed two clusters: the influents occupied the lower (herbage and shrub) vegetation layers, while the subdominants and dominants occupied the upper (shrub and arboreal) ones. The smallest number of ant species (1 or 2) in all the vegetation layers was recorded in the areas with F. rufa; areas with L. fuliginosus had twice as many species; the greatest numbers of ant species were found in the areas with L. emarginatus and in those without dominants. This pattern may result from different territoriality of the dominants: the strongest in F. rufa (defending the whole territory) and the weaker in the other two species (defending only a part of the territory). Plants of all the vegetation layers varied in their attractiveness to ants. As a result, the layers had a mosaic structure in their vertical (between-layer) and horizontal (between different plant species within one layer) arrangement. From 60 to 100% of plants of certain species were visited by ants while plants of other species were not visited at all. The main reason for visiting plants by ants was the presence of aphid colonies.     

Soil microarthropods and macrofauna in monsoon tropical forests of Cat Tien and Bi Dup-Nui Ba National Parks,southern Vietnam

The abundance, biomass, vertical distribution, and taxonomic composition of soil invertebrates (springtails, macrofauna, and termites) were studied in forest formations differing in edaphic and climatic conditions: lowland forests dominated by Lagerstroemia spp. or Dipterocarpus spp. in the Cat Tien National Park and in a mountain pine (Pinus kesiya) forest on the Da Lat Plateau, southern Vietnam. In the lowland forests, springtails had a relatively low density (10000–12000 ind./m²), but their diversity was high (41–43 species in each forest). The density of large soil invertebrates (without ants and termites) reached 500–700 ind./m² at a biomass of approximately 30 g/m² (with earthworms accounting for up to 230 ind./m² and 19–28 g/m²). Among termites, species of the genera Macrotermes and Odontotermes were dominant. Their total biomass in some areas exceeded 15–20 g/m². In the mountain pine forest, the total biomass of soil macrofauna was approximately 11 g/m², the abundance and diversity of springtails were low (7500 ind./m², 28 species), and wood-destroying species of the genera Schedorhinotermes sp. and Coptotermes sp. dominated among termites.     

Ant–aphid relations in the south of Western Siberia (Hymenoptera: Formicidae; Hemiptera: Aphididae)

Aphids, the main suppliers of energy-rich honeydew, play an important role in the life of ants. However, the data on the trophobiotic ant–aphid associations in the majority of regions are still limited. We present the first data on the ant–aphid relations in the south of Western Siberia. Investigations were carried out in the most typical biotopes of forest-steppe and steppe zones in the territory of Novosibirsk and Kurgan regions (Russia) during 1993–2014. There were revealed 35 species of ants and 198 species of aphids. Detected 456 ant–aphid associations involved 28 ant species and 134 myrmecophilous aphids. Seven ant species were found to consume honeydew of 9 non-myrmecophilous aphids, scraping it from the plant. This behaviour is typical of subdominant and subordinate ants which do not protect their foraging areas. Ants associate with various numbers of aphid species. About 36% of ants attended aphid colonies of less than 5 species. The largest number of myrmecophilous aphids is associated with L. niger (Linnaeus, 1758) (103 species), Formica pratensis Retzius, 1783 (50), Formica rufa group (25–33), F. (Serviformica) fusca Linnaeus, 1758 (26) and F. (S.) cunicularia Latreille, 1798 (27). Different ants play unequal roles in the formation of trophobiotic interactions with aphids. Due to complex territorial and foraging behaviour, including high functional specialization among honeydew collectors, dominant ants of Formica s. str. are one of the leaders in this process. The role of L. niger and Formica ants of the subgenus Serviformica requires further detailed investigation.     

Yeast communities of <Emphasis Type="Italic">Formica aquilonia</Emphasis> colonies

Yeast abundance and species diversity in the colonies of Formica aquilonia ants in birch–pine grass forest near Novosibirsk, Russia, were studied. The average yeast number in the anthill material was 10³–10⁴ CFU/g, reaching 10⁵ CFU/g in the hatching chambers. Typical litter species (Trichosporon moniliiforme and Cystofilobasidium capitatum) were predominant in soil and litter around the anthills. Apart from these species, ascomycete species of the family Debaryomycetaceae, Debaryomyces hansenii, and Schwanniomyces vanrijiae were predominant in the anthill material. Yeast population of the ant’ bodies consisted exclusively of the members of the last two species. Thus, highly specific yeast communities formed in the colonies of Formica aquilonia ants differ from the communities of surrounding soil. These differences are caused by environment-forming activity of the ants.     

Ants (Hymenoptera: Formicidae) as potential egg predators of lepidopteran pests of sweet potato, <Emphasis Type="Italic">Ipomoea batatas</Emphasis> (Solanales: Convolvulaceae), in Japan

The sweet potato Ipomoea batatas (L.) Lam., which has extrafloral nectaries, was visited by nine indigenous ant species in south Kyushu, Japan. These ant species were evaluated for their ability to remove the eggs of two lepidopteran species—Aedia leucomelas (Linnaeus) and Spodoptera litura (Fabricius)—through ant-exclusion experiments. The results showed that Tetramorium bicarinatum (Nylander), one of the dominant ant species, is an efficient predator of lepidopteran eggs. This ant species removed A. leucomelas eggs from ant-visited plants in six trials conducted from mid-July to mid-October, with high removal rates (90–100%) occurring from mid-September onward. On ant-excluded plants, only 0–18% of exposed eggs were removed by ants that accidentally invaded the plants. Tetramorium bicarinatum also removed egg masses and the hatchlings of S. litura. The mean percent area of egg-laden leaves eaten by S. litura larvae was lower on ant-visited plants (0.9%) than on ant-excluded plants (57.3%). In contrast, Formica japonica Motschulsky, another dominant ant species, was not observed removing the eggs or larvae of the lepidopteran species. These results show that certain indigenous ant species are efficient as biocontrol agents. Thus, individual ant species need to be investigated for their ability to control insect pests in various crops.     

Patterns of floral resource use by two dominant ant species in a biodiversity hotspot

Ecological dominance in ants is often fuelled by carbohydrate intake. Most studies have focused on the importance of invasive ant mutualistic associations with trophobionts whereas few studies have investigated the importance of floral nectar on invasion success. In this study, utilisation of temporarily available floral nectar by the invasive Argentine ant, Linepithema humile, was compared to that of the dominant native ant, Anoplolepis custodiens, within the Cape Floristic Region (CFR), a biodiversity hotspot. The effect of these two focal ant species on species composition and abundance of ground foraging ants as well as floral arthropod visitors in inflorescences of Proteacea species was assessed. Foraging activity, and trophic ecology inferred from the abundance of natural stable isotopes of Carbon (δ¹³C) and Nitrogen (δ¹⁵N), and the ratio of Carbon to Nitrogen (C:N) were compared between the two ant species during three flowering periods. Linepithema humile significantly reduced the abundance and species diversity of both above-ground and floral arthropod species abundance and composition. Linepithema humile increased its foraging activity with increasing nectar availability, switching its diet to a more herbivorous one. Anoplolepis custodiens did not respond as effectively to increasing floral nectar or negatively impact floral arthropod visitors. This study showed that the availability of floral nectar and ability of L. humile to more effectively utilise this temporarily available resource than native ants, can contribute significantly to the further spread and persistence of L. humile in natural environments in the CFR.     

Change in the Structure and Productivity of the Biota of Agaricoid Basidiomycetes According to the Results of Long-Term Monitoring in Pine Forests of Perm Oblast (Southern Taiga Subzone)

Results of 40-year monitoring of the agaricoid biota in four types of pine forests on Perm oblast, the southern taiga subzone, are presented. We have found that the species number of fungi in pine forests varies from 80 (sphagnum pine forest) to 194 (lichen pine forest). We have found changes in species composition and an increase in species diversity in each period of monitoring. We analyze the ecological and trophic structure and define species that are dominant in regards to the basidiome number and biomass. Despite the changes in species composition, the ecological and trophic group ratio in each forest type remains almost the same. One common feature is that mycorrhizal fungi prevail in pine forests in regards to species number (55.5–61.2% of all species in the cenosis), as well as yield (52% of all basidiomes and 94% of their total biomass). We study the relation between species diversity and productivity of microbiota with the main climatic factors. We have found that July rainfall is favorable for the fruit bearing of the agarics in a pine forest with moderate humidity in August (r_s = 0.73). In swampy cenoses, June rainfall, on the contrary, has a negative impact (r_s = ?0.70).     

Hymenoptera aculeata of raised bogs in Belarus

The long-term studies (1986–2003) of raised bogs of Belarus revealed 167 species of aculeates belonging to 7 families: Sapygidae (1), Chrysididae (11), Pompilidae (19), Eumenidae (19), Vespidae (9), Sphecidae (76), and Apidae (32). The dominant species (not counting ants) are Vespula rufa (16.6%), Bombus jonellus (11.3%), B. lucorum (7.5%), Anoplius nigerrimus (5.1%), B. pascuorum (3.9%), Passaloecus turionum (3.8%), Argogorytes mystaceus (3.2%), Dipogon bifasciatus (3.1%), Pemphredon lugens (3.0%), and Trypoxylon attenuatum (2.6%). Approximately two-thirds of the species found in raised bogs nest in the upper layer. The spatial distribution of aculeates depends primarily on the distance from the bog margins and the presence of contrasting areas, such as lake banks or glades. The highest activity of various aculeate families (except for Vespidae) was observed in June. The total abundance of aculeates was found to be correlated with solar activity, the strongest correlation being observed in Vespidae. The abundance of different wasp families varied noticeably from year to year, depending on climatic parameters.     

Ant pollination of <Emphasis Type="Italic">Syzygium occidentale</Emphasis>, an endemic tree species of tropical rain forests of the Western Ghats,India

Although mutualism between ants and flowering plants is wide spread, ant pollination has not evolved as a major pollination syndrome. So far ant pollination has been reported largely in herbaceous species, growing in warm and dry habitats. While studying pollination ecology of Syzygium species (Myrtaceae), growing in tropical forests of the Western Ghats, India, we observed one of the ant species, Technomyrmex albipes, to be the dominant floral visitor in S. occidentale (Bourd.) Chithra among a range of other insect (species of Xylocopa and Trigona, and Apis cerana) and bird visitors. We studied the role of ant species in pollination when compared to other floral visitors. The fruit set in flowers exclusively visited by T. albipes was significantly higher than those visited by any other visitor. The day and night exclusive pollination experiments allowing only T. albipes indicated diel pollination by T. albipes, which was the only active flower visitor during the night. The breeding system of the species was studied through controlled pollinations. The species is partially self-compatible and exhibits considerable autogamy.     

Ant-partners play a minor role on occurrence of the myrmecophilous butterfly <Emphasis Type="Italic">Leptotes cassius</Emphasis> in its host plant

Ant-related oviposition in facultatively myrmecophilous lycaenid butterflies is common, but not universal. In fact, our knowledge of ant-related oviposition in lycaenids is based on some common species (e.g., Rekoa marius, Allosmaitia strophius, Parrhasius polibetes), which limits generalizations about these systems. In this study, we experimentally investigated whether the oviposition pattern of the florivorous lycaenid Leptotes cassius was influenced by the presence of Camponotus ants and whether larvae were attended, rather than attacked, by ants. This might be evidence of myrmecophily. Both L. cassius and Camponotus ants occur on Bionia coriacea, an extrafloral nectaried legume shrub that grows in the Brazilian cerrado. Plants were randomly assigned to ant-present and ant-excluded treatments and were observed twice throughout the short reproductive season. Larvae of L. cassius were tended by ants, whose attendance was characterized by active antennation on the last body segments of the caterpillars. Therefore, Camponotus can be considered a partner of L. cassius. Lycaenid abundance was on average 1.9- and 1.21-fold higher in plants with ants in each sampling period, respectively, indicating a tendency of L. cassius to occur in plants with ants. Nonetheless, results were not statistically significant, suggesting that in this case ants are not a major cue for lycaenid oviposition. In many ant–lycaenid mutualisms, butterfly immatures benefit from reduced parasitism rates. However, no L. cassius immature, regardless of ant presence or absence, was parasitized. Furthermore, larvae may occur inside flower buds that may provide protection from natural enemies; thus, ants may not be required for immature protection.     

Reduced ant defenses in <Emphasis Type="Italic">Macaranga</Emphasis> myrmecophytes (Euphorbiaceae) infested with a winged phasmid <Emphasis Type="Italic">Orthomeria cuprinus</Emphasis>

Macaranga is a tree genus that includes many species of myrmecophytes, which are plants that harbor ant colonies within hollow structures known as domatia. The symbiotic ants (plant–ants) protect their host plants against herbivores; this defense mechanism is called ‘ant defense’. A Bornean phasmid species Orthomeria cuprinus feeds on two myrmecophytic Macaranga species, Macaranga beccariana and Macaranga hypoleuca, which are obligately associated with Crematogaster ant species. The phasmids elude the ant defense using specialized behavior. However, the mechanisms used by the phasmid to overcome ant defenses have been insufficiently elucidated. We hypothesized that O. cuprinus only feeds on individual plants with weakened ant defenses. To test the hypothesis, we compared the ant defense intensity in phasmid-infested and non-infested M. beccariana trees. The number of plant–ants on the plant surface, the ratio of plant–ant biomass to tree biomass, and the aggressiveness of plant–ants towards experimentally introduced herbivores were significantly lower on the phasmid-infested trees than on the non-infested trees. The phasmid nymphs experimentally introduced into non-infested trees, compared with those experimentally introduced into phasmid-infested trees, were more active on the plant surface, avoiding the plant–ants. These results support the hypothesis and suggest that ant defenses on non-infested trees effectively prevent the phasmids from remaining on the plants. Thus, we suggest that O. cuprinus feeds only on the individual M. beccariana trees having decreased ant defenses, although the factors that reduce the intensity of the ant defenses remain unclear.     

Phenotypic and phylogenetic characterization of actinomycetes isolated from <Emphasis Type="Italic">Lasius niger</Emphasis> and <Emphasis Type="Italic">Formica cunicularia</Emphasis> ants

Multiple actinomycete strains were isolated from two ant species, Lasius niger and Formica cunicularia, and their phenotypic properties and phylogenetic position were studied. Partial sequencing of 16S rRNA assigned the greater part of them to the genus Streptomyces, but only one belonged to Nocardia. However, some isolates had significant color and morphological differences from their closest phylogenetic relatives. The abundance and biodiversity of actinomycete communities isolated from L. niger ants greatly exceeded those found for F. cunicularia. All of the actinomycetes associated with F. cunicularia ants demonstrated cellulolytic activity, but only one had such ability among the strains associated with black ants.     

The beginning of the invasion of <Emphasis Type="Italic">Lasius neglectus</Emphasis> (Hymenoptera,Formicidae) in Kiev (Ukraine)

Research conducted in August and September 2016 in the city of Kiev (Ukraine). A total of 8 places of the possible location of invasive garden ant Lasius neglectus Van Loon, Boomsma & Andrasfalvy, 1990 were observed. Only one monocalic colony of L. neglectus was discovered. The forage area of L. neglectus is bounded by asphalted area on three sides. We consider that the invasion of L. neglectus at an early stage can be successful under the following conditions: (a) Favorable temperature. Mitigating of the climate during the winter months in recent years contributed to the consolidation of L. neglectus in Kiev. (b) Spatial separation of nests and foraging trails of native dominant ant species. In our case, L. neglectus colony was found on the site free of other ants. (c) Visiting trees, where native ant species are few or absent. This one partially overlaps with the previous condition.     

The population of earthworms (Lumbricidae) in the main types of dark coniferous forests in Pechora-Ilych Nature Reserve

The population of earthworms has been studied in the main types of old-growth dark coniferous forests of Pechora-Ilych Nature Reserve (Komi Republic) that have not been subject to anthropogeniс impact for a long time. Ten species of earthworms have been identified. The greatest diversity (7 species), abundance, and biomass of earthworms has been revealed in the tall-grass fir–spruce forests. P. diplotetratheca had the greatest abundance. E. nordenskioldi nordenskioldi and E. atlavinyteae had the greatest biomass. The lowest species diversity (3 species), abundance, and biomass of earthworms have been found in the largefern, blueberry–green moss, and sphagnum–horsetail fir–spruce forests. The role of deadwood in maintaining the species diversity of Lumbricidae in dark coniferous forests has been demonstrated. The complexes of Lumbricidae have been considered in anthropogenically disturbed territories, where the following species with a habitat range to the south of the northern and middle taiga have been identified: L. rubellus, A. rosea, A. caliginosa caliginosa, and E. fetida.     

Molecular phylogenetic analysis of the ant genus <Emphasis Type="Italic">Formica</Emphasis> L. (Hymenoptera: Formicidae) from Palearctic region

Sixty-five sequences of the mitochondrial DNA cytochrome b gene fragment (759 bp) and 23 sequences of the NADH dehydrogenase subunit 6 gene fragment (224 bp) were compared in ants of the genus Formica L. from different regions of the Palearctic and in Polyergus rufescens Latr. as outgroup. In total, 28 species of the genus Formica were examined. As a result, dated trees with a molecular clock were constructed showing the phylogenetic relationships of Formica ants. The topology of the obtained tree based on the Cyt-b sequences was found to be not consistent with the generally accepted opinion on the Formica rufa and F. rufibarbis groups. New data on the formation history of the present-day fauna of Formica ants of the Palearctic were obtained. It was demonstrated that a considerable fraction of the examined species (about a third) were formed in the Quaternary Period.     

Soil microbial communities buffer physiological responses to drought stress in three hardwood species

Trees possess myriad adaptations for coping with drought stress, but the extent to which their drought responses are influenced by interactions with soil microbes is poorly understood. To explore the role of microbes in mediating tree responses to drought stress, we exposed saplings of three species (Acer saccharum, Liriodendron tulipifera, and Quercus alba) to a four week experimental drought in mesocosms. Half of the pots were inoculated with a live soil slurry (i.e., a microbial inoculum derived from soils beneath the canopies of mature A. saccharum, L. tulipifera or Q. alba stands), while the other half of the pots received a sterile soil slurry. Soil microbes ameliorated drought stress in L. tulipifera by minimizing reductions in leaf water potential and by reducing photosynthetic declines. In A. saccharum, soil microbes reduced drought stress by lessening declines in leaf water potential, though these changes did not buffer the trees from declining photosynthetic rates. In Q. alba, soil microbes had no effects on leaf physiological parameters during drought stress. In all species, microbes had no significant effects on dynamic C allocation during drought stress, suggesting that microbial effects on plant physiology were unrelated to source–sink dynamics. Collectively, our results suggest that soil microbes have the potential to alter key parameters that are used to diagnose drought sensitivity (i.e., isohydry or anisohydry). To the extent that our results reflect dynamics occurring in forests, a revised perspective on plant hydraulic strategies that considers root-microbe interactions may lead to improved predictions of forest vulnerability to drought.     

Interaction between a threatened endemic plant (<Emphasis Type="Italic">Anchusa crispa</Emphasis>) and the invasive Argentine ant (<Emphasis Type="Italic">Linepithema humile</Emphasis>)

Seed dispersal mutualisms are essential to ensure the survival of diverse plant species and communities worldwide. Here, we investigated whether the invasive Argentine ant can replace native ants by fulfilling their functional role in the seed dispersal of the rare and threatened endemic myrmecochorous plant, Anchusa crispa, in Corsica (France). Our study addressed the potential of Linepithema humile to disperse elaiosome-bearing seeds of A. crispa, examining L. humile’s effects on (1) the composition of communities of ants removing seeds, (2) the number of seed removals, (3) seed preference, (4) the distance of seed dispersion, and (5) seed germination. We caught seven native species at the control site, but only the Argentine ant at invaded sites. L humile removed A. crispa seeds in greater numbers than did native ants, respectively 66 and 23%, probably due to their higher worker density. The invader was similar to native ants with respect to distance of seed transport. Finally, rates of seed germination were not significantly different between seeds previously in contact with either Argentine ants or not. Taken all together, these results suggest that the Argentine ant is unlikely to pose a threat to A. crispa population. These results have important implications for the management of this rare and threatened endemic plant and provide an example of non-negative interactions between invasive and native species.     

Mass Occurrence and Dominant Behavior of the European Ant Species <Emphasis Type="Italic">Formica fuscocinerea</Emphasis> (Forel)

Recently, masses of the ant Formica (Serviformica) fuscocinerea (Forel) have been occurring at numerous sites in Southern Germany. Although F. fuscocinerea is native to Southern Germany, these mass occurrences resemble ant invasions in density and dominance. This study aimed to investigate the underlying mechanisms that promote sudden mass occurrence of a previously inconspicuous ant species within its native range. To estimate the competitive dominance of F. fuscocinerea, species occurrence and abundance considering biotic and abiotic parameters were studied in a natural habitat where F. fuscocinerea co-occurred with two other common ant species, Myrmica ruginodis (Nylander) and Lasius niger (Linnaeus). To understand the species’ distribution in the field, laboratory experiments on interspecific competition were conducted. Finally, the colony structure of F. fuscocinerea was investigated with intraspecific aggression tests. Formica fuscocinerea dominated an area that, as indicated by strongly frequented foraging trails on the trees, provided important food sources, e.g. trophobionts, to the ants. Other ant species coexisted only at the periphery of the F. fuscocinerea range. Laboratory experiments revealed F. fuscocinerea as highly dominant species. Additionally, F. fuscocinerea showed a complete lack of intraspecific aggression between ants originating from distances up to 58 km, indicating weak or nonexistent behavioral boundaries among ants of physically separated nests. Since extraordinarily high worker densities, strong interspecific dominance and a lack of colony boundaries within supercolonies are considered to be important traits of several invasive ant species we conclude that the same traits also promote the dominance of F. fuscocinerea.     

Soil spore banks of ectomycorrhizal fungi in endangered Japanese Douglas-fir forests

Interactions between trees and ectomycorrhizal fungi are critical to the growth and survival of both partners. However, ectomycorrhizal symbiosis has barely been explored in endangered trees, and no information is available regarding soil spore banks of ectomycorrhizal fungi from forests of threatened trees. Here, we evaluated soil spore banks of ectomycorrhizal fungi from endangered Japanese Douglas-fir (Pseudotsuga japonica) forests using bioassay approaches with congeneric P. menziesii and Pinus densiflora seedlings in combination with molecular identification techniques. Rhizopogon togasawariana was predominant in soil propagule banks and was found in all remaining P. japonica forests when assayed with P. menziesii, while no colonization of this fungus was observed on Pinus seedlings. Given the observed specificity of R. togasawariana for P. menziesii and its phylogenetic position within the Pseudotsuga-specific Rhizopogon lineage, its geographical distribution is likely restricted to the remaining Japanese Douglas-fir forests, indicating a high extinction risk for this fungus as well as its endangered host. Spore banks of R. togasawariana remained highly infective after preservation for 1 year or heat treatment at 70 °C, suggesting an ecological strategy of establishing ectomycorrhizal associations on regenerating Japanese Douglas-fir seedlings after disturbance, as observed in other Rhizopogon–Pinaceae combinations. Therefore, the regeneration of Japanese Douglas-fir seedlings may depend largely on the soil spore banks dominated by R. togasawariana, which has co-evolved with the Japanese Douglas-fir for over 30 million years. More attention must be paid to underground ectomycorrhizal fungi for the conservation of endangered tree species, especially in the era of human-induced mass extinction.