The black truffle Tuber melanosporum was disseminated all over the world, propelled by the development of a wide variety of empirical practices. A widespread practice, called 'truffle trap', consists of placing pieces of truffles into excavations dug under host trees, and of collecting truffle in these traps in the next years. This research aims at (1) evaluating the effect of this practice on fruitbody production based on the analysis of 9924 truffle traps installed in 11 orchards across T. melanosporum native area in France and (2) exploring the mechanisms involved in fruitbody emergence using traps where the genotypes of introduced truffles were compared with those of fruitbodies collected in the same traps. We confirmed that truffle traps provide a major and highly variable part of truffle ground production, representing up to 89% of the collected fruitbodies. We evidenced a genetic link between introduced spores and collected fruitbodies, and then demonstrated that truffle growers provide paternal partners for mating with local maternal mycelia. We also highlighted that soil disturbance stimulate the vegetative development of established maternal mycelia. This research supports that a widely used traditional practice enhances fruitbody production by shaping favorable conditions and providing sexual partners required for fruiting.

Tuber melanosporum is an ectomycorrhizal (ECM) fungus from Mediterranean transitional ecosystems where ECM trees start to dominate among arbuscular-mycorrhizal (AM) shrubs and herbs (companion plants). Its presence entails the development of 'brûlés', where vegetation is scarce for unknown reasons. Current T. melanosporum production comes from plantations where management often suppresses the understory vegetation, although empirical knowledge advocates a positive role of some companion plants in truffle production. This study aimed at (i) experimentally testing the reciprocal interaction between T. melanosporum and companion plants and (ii) examining T. melanosporum-mediated soil feedback involved in the dynamics of truffle ground vegetation. A three-year experiment was set up with Quercus ilex associated with T. melanosporum (or not, as control), grown in association (or not, as control) with a companion plant. Six companion plant species were chosen based on different empirical criteria including those indicated by local truffle growers' knowledge. A trait-based approach was applied to plants and associated fungi (abundance of T. melanosporum and AM fungi mycelium). Companion plants promoted the development of truffle mycelium. In the presence of T. melanosporum, companion plant growth and nutrition and AM fungi abundance decreased, while the nutrition status of its host increased. The truffle inhibited germination of weed seeds. These results highlight the role of T. melanosporum in mediating plant-plant interactions, possible mechanisms underlying brûlé formation and a potential successional role for T. melanosporum.

The ectomycorrhizal Tuber melanosporum forms edible fruit-bodies after mating between two haploid parents: a maternal individual, which feeds the fruit-body and colonizes surrounding ectomycorrhizas, and a paternal one hitherto undetected around fruit-bodies. Several other aspects of T melanosporum ecology remain unclear, such as the formation of a bride around the mycelium, i.e. patches where plant growth is impaired. Here, we test the hypothesis that T melanosporum, and specifically paternal individuals, is present on the roots of herbaceous, non-ectomycorrhizal plants of the bride. Specific primers detected T. melanosporum on 78.9% of the plant individuals investigated (89.7% of the 80 species considered), and metabarcoding confirmed its presence on 46 of the 70 plant individuals investigated. Genotyping with 13 microsatellites detected maternal individuals of nearby fruit-bodies, but not paternal individuals, whose niche thus remains elusive. We discuss the possibility that T melanosporum is also a root endophyte in non-ectomycorrhizal plants. (C) 2017 Elsevier Ltd and British Mycological Society. All rights reserved.

Abstract Speciation is typically accompanied by the formation of isolation barriers between lineages. Commonly, reproductive barriers are separated into pre‐ and post‐zygotic mechanisms that can evolve with different speed. In this study, we measured the strength of different reproductive barriers in two closely related, sympatric orchids of the Ophrys insectifera group, namely Ophrys insectifera and Ophrys aymoninii to infer possible mechanisms of speciation. We quantified pre‐ and post‐pollination barriers through observation of pollen flow, by performing artificial inter‐ and intraspecific crosses and analyzing scent bouquets. Additionally, we investigated differences in mycorrhizal fungi as a potential extrinsic factor of post‐zygotic isolation. Our results show that floral isolation mediated by the attraction of different pollinators acts apparently as the sole reproductive barrier between the two orchid species, with later‐acting intrinsic barriers seemingly absent. Also, the two orchids share most of their fungal mycorrhizal partners in sympatry, suggesting little or no importance of mycorrhizal symbiosis in reproductive isolation. Key traits underlying floral isolation were two alkenes and wax ester, present predominantly in the floral scent of O. aymoninii. These compounds, when applied to flowers of O. insectifera, triggered attraction and a copulation attempt of the bee pollinator of O. aymoninii and thus led to the (partial) breakdown of floral isolation. Based on our results, we suggest that adaptation to different pollinators, mediated by floral scent, underlies species isolation in this plant group. Pollinator switches may be promoted by low pollination success of individuals in dense patches of plants, an assumption that we also confirmed in our study.

Aim Pleistocene climatic oscillations induced range fluctuations in African rain forest organisms and may have shaped species diversification through allopatric speciation events. We compared the spatial genetic structure of two forest species that live in obligate symbiosis and thus must have experienced the same range fluctuations, as a means to discriminate incipient speciation from transient differentiation simply resulting from past divergence.Location Western central Africa.Methods We genotyped 765 individuals of the tree Barteria fistulosa and 605 colonies of its symbiotic ant Tetraponera aethiops at 12 and 13 microsatellite loci, respectively. We compared the spatial genetic structure of the two symbionts by using Bayesian clustering algorithms, isolation-by-distance analyses and clines of synthetic alleles. We used species niche modelling (climatic and soil variables) to investigate ecological variables associated with genetic discontinuities in tree populations.Results The trees and the ants showed congruent patterns of spatial genetic structure. However, the trees showed a very steep genetic discontinuity between groups north and south of latitude 1 degrees N, which was much weaker in the ants. There was no evidence for effective gene flow between the two tree lineages in contact at the transition zone, despite the presence of a few hybrids. Niche modelling did not predict the occurrence of northern trees south of this genetic transition, and vice versa.Main conclusions The genetic discontinuity near latitude 1 degrees N is inferred to be a tension zone resulting from reproductive incompatibilities between previously allopatric tree lineages. This tension zone may have stabilized at a climatic transition (between boreal and austral seasonal regimes), and matches patterns of genetic structure previously observed in other forest plant species. Our results illustrate independent speciation between two species that live in specific and obligate symbiosis and suggest that a tension zone may separate lineages of several central African forest plants near the thermal equator.

Ectomycorrhizal networks may facilitate the establishment and survival of seedlings regenerating under the canopies of tropical forests and are often invoked as a potential contributor to monodominance. We identified ectomycorrhizal fungi in a monodominant Gilbertiodendron dewevrei (Fabaceae) rain forest in Cameroon, using sporocarps and ectomycorrhizae of three age categories (seedlings, intermediate trees, and large trees) and tentatively revealed nutrient transfer through ectomycorrhizal networks by measuring spontaneous isotopic (C-13 and N-15) abundances in seedlings. Sporocarp surveys revealed fewer ectomycorrhizal fungal taxa (59 species from 1030 sporocarps) than molecular barcoding of ectomycorrhizal roots (75 operational taxonomic units from 828 ectomycorrhizae). Our observations suggest that ectomycorrhizal fungal diversity is similar to that in other mixed tropical forests and provide the first report of the Tuber-Helvella lineage in a tropical forest. Despite some differences, all age categories of G.dewevrei had overlapping ectomycorrhizal fungal communities, with families belonging to Thelephoraceae, Russulaceae, Sebacinaceae, Boletaceae, and Clavulinaceae. Of the 49 operational taxonomic units shared by the three age categories (65.3% of the ectomycorrhizal fungal community), 19 were the most abundant on root tips of all categories (38.7% of the shared taxa), supporting the likelihood of ectomycorrhizal networks. However, we obtained no evidence for nutrient transfer from trees to seedlings. We discuss the composition of the ectomycorrhizal fungal community among the G.dewevrei age categories and the possible role of common ectomycorrhizal networks in this rain forest.Resume Les reseaux mycorhiziens pourraient faciliter l'etablissement et la survie des semis sous la canopee des forets tropicales, en particulier dans les forets monodominantes. Pour etudier ce mecanisme, des champignons ont ete identifies a partir de sporophores et d'ectomycorhizes sur trois categories d'age (semis, arbres intermediaires, arbres adultes) et leur role potentiel dans le transfert de nutriments au benefice des semis a ete teste par des mesures de teneurs isotopiques spontanees (C-13 et N-15) dans une foret monodominante a Gilbertiodendron dewevrei (Fabaceae) du Cameroun. Les sporophores ont ete moins diversifies (59 especes parmi 1030 sporophores) que les ectomycorhizes (75 unites taxonomiques operationnelles parmi 828 ectomycorhizes). La diversite fongique ne differe pas de celle des forets tropicales mixtes. Nous decrivons la lignee Tuber-Helvella pour la premiere fois en foret tropicale. Malgre des differences, les categories d'age, G.dewevrei, partagent des especes de Thelephoraceae, Russulaceae, Sebacinaceae, Boletaceae et Clavulinaceae. Parmi les 49 unites taxonomiques operationnelles (65,3% de la communaute fongique) partages par les trois categories d'age, 19 d'entre eux (38,7% de la communaute fongique partagee) formeraient un reseau mycorhizien commun entre les arbres et les semis. Toutefois, les approches isotopiques n'ont pas permis de detecter de transfert de nutriments des arbres aux semis. Nous discutons la composition et le role des communautes fongiques des trois categories d'age de G.dewevrei dans cette foret tropicale humide.

Elaphomyces ('deer truffles') is one of the most important ectomycorrhizal fungal genera in temperate and subarctic forest ecosystems, but also one of the least documented in public databases. The current systematics are mainly based on macromorphology, and is not significantly different from that proposed by Vittadini (1831). Within the 49 species recognised worldwide, 23 were originally described from Europe and 17 of these were described before the 20th century. Moreover, very recent phylogenetic treatments of the genus are mainly based on a few extra-European species and most common European species are still poorly documented. Based on an extensive taxonomic sampling mainly made in the biogeographically rich Cantabrian area (Spain), complemented with collections from France, Greece, Italy, Norway, Portugal and Sweden, all currently recognized species in Europe have been sequenced at the ITS and 28S of the rDNA. Combined phylogenetic analyses yielded molecular support to sections Elaphomyces and Ceratogaster (here emended), while a third, basal lineage encompasses the sections Malacodermei and Ascoscleroderma as well as the tropical genus Pseudotulostoma. Species limits are discussed and some taxa formerly proposed as genuine species based on morphology and biogeography are re-evaluated as varieties or forms. Spore size and ornamentation, features of the peridial surface, structure of the peridium, and the presence of mycelium patches attached to the peridial surface emerge as the most significant systematic characters. Four new species: E. barrioi, E. quercicola, E. roseolus and E. violaceoniger, one new variety: E. papillatus var. sulphureopallidus, and two new forms: E. granulatus forma pallidosporus and E. anthracinus forma talosporus are introduced, as well as four new combinations in the genus: E. muricatus var. reticulatus, E. muricatus var. variegatus, E. papillatus var. striatosporus and E. morettii var. cantabricus. Lectotypes and epitypes are designated for most recognised species. For systematic purposes, new infrageneric taxa are introduced: E. sect. Ascoscleroderma stat. nov., E. subsect. Sclerodermei stat. nov., E. subsect. Maculati subsect. nov., E. subsect. Muricati subsect. nov., and E. subsect. Papillati subsect. nov. Lastly, E. laevigatus, E. sapidus, E. sulphureopallidus and E. trappei are excluded from the genus and referred to Rhizopogon roseolus, Astraeus sapidus comb. nov., Astraeus hygrometricus and Terfezia trappei comb. nov. (syn.: Terfezia cistophila), respectively.

The life cycles and dispersal of edible fungi are still poorly known, thus limiting our understanding of their evolution and domestication. The prized Tuber melanosporum produces fruitbodies (fleshy organs where meiospores mature) gathered in natural, spontaneously inoculated forests or harvested in plantations of nursery-inoculated trees. Yet, how fruitbodies are formed remains unclear, thus limiting yields, and how current domestication attempts affect population genetic structure is overlooked. Fruit-bodies result from mating between two haploid individuals: the maternal parent forms the flesh and the meiospores, while the paternal parent only contributes to the meiospores. We analyzed the genetic diversity of T. melanosporum comparatively in spontaneous forests vs. plantations, using SSR polymorphism of 950 samples from South-East France. All populations displayed strong genetic isolation by distance at the metric scale, possibly due to animal dispersal, meiospore persistence in soil, and/or exclusion of unrelated individuals by vegetative incompatibility. High inbreeding was consistently found, suggesting that parents often develop from meiospores produced by the same fruitbody. Unlike maternal genotypes, paternal mycelia contributed to few fruitbodies each, did not persist over years, and were undetectable on tree mycorrhizae. Thus, we postulate that germlings from the soil spore bank act as paternal partners. Paternal genetic diversity and outbreeding were higher in plantations than in spontaneous truffle-grounds, perhaps because truffle growers disperse fruitbodies to maintain inoculation in plantations. However, planted and spontaneous populations were not genetically isolated, so that T. melanosporum illustrates an early step of domestication where genetic structure remains little affected.

Background: Pleistocene climatic oscillations led to range fluctuations in African rain forest organisms. Isolation of populations during the driest climatic phases resulted in genetic differentiation through mutation and drift. Recent re-expansion of the forest brought differentiated groups into secondary contact. We investigated whether past climate changes may have led to sufficient differentiation to trigger speciation in a central African rainforest tree, Barteria fistulosa (Passifloraceae). Method: We genotyped 765 individuals of B. fistulosa at 12 microsatellite loci and characterized the spatial genetic structure by using Bayesian clustering algorithms, isolation-by-distance analyses and clines of synthetic alleles. We used species niche modelling (environmental and soil variables) to investigate ecological variables associated with genetic discontinuities. Results: Trees showed a very steep genetic discontinuity between groups north and south of latitude 1°N. There was no evidence for effective gene flow between the two tree lineages in contact at the transition zone, despite the presence of a few hybrids. Niche modelling did not predict the occurrence of northern trees south of this genetic transition, and vice versa. The variable that contributed the most to niche differentiation was precipitation during the driest quarter of the year. Discussion: The genetic discontinuity near latitude 1°N is inferred to be a tension zone resulting from reproductive incompatibilities between previously allopatric tree lineages. This tension zone may have stabilized at a climatic transition (between boreal and austral seasonal regimes), and matches patterns of genetic structure previously observed in other forest plant species of the region, suggesting that a tension zone may separate distinct lineages of several central African forest plants near the thermal equator. Our results suggest that northern and southern lineages could be locally adapted to climatic parameters, even for species with a continuous distribution in this area, and thus may respond differently to climate change. Understanding spatial genetic structure may thus help refine prediction of species distribution under future climate conditions. (Texte integral)

Summary In the M editerranean region, patches of vegetation recovering from disturbance and transiently dominated by shrubs produce one of the world's most prized fungi, the black truffle ( T uber melanosporum ). In these successional plant communities, we have fragmentary knowledge of the distribution of T . melanosporum in space among ectomycorrhizal ( ECM ) host species and in time. Molecular identification of hosts (Restriction Fragment Length Polymorphism) and fungi (Internal Transcribed Spacer sequencing) and quantification of T . melanosporum mycelium (quantitative Polymerase Chain Reaction) were employed to evaluate the presence of T . melanosporum on four dominant ECM host species ( Q uercus ilex , Q uercus coccifera , A rbutus unedo , C istus albidus ) and the extent to which their respective ECM communities shared fungal diversity, over the course of development of truffle grounds, from recent unproductive brûlés to senescent ones where production has stopped. We found that truffle grounds host rich communities in which multi‐host fungal species dominate in frequency. When considering both ECM tips and soil mycelia, we documented a dynamic and spatially heterogeneous pattern of T . melanosporum distribution in soils and a presence of ECM tips restricted to Q . ilex roots. This study advances our knowledge of the ecology of T . melanosporum , and provides insight into the extent of ECM fungal sharing among plant species that dominate M editerranean landscapes.

Summary Some forest understorey plants recover carbon (C) not only from their own photosynthesis, but also from mycorrhizal fungi colonizing their roots. How these mixotrophic plants use the resources obtained from mycorrhizal and photosynthetic sources remains unknown. We investigated C sources and allocation in mixotrophic perennial orchids from the genus Epipactis. Based on the assumption that fungal biomass has high δ13C and N content, while photosynthetic biomass has lower δ13C and N content, we indirectly estimated the respective contributions of these two resources to various organs, at various times over the growth season. Fully heterotrophic and fully autotrophic plants from the same sites were used as references for δ13C and N content of biomass purely issuing from fungi and photosynthesis, respectively. In four investigated populations, the biomass shifted from fully heterotrophic in young spring shoots to 80–100% autotrophic in leaves and fruits at fruiting time, suggesting that photosynthesis supported mostly fruiting costs. In addition, fungal colonization decreased in roots over this period. Based on δ13C and N content, below‐ground organs and young spring shoots from green (mixotrophic) individuals and spontaneous achlorophyllous variants (fully heterotrophic) displayed similar fungal C contributions. Similar fungal contributions were also found in shoots of individuals that were either sprouting (and thus partially photosynthetic) or dormant (and thus fully heterotrophic) in the previous years. Therefore, fungal C supported mostly young spring shoots and below‐ground organs. Although experimentally shaded plants had decreased contributions of photosynthetic C in shoots, experimentally defoliated plants showed no increase in fungal C contribution as compared with non‐defoliated controls. Strikingly, these defoliated plants maintained the same seed production: they likely compensated defoliation by increasing stem and fruit photosynthesis. Synthesis. We propose a falsifiable model of C resource allocation in mixotrophic orchids, where mycorrhizal fungi mostly support below‐ground organs and survival, while photosynthesis mostly supports above‐ground sexual reproduction, but not below‐ground reserves. We discuss how this allocation pattern, where seed production depends on photosynthesis, complicates the evolutionary route to full heterotrophy in mixotrophic orchids.

Applying early names, with or without original material, to genealogical species is challenging. For morels this task is especially difficult because of high morphological stasis and high plasticity of apothecium color and shape. Here we propose a nomenclatural revision of true morels (Morchella, Pezizales) from Europe and North America, based on molecular phylogenetic analyses of portions of the genes for RNA polymerase II largest subunit (RPB1) and second largest subunit (RPB2), translation elongation factor-1α (TEF1), the nuc rDNA region encompassing the internal transcribed spacers 1 and 2, along with the 5.8S rDNA (ITS), and partial nuc 28S rDNA D1-D2 domains (28S). The 107 newly sequenced collections were from both continents, including 48 types, together with previously published sequences. Names are applied to 30 of the 65 currently recognized genealogical species. Results of the present study revealed that the number of Morchella species in Europe (n = 21) is nearly identical to that in North America (n = 22). Only seven species were found on both continents, consistent with previous reports of high continental endemism within the genus. Presently it is not possible to tell whether the transoceanic disjunctions were due to human activities, migration across a Bering land bridge or long-distance dispersal. In an effort to stabilize the taxonomy, due in part to the recent publication of synonyms for 11 of the species, accepted names are presented together with their corresponding later synonyms. A new subclade that includes holotypes of M. castanea and M. brunneorosea is identified in sect. Morchella (Esculenta Clade). Lectotypes for Morchella deliciosa, M. eximia and M. tridentina are designated here, as well as epitypes for M. dunalii, M. eximia, M. purpurascens and M. vulgaris. Morchella conica was determined to be illegitimate, and further research is required to determine the identity of M. elata and M. inamoena.

BACKGROUND AND AIMS:Adansonia comprises nine species, six of which are endemic to Madagascar. Genetic relationships between the Malagasy species remain unresolved due to conflicting results between nuclear and plastid DNA variation. Morphologically intermediate individuals between distinct species have been identified, indicative of interspecific hybridization. In this paper, microsatellite data are used to identify potential cases of hybridization and to provide insights into the evolutionary history of the genus on Madagascar.METHODS:Eleven microsatellites amplified with new primers developed for Adansonia rubrostipa were used to analyse 672 individuals collected at 27 sites for the six Malagasy species and morphologically intermediate individuals. Rates of individual admixture were examined using three Bayesian clustering programs, STRUCTURE, BAPS and NewHybrids, with no a priori species assignment.KEY RESULTS:Population differentiation was coherent, with recognized species boundaries. In the four Malagasy species of section Longitubae, 8·0, 9·0 and 9·5 % of individuals with mixed genotypes were identified by BAPS, NewHybrids and STRUCTURE, respectively. At sites with sympatric populations of A. rubrostipa and A. za, NewHybrids indicated these individuals to be F2 and, predominantly, backcrosses with both parental species. In northern Madagascar, two populations of trees combining A. za and A. perrieri morphology and microsatellite alleles were identified in the current absence of the parental species.CONCLUSIONS:The clear genetic differentiation observed between the six species may reflect their adaptation to different assortments of climate regimes and habitats during the colonization of the island. Microsatellite variation reveals that hybridization probably occurred in secondary contact between species of section Longitubae. This type of hybridization may also have been involved in the differentiation of a local new stabilized entity showing specific microsatellite alleles and morphological characters, suggesting a potential role of hybridization in the recent history of diversification on Madagascar.

Dust seeds that germinate by obtaining nutrients from symbiotic fungi have evolved independently in orchids and 11 other plant lineages. The fungi involved in this 'mycoheterotrophic' germination have been identified in some orchids and non-photosynthetic Ericaceae, and proved identical to mycorrhizal fungi of adult plants. We investigated a third lineage, the Pyroleae, chlorophyllous Ericaceae species whose partial mycoheterotrophy at adulthood has recently attracted much attention. We observed experimental Pyrola asarifolia germination at four Japanese sites and investigated the germination pattern and symbiotic fungi, which we compared to mycorrhizal fungi of adult plants. Adult P. asarifolia, like other Pyroleae, associated with diverse fungal species that were a subset of those mycorrhizal on surrounding trees. Conversely, seedlings specifically associated with a lineage of Sebacinales clade B (endophytic Basidiomycetes) revealed an intriguing evolutionary convergence with orchids, some of which also germinate with Sebacinales clade B. Congruently, seedlings clustered spatially together, but not with adults. This unexpected transition in specificity and ecology of partners could support the developmental transition from full to partial mycoheterotrophy, but probably challenges survival and distribution during development. We discuss the physiological and ecological traits that predisposed to the repeated recruitment of Sebacinales clade B for dust seed germination.

The four species of the central African genus Barteria show variation in habitat and in degree of association with ants. Whereas B. solida, restricted to submontane forests, attracts opportunistic ants to extrafloral nectar, the three other species, found in lowland rainforests (B. fistulosa, B. dewevrei) and in littoral scrub (B. nigritana), possess stem domatia of varying shapes and degrees of specialisation, hosting either non-specific arboreal ants (B. nigritana, some B. dewevrei) or two large species of ants of the genus Tetraponera Smith, 1852 that are specific to some species of Barteria (B. fistulosa, some B. dewevrei). We aimed to investigate whether this variation represents an evolutionary trend toward increasing specialisation of mutualism or the reduction or loss of myrmecophytic traits. For this, we determined phylogenetic relationships within the genus using DNA sequences (primarily nuclear ITS) and microsatellite genotypes (11 loci) on a large sample of individuals, mostly from Cameroon and Gabon. The two types of markers support an initial dichotomy that groups B. dewevrei with B. nigritana and B. fistulosa with B. solida respectively. Within these pairs, species do not appear reciprocally monophyletic. At microsatellite loci, B. nigritana forms a clade embedded within B. dewevrei; and within both B. solida and B. fistulosa, geographical populations show levels of differentiation similar to that observed between populations of B. solida and B. fistulosa. Geographic distance alone does not account for genetic differentiation between species, which indicates reproductive isolation. Divergence in each of the two pairs implies evolutionary transitions in habitat and in myrmecophytism. Specialised mutualism with specific ant species of the genus Tetraponera has been lost in species found in more marginal habitats.

This article documents the addition of 299 microsatellite marker loci and nine pairs of single‐nucleotide polymorphism (SNP) EPIC primers to the Molecular Ecology Resources (MER) Database. Loci were developed for the following species: Alosa pseudoharengus, Alosa aestivalis, Aphis spiraecola, Argopecten purpuratus, Coreoleuciscus splendidus, Garra gotyla, Hippodamia convergens, Linnaea borealis, Menippe mercenaria, Menippe adina, Parus major, Pinus densiflora, Portunus trituberculatus, Procontarinia mangiferae, Rhynchophorus ferrugineus, Schizothorax richardsonii, Scophthalmus rhombus, Tetraponera aethiops, Thaumetopoea pityocampa, Tuta absoluta and Ugni molinae. These loci were cross‐tested on the following species: Barilius bendelisis, Chiromantes haematocheir, Eriocheir sinensis, Eucalyptus camaldulensis, Eucalyptus cladocalix, Eucalyptus globulus, Garra litaninsis vishwanath, Garra para lissorhynchus, Guindilla trinervis, Hemigrapsus sanguineus, Luma chequen. Guayaba, Myrceugenia colchagüensis, Myrceugenia correifolia, Myrceugenia exsucca, Parasesarma plicatum, Parus major, Portunus pelagicus, Psidium guayaba, Schizothorax richardsonii, Scophthalmus maximus, Tetraponera latifrons, Thaumetopoea bonjeani, Thaumetopoea ispartensis, Thaumetopoea libanotica, Thaumetopoea pinivora, Thaumetopoea pityocampa ena clade, Thaumetopoea solitaria, Thaumetopoea wilkinsoni and Tor putitora. This article also documents the addition of nine EPIC primer pairs for Euphaea decorata, Euphaea formosa, Euphaea ornata and Euphaea yayeyamana.