Insect wood borers on commercial North American tree species growing in China: review of Chinese peer-review and grey literature.

Invasive insect wood borers are a threat to global forests and tree-related industries as they can damage trees and spread plant pathogens. Reports of damages by wood borers on plants that were planted overseas may facilitate the identification of potential invaders and speed up risk assessment. However, much of this information remains unavailable to the international plant protection community due to language barriers, lack of digitization, or limited circulation of regional literature. Here, we investigated reports of wood borers on 7 important North American commercial tree species planted in China (Carya illinoinensis, Liquidambar styraciflua, Pinus elliottii, Pinus taeda, Quercus texana, Quercus rubra, and Quercus virginiana) in peer-reviewed as well as "grey" (nonpeer-reviewed) Chinese literature. A total of 60 unique wood borer records were found, yielding reports of 4 orders, 39 genera, and 44 species of insect wood borers. Among Coleoptera, longhorned beetles (Cerambycidae) were the most commonly reported colonizers of North American trees in China. Chinese peer-reviewed reports of pests on alien plants are a valuable tool to survey for potential wood-boring invaders of North America, and wherever North American trees are planted and have the potential to encounter Asian invasive insects. Digitization and dissemination of non-English literature are essential for contemporary risk assessment. On the other hand, the nonpeer reviewed "grey" literature, primarily agency reports and student theses, provided only 5% of the records; many incidental observations were unreliable.

Four New Species of Harringtonia: Unravelling the Laurel Wilt Fungal Genus.

Symbiosis between beetles and fungi arose multiple times during the evolution of both organisms. Some of the most biologically diverse and economically important are mutualisms in which the beetles cultivate and feed on fungi. Among these are bark beetles and Harringtonia, a fungal genus that produces Raffaelea-like asexual morph and hosts the causal agent of laurel wilt, H. lauricola (formerly Raffaelea lauricola). In this study, we propose four new species of Harringtonia associated with beetles from Belize and Florida (USA). We hope to contribute towards a more robust and inclusive phylogenetic framework for future studies on these beetle-fungi relationships and their potential impact in crops and forests worldwide.

Field Response of Black Turpentine Beetle to Pine Resin Oxidation and Pheromone Displacement.

The black turpentine beetle, Dendroctonus terebrans, is an economically important pest of pines in the Southeastern U.S., with a high potential for invasion to other pine-rich regions. Dendroctonus terebrans attraction to an injured host tree lessens over time as the host material degrades. Likewise, kairomonal volatiles emitted from the host change as constituents of the defensive resin oxidize. Therefore we hypothesized that volatiles associated with a fresh host would be more attractive to D. terebrans than those associated with a dead or dying host. We replicated the natural oxidation process of turpentine, fractionated the distilled products to isolate the oxidized products, and deployed the complex mixtures to measure field attraction based on the amount of oxidation performed. Contrasting with previous studies, our results suggest that D. terebrans attraction is not primarily based on host tree degradation. In a second experiment incorporating Dendroctonus pheromones, we demonstrate D. terebrans has a displacement-dependent response to endo-brevicomin, a pheromone associated with the sympatric southern pine beetle, D. frontalis. This has implications not only for possible interspecific signaling, but also for the role of endo-brevicomin in D. terebrans colonization behavior. The results from this study broaden the understanding of D. terebrans chemical ecology and directly contribute to the development of an effective lure-based monitoring system that will benefit future research and management efforts. This may become important if the species is established outside its native range, as in the closely related red turpentine beetle, Dendroctonus valens, which caused mass pine tree mortality following its introduction to Asia.

Collecting and preserving bark and ambrosia beetles (Coleoptera: Curculionidae: Scolytinae & Platypodinae).

This protocol describes the different methods to collect and preserve bark and ambrosia beetles, detailing collecting tools, recording relevant data, and optimizing step-by-step methods to extract beetles from twigs, branches, bark, and trunks. It elaborates on trapping techniques, tools, lures, baits, and beetle preservation. The main rule of manual collecting is to not attempt to pry the insect out of the wood or bark, but instead, remove the wood/bark away from the beetle: gently and systematically. The main rule of trapping is that there is no general attractant; instead, attractants and traps should reflect the ecology of the targeted beetle taxa.

Geosmithia Species Associated With Bark Beetles From China, With the Description of Nine New Species.

Fungi of the genus Geosmithia are frequently associated with bark beetles that feed on phloem on various woody hosts. Most studies on Geosmithia were carried out in North and South America and Europe, with only two species being reported from Taiwan, China. This study aimed to investigate the diversity of Geosmithia species in China. Field surveys in Fujian, Guangdong, Guangxi, Hunan, Jiangsu, Jiangxi, Shandong, Shanghai, and Yunnan yielded a total of 178 Geosmithia isolates from 12 beetle species. The isolates were grouped based on morphology. The internal transcribed spacer, ß-tubulin, and elongation factor 1-α gene regions of the representatives of each group were sequenced. Phylogenetic trees were constructed based on those sequences. In total, 12 species were identified, with three previously described species (Geosmithia xerotolerans, G. putterillii, and G. pallida) and nine new species which are described in this paper as G. luteobrunnea, G. radiata, G. brevistipitata, G. bombycina, G. granulata (Geosmithia sp. 20), G. subfulva, G. pulverea (G. sp. 3 and Geosmithia sp. 23), G. fusca, and G. pumila sp. nov. The dominant species obtained in this study were G. luteobrunnea and G. pulverea. This study systematically studied the Geosmithia species in China and made an important contribution to filling in the gaps in our understanding of global Geosmithia species diversity.

Ambrosia beetles.

Marek Dzurenko and Jiri Hulcr introduce the fungus farming ambrosia beetles.

Preinvasion Assessment of Exotic Bark Beetle-Vectored Fungi to Detect Tree-Killing Pathogens.

Exotic diseases and pests of trees have caused continental-scale disturbances in forest ecosystems and industries, and their invasions are considered largely unpredictable. We tested the concept of preinvasion assessment of not yet invasive organisms, which enables empirical risk assessment of potential invasion and impact. Our example assesses fungi associated with Old World bark and ambrosia beetles and their potential to impact North American trees. We selected 55 Asian and European scolytine beetle species using host use, economic, and regulatory criteria. We isolated 111 of their most consistent fungal associates and tested their effect on four important southeastern American pine and oak species. Our test dataset found no highly virulent pathogens that should be classified as an imminent threat. Twenty-two fungal species were minor pathogens, which may require context-dependent response for their vectors at North American borders, while most of the tested fungi displayed no significant impact. Our results are significant in three ways; they ease the concerns over multiple overseas fungus vectors suspected of heightened potential risk, they provide a basis for the focus on the prevention of introduction and establishment of species that may be of consequence, and they demonstrate that preinvasion assessment, if scaled up, can support practical risk assessment of exotic pathogens.

Diversity and Evolution of Entomocorticium (Russulales, Peniophoraceae), a Genus of Bark Beetle Mutualists Derived from Free-Living, Wood Rotting Peniophora.

Symbiosis between insects and fungi arose multiple times during the evolution of both groups, and some of the most biologically diverse and economically important are mutualisms in which the insects cultivate and feed on fungi. Among these are bark beetles, whose ascomycetous cultivars are better known and studied than their frequently-overlooked and poorly understood basidiomycetous partners. In this study, we propose five new species of Entomocorticium, fungal mutualists in the Russulales (Basidiomycota) that are mutualistic symbionts of scolytine beetles. We have isolated these fungi from the beetle mycangia, which are structures adapted for the selective storage and transportation of fungal mutualists. Herein, we present the most complete phylogeny of the closely related genera Entomocorticium and Peniophora and provide insights into how an insect-associated taxon (Entomocorticium) evolved from within a wood-decaying, wind-dispersed lineage (Peniophora). Our results indicate that following a transition from angiosperms to gymnosperms, fungal domestication by beetles facilitated the evolution and diversification of Entomocorticium. We additionally propose four new species: Entomocorticium fibulatum Araújo, Li & Hulcr, sp. nov.; E. belizense Araújo, Li & Hulcr, sp. nov.; E. perryae Araújo, Li & Hulcr, sp. nov.; and E. macrovesiculatum Araújo, Li, Six & Hulcr, sp. nov. Our findings highlight the fact that insect-fungi associations remain an understudied field and that these associations harbor a large reservoir of novel fungal species.

Species-rich bark and ambrosia beetle fauna (Coleoptera, Curculionidae, Scolytinae) of the Ecuadorian Amazonian Forest Canopy.

Canopy fogging was used to sample the diversity of bark and ambrosia beetles (Coleoptera, Curculionidae, Scolytinae) at two western Amazonian rainforest sites in Ecuador. Sampling was conducted by Dr Terry Erwin and assistants from 1994-2006 and yielded 1158 samples containing 2500 scolytine specimens representing more than 400 morphospecies. Here, we analyze a subset of these data representing two ecological groups: true bark beetles (52 morphospecies) and ambrosia beetles (69 morphospecies). A high percentage of these taxa occurred as singletons and doubletons and their species accumulation curves did not reach an asymptote. Diversity estimates placed the total scolytine species richness for this taxon subset present at the two sites between 260 and 323 species. The α-diversity was remarkably high at each site, while the apparently high ß-diversity was an artifact of undersampling, as shown by a Monte Carlo resampling analysis. This study demonstrates the utility of canopy fogging for the discovery of new scolytine taxa and for approximate diversity assessment, but a substantially greater sampling effort would be needed for conclusive alpha as well as beta diversity estimates.

Two new invasive Ips bark beetles (Coleoptera: Curculionidae) in mainland China and their potential distribution in Asia.

BACKGROUND: Ips is a bark beetle genus of 45 species, many of which are pests of conifer forests and plantations under stress. Twelve Ips species are recorded from China and presumably native there. From 2016 to 2018, specimens suspected to be Ips calligraphus and Ips grandicollis, were collected from traps with ethanol as a sole lure in Zhuhai, Guangdong, China. Both species originate in North America and infest various species of native or introduced pines. Since Ips species are known to cause or exacerbate problems in pine plantations, and a regional survey using traps baited with attractants were implemented in this study to investigate the extent of the introduction. RESULTS: Both I. calligraphus and I. grandicollis have been collected repeatedly from several traps with Ips attractants in Zhuhai, Guangdong, China since 2016. Potential distributions of these two species in Asia, inferred using MaxEnt, is extensive, given the high projected environmental suitability in North America, South America, Mediterranean Europe, Northern Africa, and Eastern Asia. The host plant of I. calligraphus from Zhuhai was identified as slash pine Pinus elliottii using DNA barcoding of gut contents from trapped individuals. CONCLUSION: This is the first report of the establishment of two American pine bark beetles, I. calligraphus and I. grandicollis in continental Asia. The gut content of both species suggests that these pest feeds on a non-native host. Whether the two species present high-risk to Asian forests will become clear with more research on their interactions with native pines.

Esteya floridanum sp. nov.: An Ophiostomatalean Nematophagous Fungus and Its Potential to Control the Pine Wood Nematode.

The nematophagous fungal genus Esteya is reported as a natural enemy of the pine wood nematode Bursaphelenchus xylophilus, which causes pine wilt in Asia and Europe. During a survey of fungi associated with ambrosia beetles in Florida, an undescribed Esteya species was found. A phylogenetic analysis based on nuclear large subunit and ß-tubulin DNA sequences supported this isolate as a new species, E. floridanum. Morphological and phylogenetic characteristics and a species description are provided here. The fungus was observed to kill the pine wood nematode in vitro. To evaluate the ability of E. floridanum to protect trees against the pine wood nematode in vivo, the effect of prophylactic inoculation was tested on Pinus koraiensis and Larix olgensis in Liaoning, China. The results suggest that the fungus is not a plant pathogen and that it delays wilt and postpones death of two conifer trees. This presents a potential new avenue for research on biocontrol of pine wilt disease and stresses the value of research on pest organisms in their native regions.

East Asian Cryphalus Erichson (Curculionidae, Scolytinae): new species, new synonymy and redescriptions of species.

Cryphalus Erichson, 1836 is a taxonomically challenging genus. It is particularly speciose in Asia. Many species are minor pests of fruit tree crops and forest products. We review collections from East Asia, using external morphology, internal morphology and genetic markers with a focus on sub-tropical species from fruit trees. Four new species are described; Cryphalus gnetivorus Johnson, sp. nov., C. itinerans Johnson, sp. nov., C. morivorus Johnson, sp. nov., and C. paramangiferae Johnson, sp. nov. Ten species are redescribed to enable accurate identification: C. artocarpus (Schedl, 1939), C. dilutus Eichhoff, 1878, C. dorsalis (Motschulsky, 1866), C. exiguus Blandford, 1894, C. kyotoensis Nobuchi, 1966, C. lipingensis Tsai & Li, 1963 (= C. kesiyae Browne, 1975, syn. nov.), C. mangiferae Stebbing, 1914 (= C. artestriatus Browne, 1970, syn. nov.), C. meridionalis (Nobuchi, 1975), C. scopiger Berger, 1917, and C. viburni Stark, 1936. Additional records from new localities and new hosts are also presented.

The Ambrosia Beetle Sueus niisimai (Scolytinae: Hyorrhynchini) is Associated with the Canker Disease Fungus Diatrypella japonica (Xylariales).

Ambrosia beetles in the subtribe Hyorrhynchini are one example of an entire ambrosia beetle lineage whose fungi have never been studied. Here, we identify one dominant fungus associated with a widespread Asian hyorrhynchine beetle Sueus niisimai. This fungus was consistently isolated from beetle galleries from multiple collections. Phylogenetic analyses of combined ITS rDNA and ß-tubulin sequences identified the primary fungal symbiont as Diatrypella japonica Higuchi, Nikaido & Hattori (Diatrypaceae, Xylariales, Sordariomycetes), which was recently described as a pathogen of sycamore (Platanus spp.) in Japan. To assess the invasion potential of this beetle-fungus interaction into the U.S., we have investigated the pathogenicity of two D. japonica strains on four species of healthy landscape trees native to the southeastern United States. Only Shumard oak (Quercus shumardii) responded with lesions significantly greater than the control inoculations, but there was no observable dieback or tree mortality. Although disease symptoms were not as prominent as in previous studies of the same fungus in Japan, routine reisolation from the inoculation point suggests that this species is capable of colonizing healthy sapwood of several tree species. Our study shows that the geographical area of its distribution is broader in Asia and potentially includes many hosts of its polyphagous vector. We conclude that the Sueus-Diatrypella symbiosis has high invasion potential but low damage potential, at least on young trees during the growing season.

Peering into the Cuba phytosanitary black box: An institutional and policy analysis.

To mitigate the movement of non-native organisms with trade, phytosanitary systems have been implemented within and between countries. In some countries such as Cuba, little is known about the within-state plant health system. To facilitate the development of future trade partnership between Cuba and the United States, agencies need to understand the organizational structure and diagnostic capacity of the Cuban Plant Protection System, identify potential synergies between the United States and Cuban systems, and identify steps towards cooperation. This paper fills this critical void by presenting a descriptive analysis of the plant health system in Cuba. Information was integrated from available literature, informal interviews with Cuban experts, and workshops focused on Cuban policies, risk, and potential collaboration attended by Cuban and American experts. We identify the next practical steps in improving cooperation, including building trust and capacity. Mutual understanding of phytosanitary systems will be crucial for the regional economic and environmental stability of a post-embargo United States-Cuban relationship.

Lipids and small metabolites provisioned by ambrosia fungi to symbiotic beetles are phylogeny-dependent, not convergent.

Long-term symbiotic associations often lead to reciprocal adaptation between the involved entities. One of the main challenges for studies of such symbioses is differentiating adaptation from neutral processes and phylogenetic background. Ambrosia fungi, cultivated by ambrosia beetles as their sole food source, provide an excellent model to study evolutionary adaptation in a comparative framework because they evolved many times, and each origin bears features seemingly convergently adapted to the symbiosis. We tested whether the symbiotic lifestyle of unrelated ambrosia fungi has led to convergence in the key feature of the symbiotic phenotype-nutrition provisioning to the vector beetles. We compared conidia and mycelium content in three phylogenetic pairs of ambrosia fungi and their closely related nonambrosia relatives using an untargeted metabolomic assay. Multivariate analysis of 311 polar metabolites and 14063 lipid features revealed no convergence of nutrient content across ambrosia lineages. Instead, most variation of the metabolome composition was explained by phylogenetic relationships among the fungi. Thus the overall metabolome evolution of each ambrosia fungus is mostly driven by its inherited metabolism rather than the transition toward symbiosis. We identified eight candidate lipid compounds with expression levels different between the swollen ambrosia spores and other tissues, but they were not consistently elevated across ambrosia fungi. We conclude that ambrosia provisions consist either of nonspecific nutrients in elevated amounts, or of metabolites that are specific to each of the ambrosia symbioses.

Relationships among wood-boring beetles, fungi, and the decomposition of forest biomass.

A prevailing paradigm in forest ecology is that wood-boring beetles facilitate wood decay and carbon cycling, but empirical tests have yielded mixed results. We experimentally determined the effects of wood borers on fungal community assembly and wood decay within pine trunks in the southeastern United States. Pine trunks were made either beetle-accessible or inaccessible. Fungal communities were compared using culturing and high-throughput amplicon sequencing (HTAS) of DNA and RNA. Prior to beetle infestation, living pines had diverse fungal endophyte communities. Endophytes were displaced by beetle-associated fungi in beetle-accessible trees, whereas some endophytes persisted as saprotrophs in beetle-excluded trees. Beetles increased fungal diversity several fold. Over forty taxa of Ascomycota were significantly associated with beetles, but beetles were not consistently associated with any known wood-decaying fungi. Instead, increasing ambrosia beetle infestations caused reduced decay, consistent with previous in vitro experiments that showed beetle-associated fungi reduce decay rates by competing with decay fungi. No effect of bark-inhabiting beetles on decay was detected. Platypodines carried significantly more fungal taxa than scolytines. Molecular results were validated by synthetic and biological mock communities and were consistent across methodologies. RNA sequencing confirmed that beetle-associated fungi were biologically active in the wood. Metabarcode sequencing of the LSU/28S marker recovered important fungal symbionts that were missed by ITS2, though community-level effects were similar between markers. In contrast to the current paradigm, our results indicate ambrosia beetles introduce diverse fungal communities that do not extensively decay wood, but instead reduce decay rates by competing with wood decay fungi.

Reassessment of the Species in the Euwallacea Fornicatus (Coleoptera: Curculionidae: Scolytinae) Complex after the Rediscovery of the "Lost" Type Specimen.

Ambrosia beetles of the Euwallacea fornicatus (Eichhoff, 1868) species complex are emerging tree pests, responsible for significant damage to orchards and ecosystems around the world. The species complex comprises seven described species, all of which are nearly identical. Given that the morphology-defined species boundaries have been ambiguous, historically, there has been much disagreement on species validity, which was compounded by the presumed loss of the type series of E. fornicatus. The species complex was recently reviewed using morphometrics to associate the type specimens to the clades delineated with molecular data under the assumption of the lost type series. We rediscovered a syntype of Xyleborus fornicatus, and reevaluated the species in the complex using morphometrics. We propose the following taxonomic changes to the species complex: Euwallacea fornicatus (=E. tapatapaoensis (Schedl, 1951); = E. whitfordiodendrus (Schedl, 1942)) syn. res.); E. fornicatior (Eggers, 1923) (=E. schultzei (Schedl, 1951) syn. nov.); E. kuroshio (Gomez and Hulcr, 2018) and E. perbrevis (Schedl, 1951) stat. res. These taxonomic changes shift the species name associated with the widely used common names for two taxa, namely: Euwallacea fornicatus should be used for the "Polyphagous Shot Hole Borer", and E. perbrevis for the "Tea Shot Hole Borer clade a". A lectotype is designated for X. fornicatus in order to stabilize the use of the name.

Cryphalus eriobotryae sp. nov. (Coleoptera: Curculionidae: Scolytinae), a New Insect Pest of Loquat Eriobotrya japonica in China.

A previously unknown bark beetle species, Cryphalus eriobotryae sp. nov. Johnson, 2019 has emerged as a lethal pest of loquat (Eriobotrya japonica) in China. The description of new species has been provided. The new species is distinguished from the other Cryphalus by the weakly aciculate frons, by the antennae, with unevenly spaced procurved sutures, by the short pronotal disc, with hair-like setae, and by the widely spaced mesocoxae. The survey of plantation records from around Suzhou suggests that this beetle was introduced from another area not long before 2017. In the surveyed loquat plantation in 2018, 20-90% of trees showed signs of infestation, and 5% were killed in 2018, resulting in the death of over 1000 trees. Outbreaks of the apparently loquat-specific Cryphalus eriobotryae can be diagnosed by hundreds of cankers on the trunk, and wilted foliage. This pest is of concern as a loquat plantation pest and as a pest of fruit production and ornamental trees within Suzhou, and globally.

A selective fungal transport organ (mycangium) maintains coarse phylogenetic congruence between fungus-farming ambrosia beetles and their symbionts.

Thousands of species of ambrosia beetles excavate tunnels in wood to farm fungi. They maintain associations with particular lineages of fungi, but the phylogenetic extent and mechanisms of fidelity are unknown. We test the hypothesis that selectivity of their mycangium enforces fidelity at coarse phylogenetic scales, while permitting promiscuity among closely related fungal mutualists. We confirm a single evolutionary origin of the Xylosandrus complex-a group of several xyleborine genera that farm fungi in the genus Ambrosiella. Multi-level co-phylogenetic analysis revealed frequent symbiont switching within major Ambrosiella clades, but not between clades. The loss of the mycangium in Diuncus, a genus of evolutionary cheaters, was commensurate with the loss of fidelity to fungal clades, supporting the hypothesis that the mycangium reinforces fidelity. Finally, in vivo experiments tracked symbiotic compatibility throughout the symbiotic life cycle of Xylosandrus compactus and demonstrated that closely related Ambrosiella symbionts are interchangeable, but the probability of fungal uptake in the mycangium was significantly lower in more phylogenetically distant species of symbionts. Symbiont loads in experimental subjects were similar to wild-caught beetles. We conclude that partner choice in ambrosia beetles is achieved in the mycangium, and co-phylogenetic inferences can be used to predict the likelihood of specific symbiont switches.

Genetic Variability Among Xyleborus glabratus Populations Native to Southeast Asia (Coleoptera: Curculionidae: Scolytinae: Xyleborini) and the Description of Two Related Species.

The redbay ambrosia beetle, Xyleborus glabratus Eichhoff, is native to Southeast Asia, where it specializes on Lauraceae trees. It forms a symbiosis with the ambrosia fungus Raffaelea lauricola T.C. Harr., Fraedrich & Aghayeva, which can act as a pathogen in living host trees. The beetle and fungus were recently introduced into the United States, where they have killed millions of native Lauraceae trees and threaten the avocado industry. These introduced populations have limited genetic variation. In the native range, the fungi are genetically variable, but the native genetic variability of the beetles is unknown. It is important to assess the beetle's native genetic variation because different lineages may vary in the capacity to vector this fungus, which may affect disease etiology. Here, we analyzed genetic variation in several Chinese, Taiwanese, and Vietnamese populations of X. glabratus using mitochondrial (COI) and nuclear DNA (CAD) markers. Phylogenetic analysis revealed nine COI haplotypes and four CAD genotypes. Uncorrected 'p' distance for intrapopulation comparisons ranged from 0 to 0.1 and 0 to 0.013 and interpopulation comparisons ranged from 0.137 to 0.168 and 0.015 to 0.032 for COI and CAD, respectively. Two populations exceeded the range of intraspecific nucleotide differences for both genes. Given that individuals from these populations also exhibited consistent morphological differences, they are described as two new species: Xyleborus insidiosus Cognato & Smith, n. sp. and Xyleborus mysticulus Cognato & Smith, n. sp. Xyleborus glabratus was redescribed and a lectotype was designated to facilitate its recognition in light of these new species. These results indicate that X. glabratus is genetically variable and is related to two morphologically similar species. Whether these new species and X. glabratus lineages associate with different fungal strains is unknown. Given that the biology and host colonization of these new species are unknown, preventing their introduction to other regions is prudent.