Can the invasive ambrosia beetle Xylosandrus germanus withstand an unusually cold winter in the West Carpathian forest in Central Europe?

The capability of a non-native species to withstand adverse weather is indicative of its establishment in a novel area. An unusually cold winter of 2016/2017 that occurred in the West Carpathians of Slovakia and other regions within Europe provided an opportunity to indirectly assess survival of the invasive ambrosia beetle Xylosandrus germanus (Coleoptera: Curculionidae, Scolytinae). We compared trap captures of this species in the year preceding and succeeding the respective cold winter. Ethanol-baited traps were deployed in 24 oak dominated forest stands within the southern and central area from April to August 2016, and again from April to August 2017 to encompass the seasonal flight activity of X. germanus and to get acquainted with temporal changes in the abundance of this species in these two distant areas. Dispersing X. germanus were recorded in all surveyed stands before and after the aforementioned cold winter. Their total seasonal trap captures were lower in the southern area following low winter temperatures, but remained similar in the central area. Our results suggest that X. germanus can withstand adverse winter weather in oak dominated forests of the West Carpathians within altitudes of 171 and 450 m asl. It is likely that minimum winter temperatures will not reduce the establishment or further spread of this successful invader in forests in Central Europe.

Stability of Nuclear and Mitochondrial Reference Genes in Selected Tissues of the Ambrosia Beetle Xylosandrus germanus

The fungus-farming ambrosia beetle Xylosandrus germanus (Blandford) uses a pouch-like structure (i.e., mycangium) to transport spores of its nutritional fungal mutualist. Our current study sought to identify reference genes necessary for future transcriptome analyses aimed at characterizing gene expression within the mycangium. Complementary DNA was synthesized using selected tissue types from laboratory-reared and field-collected X. germanus consisting of the whole body, head + thorax, deflated or inflated mycangium + scutellum, inflated mycangium, and thorax + abdomen. Quantitative reverse-transcription PCR reactions were performed using primers for 28S ribosomal RNA (28S rRNA), arginine kinase (AK), carbamoyl-phosphate synthetase 2-aspartate transcarbamylase-dihydroorotase (CAD), mitochondrial cytochrome oxidase 1 (CO1), and elongation factor-1α (EF1α). Reference gene stability was analyzed using GeNorm, NormFinder, BestKeeper, ΔCt, and a comprehensive final ranking by RefFinder. The gene CO1 was identified as the primary reference gene since it was generally ranked in first or second position among the tissue types containing the mycangium. Reference gene AK was identified as a secondary reference gene. In contrast, EF1α was generally ranked in the last or penultimate place. Identification of two stable reference genes will aid in normalizing the expression of target genes for subsequent gene expression studies of X. germanus’ mycangium.

Revision of the xyleborine ambrosia beetle genus Microperus Wood, 1980 (Curculionidae, Scolytinae, Xyleborini) of Thailand with four new species and four newly recorded species

Microperus Wood, 1980 ambrosia beetles in Thailand are reviewed. Four species, M. bidentatussp. nov., M. bucolicussp. nov., M. globodeclivissp. nov., and M. serratussp. nov. are described. Four new combinations are given: Microperus armaticeps (Schedl, 1942) comb. nov., Microperus exsculptus (Eggers, 1927) comb. nov., Microperus pedellus (Schedl, 1969) comb. nov., and Microperus spicatulus (Browne, 1986) comb. nov., stat. res., all from Xyleborus. Two new synonyms are proposed: Microperus cruralis (Schedl, 1975) (= Xyleborus myllus Browne, 1986 syn. nov.), Microperus exsculptus (Eggers, 1927) (= Xyleborus dentipennis Browne, 1983 syn. nov.). Four species are reported from Thailand for the first time: Microperus chrysophylli (Eggers, 1930), Microperus exsculptus, Microperus nanus (Browne, 1949) and Microperus quercicola (Eggers, 1926). With the inclusion of the Microperus species described and recorded herein, the diversity of Microperus is increased to 35 species, of which 18 are recorded in Thailand. An updated key to the Microperus of the Indochinese Peninsula and China is provided. The taxonomy, diagnostic characters, and distribution of species are briefly discussed.

Microbiome Associated with the Mycangia of Female and Male Adults of the Ambrosia Beetle Platypus cylindrus Fab. (Coleoptera: Curculionidae)

The ambrosia beetle Platypus cylindrus Fab. (Coleoptera: Curculionidae) is a major cork oak pest in Portugal. Female and male beetles have different roles in host tree colonization and are both equipped with prothoracic mycangia for fungal transportation. Despite a known beneficial role of bacteria in ambrosia beetles, information on bacterial composition associated with prothoracic mycangia structures is scarce. Bacterial community from mycangia of P. cylindrus male and female beetles collected from cork oak galleries was investigated by means of 16S metagenomics. Mycangia anatomical structure was also explored with histological techniques and X-ray computed microtomography to highlight evidence supporting biological sexual dimorphism. A bacterial community with highly diverse bacterial taxa with low abundances at the genus level was revealed. Lactobacillales, Leptotrichia, Neisseria, Rothia, and Sphingomonadaceae were significantly more abundant in males, while Acinetobacter, Chitinophagaceae, Enterobacteriaceae, Erwiniaceae, Microbacteriaceae, and Pseudoclavibacter were more abundant in females. Additionally, a core bacteriome of five genera was shared by both sexes. Histological examination revealed visible connections linking external and internal tissues in females, but none in males. Overall, these results provide the first insights into sexual differentiation for bacteria in a Platypodinae beetle species, identifying key patterns of bacteria distribution in the context of beetle ecology and functional behavior.

Novel Symbiotic Association Between Euwallacea Ambrosia Beetle and Fusarium Fungus on Fig Trees in Japan

Ficus carica plantations in Japan were first reported to be infested by an ambrosia beetle species, identified as Euwallacea interjectus, in 1996. The purpose of this study was to determine the symbiotic fungi of female adults of E. interjectus emerging from F. carica trees infected with fig wilt disease (FWD). Dispersal adults (51 females) of E. interjectus, which were collected from logs of an infested fig tree in Hiroshima Prefecture, Western Japan, were separated into three respective body parts (head, thorax, and abdomen) and used for fungal isolation. Isolated fungi were identified based on the morphological characteristics and DNA sequence data. Over 13 species of associated fungi were detected, of which a specific fungus, Fusarium kuroshium, was dominant in female head (including oral mycangia). The plant-pathogenic fungus of FWD, Ceratocystis ficicola, was not observed within any body parts of E. interjectus. We further discussed the relationship among E. interjectus and its associated fungi in fig tree.

Xylosandrus germanus (Blandford, 1894) on Grapevines in Italy with a Compilation of World Scolytine Weevils Developing on Vitaceae

The invasive ambrosia beetle Xylosandrus germanus (Curculionidae: Scolytinae: Xyleborini) is recorded for the first time infesting wine grapes in Italy. The type of the attack is illustrated and the possible causes of the onset of the infestation are discussed. Furthermore, given the continuously increasing number of alien wood-borer beetles introduced worldwide, we provide and discuss the updated world checklist of Scolytinae attacking Vitaceae, and Vitis sp. in particular.

XYLOSANDRUS COMPACTUS AND LIPARTHRUM COLCHICUM (COLEOP- TERA SCOLYTINAE) IN TUSCANY: A PRELIMINARY SCREENING OF ASSOCIATED FUNGI

Xylosandrus compactus and Liparthrum colchicum are two Scolytinae recorded for the first time in Italy in 2011 and 2019, respectively. X. compactus is an “ambrosia beetle” causing damages to several plants typical of the Mediterranean maquis through its digging activity on twigs and branches of the host plants. L. colchicum is a “bark beetle”, monophagous on laurel shrubs and trees. During a survey performed in summer of 2020, in “Tenuta Salviati” located in the Migliarino Natural Park (Pisa, Tuscany), adults of X. compactus and L. colchicum were collected from the same branches and apical twigs of Laurus nobilis hedges. The study aimed at characterizing fungi isolated from the external surface of the two scolytids bodies, to evaluate similarities and even their role in fungal spreading in the environment.

Ambrosia Beetles Prefer Closed Canopies: A Case Study in Oak Forests in Central Europe

Research Highlights: The percentage of canopy closure was found to be the main factor associated with ambrosia beetle abundance and species richness. The latter two variables increased as canopy closure increased, probably because a high percentage of canopy closure provides a stable and humid environment suitable for the growth of ambrosia fungi. Objectives: Oak is a common host tree for ambrosia beetles (Coleoptera: Curculionidae: Scolytinae), which have independently evolved a nutritional mutualism with fungi. We suspected that ambrosia beetles might have specific habitat preferences that are different from those of other saproxylic beetles and that reflect the specific habitat preferences of their food, i.e., ambrosia fungi. Methods: We assessed ambrosia beetle abundance with ethanol-lured traps in five old-growth oak dominated forests and five managed oak dominated forests (one trap per forest) during the vegetation period in 2020. We determined whether ambrosia beetle abundance and species richness depend on forest type (managed vs. unmanaged), degree of canopy closure, abundance of oak trees, abundance of coarse deadwood, and abundance of dead oak branches. Results: In total, 4137 individuals of six species of ambrosia beetles associated with oaks were captured. The native ambrosia beetle Anisandrus dispar represented the majority of trapped ambrosia bark beetles. A. dispar along with another ambrosia beetle, Xyleborinus saxesenii, represented 99% of all captured beetles. Conclusions: In addition to canopy closure, the abundance of oak trees and the abundance of dead oak branches were significantly associated with ambrosia beetle abundance and species richness. The abundance of A. dispar was mainly correlated with dead oak branch abundance and the degree of canopy closure, whereas the abundances of X. saxesenii and of the invasive species Xyleborinus attenuatus and Cyclorhipidion bodoanum were mainly correlated with the net area occupied by oak trees.

Co-occurrence variation explains the low host dependence of ambrosia beetles along altitude gradients in SW China

Background Separation of biotic and abiotic impacts on species diversity distribution patterns across a significant climatic gradient is a challenge in the study of diversity maintenance mechanisms. The basic task is to reconcile scale-dependent effects of abiotic and biotic processes on species distribution models. However, Eltonian noise hypothesis predicted that the effects of biotic interactions will be averaged out at macroscales, and there are many empirical observations that biotic interactions would constrain species distributions at micro-ecological scales. Here, we used a hierarchical modeling method to detect the host specificities of ambrosia beetles (Scolytinae and Platypodinae) with their dependent tree communities across a steep climatic gradient, which was embedded within a relatively homogenous spatial niche. Results Species turnover of both trees and ambrosia beetles have a relatively similar pattern, characterized by the climatic proxy at a regional scale, but not at local scales. This pattern confirmed the Eltonian noise hypothesis wherein emphasis was on influences of macro-climate on local biotic interactions between trees and hosted ambrosia beetle communities, whereas local biotic relations, represented by host specificity dependence, were regionally conserved. Conclusions At a confined spatial scale, cross-taxa comparisons of co-occurrence highlighted the importance of the organism’s dispersal. The effects of tree abundance and phylogeny diversity on ambrosia beetle diversity were, to a large extent, indirect, operating via changes in ambrosia beetle abundance through spatial and temporal dynamics of resources distribution. Tree host dependence plays a minor role on the hosted ambrosia beetle community in this concealed wood decomposing interacting system.