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.

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.

Electrophysiological and Behavioral Responses of an Ambrosia Beetle to Volatiles of its Nutritional Fungal Symbiont

Ambrosia beetles (Coleoptera: Scolytinae) cultivate their fungal symbiont within host substrates as the sole source of nutrition on which the larvae and adults must feed. To investigate a possible role for semiochemicals in this interaction, we characterized electrophysiological and behavioral responses of Xylosandrus germanus to volatiles associated with its fungal symbiont Ambrosiella grosmanniae. During still-air walking bioassays, X. germanus exhibited an arrestment response to volatiles of A. grosmanniae, but not antagonistic fungi Beauveria bassiana, Metarhizium brunneum, Trichoderma harzianum, the plant pathogen Fusarium proliferatum, or malt extract agar. Solid phase microextraction-gas chromatography-mass spectrometry identified 2-ethyl-1-hexanol, 2-phenylethanol, methyl benzoate and 3-methyl-1-butanol in emissions from A. grosmanniae; the latter two compounds were also detected in emissions from B. bassiana. Concentration-responses using electroantennography documented weak depolarizations to A. grosmanniae fungal volatiles, unlike the comparatively strong response to ethanol. When tested singly in walking bioassays, volatiles identified from A. grosmanniae elicited relatively weak arrestment responses, unlike the responses to ethanol. Xylosandrus germanus also exhibited weak or no long-range attraction to the fungal volatiles when tested singly during field trials in 2016–2018. None of the fungal volatiles enhanced attraction of X. germanus to ethanol when tested singly; in contrast, 2-phenylethanol and 3-methyl-1-butanol consistently reduced attraction to ethanol. Volatiles emitted by A. grosmanniae may represent short-range olfactory cues that could aid in distinguishing their nutritional fungal symbiont from other fungi, but these compounds are not likely to be useful as long-range attractants for improving detection or mass trapping tactics.

Colonization of Trees by Ambrosia Beetles (Coleoptera: Curculionidae: Scolytinae) Is Influenced by Duration of Flood Stress

The ambrosia beetles Xylosandrus germanus (Blandford) and Xylosandrus crassiusculus (Motschulsky) bore into flood-stressed trees to establish colonies, but the influence of flooding duration on colonization is unknown. This relationship was examined by flooding trees for various time periods and evaluating colonization. In one experiment, X. germanus bored into 20 dogwood (Cornus florida L.) trees during a 3-d flood treatment. Ten trees dissected that season had no offspring present in tunnels; the remaining trees appeared healthy and bloomed the following spring. In another experiment, dogwood trees were flooded for 3 or 7 d and then dissected to assess colonization. The incidence of superficial (short unbranched) and healed (callus tissue in entrance) tunnels was greater in the 3-d trees, while the incidence of tunnels with X. germanus or offspring was greater in the 7-d trees. Four experiments (three in Ohio and one in Virginia) had flood treatments of 0 (nonflooded), 3, 5, 7, and 10 d. Numbers of tunnel entrances, tunnels with X. germanus, and incidence of tunnels with offspring or live foundresses tended to increase as flood duration increased on apple (Malus × domestica Borkh.), dogwood, and redbud (Cercis canadensis L.) in Ohio and redbud in Virginia. Nonflooded trees in Ohio had no boring activity, but ambrosia beetles bored into three nonflooded trees in Virginia. Indicators of unsuccessful colonization, such as superficial tunnels and healing, decreased as flood duration increased. These results suggest tree crops may recover from boring by ambrosia beetles following short-duration flood events, and not necessarily require culling.

Semiochemical-mediated host selection by Xylosandrus spp. ambrosia beetles (Coleoptera: Curculionidae) attacking horticultural tree crops: a review of basic and applied science

Exotic ambrosia beetles (Curculionidae: Scolytinae) in the tribe Xyleborini include destructive pests of trees growing in horticultural cropping systems. Three species are especially problematic: Xylosandrus compactus (Eichhoff), Xylosandrus crassiusculus (Motschulsky), and Xylosandrus germanus (Blandford). Due to similarities in their host tree interactions, this mini-review focuses on these three species with the goal of describing their host-selection behaviour, characterising associated semiochemicals, and assessing how these interactions relate to their management. All three of these Xylosandrus spp. attack a broad range of trees and shrubs. Physiologically stressed trees are preferentially attacked by X. crassiusculus and X. germanus, but the influence of stress on host selection by X. compactus is less clear. Ethanol is emitted from weakened trees in response to a variety of stressors, and it represents an important attractant for all three species. Other host-derived compounds tested are inconsistent or inactive. Verbenone inhibits attraction to ethanol, but the effect is inconsistent and does not prevent attacks. Integrating repellents and attractants into a push–pull management strategy has been ineffective for reducing attacks but could be optimised further. Overall, maintaining host vigour and minimising stress-induced ethanol are keys for managing these insects, particularly X. crassiusculus and X. germanus.