Thousand cankers disease, caused by the pathogen Geosmithia morbida vectored by the bark beetle Pityophthorus juglandis, has emerged as an important disease of walnut trees in Europe. The present ...study was performed to evaluate the efficacy of trunk injections of four commercial fungicides and one insecticide for control of the fungus and its vector. Laboratory tests indicated that fungicides containing prochloraz + tetraconazole were the most effective. Field trials on non-infected trees allowed for the selection of a mixture containing prochloraz and tetraconazole (Binal Pro), the insecticide abamectin (Vertimec EC) and the adjuvant 2-(2-ethoxyethoxy) ethanol (CarbitolTM) as having rapid host uptake. Injections of this formulation in naturally infected black walnut trees reduced the presence of G. morbida, supporting trunk injection as an efficient and low impact technique to manage fungal damage on infected trees.
Thousand cankers disease (TCD) results from the combined activity of the fungal pathogen, Geosmithia morbida Kolařík, Freeland, Utley, and Tisserat and its principle vector, Pityophthorus juglandis ...(Blackman) (Coleoptera: Curculionidae: Scolytinae) in Juglans L. spp. and Pterocarya Kunth spp. host plants. TCD has been reported from the eastern and western United States. To evaluate potential for other beetle species to vector the fungus in east Tennessee, specimens were collected using ethanol-baited traps that were suspended beneath crowns of TCD-symptomatic trees. Associations of G. morbida with insect species collected in traps were assessed in an unsuccessful, preliminary culture-based fungal assay, and then with a molecular-based detection method. For culture-based assays, rinsate from washed, individual insects was plated on nutrient media and growing colonies were subcultured to obtain axenic G. morbida cultures for identification. For the molecular-based method, G. morbida presence was detected by amplifying the previously developed, species-specific microsatellite locus GS004. Capillary electrophoresis was used to detect the amplified amplicons and representative reactions were validated using Sanger sequencing. Eleven beetle species were found to carry G. morbida, including Cnestus mutilatus (Blandford), Dryoxylon onoharaensum (Murayama), Hylocurus rudis (LeConte), Monarthrum fasciatum (Say), Monarthrum mali (Fitch), Xyleborinus saxesenii (Ratzeburg), Xylosandrus crassiusculus (Motschulsky), Xylosandrus germanus (Blandford) (all Coleoptera: Curculionidae: Scolytinae), Stenomimus pallidus (Boheman) (Coleoptera: Curculionidae: Cossoninae), Oxoplatypus quadridentatus (Olivier) (Coleoptera: Curculionidae: Platypodinae), and Xylops basilaris (Say) (Coleoptera: Bostrichidae). These findings raise concerns that alternative subcortical insect species that already occur within quarantined habitats can sustain incidence of introduced G. morbida and contribute to spread within the native range of black walnut, Juglans nigra L., in the eastern United States.
Thousand Cankers Disease (TCD) of walnut trees is caused by the pathogenic fungus Geosmithia morbida vectored by the walnut twig beetle (WTB) Pityophthorus juglandis. Monitoring efforts for WTB rely ...on pheromone‐baited traps, but lures are likely effective at attracting beetles only over short distances. Fungal‐derived kairomones may increase the efficacy of current lures, while additional volatiles may repel beetles from valuable trees.
The objective of this study was to determine the extent to which fungal, host and non‐host volatiles modify the attraction of WTB to pheromone‐baited traps. A trapping study that combined fungal, host‐associated and non‐host compounds with WTB‐pheromone lures was conducted over three years in black walnut plantations experiencing a TCD outbreak in Walla Walla, WA.
Traps baited with pheromone and G. morbida volatiles (i.e., isoamyl and isobutyl alcohol) consistently attracted more WTB, while other fungal volatiles inconsistently increased attraction compared to those baited with pheromone lure alone. This is the first field study that demonstrates fungal volatiles can increase the attraction of a bark beetle to its pheromone in a hardwood system.
One fungal (benzyl alcohol) and two additional volatiles (limonene, piperitone) repelled WTB from pheromone‐baited traps. Although limonene is known to repel WTB, this is the first demonstration that benzyl alcohol and piperitone repel a bark beetle.
Fungal volatiles may increase the efficacy of monitoring efforts and may play an important role in management tactics for WTB, especially in detecting the introduction and establishment of nascent populations and protecting trees from colonizing beetles.
Thousand cankers disease (TCD), first documented in the western United States in the early 2000s, has spread into nine western and seven eastern states in the United States and northern Italy. TCD ...incidence and severity differ between eastern and western United States outbreak localities. Black walnut (
Juglans nigra
) trees, introduced into both urban and plantation settings in the western United States, have been severely impacted as evident by the documented high disease incidence and mortality. However, in eastern United States localities, where
J. nigra
is native, host-pathogen-vector interactions resulted in two different outcomes: trees either die or partly recover followed by infection. Recent genetic studies on the TCD causal agent,
Geosmithia morbida
, indicate the spatial genetic structure and high levels of genetic diversity among United States populations. Using detached branch inoculation assays, we reported differential virulence among 25
G. morbida
isolates collected across the current distribution range of the disease. As a proxy for virulence, the canker area was measured to 7 days after inoculation. Varying degrees of virulence were observed among tested
G. morbida
isolates, which was partly explained by their genetic provenance (genetic clusters). Isolates that grouped within genetic cluster 2 (
n
= 7 from the eastern United States and
n
= 6 from the western United States; mean = 210.34 mm
2
) induced significantly larger cankers than isolates that grouped within genetic cluster 1 (
n
= 12; all western United States isolates; mean = 153.76 mm
2
). Canker sizes varied among isolates within each genetic cluster and were not correlated with a geographic region (eastern vs. western United States) but rather to the isolated state of origin. Mean canker size also differed in response to isolates that originated from different tree host species.
G. morbida
isolates collected from
Juglans major
induced statistically smaller cankers when compared to isolates recovered from undetermined
Juglans
species but not from
J. nigra
. In sum, the increased mortality reported for western United States walnut tree populations cannot be explained by a higher virulence of local
G. morbida
. Plausible explanations for the observed disparity include environmental conditions, such as prolonged drought, greater population densities of walnut twig beetle causing a higher number of inoculation events to individual trees, and multiple introductions of
G. morbida
originating from multiple locations. Future experimental evaluation should be undertaken to quantify the influence of these factors on the local epidemics.
A phenotype is the expression of interactions between species genotype and environment. We quantified the contributions of ecological and phylogenetic associations to phenotypic variation in ...Geosmithia fungi. Geosmithia are symbiotic beetle-associated saprotrophs with a range of life histories and host specificities, including obligate nutritional beetle mutualists (ambrosia fungi) and phytopathogens. We hypothesized that: (1) species phenotypes are better explained by their ecology than by their phylogenetic relationships; (2) niche specialization was accompanied by enzymatic capability losses; and (3) ambrosia Geosmithia species have higher nutritional quality and antibiotic capabilities than species with facultative symbioses. Our results confirmed that long-term co-evolved specialists have reduced metabolic breadth in comparison to generalists. Phytopathogenic G. morbida produces unique enzyme suites with affinity to ligno-cellulose. Mycelia of ambrosia fungi contain large amounts of oleic fatty acid with nutritive and possibly allelopathic function. Overall, our results indicate that Geosmithia ecology have greater effect on species phenotype than their phylogenetic relationships.
•Enzymatic profiles, biochemical composition and antibiosis in Geosmithia analysed.•Adaptive traits related to the various ecological strategies tracked.•Potential virulence factors of Geosmithia morbida identified.•Specialists exhibit unique enzyme suites and reduced metabolic breadth.•Obligatory symbiotic ambrosia exhibits specific profiles of fatty acids.
Logs of high-value eastern black walnut (Juglans nigra L.) are commonly exported from the United States for production of veneer and lumber. Veneer logs are not debarked to minimize degradation of ...wood quality and reduce moisture loss. Thousand cankers disease (TCD) is caused by the walnut twig beetle (Pityophthorus juglandis Blackman) and the fungal pathogen, Geosmithia morbida M. Kolarik, E. Freeland, C. Utley and N. Tisserat sp. nov., which colonize the inner bark of Juglans species. Effective eradication of these organisms by heat or chemical fumigation treatment is required for walnut logs prior to export. Because vacuum steam is an effective and efficient means of heating round wood, its use in eliminating the TCD causal agents was evaluated using Juglans logs (12- to 44-cm small end diameter and 1.7- to 1.9-m length) fromTCD-symptomatic trees in Oregon and Washington State. Five replicate trials with three logs per load were conducted in a portable vacuum chamber to test two treatment schedules: 60°C for 60 min and 56°C for 30 min. Complete elimination of P. juglandis and G. morbida was achieved when using a minimum of 56°C at 5-cm targeted depth from bottom of bark furrow into the sapwood and held for 30 min. Treatment cycle time ranged from 298 to 576 min depending on log diameter and initial log temperature. Artificial inoculation of J. nigra trees with G. morbida within the TCD range in Pennsylvania was minimally successful in producing adequately colonized logs for experimental trials.
The cultivation of walnuts (Juglans sp.) in Europe retains high economic, social, and environmental value. The recent reporting of the Thousand Cankers Disease (TCD) fungus, Geosmithia morbida, and ...of its vector, Pityophthorus juglandis, in walnut trees in Italy is alarming the whole of Europe. Although Italy is at present the only foothold of the disease outside North America, given the difficulties inherent in traditional identification of both members of this beetle/fungus complex, a rapid and effective protocol for the early detection and identification of TCD organisms is an absolute priority for Europe. Here we report the development of an effective and sensitive molecular tool based on simplex/duplex qPCR assays for the rapid, accurate and highly specific detection of both the bionectriaceous fungal pathogen and its bark-beetle vector. Our assay performed excellently, detecting minute amounts of target DNA without any non-specific amplification. Detection limits from various and heterogeneous matrices were lower than other reported assays. Our molecular protocol could assist in TCD organism interception at entry points, territory monitoring for the early identification and eradication of outbreaks, delineation of quarantine areas, and tracing back TCD entry and dispersal pathways.
The walnut twig beetle, Pityophthorus juglandis Blackman, and associated fungus Geosmithia morbida Kolařík, Freeland, Utley, & Tisserat constitute the insect–fungal complex that causes thousand ...cankers disease in walnut, Juglans spp., and wingnut, Pterocarya spp. Thousand cankers disease is responsible for the decline of Juglans species throughout the western United States and more recently, the eastern United States and northern Italy. We examined the flight capacity of P. juglandis over 24-h trials on a flight mill in the laboratory. The maximum total flight distance observed was ~3.6 km in 24 h; however, the mean and median distances flown by beetles that initiated flight were ~372 m and ~158 m, respectively. Beetles flew for 34 min on average within a 24-h flight trial. Male and female flight capacities were similar, even though males were larger than females (0.64 vs. 0.57 mm pronotal width). Age postemergence had no effect on flight distance, flight time, or mean flight velocity. The propensity to fly, however, decreased with age. We integrated results of flight distance with propensity to fly as beetles aged in a Monte Carlo simulation to estimate the maximum dispersal capacity over 5 d, assuming no mortality. Only 1% of the insects would be expected to fly >2 km, whereas one-third of the insects were estimated to fly <100 m. These results suggest that nascent establishments remain relatively localized without anthropogenic transport or wind-aided dispersal, which has implications for management and sampling of this hardwood pest.
Fungi in the genus Geosmithia (Ascomycota: Hypocreales) are frequent associates of bark beetles and woodborers that colonize hardwood and coniferous trees. One species, Geosmithia morbida, is an ...economically damaging invasive species. The authors surveyed the Geosmithia species of California and Colorado, USA, to (i) provide baseline data on taxonomy of Geosmithia and beetle vector specificity across the western USA; (ii) investigate the subcortical beetle fauna for alternative vectors of the invasive G. morbida; and (iii) interpret the community composition of this region within the emerging global biogeography of Geosmithia. Geosmithia was detected in 87% of 126 beetle samples obtained from 39 plant species. Twenty-nine species of Geosmithia were distinguished, of which 13 may be new species. Bark beetles from hardwoods, Cupressus, and Sequoia appear to be regular vectors, with Geosmithia present in all beetle gallery systems examined. Other subcortical insects appear to vector Geosmithia at lower frequencies. Overall, most Geosmithia have a distinct level of vector specificity (mostly high, sometimes low) enabling their separation to generalists and specialists. Plant pathogenic Geosmithia morbida was not found in association with any other beetle besides Pityophthorus juglandis. However, four additional Geosmithia species were found in P. juglandis galleries. When integrated with recent data from other continents, a global pattern of Geosmithia distribution across continents, latitudes, and vectors is emerging: of the 29 Geosmithia species found in the western USA, 12 have not been reported outside of the USA. The most frequently encountered species with the widest global distribution also had the broadest range of beetle vectors. Several Geosmithia spp. with very narrow vector ranges in Europe exhibited the similar degree of specialization in the USA. Such strong canalization in association could reflect an ancient origin of each individual association, or a recent origin and a subsequent diversification in North America.
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