Search

Laurel wilt, a fungal disease (Harringtonia lauricola T.C. Harr., Fraedrich and Aghayeva) spread by the Asian redbay ambrosia beetle [Xyleborus glabratus Eichhoff (Coleoptera: Curculionidae: Scolytinae)], presents an imminent threat to North American members of the Lauraceae family, having caused extensive mortality in several species, especially redbay (Persea borbonia L. Spreng). Varying levels of disease resistance have been recorded in redbay under controlled conditions. To investigate if previously monitored putatively resistant field redbays have influenced the regeneration and survival of conspecifics within the surrounding 0.08 ha, a survey was conducted in 2018-19 and compared to similar data collected years prior (2008-09, 2013) along the coasts of Florida, Georgia, and South Carolina, United States. Plots were originally established at six disease-infested study sites around large redbay (>7.5-cm diameter at breast height (1.37 m) (DBH)) that had survived the initial laurel wilt disease epidemic that began in approximately 2007. In 2018-19, a subset of 61 plots within 16 m of the original "survivor" redbay were recorded and compared to previous surveys. Among the original redbay selected for resistance, 22 of 61 (36%) survivors across all sites were alive in 2018 with survival rates varying from 0 to 70% between survey periods (average mortality 3.6%/year). Trees that died in years since 2008-09 had their plots reclassified as susceptible or "suscepts". Changes in mean quadratic diameter at 1.37 m of redbay plots near survivors were significantly greater than those near suscepts, and in 2018-19, the average diameter of redbay near survivors was 7.62 cm vs. 4.90 cm for suscepts. The diameter distribution of dead and live redbay in the whole population showed a decrease in live individuals surviving past 8 cm DBH in 2018–2019, but 20 of 22 survivor candidate trees were larger than 8 cm DBH. Regeneration was occurring both clonally and sexually and tended to be greater near suscepts, but midstory resprouts per hectare and understory seedlings interacted significantly with the site and the latter differed between sites. These findings indicate that redbay is regenerating in these ecosystems, and disease resistance may allow for increased average tree size for some individuals, but an upper size threshold of around 8 cm DBH may still exist for much of the population. In addition, the importance of site variables in regeneration was apparent, making either local genetic or environmental effects an important topic for future research. Continuing to monitor these survivors while locating new candidates for disease screenings and breeding, preventing the introductions of new strains of H. lauricola, shedding light on the nature of resistance and its heritability, and initiating outplanting trials with resistant germplasm are instrumental steps in bringing redbay back to prominence in its historical range. [ABSTRACT FROM AUTHOR]

The redbay ambrosia Xyleborus glabratus beetle has emerged as a significant pest of Laurel trees, Persea spp. [Laurales: Lauraceae], in the southeastern USA due to its symbiotic association with the pathogenic fungus Harringtonia lauricola, the causal agent of the laurel wilt disease. We evaluated the interaction of different species of ambrosia beetles with redbay trees Persea barbonia infected by H. lauricola. We measured the landing and emergence rates of different ambrosia beetle species throughout the wilting process at different heights. We assessed landing height passively in two redbay stands by placing staggered, unbaited white sticky traps at three height levels (low: 0–1 m; middle: 1–1.5 m; high: 1.5–2 m). We collected nine other Scolytinae species including Xyleborus volvulus, Xyleborinus saxesenii, Euplatypus compositus, Xyleborus bispinatus, and Xyleborus affinis. In the three experiments and both locations, the primary vector of H. lauricola, X. glabratus, was the most frequently collected species both in the landing captures and in the emergence rate, independently of the wilting status of the host. However, the numbers and diversity of ambrosia beetles landing on trees and emerging from logs increased when trees were in an advanced stage of wilting. Our results indicate that other ambrosia beetles benefit marginally from laurel wilt and that the contribution of secondary ambrosia beetle vectors in the spread of H. lauricola through lateral infection in the redbay system is most likely minimal as compared to the contribution of X. glabratus. [ABSTRACT FROM AUTHOR]

The redbay ambrosia beetle, harmless in its native Asia, has become a formidable pest since its introduction to the United States in 2002. The beetle spreads the fungus Harringtonia lauricola, a lethal pathogen of North American Lauraceae trees, including avocados. The fungus infection triggers a rapid onset of laurel wilt disease, which has nearly eliminated laurel trees across the southeastern United States within a few years. The beetle has spread across nearly 300 counties in the Southeast, facilitated by trade in wood products. Effective control in natural environments remains an unresolved challenge; in avocado groves, infected trees must be removed and destroyed. Introduction of X. glabratus into Mexico, California, Central America, or South America, regions with substantial avocado industries and diverse native Lauraceae species, would be disastrous. International cooperation is necessary to address this threat.

Little is known concerning terpenoids produced by members of the fungal order Ophiostomales, with the member Harringtonia lauricola having the unique lifestyle of being a beetle symbiont but potentially devastating tree pathogen. Nine known terpenoids, including six labdane diterpenoids (1–6) and three hopane triterpenes (7–9), were isolated from H. lauricola ethyl acetate (EtOAc) extracts for the first time. All compounds were tested for various in vitro bioactivities. Six compounds, 2, 4, 5, 6, 7, and 9, are described functionally. Compounds 2, 4, 5, and 9 expressed potent antiproliferative activity against the MCF-7, HepG2 and A549 cancer cell lines, with half-maximal inhibitory concentrations (IC50s) ~12.54–26.06 μM. Antimicrobial activity bioassays revealed that compounds 4, 5, and 9 exhibited substantial effects against Gram-negative bacteria (Escherichia coli and Ralstonia solanacearum) with minimum inhibitory concentration (MIC) values between 3.13 and 12.50 μg/mL. Little activity was seen towards Gram-positive bacteria for any of the compounds, whereas compounds 2, 4, 7, and 9 expressed antifungal activities (Fusarium oxysporum) with MIC values ranging from 6.25 to 25.00 μg/mL. Compounds 4, 5, and 9 also displayed free radical scavenging abilities towards 2,2-diphenyl-1-picrylhydrazyl (DPPH) and superoxide (O2−), with IC50 values of compounds 2, 4, and 6 ~3.45–14.04 μg/mL and 22.87–53.31 μg/mL towards DPPH and O2−, respectively. These data provide an insight into the biopharmaceutical potential of terpenoids from this group of fungal insect symbionts and plant pathogens. [ABSTRACT FROM AUTHOR]

The rate at which invading organisms disperse into novel habitats is fundamental to their distribution and abundance. Forecasts of spread often assume that invasion speed is constant through time and among directions but, depending on the extent to which this assumption is violated, the efficacy of delimitation surveys and eradication programs could suffer. Knowledge of the mechanisms underlying spatiotemporal variation in spread could help refine forecasts and guide management, particularly in the early stages of invasions. We investigated rates of spread by laurel wilt, one of the most damaging non-native forest pests in North America, using three standard approaches (effective range radius, distance regression, and boundary displacement) and evaluated the strength and drivers of variation in directional spread (i.e., anisotropy). Estimates of mean annual spread varied from 24 to 40 km/yr, but spread was highly anisotropic with invasion speeds reaching approximately 100 km/yr south, 80 km/yr west, and 50 km/yr north, a pattern that we attribute to the abundance of host redbay trees and warmer temperatures fostering rapid southern and western spread. This pattern—quicker spread of laurel wilt from the point of introduction into areas forecasted as highly suitable for its persistence—suggests that establishment location might have a major influence on rates of anisotropy. Our findings underscore the utility of habitat suitability modeling—in which host availability and suitable climate are widely used to forecast establishment risk—for identifying areas into which spread will proceed most rapidly following establishment of a new invader and/or a satellite population via a long-distance dispersal event. [ABSTRACT FROM AUTHOR]

The redbay ambrosia beetle, Xyleborus glabratus Eichhoff (Coleoptera: Curculionidae: Scolytinae), vectors the mycopathogen that causes laurel wilt, a lethal vascular disease of trees in the Lauraceae. Since being detected in Georgia, USA in 2002, this invasive pest has become established in 11 additional states.With continued spread, X. glabratus will likely enter Mexico. In advance of this event, this study was initiated to assess the risk posed to eight native laurels and Hass avocados, the predominant cultivar grown in Mexico.Wood bolts from each species were used in (a) field tests to determine the relative attraction of female X. glabratus, (b) laboratory bioassays to evaluate boring preferences, and (c) GC–MS analyses to identify host kairomones. For comparison, tests included control bolt treatments consisting of silkbay (an attractive U.S. laurel) and Simmonds avocado (a Florida cultivar susceptible to laurel wilt).Hass avocado and two native laurels (Persea schiedeana and Ocotea heribertoi vel aff.) were highly attractive to females and elicited strong boring responses. These species were high in sesquiterpene hydrocarbons, including α‐copaene and α‐cubebene.Results of this study suggest that X. glabratus could become a serious agricultural and forest pest upon incursion into Mexico, with severe economic and ecological impacts. [ABSTRACT FROM AUTHOR]

Introduction: Ambrosia beetles maintain strict associations with specific lineages of fungi. However, anthropogenic introductions of ambrosia beetles into new ecosystems can result in the lateral transfer of their symbionts to other ambrosia beetles. The ability of a Florida endemic ambrosia beetle, Xyleborus bispinatus, to feed and establish persistent associations with two of its known symbionts (Raffaelea subfusca and Raffaelea arxii) and two other fungi (Harringtonia lauricola and Fusarium sp. nov.), which are primary symbionts of invasive ambrosia beetles, was investigated. Methods: The stability of these mutualisms and their effect on the beetle's fitness were monitored over five consecutive generations. Surface-disinfested pupae with non-developed mycangia were reared separately on one of the four fungal symbionts. Non-treated beetles (i.e., lab colony) with previously colonized mycangia were used as a control group. Results: Xyleborus bispinatus could exchange its fungal symbionts, survive, and reproduce on different fungal diets, including known fungal associates and phylogenetically distant fungi, which are plant pathogens and primary symbionts of other invasive ambrosia beetles. These changes in fungal diets resulted in persistent mutualisms, and some symbionts even increased the beetle's reproduction. Females that developed on Fusarium sp. nov. had a significantly greater number of female offspring than non-treated beetles. Females that fed solely on Harringtonia or Raffaelea symbionts produced fewer female offspring. Discussion: Even though some ambrosia beetles like X. bispinatus can partner with different ambrosia fungi, their symbiosis under natural conditions is modulated by their mycangium and possibly other environmental factors. However, exposure to symbionts of invasive beetles can result in stable partnerships with these fungi and affect the population dynamics of ambrosia beetles and their symbionts. [ABSTRACT FROM AUTHOR]

Context: Timely responses to mitigate economic and environmental impacts from invading species are facilitated by knowledge of the speed and drivers of invasions. Objective: Quantify changes in invasion patterns through time and factors that governed time-to-invasion by laurel wilt, one of the most damaging, non-native disturbance agents invading forests of the United States. Methods: We analyzed county-level occurrence data (2004–2021) for laurel wilt across the southeastern United States. A Cox proportional hazards modeling framework was used to elucidate drivers of invasion. Results: As of 2021, laurel wilt had been detected in 275 counties and made 72 discrete jumps (averaging 164 km ± 16 SE) into counties that did not share a border with a previously invaded county. Spread decelerated from 40 km/yr to 24 km/yr after 5 years, with a marked decline in the number of counties invaded in 2021 (16) compared with 2020 (33). The Cox proportional hazards model indicated that proxies for anthropogenic movement and habitat invasibility increased invasion risk. Conclusion: The recent decline in number of counties invaded could be due to disruptions to travel and/or surveys from the coronavirus pandemic, but exhaustion of the most suitable habitat, such as counties in the southeastern US with warm annual temperatures and high densities of host trees, could have also contributed to this trend. This work suggests that without a shift in spread driven by additional insect vectors, that rates of range expansion by laurel wilt might have peaked in 2020 and could continue decelerating. [ABSTRACT FROM AUTHOR]

Some tree species respond to disease by producing basal sprouts from the base and root system of a dying tree, which can alter disease dynamics by altering demography. In the case of many lethal, airborne tree diseases, the production of basal sprouts can be a key contributor to population resurgence post-epidemic, but the effect in lethal, vector-borne tree diseases has not yet been studied. To determine the role of basal sprout production and secondary infection via the root system of infected parent trees in lethal, vector-borne tree diseases, we develop a stage-structured SI-X mathematical model and use laurel wilt, a vector-borne tree disease in which infected trees provide suitable material for vector reproduction, as our model system. The mathematical model shows that the production and secondary infection of basal sprouts do not affect the short-term dynamics of laurel wilt but profoundly alter the long-term dynamics of the laurel wilt epidemic. In particular, in the absence of basal sprout infection, basal sprout production yields a larger host population after disease establishment, but as secondary infection increases, the utility of basal sprouts to maintain the host population decreases. Results suggest management strategies for lethal, vector-borne diseases should depend on the ratio of the basal sprout production rate to the secondary infection rate. [ABSTRACT FROM AUTHOR]

Little is known concerning terpenoids produced by members of the fungal order Ophiostomales, with the member Harringtonia lauricola having the unique lifestyle of being a beetle symbiont but potentially devastating tree pathogen. Nine known terpenoids, including six labdane diterpenoids (1–6) and three hopane triterpenes (7–9), were isolated from H. lauricola ethyl acetate (EtOAc) extracts for the first time. All compounds were tested for various in vitro bioactivities. Six compounds, 2, 4, 5, 6, 7, and 9, are described functionally. Compounds 2, 4, 5, and 9 expressed potent antiproliferative activity against the MCF-7, HepG2 and A549 cancer cell lines, with half-maximal inhibitory concentrations (IC50s) ~12.54–26.06 μM. Antimicrobial activity bioassays revealed that compounds 4, 5, and 9 exhibited substantial effects against Gram-negative bacteria (Escherichia coli and Ralstonia solanacearum) with minimum inhibitory concentration (MIC) values between 3.13 and 12.50 μg/mL. Little activity was seen towards Gram-positive bacteria for any of the compounds, whereas compounds 2, 4, 7, and 9 expressed antifungal activities (Fusarium oxysporum) with MIC values ranging from 6.25 to 25.00 μg/mL. Compounds 4, 5, and 9 also displayed free radical scavenging abilities towards 2,2-diphenyl-1-picrylhydrazyl (DPPH) and superoxide (O2−), with IC50 values of compounds 2, 4, and 6 ~3.45–14.04 μg/mL and 22.87–53.31 μg/mL towards DPPH and O2−, respectively. These data provide an insight into the biopharmaceutical potential of terpenoids from this group of fungal insect symbionts and plant pathogens.

IntroductionAmbrosia beetles maintain strict associations with specific lineages of fungi. However, anthropogenic introductions of ambrosia beetles into new ecosystems can result in the lateral transfer of their symbionts to other ambrosia beetles. The ability of a Florida endemic ambrosia beetle, Xyleborus bispinatus, to feed and establish persistent associations with two of its known symbionts (Raffaelea subfusca and Raffaelea arxii) and two other fungi (Harringtonia lauricola and Fusarium sp. nov.), which are primary symbionts of invasive ambrosia beetles, was investigated.MethodsThe stability of these mutualisms and their effect on the beetle’s fitness were monitored over five consecutive generations. Surface-disinfested pupae with non-developed mycangia were reared separately on one of the four fungal symbionts. Non-treated beetles (i.e., lab colony) with previously colonized mycangia were used as a control group.ResultsXyleborus bispinatus could exchange its fungal symbionts, survive, and reproduce on different fungal diets, including known fungal associates and phylogenetically distant fungi, which are plant pathogens and primary symbionts of other invasive ambrosia beetles. These changes in fungal diets resulted in persistent mutualisms, and some symbionts even increased the beetle’s reproduction. Females that developed on Fusarium sp. nov. had a significantly greater number of female offspring than non-treated beetles. Females that fed solely on Harringtonia or Raffaelea symbionts produced fewer female offspring.DiscussionEven though some ambrosia beetles like X. bispinatus can partner with different ambrosia fungi, their symbiosis under natural conditions is modulated by their mycangium and possibly other environmental factors. However, exposure to symbionts of invasive beetles can result in stable partnerships with these fungi and affect the population dynamics of ambrosia beetles and their symbionts.

Most beetle–fungus symbioses do not represent a threat to agricultural and natural ecosystems; however, a few beetles are able to inoculate healthy hosts with disease-causing fungal symbionts. Here, we report the putative nutritional symbionts associated with five native species of ambrosia beetles colonizing commercial avocado trees in four locations in Michoacán. Knowing which beetles are present in the commercial orchards and the surrounding areas, as well as their fungal associates, is imperative for developing a realistic risk assessment and an effective monitoring system that allows for timely management actions. Phylogenetic analysis revealed five potentially new, previously undescribed species of Raffaelea, and three known species (R. arxi, R. brunnea, R. fusca). The genus Raffaelea was recovered from all the beetle species and across the different locations. Raffaelea lauricola (RL), which causes a deadly vascular fungal disease known as laurel wilt (LW) in Lauraceae species, including avocado, was not recovered. This study points to the imminent danger of native ambrosia beetles spreading RL if the pathogen is introduced to Mexico's avocado orchards or natural areas given that these beetles are associated with Raffaelea species and that lateral transfer of RL among ambrosia beetles in Florida suggests that the likelihood of this phenomenon increases when partners are phylogenetically close. Therefore, this study provides important information about the potential vectors of RL in Mexico and other avocado producing regions. Confirming beetle–fungal identities in these areas is especially important given the serious threat laurel wilt disease represents to the avocado industry in Mexico. [ABSTRACT FROM AUTHOR]

Abstract Background Laurel wilt caused by Raffaelea lauricola is a lethal vascular disease of North American members of the Lauraceae plant family. This fungus and its primary ambrosia beetle vector Xyleborus glabratus originated from Asia; however, there is no report of laurel wilt causing widespread mortality on native Lauraceae trees in Asia. To gain insight into why R. lauricola is a tree-killing plant pathogen in North America, we generated and compared high quality draft genome assemblies of R. lauricola and its closely related non-pathogenic species R. aguacate. Results Relative to R. aguacate, the R. lauricola genome uniquely encodes several small-secreted proteins that are associated with virulence in other pathogens and is enriched in secondary metabolite biosynthetic clusters, particularly polyketide synthase (PKS), non-ribosomal peptide synthetase (NRPS) and PKS-NRPS anchored gene clusters. The two species also exhibit significant differences in secreted proteins including CAZymes that are associated with polysaccharide binding including the chitin binding CBM50 (LysM) domain. Transcriptomic comparisons of inoculated redbay trees and in vitro-grown fungal cultures further revealed a number of secreted protein genes, secondary metabolite clusters and alternative sulfur uptake and assimilation pathways that are coordinately up-regulated during infection. Conclusions Through these comparative analyses we have identified potential adaptations of R. lauricola that may enable it to colonize and cause disease on susceptible hosts. How these adaptations have interacted with co-evolved hosts in Asia, where little to no disease occurs, and non-co-evolved hosts in North America, where lethal wilt occurs, requires additional functional analysis of genes and pathways.

Invasive ambrosia beetles and their symbiotic fungi are responsible for destructive diseases like Laurel Wilt and Fusarium Dieback. Some biological treatments and chemical pesticides (e.g. malathion) have been proposed as alternatives to control these pests, however, to date there are no effective treatments. Here we evaluated the susceptibility of the ambrosia beetle Xyleborus affinis Eichhoff to the entomopathogenic fungus Metarhizium robertsii strain Xoch 8.1, and the efficacy of the mycopathogenic fungus Trichoderma asperellum strain Th-T4 (3) to inhibit the growth of the pathogenic ambrosia fungus Fusarium euwallaceae strain 744. Also, we compared the efficacy of a combination of M. robertsii Xoch 8.1 and T. asperellum Th-T4 (3) against X. affinis, and examined the feasibility of using this combination as an alternative to reduce the malathion doses that have been proposed to control ambrosia beetles. The treatment of X. affinis with M. robertsii (1 × 107 con/mL) alone or in combination with T. asperellum (1 × 106 con/mL) resulted in an average mortality of 76% (±7.5), which was statistically similar to that obtained with malathion at 600 ppm (96% ± 4.0). The combination of a lower concentration of malathion (60 ppm or 200 ppm) and both fungi was as efficient as malathion at 600 ppm. Mortality in negative controls (without fungal conidia or malathion) resulted in an average of 16% (±7.5). The proposed combined treatment reduces by up to one order of magnitude the required amount of malathion, and thus represents a potential alternative to control ambrosia beetle pests. [ABSTRACT FROM AUTHOR]

Fungus farming ambrosia beetles carry their nutritional mutualistic fungi in specialized structures called mycetangia. Fungal propagules are also dispersed phoretically on the beetle's exoskeleton. We determined the phoretic presence and abundance of Raffaelea lauricola, the causal agent of the laurel wilt disease in avocado, on five ambrosia beetle species: Xyleborus bispinatus, Xyleborus volvulus, Xyleborus affinis, Xyleborinus saxesenii and Xylosandrus crassiusculus. Beetles were captured while in flight, excavated from logs, and from logs placed in emergence chambers. Beetles collected by the three methods were assayed for the presence of internal (gut and mycetangium) and external (attached to the exoskeleton) colony forming units (CFUs) of R. lauricola. The pathogen was recovered from the exoskeleton of all beetle species. The collection method significantly influenced the frequency of pathogen recovery, and the abundance of both internal and phoretic R. lauricola was species-specific. Internal CFUs recovery was greater than phoretic recovery. Besides R. lauricola, other cycloheximide tolerant fungi, including mutualistic and entomopathogenic fungi, were isolated from the beetle's exoskeleton. However, phoretic CFUs of R. lauricola were more prevalent and abundant than any other mutualistic phoretic fungi across the beetle species. Our results suggest that phoresy is a common mechanism of transportation of wood-inhabiting fungi and that phoretic transmission of R. lauricola may potentially contribute to the infection in avocado. [ABSTRACT FROM AUTHOR]

Laurel wilt (LW) is a vascular disease caused by a fungal pathogen transmitted to avocado trees by several ambrosia beetle species and through root grafts among adjacent avocado trees. A critical part of preventing and controlling plant diseases is determining the causal agent so that the appropriate management practices can be implemented to eradicate or contain the outbreak. Proper sampling is a critical step in disease diagnosis and in the determination of the causal agent of disease. This new 3-page publication of the UF/IFAS Horticultural Sciences Department was written by Jonathan Crane, Romina Gazis, Jeff Wasielewski, Daniel Carrillo, Bruce Schaffer, Fredy Ballen, and Edward Evans. https://edis.ifas.ufl.edu/hs1394

Avocado trees are a popular choice for homeowners in Florida, with over 600,000 growing in Florida home landscapes. However, avocado trees as well as others in the Lauraceae family are susceptible to laurel wilt disease, which can kill a tree in as few as three weeks. This new publication of the UF/IFAS Horticultural Sciences Department provides home owners recommendations for identifying and mitigating laurel wilt disease in the home landscape. Written by Jonathan H. Crane, Jeff Wasielewski, Daniel Carrillo, Romina Gazis, Bruce Schaffer, Fredy Ballen, and Edwards Evans. https://edis.ifas.ufl.edu/hs1358

The fungal pathogen, Raffaelea lauricola, is the causative agent of laurel wilt, a devastating disease affecting the Lauraceae family. The fungus is vectored by ambrosia beetles that carry the fungus in specialized structures (mycangia), with the fungus acting as a symbiont and food source for beetle larvae growing in tree galleries. In order to probe the molecular basis for plant pathogenicity and insect symbiosis of the laurel wilt fungus, molecular tools including establishment of efficient transformation protocols are required. Resistance marker profiling revealed susceptibility of R. lauricola to phosphinothricin, chlorimuron ethyl, hygromycin, and benomyl. Agrobacterium-mediated transformation using either the bar or sur marker resulted in 1–200 transformants/105 spores. A second protocol using lithium acetate-polyethylene glycol (LiAc-PEG) treatment of fungal blastospores yielded 5–60 transformants/μg DNA/108 cells. Transformants were mitotically stable (at least 5 generations), and > 95% of transformants showed a single integration event. R. lauricola strains expressing green and red fluorescent proteins (EGFP and RFP), as well as glucuronidase (GUS), were constructed. Using the Agrobacterium-mediated method, a random T-DNA insertion library was constructed, and genetic screens led to the isolation of developmental mutants as well as mutants displaying enhanced resistance to sodium dodecyl sulfate (SDS) or fluconazole, and those showing decreased susceptibility to biphenol. These results establish simple and reliable genetic tools for transformation of R. lauricola needed for genetic dissection of the symbiotic and virulent lifestyles exhibited by this fungus and establish a library of insertion mutants that can be used in various genetic screens to dissect molecular pathways. Key points: • Vectors and transformation protocols were developed for Raffaelea lauricola. • Method was used for construction of a random insertion mutant library. • Mutant library was validated by phenotypic screens for resistance and susceptibility to various agents. [ABSTRACT FROM AUTHOR]

Laurel wilt and Fusarium dieback are vascular diseases caused by fungal symbionts of invasive ambrosia beetles (Coleoptera: Curculionidae: Scolytinae). Both diseases threaten avocado trees in Florida. Redbay ambrosia beetle, Xyleborus glabratus , is the primary vector of the laurel wilt pathogen, Raffaelea lauricola , but in recent years this symbiont has been transferred laterally to at least nine other species of ambrosia beetle, which now comprise a community of secondary vectors. Dieback disease, caused by Fusarium spp. fungi, is spread by shot hole borers in the Euwallacea fornicatus species complex. In this study, we conducted field tests in Florida avocado groves to compare efficacy of four trap designs for detection of Scolytinae. Treatments included an 8-funnel Lindgren trap, black 3-vane flight interception trap, green 3-vane interception trap, white sticky panel trap, and an unbaited sticky panel (control). In two tests targeting E. nr. fornicatus and X. glabratus , traps were baited with a two-component lure (α-copaene and quercivorol). In a test targeting other species, traps were baited with a low-release ethanol lure. For E. nr. fornicatus, sticky panels and black interception traps captured significantly more beetles than Lindgren traps; captures with green traps were intermediate. With ethanol-baited traps, 20 species of bark/ambrosia beetle were detected. Trap efficacy varied by species, but in general, sticky traps captured the highest number of beetles. Results indicate that sticky panel traps are more effective for monitoring ambrosia beetles than Lindgren funnel traps, the current standard, and may provide an economical alternative for pest detection in avocado groves. [ABSTRACT FROM AUTHOR]

Raffaelea lauricola is the causative agent of laurel wilt, a devastating disease of lauraceous trees. R. lauricola is also an obligate nutritional symbiont of several ambrosia beetle species who act as vectors for the pathogen. Here, we sought to establish the baseline “phenome” of R. lauricola with knowledge concerning its metabolic capability, expanding our understanding of how these processes are impacted by environmental and host nutrients. Phenotypic screening using a microarray of over one thousand compounds was used to generate a detailed profile of R. lauricola substrate utilization and chemical sensitivity. These data revealed (i) relatively restricted carbon utilization, (ii) broad sulfur and phosphate utilization, and (iii) pH and osmotic sensitivities that could be rescued by specific compounds. Additional growth profiling on fatty acids revealed toxicity on C10 substrates and lower, with robust growth on C12–C18 fatty acids. Conditions for lipid droplet (LD) visualization and LD dynamics were examined using a series of lipid dyes. These data provide unique insights regarding R. lauricola metabolism and physiology, and identify distinct patterns of substrate usage and sensitivity which likely reflect important aspects of the host-microbe interface and can be exploited for the development of strategies for mitigating the spread of laurel wilt.

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. [ABSTRACT FROM AUTHOR]

Laurel wilt disease (LWD) is a highly virulent disease vectored by an invasive beetle. It was first observed in South Carolina in 2003, and it is now found in all Florida counties and nine southeastern US states. This disease causes extensive mortality of Lauraceae plants, an ecologically and economically important group. Many arthropods are associated with Lauraceae species, including palamedes (Papilio palamedes (Drury)) and spicebush (Papilio troilus (L.)) swallowtail butterflies, which host on swamp bay (Persea palustris) and redbay (Persea borbonia). It is thought that P. palamedes are at greatest risk from LWD. Limited information exists on the impacts on these butterflies after LWD becomes established. Therefore, we use a unique long-term tree and butterfly dataset in central Florida that spanned the introduction of LWD. We found that over two decades, swamp bay declined 90% in abundance. Interestingly, the majority (70%) of the swamp bays' decline preceded the introduction of LWD. We suspect that a 5 year severe drought was the primary cause of mortality, since there was a 45% reduction in overall tree abundances. Despite these large reductions, a small number of swamp bay trees persisted, and others sprouted from basal roots. Butterfly monitoring began after 70% of the swamp bays had died. We found that, even though there was extensive loss of swamp bays, P. palamedes and P. troilus populations were relatively stable, and potentially were increasing. However, after LWD was confirmed in this area, these butterflies declined slightly. We also found that the reduction was slightly greater in P. palamedes than P. troilus. P. palamedes are more dependent on swamp bay whereas P. troilus are more adaptable at host-switching, perhaps to a Lauraceae tree Cinnamomum camphora (L.) J. Pesl. (Camphortree), which is common in our area and does not appear to be locally affected by LWD. Thus, while we found that swamp bay declined dramatically, dependent butterfly populations declined far less dramatically. Butterflies may be surviving because of host-switching, or because the remaining trees provide enough resources. Future research should focus on the persistence of these species, especially as this disease continues to spread throughout the southeastern US. [ABSTRACT FROM AUTHOR]

This review highlights current advances in the management of the redbay ambrosia beetle, Xyleborus glabratus, a primary vector of the pathogenic fungus, Raffaelea lauricola, that causes laurel wilt. Laurel wilt has a detrimental effect on forest ecosystems of southeastern USA, with hundreds of millions of Lauraceae deaths. Currently, preventive measures mostly focus on infected-tree removal to potentially reduce local beetle populations and/or use of preventative fungicide applications in urban trees. Use of semiochemicals may offer an opportunity for the management of X. glabratus. Research on attractants has led to the development of α-copaene lures that are now the accepted standards for X. glabratus sampling. Research conducted on repellents first included methyl salicylate and verbenone and attained significant reduction in the number of X. glabratus captured on redbay and swamp bay trees treated with verbenone. However, the death rate of trees protected with verbenone, while lower compared to untreated trees, is still high. This work underscores the necessity of developing new control methods, including the integration of repellents and attractants into a single push-pull system.

Raffaelea lauricola is the causal agent of laurel wilt and a fungal symbiont of its vector, the redbay ambrosia beetle, Xyleborus glabratus. Beetle populations increase rapidly where redbays have died from laurel wilt. In a series of experiments using bolts from healthy and diseased trees, X. glabratus reproduced better in bolts from diseased redbay; ratio of beetles emerged to entrance holes (BE/EH ratio) ranged from 4.1 to 6.0 in diseased bolts versus 0.3 to 1.9 in healthy bolts. Emergence was greater from redbay bolts when their ends were treated with paraffin wax compared to untreated bolts. Although sassafras sapwood has been considered poor brood material for X. glabratus, bolts from a diseased sassafras had BE/EH ratios comparable to those from diseased redbay bolts. It was hypothesized that better brood production in bolts from diseased trees was due to precolonization of the bolts by R. lauricola, which would allow for better establishment of the pathogen in beetle tunnels. However, the frequency of R. lauricola isolation and colony forming units (CFU) per beetle did not differ between beetles emerged from diseased versus healthy redbay bolts. Changes in the xylem of trees with laurel wilt appear to provide conditions favorable for brood production. [ABSTRACT FROM AUTHOR]

The ascomycete Raffaelea lauricola causes laurel wilt, a lethal vascular disease of avocado, Persea americana, and other members of the Lauraceae plant family. Few effective control measures for laurel wilt exist and new measures are needed. In this study, biological control of the disease with endophytic fungi from avocado was examined. Thirty‐two endophytes (24 operational taxonomic units or OTUs) isolated from the xylem of healthy trees (the infection court of R. lauricola) were evaluated against R. lauricola with in vitro dual‐culture assays. Nine OTUs that showed strong in vitro antagonism of the pathogen were tested in planta against laurel wilt. In three greenhouse experiments, grafted avocado plants of Simmonds or Russell cultivars, which are both susceptible to laurel wilt, were inoculated with endophytes and, after 10–16 days, inoculated with the same isolate of R. lauricola that was used in the in vitro assays. Within 14 days of inoculation with R. lauricola, laurel wilt developed in plants that were not treated with endophytes (positive controls) but also developed in endophyte‐treated plants to the extent observed in the positive controls (P = 0.05). The pathogen colonized plants rapidly and systemically, but endophytes generally did not colonize xylem more than 2 cm above the point at which plants were inoculated. Although the tested endophytes strongly antagonized the pathogen in vitro, this did not translate to an ability to reduce development of laurel wilt. The management of laurel wilt and other plant diseases with fungal endophytes is discussed. [ABSTRACT FROM AUTHOR]

Abstract Survival of ambrosia beetles relies on obligate nutritional relationships with fungal symbionts that are cultivated in tunnels excavated in the sapwood of their host trees. The dynamics of fungal associates, along with the developmental biology, and gallery construction of the ambrosia beetle Xyleborus bispinatus were elaborated. One generation of this ambrosia beetle was reared in an artificial medium containing avocado sawdust. The developmental time from egg to adult ranged from 22 to 24 d. The mean total gallery length (14.4 cm and 13 tunnels) positively correlated with the number of adults. The most prevalent fungal associates were Raffaelea arxii in the foundress mycangia and new galleries, and Raffaelea subfusca in the mycangia of the F 1 adults and the final stages of the galleries. Raffaelea sp. PL1001, Raffaelea subalba and seven yeast species were also recovered. These results indicate flexibility in the association between X. bispinatus and its symbiont species, and that this beetle may use more than one Raffaelea species as a food source. These results are important, as X. bispinatus has been associated with the transmission of Raffaelea lauricola , the causal agent of laurel wilt, a lethal disease affecting avocado trees. [ABSTRACT FROM AUTHOR]

Ambrosia beetles subsist on fungal symbionts that they carry to, and cultivate in, their natal galleries. These symbionts are usually saprobes, but some are phytopathogens. Very few ambrosial symbioses have been studied closely, and little is known about roles that phytopathogenic symbionts play in the life cycles of these beetles. One of the latter symbionts, Raffaelea lauricola, causes laurel wilt of avocado, Persea americana, but its original ambrosia beetle partner, Xyleborus glabratus, plays an uncertain role in this pathosystem. We examined the response of a putative, alternative vector of R. lauricola, Xyleborus bispinatus, to artificial diets of R. lauricola and other ambrosia fungi. Newly eclosed, unfertilized females of X. bispinatus were reared in no-choice assays on one of five different symbionts or no symbiont. Xyleborus bispinatus developed successfully on R. lauricola, R. arxii, R. subalba and R. subfusca, all of which had been previously recovered from field-collected females of X. bispinatus. However, no development was observed in the absence of a symbiont or on another symbiont, Ambrosiella roeperi, recovered from another ambrosia beetle, Xylosandrus crassiusculus. In the no-choice assays, mycangia of foundress females of X. bispinatus harbored significant colony-forming units of, and natal galleries that they produced were colonized with, the respective Raffaelea symbionts; with each of these fungi, reproduction, fecundity and survival of the beetle were positively impacted. However, no fungus was recovered from, and reproduction did not occur on, the A. roeperi and no symbiont diets. These results highlight the flexible nature of the ambrosial symbiosis, which for X. bispinatus includes a fungus with which it has no evolutionary history. Although the “primary” symbiont of the neotropical X. bispinatus is unclear, it is not the Asian R. lauricola. [ABSTRACT FROM AUTHOR]

Native Lauraceae (e.g. sassafras, redbay) in the southeastern USA are being severely impacted by laurel wilt disease, which is caused by the pathogen Raffaelea lauricola T. C. Harr., Fraedrich and Aghayeva, and its symbiotic vector, the redbay ambrosia beetle (Xyleborus glabratus Eichhoff). Cold temperatures are currently the only viable limitation to the establishment of X. glabratus in northern populations of sassafras. The observed lower lethal temperature of X. glabratus (− 10.0 °C) is warmer than its supercooling point (− 22.0 °C), indicating the beetle is a freeze intolerant and chill susceptible species. Empirically derived X. glabratus lower lethal temperature thresholds were combined with host distribution and microhabitat-corrected climate data to produce species distribution models for X. glabratus in the eastern USA. Macroclimate data (30-year mean annual minimum temperature) were corrected (− 1.2 °C) to account for thermal buffering afforded to X. glabratus while living inside sassafras trees. Only 0.1% of the current US sassafras spatial extent experiences sufficiently harsh winters (locales where mean annual minimum winter temperatures ≤ − 6.2 °C for ≥ 12 h) to exclude X. glabratus establishment in our species distribution model. Minimum winter temperatures will likely cause some X. glabratus mortality in ~ 52% of the current spatial extent of sassafras, although current data do not allow a quantification of X. glabratus mortality in this zone. Conversely, ~ 48% of the current spatial extent of sassafras is unlikely to experience sufficiently cold winter temperatures to cause any significant impediment to X. glabratus spread or establishment. A modest climate change scenario (RCP4.5) of + 1.4 °C would result in 91% of the current spatial extent of sassafras in the eastern USA occurring where winter minimum temperatures are unlikely to cause any mortality to X. glabratus. [ABSTRACT FROM AUTHOR]

Laurel wilt (Lw) is a very destructive disease and poses a serious threat to the commercial production of avocado in Florida, USA. External symptoms of Lw are similar to those that are caused by other diseases and disorders. A rapid technique to distinguish Lw infected avocado from healthy trees and trees with other abiotic stressors is presented in this paper. A novel method was developed to analyze data from hyperspectral data using finite difference approximation (FDA) and bivariate correlation (BC) to discriminate Lw, Nitrogen (N), and Iron (Fe) deficiencies from healthy avocado plants. Several combinatorial methods were used in preprocessing the data, such as standard normal transformation of data, smoothing of the data, and polynomial fit. The FDA technique was derived using a Taylor Polynomial finite difference approximation. This FDA accentuates inflection points in the spectrum. These, in turn, reveal variance in the data that can be used to identify spectral signature associated with healthy and diseased states. By statistical correlation using the bivariate correlation coefficient of these enhanced spectral patterns, an algorithm (FDA-BC) for distinguishing Lw avocado leaves from all other categories of healthy or mineral deficient avocado leaves is achieved with an overall accuracy of 100%.

Like other ambrosia beetles, Xyleborus volvulus Fabricius (Coleoptera: Curculionidae) lives in a mutualistic symbiotic relationship with fungi that serve as food source. Until recently, X. volvulus was not considered a pest, and none of its symbionts were considered plant pathogens. However, recent reports of an association between X. volvulus and Raffaelea lauricola T.C. Harr., Fraedrich & Aghayeva (Ophiostomatales: Ophiostomataceae), the cause of the laurel wilt disease of avocado (Persea americana Mill. [Laurales: Lauraceae]), and its potential role as vector of the pathogen merit further investigation. The objective of this study was to evaluate three artificial media containing sawdust obtained from avocado or silkbay (Persea humilis Nash) for laboratory rearing of X. volvulus. The effect of R. lauricola in the media on the beetle's reproduction was also evaluated. Of the three media, the one with the lowest content of sawdust and intermediate water content provided the best conditions for rearing X. volvulus. Reproduction on this medium was not affected by the sawdust species or the presence of R. lauricola. On the other two media, there was a significant interaction between sawdust species and R. lauricola. The presence of R. lauricola generally had a negative effect on brood production. There was limited colonization of the mycangia of X. volvulus by R. lauricola on media inoculated with the pathogen. From galleries formed within the best medium, there was 50% recovery of R. lauricola, but recovery was much less from the other two media. Here, we report the best artificial substrate currently known for X. volvulus. [ABSTRACT FROM AUTHOR]

The redbay ambrosia beetle, Xyleborus glabratus, is the vector of the laurel wilt disease fungal pathogen, Raffaelea lauricola. Since the vector's initial detection in the USA in the early 2000s, laurel wilt has killed millions of redbay, Persea borbonia, trees and other members of the plant family Lauraceae. To protect host trees from beetle attack and laurel wilt infection, we tested the efficacy of host- and non-host-derived and commercial compounds as X. glabratus repellents in field experiments. In our first trial, the major constituents of the non-host tree, longleaf pine, Pinus palustris, and SPLAT Verb (verbenone 10%) were paired with manuka oil attractants and beetle captures were counted. Verbenone and a 1 : 1 blend of myrcene and camphene were intermediate to both the manuka positive and blank negative controls. Subsequently, we tested different blends of methyl salicylate (MeSA), a host defence and signalling compound, and verbenone in SPLAT dispensers using freshly cut redbay bolts as an attractant. All treatments reduced X. glabratus captures and boring holes as compared to the redbay (-) repellent positive control; however, SPLAT Verb and SPLAT MeSA-Verb (5% each) achieved the highest repellency, with results comparable to that of the non-host (laurel oak). These trials establish that host-derived and commercially available repellent compounds can reduce X. glabratus attacks and therefore have potential as part of an integrated management strategy against laurel wilt and its vector. [ABSTRACT FROM AUTHOR]

Laurel wilt is caused by Raffaelea lauricola, a nutritional symbiont of an Asian ambrosia beetle, Xyleborus glabratus. American members of the Lauraceae plant family are most susceptible and 300 million trees have been killed by the disease in the southeastern USA since 2003. Recently, commercial production of an important crop in the laurel family, avocado ( Persea americana), has been affected in southern Florida. We summarize studies in which diverse measures were tested for managing the disease. In all studies, trees were treated with potential laurel wilt control measures and subsequently inoculated with R. lauricola. On potted plants in greenhouse experiments, commercial nutritional products (Greenstim and Keyplex 350) and SAR products (Agri-Fos and Nutri-Phite), when applied as soil drenches or foliar sprays, had either no impact on, or increased laurel wilt symptom development compared to non-treated control treatments. Bark applications of Tilt (a propiconazole fungicide for which emergency registration had been obtained in 2010) in a surfactant (Pentrabark) enabled significant laurel wilt protection in greenhouse studies on small potted plants, but Pentrabark and other surfactants moved little propiconazole into the xylem of fruit-bearing trees in field studies. In efficacy studies in the field, Propiconazole Pro (an injectable formulation of propiconazole), Tilt, and two experimental formulations of another triazole fungicide, tebuconazole, decreased the development of laurel wilt compared to nontreated control trees when applied as undiluted injections into branches and scaffold limbs (microinjection), or injection of dilute fungicide into tree flare roots (macroinfusion). However, symptoms developed in all treated trees by 10-11 months after inoculation with R. lauricola, indicating that trees would need to be re-treated at least on an annual basis. Regardless of how fungicides were applied, insignificant levels of different active ingredients entered fruit. Although fungicide treatment of fruit-bearing avocado trees is not a concern for food safety and several triazole and DMI fungicides can protect avocado trees from laurel wilt, cost-effective measures with which the xylem could be loaded with and protected by these products remains a challenge. Management of laurel wilt in commercial avocado production areas is discussed. [ABSTRACT FROM AUTHOR]

The invasive redbay ambrosia beetle, Xyleborus glabratus Eichhoff (Coleoptera: Curculionidae: Scolytinae), vectors the fungal pathogen (Raffaelea lauricola) that causes laurel wilt, a disease responsible for widespread mortality of trees in the Lauraceae in the southeastern United States. Early detection of incipient vector populations may allow for management practices that could successfully mitigate damage. Developing new, highly effective attractants is a priority for improving sensitivity of early detection efforts. In this study, two field tests were conducted to evaluate combinations of commercially available bark and ambrosia beetle lures for enhanced attraction of host-seeking female X. glabratus. In addition, lures were compared for capture of nontarget scolytine beetles. In the first experiment, traps baited with a combination of cubeb oil, conophthorin, chalcogran, and ethanol captured greater numbers of X. glabratus than cubeb oil alone, the current standard attractant. However, this combination lure resulted in higher nontarget scolytine captures than with the cubeb lure. In the second field test, an oil enriched in the sesquiterpene α-copaene caught significantly more X. glabratus than other lures currently available for monitoring this pest. There were no differences in efficacy between cubeb oil lures produced by two different manufacturers, and a combination lure containing copaiba and cubeb oils did not increase captures over the cubeb lure alone. Results of these two tests suggest that increased sensitivity for detection of X. glabratus may be achieved with a multicomponent lure that incorporates α-copaene, spiroketals, and low release of ethanol. [ABSTRACT FROM AUTHOR]

Laurel wilt is an extraordinarily destructive exotic tree disease in the southeastern United States that involves new-encounter hosts in the Lauraceae, an introduced vector ( Xyleborus glabratus) and pathogen symbiont ( Raffaelea lauricola). USDA Forest Service Forest Inventory and Analysis data were used to estimate that over 300 million trees of redbay ( Persea borbonia sensu lato) have succumbed to the disease since the early 2000s (ca 1/3 of the pre-invasion population). In addition, numerous native shrub and tree species in the family are susceptible and threatened in the Western Hemisphere. Genetic markers were used to test the hypothesis that the vector and pathogen entered North America as a single introduction. With a portion of the c ytochrome oxidase I gene, a single X. glabratus haplotype was detected in the USA. Similarly, Amplified Fragment Length Polymorphisms indicated that 95% (54 of 57) of the isolates of R. lauricola that were examined were of a single clonal genotype; only minor variation was detected in three polymorphic isolates. Similar levels of disease developed after swamp bay ( P. palustris) was inoculated with each of the four genotypes of R. lauricola. It is proposed that a single founding event is responsible for the laurel wilt epidemic in the United States. [ABSTRACT FROM AUTHOR]

Raffaelea lauricola , a fungus causing a vascular wilt (laurel wilt) in Lauraceae trees, was introduced into the United States in the early 2000s. It has devastated forests in the Southeast and has now moved into the commercial avocado groves in southern Florida. Trained detection canines are currently one of the few successful methods for early detection of pre-symptomatic diseased trees. In order to achieve the universal and frequent training required to have successful detection canines, it is desirable to create accessible, safe, and long-lasting training aids. However, identification of odorants and compounds is limited by several factors, including both the availability of chemicals and the need to present chemicals individually and in combination to detection canines. A method for the separation and identification of volatile organic compounds (VOCs) from environmental substances for the creation of such a canine training aid is presented here. Headspace solid phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS) was used to identify the odors present in avocado trees infected with the R. lauricola phytopathogen. Twenty-eight compounds were detected using this method, with nine present in greater than 80% of samples. The majority of these compounds were not commercially available as standard reference materials, and a canine trial was designed to identify the active odors without the need of pure chemical compounds. To facilitate the creation of a canine training aid, the VOCs above R. lauricola were separated by venting a 0.53 mm ID solgel-wax gas chromatography column to the atmosphere. Ten minute fractions of the odor profile were collected on cotton gauze in glass vials and presented to the detection canines in a series of field trials. The canines alerted to the VOCs from the vials that correspond to a portion of the chromatogram containing the most volatile species from R. lauricola. This innovative fractionation and collection method can be used to develop reliable and cost effective canine training aids. [ABSTRACT FROM AUTHOR]

We compared the flight activity of Xyleborus glabratus Eichhoff, vector and symbiont of the causal agent of laurel wilt disease (Raffaelea lauricola), with a native species Monarthrum mali (Fitch) using flight mills. Flight mills were operated either for 24h or for three 3-h time intervals. During the 3-h interval experiment, the shortest time to flight initiation for X. glabratus occurred at 1600-1900 hours. The average flight time and total flying distance during 1600-2100hours were also higher than those quantified during the other two recording times investigated. However, total flight duration and proportion of fliers was highest at 1000-1300 hours. We compared several flight parameters. About 64.0% of tested X. glabratus flew <20 m. During 24-h recording periods, M. mali flew longer distances than X. glabratus. Over 50.0%of M. mali flew over 100 mon the flight mill. Xyleborus glabratus flight activity was greatest between 1200 and 1800hours, while M. mali flew most frequently between 1500 and 2100 hours. Monarthrum mali flew more than five times more frequently than X. glabratus, and their longest single flight distance (37.5612.5 m) and total flight distance (213.7685.5 m) were greater than those of X. glabratus. These data will be useful for development of species-specific control and monitoring protocols for these ambrosia beetles based on greater understanding of their flight capacities and associated invasion distance. [ABSTRACT FROM AUTHOR]

The redbay ambrosia beetle Xyleborus glabratus is the vector of the symbiotic fungus, Raffaelea lauricola that causes laurel wilt, a highly lethal disease to members of the Lauraceae family. Pioneer X. glabratus beetles infect live trees with R. lauricola, and only when tree health starts declining more X. glabratus are attracted to the infected tree. Until now this sequence of events was not well understood. In this study, we investigated the temporal patterns of host volatiles and phytohormone production and vector attraction in relation to laurel wilt symptomology. Following inoculations with R. lauricola, volatile collections and behavioral tests were performed at different time points. Three days after infection (DAI), we found significant repellency of X. glabratus by leaf odors of infected swamp bay Persea palustris as compared with controls. However, at 10 and 20 DAI, X. glabratus were more attracted to leaf odors from infected than non-infected host plants. GC-MS analysis revealed an increase in methyl salicylate (MeSA) 3 DAI, whereas an increase of sesquiterpenes and leaf aldehydes was observed 10 and 20 DAI in leaf volatiles. MeSA was the only behaviorally active repellent of X. glabratus in laboratory bioassays. In contrast, X. glabratus did not prefer infected wood over healthy wood, and there was no associated significant difference in their volatile profiles. Analyses of phytohormone profiles revealed an initial increase in the amount of salicylic acid (SA) in leaf tissues following fungal infection, suggesting that the SA pathway was activated by R. lauricola infection, and this activation caused increased release of MeSA. Overall, our findings provide a better understanding of X. glabratus ecology and underline chemical interactions with its symbiotic fungus. Our work also demonstrates how the laurel wilt pathosystem alters host defenses to impact vector behavior and suggests manipulation of host odor by the fungus that attract more vectors. [ABSTRACT FROM AUTHOR]

Volatile organic compounds (VOCs) in the headspace of the fungus Raffaelea lauricola have been monitored and identified over a twenty-eight day growth period. R. lauricola is an invasive and phytopathogenic fungus that was first identified in the United States in the mid-2000s. It is believed to be spread by a host beetle, Xyleborus glabratus , and is detrimental both to wild members of the Lauraceae family and to commercial avocado groves particularly in the Southeastern region of the country. The fungus causes the fatal laurel wilt disease, a result of the host tree shutting down its vascular system in order to halt the spread of the fungus. The current study identified the VOCs present in the headspace of R. lauricola over the initial growth stage using headspace solid phase microextracion-gas chromatography-mass spectrometry (HS-SPME-GC–MS). Results revealed the VOC dynamics of the fungus in culture, indicating that the initial growth period of the fungus may coincide with potential responses from the host trees that may recognize and respond to the pathogen when the fungal VOCs are produced as a result of primary metabolic processes. As fungal growth progresses past initial growth phases, the predominant compounds seen in the odor profile are hydrocarbons and terpenes, produced from secondary metabolic processes. The odor profile pattern for the twenty-eight day growth period did change with the stages of growth. Based on the information learned from this pilot study, a discussion is presented of possible host tree reactions to R. lauricola and implications for future experiments. [ABSTRACT FROM AUTHOR]

Laurel wilt kills members of the Lauraceae plant family in the southeastern United States. It is caused by Raffaelea lauricola T.C. Harr., Fraedrich and Aghayeva, a nutritional fungal symbiont of an invasive Asian ambrosia beetle, Xyleborus glabratus Eichhoff, which was detected in Port Wentworth, Georgia, in 2002. The beetle is the primary vector of R. lauricola in forests along the southeastern coastal plain of the United States, but other ambrosia beetle species that obtained the pathogen after the initial introduction may play a role in the avocado (Persea americana Miller) pathosystem. Susceptible taxa are naïve (new-encounter) hosts that originated outside Asia. In the southeastern United States, over 300 million trees of redbay (P. borbonia (L.) Spreng.) have been lost, and other North American endemics, non-Asian ornamentals and avocado--an important crop that originated in MesoAmerica--are also affected. However, there are no reports of laurel wilt on the significant number of lauraceous endemics that occur in the Asian homeland of R. lauricola and X. glabratus; coevolved resistance to the disease in the region has been hypothesized. The rapid spread of laurel wilt in the United States is due to an efficient vector, X. glabratus, and the movement of wood infested with the insect and pathogen. These factors, the absence of fully resistant genotypes, and the paucity of effective control measures severely constrain the disease's management in forest ecosystems and avocado production areas. [ABSTRACT FROM AUTHOR]