Ophiostomatoid fungi associated with bark beetles onAbies veitchii in wave-regenerated forests

Relationships between tree mortality and bark beetle infestation onAbies veitchii at the wave-regenerated forest in Mt. Asahi, Okuchichibu area were investigated. Most of the firs with green needles and newly developed current year’s shoots in the dieback zone were heavily infested by bark beetles before the death of the trees. After heavy infestation of beetles, about half of the infested firs died within the year, and the other half died in the next year. When the species composition of bark beetles and associated ophiostomatoid fungi were investigated in Mt. Asahi and also at a typical wave-regenerated forest in Mt. Shimagare, Yatsugatake area,Cryphalus montanus andC. piceae were dominant beetle species for Mt. Asahi and Mt. Shimagare, respectively.Ophiostoma subalpinum andO. europhioides were dominant fungal species at both wave-generated forests. BecauseO. subalpinum was detected more frequently from deeper areas of sapwood thanO. europhioides, it was suggested that the fungal species may accelerate the death of stressed firs in wave-regenerated forests. Contribution No. 169, Laboratory of Plant Parasitic Mycology, Institute of Agriculture and Forestry, University of Tsukuba.     

Phoretic mites of three bark beetles (<Emphasis Type="Italic">Pityokteines</Emphasis> spp.) on Silver fir

The species composition and abundance of phoretic mites of the bark beetles Pityokteines curvidens, P. spinidens, and P. vorontzowi on Silver fir (Abies alba) were investigated in 2003 at two locations (Trakoscan and Litoric) in Croatia. Stem sections and branches from A. alba trees infested by Pityokteines ssp. were collected and incubated in rearing cages. Bark beetles emerging from the stem sections and branches were examined for photetic mites. A total of ten mite species were documented for the first time as associates of Pityokteines spp. on A. alba. These included Dendrolaelaps quadrisetus, Ereynetes scutulis, Histiostoma piceae, Paraleius leontonychus, Pleuronectocelaeno japonica, Proctolaelaps hystricoides, Schizostethus simulatrix, Tarsonemus minimax, Trichouropoda lamellose, and Uroobovella ipidis. T. minimax was the most frequent phoretic mite of all the three scolytines and U. ipidis was also common, whereas, the other mite species occurred less frequently. The species spectrum and relative abundance of mite associates were similar for all three Pityokteines species. Another species, Pleuronectocelaeno barbara was commonly found phoretic on P. curvidens, captured in pheromone traps in 2005 at the location Litoric. Furthermore, two previously collected mite specimens from Switzerland, phoretic on P. curvidens, were identified as Nanacarus sp. and Bonomia sp. The records from Croatia and Switzerland in the present study increase the number of known mite associates of Pityokteines spp. from one previously documented species to 14 species. None of the phoretic mites found in the survey in Croatia appear to have the potential to be used for biological control of Pityokteines spp., although the feeding habits are unknown for many species recorded.     

Bioremediation of CCA-treated wood by brown-rot fungi <Emphasis Type="Italic">Fomitopsis palustris</Emphasis>, <Emphasis Type="Italic">Coniophora puteana</Emphasis>, and <Emphasis Type="Italic">Laetiporus sulphureus</Emphasis>

This study evaluated oxalic acid accumulation and bioremediation of chromated copper arsenate (CCA)-treated wood by three brown-rot fungi Fomitopsis palustris, Coniophora puteana, and Laetiporus sulphureus. The fungi were first cultivated in a fermentation broth to accumulate oxalic acid. Bioremediation of CCA-treated wood was then carried out by leaching of heavy metals with oxalic acid over a 10-day fermentation period. Higher amounts of oxalic acid were produced by F. palustris and L. sulphureus compared with C. puteana. After 10-day fermentation, oxalic acid accumulation reached 4.2 g/l and 3.2 g/l for these fungi, respectively. Fomitopsis palustris and L. sulphureus exposed to CCA-treated sawdust for 10 days showed a decrease in arsenic of 100% and 85%, respectively; however, C. puteana remediation removed only 18% arsenic from CCA-treated sawdust. Likewise, chromium removal in F. palustris and L. sulphureus remediation processes was higher than those for C. puteana. This was attributed to low oxalic acid accumulation. These results suggest that F. palustris and L. sulphureus remediation processes can remove inorganic metal compounds via oxalic acid production by increasing the acidity of the substrate and increasing the solubility of the metals.An erratum to this article can be found at     

Relative dominance of the mold fungus,<Emphasis Type="Italic">Raffaelea</Emphasis> sp., in the mycangium and proventriculus in relation to adult stages of the oak platypodid beetle,<Emphasis Type="Italic">Platypus quercivorus</Emphasis> (Coleoptera; Platypodidae)

Microorganisms associated with the oak platypodid beetle,Platypus quercivorus (Murayama), were isolated from the mycangium of adult females, from the proventriculus of adults of both sexes, and from galleries in the period before dispersal. Fungi isolated from female mycangia were separated into three groups:Raffaelea sp., yeasts (mainlyCandida spp.), and other species.Raffaelea sp. was isolated predominantly from the teneral stage to the dispersal stage, but was not isolated from the mycangia of mother beetles in new galleries. Yeasts were isolated in every beetle stage tested and their isolation rate was over 80% from the teneral stage to the dispersal stage. All three fungal groups were found in the female proventriculus.Raffaelea sp. was isolated only in the dispersal stage at a rate of 40%, whereas the isolation rate of yeasts gradually increased beginning in the mature stage and reached 100% in the rearing stage. In contrast, in the male proventriculus, onlyRaffaelea sp. and yeasts were isolated in the dispersal and rearing stages. The isolation rate in the rearing stage ofRaffaelea sp. was less than 20%, but that of yeasts was 100%, although these rates were almost the same in the dispersal stage.Raffaelea sp. and yeasts were also isolated from cradles containing eclosing pupae. These results suggest thatP. quercivorus acquiresRaffaelea sp. and yeasts in their mycangia from the cradles immediately after eclosion, and maintain them to the dispersal stage. Then, they transmit these fungi from old to new galleries by way of mycangia, and possibly the digestive system.     

<Emphasis Type="Italic">Bursaphelenchus mucronatus</Emphasis> (Nematoda: Aphelenchoididae) vectored by <Emphasis Type="Italic">Monochamus </Emphasis><Emphasis Type="Italic">urussovi</Emphasis> (Coleoptera: Cerambycidae) in Hokkaido,Japan

Bursaphelenchus mucronatus Mamiya et Enda has been recovered for the first time from adults of the cerambycid beetle, Monochamus urussovi (Fischer), in Hokkaido, Japan. The nematode was also recovered from the inner bark of Picea jezoensis (Siebold et Zuccarini) Carrière and Abies sachalinensis (Fr Schmidt) Masters infested with M. urussovi larvae. PCR–RFLP analysis indicated that B. mucronatus in Hokkaido is the European type.     

Occurrence and pathogenicity of fungi in necrotic and non-symptomatic shoots of declining common ash (<Emphasis Type="Italic">Fraxinus excelsior</Emphasis>) in Sweden

Currently, massive dieback of Fraxinus excelsior is observed in countries of eastern, northern and central Europe, and the reasons for it are unclear. The aims of the present work were (a) to study fungal communities in declining F. excelsior crowns; (b) to clarify role of fungi in the decline. Shoots from symptomatic crowns were collected in four localities in central Sweden, and distributed into the following categories: (a) visually healthy; (b) initial necroses; (c) advanced necroses; (c) dead tops. The most frequently isolated fungi were Gibberella avenacea, Alternaria alternata, Epicoccum nigrum, Botryosphaeria stevensii, Valsa sp., Lewia sp., Aureobasidium pullulans and Phomopsis sp., and these taxa were consistently found in shoots of all four symptomatic categories. Forty-eight taxa of other fungi were isolated, and fungal diversity was not exhausted by the sampling effort. The same taxa of fungi were dominant in F. excelsior shoots of different symptomatic categories, and moderate to high similarity of fungal communities was observed in shoots despite the symptoms. Forty-four isolates from 24 fungal taxa were used for artificial inoculations of 277 1-year-old F. excelsior seedlings in bare root nursery. After 2 years, only four fungi caused symptomatic necroses of bark and cambium: A. alternata, E. nigrum, Chalara fraxinea and Phomopsis sp. The most pathogenic was C. fraxinea, inducing symptoms on 50% of inoculated trees, while three other fungi caused necroses on 3–17% of inoculated trees. Infection biology of C. fraxinea and environmental factors determining susceptibility of F. excelsior to decline deserve future investigations.     

A survey of Ophiostoma species vectored by Hylastes ater to pine seedlings in New Zealand

Ecological studies in New Zealand require consideration of the impact of exotic (introduced) organisms from disparate environments. A survey of Ophiostoma species isolated and identified from Pinus radiata stumps, the bark beetle Hylastes ater collected from these stumps and from P. radiata seedlings following sublethal damage by H. ater, was undertaken in 10 reforestation sites in New Zealand. Nine Ophiostoma species were isolated and identified in this survey. Ophiostoma galeiformis and Ophiostoma huntii were isolated most frequently from all the substrates sampled. Ophiostoma floccosum, Ophiostoma setosum, Leptographium procerum and Leptographium truncatum were also isolated from stumps, damaged seedlings and beetles. Ophiostoma quercus was only isolated from stumps and beetles and Ophiostoma ips only from beetles. A strong relationship between the severity of H. ater feeding damage, and the presence and number of species of Ophiostoma was found. Eight Ophiostoma species were transmitted by H. ater to P. radiata seedlings in laboratory transmission experiments. From the results of this field survey and the laboratory experiments we confirm that H. ater is a vector of Ophiostoma species to P. radiata seedlings in New Zealand.     

Antifeedant activities of secondary metabolites from <Emphasis Type="Italic">Ajuga nipponensis</Emphasis> against adult of striped flea beetles, <Emphasis Type="Italic">Phyllotreta striolata</Emphasis>

Antifeedant activities of the isolated chemical compounds from Ajuga nipponensis, were studied against adult of striped leaf beetles. The methanol, petroleum ether, ethyl acetate and n-butanol extracts at 1.0 mg/ml, were used in this study. All four extracts exhibited more than 65 antifeedant index at 24 h and ethyl acetate extract showed significant activity against striped leaf beetles with 83.12 antifeedant index. Six compounds and one fraction were isolated by chromatography and their structures were identified by NMR, MS and FTIR spectra. At 2.0 mg/ml for 24 h the three compounds 20-hydroxyecdysone, acacetin and apigenin showed considerable activities with antifeedant indexes 59.29, 51.22 and 61.55, respectively. In contrast to this the antifeedant indexes of acacetin and apigenin, were sharply reduced as the time extended and that of 20-hydroxyecdysone remained unchanged. In addition, the synergistic effects of two mixtures of secondary metabolites, were studied and no sharp difference was observed.     

Interactions between<Emphasis Type="Italic"> Orthotomicus erosus</Emphasis> (Woll.) (Col., Scolytidae) and the Argentine ant<Emphasis Type="Italic"> Linepithema humile</Emphasis> (Mayr) (Hym., Formicidae)

Interactions occurring between Orthotomicus erosus, an economically important pine bark beetle in Portugal, and the aggressive Argentine ant Linepithema humile were investigated. Tri-trophic experimental enclosures consisting of Pinus pinaster logs, O. erosus adults and L. humile workers were set up in the field and in the laboratory. Interactions between ants and scolytids were observed and quantified in terms of the bark beetles success in colonizing the logs. The presence of L. humile did not influence the ability of O. erosus to colonize the logs, apparently due to the difficulties met by the ants in handling the bark beetles and to their inability to injure them. Thus, although abundant in many pine stands in Portugal and in spite of its aggressiveness, L. humile does not seem to affect populations of O. erosus.     

Successful <Emphasis Type="Italic">Agrobacterium</Emphasis>-mediated transformation of <Emphasis Type="Italic">Populus tomentosa</Emphasis> with apple <Emphasis Type="Italic">SPDS</Emphasis> gene

The problem of salinized soils has become one of the most serious constraints to agricultural and forest productivity. With the purpose of enhancing salt stress tolerance of Populus tomentosa, we transformed this tree species with spermidine synthase （SPDS） genes derived from an apple by an Agrobacterium-mediated method. Four transgenic clones were confu＇med by PCR and Southern blot analysis. As well, the expression of introduced SPDS genes was analyzed by real-time quantitative PCR.     

The influence of Desert False Indigo, <Emphasis Type="Italic">Amorpha fruticosa</Emphasis>, and three arbuscular mycorrhizae fungi on the growth of <Emphasis Type="Italic">Populus ussuriensis</Emphasis> seedlings

The influence of Desert False Indigo, Amorpha fruticosa, on the growth of Populus ussuriensis seedlings inoculated with three arbuscular mycorrhizae (AM) fungi (Glomus mosseae, Glomus intraradices, Glomus sinuosa) was studied using the nylon net method. The results showed that all three AM fungi infected P. ussuriensis seedlings and G. intraradices and also G. mosseae infected A. fruticosa. The AM fungi promoted growth of P. ussuriensis and Desert False indigo seedlings. Moreover, under co-cultivation with A. fruticosa, the biomass of P. ussuriensis increased significantly. The concentration of nitrogen in P. ussuriensis grown with A. fruticosa and the concentration of soluble nitrogen in the rhizosphere were also higher than when grown alone. Hypha were found on the two plant seedlings inoculated with G. mosseae and G. intraradices, suggesting that AM fungi may transport nutrients from seedlings of A. fruticosa to the rhizosphere of P. ussuriensis seedlings, which may have promoted the growth of P. ussuriensis. The AM fungi played a critical role on the effect of A. fruticosa on growth of P. ussuriensis.     

Adventitious root formation of two <Emphasis Type="Italic">Abies</Emphasis> species on log and soil in an old-growth subalpine forest in central Japan

We assessed stem burial and adventitious root formation of two late-successional species, Abies mariesii and A. veitchii, in central Japan. In a plot (5 × 5 m), all seedlings between 8 and 24 cm tall were excavated: six A. mariesii seedlings in soil, and six and four A. veitchii seedlings in soil and on logs, respectively. For each sampled seedling, the number of terminal bud scars (TBS) was counted on the aboveground and belowground stems. Stem length was measured, and divided into aboveground and belowground stems. Among the three groups (A. mariesii seedlings in soil and A. veitchii seedlings in soil or on logs), there was no significant difference in height or total root weight (sum of adventitious roots and primary roots), but diameter at ground level and number of TBS were significantly different. Counting TBS on the aboveground stem of seedlings in soil underestimated seedling age, whereas the estimate was much closer to the true age for seedlings on logs. Seedlings in soil formed more adventitious roots than seedlings on logs. A large proportion of the stem was buried in humus for seedlings in soil, while most of the stem was not buried in humus for seedlings on logs. These results suggest that substrate affects adventitious root formation, the formation of which is important to shade tolerance. Thus, our preliminary results suggest that consideration of adventitious root formation is necessary to understand seedling bank dynamics and estimate seedling ages of these Abies species in spatially heterogeneous old-growth subalpine forests.     

Spatial pattern of beech (<Emphasis Type="Italic">Fagus sylvatica</Emphasis> L.) and oak (<Emphasis Type="Italic">Quercus pubescens</Emphasis> Mill.) seedlings in natural pine (<Emphasis Type="Italic">Pinus sylvestris</Emphasis> L.) woodlands

Spatial pattern of recruitment is an important factor influencing population dynamics of plant communities. The spatial pattern is determined by seed dispersal and by the spatial variability of germination and initial survival. In the process of forest expansion following farmland abandonment, mid- and late-successional species are often dispersed in pioneer forests by birds. This could result in an aggregated spatial pattern of seeds that could influence the dynamics of these species in mixed pioneer forests. In the sub-Mediterranean area, mid- and late-successional species such as Quercus pubescens (downy oak) and Fagus sylvatica (European beech) are expected to replace pioneer Pinus species. Using a point sampling method we demonstrated that beech and oak seedlings (height <2 m) have a clumped distribution in the understorey of pine. This could result from an aggregated dispersal by jays (dispersal effect) or from preferential recruitment in particular habitats (habitat effect). To test these hypotheses we proposed a statistical analysis of spatial patterns of regeneration of beech and oak. Ground cover variables (i.e. cover by rock outcrops, herbs, box shrubs, mosses or pine) did not differ significantly around seedlings as compared with random sample plots. Likewise, clumped seedlings had growth similar to isolated seedlings, thus refuting the hypothesis of preferential location in the most favourable microsites. Aggregated dispersal seems to be involved in the process of regeneration. Since beech and oak seedlings have contrasting ecological demands, we discuss the implication of this pattern for the replacement dynamics of pine by these species.     

Distinction of resin compounds between the healthy bark and the resinous stem canker of <Emphasis Type="Italic">Thujopsis dolabrata</Emphasis> var. <Emphasis Type="Italic">hondae</Emphasis>

Methyl esters of higher fatty acids were detected from the healthy bark of Thujopsis dolabrata var. hondae but not from the bark of the resinous stem canker of T. dolabrata var. hondae. This difference enabled us to distinguish healthy trees from diseased ones. Fourteen diterpenes were also isolated from the n-hexane extracts of the bark-glued resin taken from the resinous stem canker of T. dolabrata var. hondae. Of these diterpenes, abietane diterpenes [abieta-7,13-diene (1), abietinol (2), dehydroabietinol (4)], pimarane diterpene [sandaracopimaric acid (8)], and labdane diterpenes [manool (10), torulosol (11), torulosal (12), cupressic acid (13)] were first isolated from T. dolabrata var. hondae.Part of this paper was presented at the 47th Annual Meeting of the Japan Wood Research Society, Tokyo, April 1999     

Ophiostomatoid fungi isolated from three different pine species in Argentinian Patagonia

Pine plantations in Argentinian Patagonia cover ca. 95,000 ha in Chubut, Río Negro and Neuquén provinces. Exotic bark beetles (Orthotomicus laricis, Hylastes ater and Hylurgus ligniperda) commonly occur in freshly cut logs, stumps and slash. These beetles are vectors of “ophiostomatoid” fungi which include primary tree pathogens as well as important agents of blue stain. The aim of this study was to identify these beetle‐associated fungi. Sawing mills and pine plantations were surveyed three consecutive years. Fungal isolates from stained logs, processed wood and insect galleries were identified based on morphological and DNA sequence comparisons of ITS and β‐tubulin gene regions. Two Grosmannia, one Graphilbum and three Ophiostoma species were identified. Ophiostoma piliferum and O. peregrinum sp. nov. were the most frequently isolated taxa. O. peregrinum occurred in all provinces, colonizing different conifer species and, interestingly, also the native broadleaved species Nothofagus dombeyi. Pine plantation forestry in southern South America includes Argentina, Brazil, Chile, Paraguay and Uruguay. Emerging data from Argentina, Chile and Uruguay revealed some coincidences between these countries, but also several differences, probably, as a result of multiple introduction events.     

Population dynamics of <Emphasis Type="Italic">Monochamus galloprovincialis</Emphasis> Olivier (Coleoptera: Cerambycidae) in two pine species under laboratory conditions

The study of the life history dynamics of the pine sawyer beetle Monochamus galloprovincialis (Olivier) is important because the beetles vector the pinewood nematode. The objectives of this study were to investigate differences in beetle life history parameters between the two common host tree species in Turkey, Pinus sylvestris and Pinus nigra. Relationships between the number of oviposition sites, holding time (time between log cut and oviposition), log moisture content, log volume and area, and adult survivorship were evaluated. No significant differences were found between the two tree species except adult density per m². The holding time did not influence the number of oviposition sites, larval entry holes or adults for P. sylvestris. There was a negative correlation between the holding time and the number of oviposition sites for P. nigra. The number of adults emerged per log was correlated with the number of larval entry holes only for P. sylvestris. Only 12 and 15% of the initial cohort completed development and emerged as adults for P. sylvestris and P. nigra, respectively. Both tree species are equally suitable hosts for M. galloprovincialis development. We conclude that the high within-log mortality of beetles is a combination of resource quality and quantity factors and intraspecific competition and cannibalism during larval development.     

Laboratory and field evaluation of <Emphasis Type="Italic">Beauveria bassiana</Emphasis> for controlling Mulberry whitefly <Emphasis Type="Italic">Pealius mori</Emphasis> Takahashi (Homoptera: Aleyrodidae) in mulberry (<Emphasis Type="Italic">Morus alba</Emphasis> Linn)

In a laboratory screening of 12 isolates of entomopathogenic fungi against nymphs of the mulberry whitefly (Pealius mori Takahashi), Beauveria bassiana (Balsamo-Crivelli) Vuillemin CKB-048 was the most virulent, causing 87 ± 3% mortality at 1 × 10⁶ conidia/ml. Infection was confirmed by growth of the fungus from cadavers and by scanning electron microscopy of treated nymphs. Beauveria bassiana CKB-048 was formulated as a wettable powder (1 × 10⁹ conidia/g) and tested in two mulberry (Morus alba Linn) plantations in central and northeastern Thailand. In both locations, two spray applications of B. bassiana CKB-048 at 3.75 × 10¹² to 6.25 × 10¹² conidia/ha and at 14 day intervals provided good control of whitefly nymphs; control with B. bassiana CKB-048 was comparable to that with the pesticide buprofezin at 250 g of active ingredient/ha. In addition, no mortality of silkworm larvae occurred when the larvae were fed with mulberry leaves sprayed with B. bassiana CKB-048 7, 14, or 21 days earlier.     

Effects of organic acid treatments on small hive beetles, <Emphasis Type="Italic">Aethina tumida</Emphasis>, and the associated yeast <Emphasis Type="Italic">Kodamaea ohmeri</Emphasis>

In honeybee, Apis mellifera, colonies infested with larval and adult small hive beetles (SHB), hive material, and in particular honey, tends to ferment, probably due to SHB-associated yeasts such as the predominant Kodamaea ohmeri. Here, we test the effects of organic acids on K. ohmeri and on SHB-infested honey/pollen combs. Organic acids were applied at standard concentrations used by beekeepers to control other pests. In laboratory tests, the growth of K. ohmeri was significantly inhibited by lactic, formic and acetic acids. Treatments of SHB-infested honey/pollen combs (N = 18 colonies) with acetic acid significantly increased mortality of adult SHB and treatments with formic acid significantly reduced larval infestation. Our data suggest that treatment of honeybee colonies and storage rooms with organic acids could also help in reducing SHB damage.     

Stereochemistry of matairesinol formation by <Emphasis Type="Italic">Daphne</Emphasis> secoisolariciresinol dehydrogenase

Secoisolariciresinol dehydrogenase activity was detected for the first time from Daphne odora and Daphne genkwa (Thymelaeaceae), which are known to produce optically pure (+)-matairesinol. In sharp contrast, (–)-matairesinol was formed selectively over the (+)-antipode by the secoisolariciresinol dehydrogenase preparation from both D. odora callus and D. genkwa shoots.     

Diversity of Ophiostomatales species associated with conifer-infesting beetles in the Western Carpathians

Bark beetles (Coleoptera: Curculionidae, Scolytinae) are commonly recognised as important agents of tree mortality in coniferous forests of the Western Carpathians. They, together with weevils, are consistently associated with ophiostomatoid fungi. Information regarding conifer beetle-associated fungi in the Western Carpathians remains incomplete and unreliable, particularly with respect to fir-infesting bark beetles. This study aims to clarify associations between fungi in the genera Graphilbum, Leptographium, Ophiostoma and Sporothrix (Ophiostomatales) and their beetle vectors in Norway spruce (Picea abies), European larch (Larix decidua) and silver fir (Abies alba). Samples associated with 20 bark beetle species and weevils were collected from nine stands in Poland and the Czech Republic. Fungi were isolated from adult beetles and galleries. Isolates were identified based on morphology, DNA sequence comparisons for four gene regions (ITS, LSU, ß-tubulin, TEF 1-α) and phylogenetic analyses. In total, 46 distinct taxa were identified, including 25 known and 21 currently unknown species. Several associations between fungi and subcortical insects were recorded for the first time. In addition, O. borealis and O. quercus were detected from A. alba for the first time. The composition of the fungal communities varied among the studied tree species and to a lesser degree among the beetle species. The spruce-infesting bark beetles were commonly associated with species of Leptographium s. l. and Ophiostoma s. str.; the larch-infesting bark beetles were often associated with Ophiostoma s. str. and Sporothrix, while the fir-infesting bark beetles were commonly associated with Ophiostoma s. str. and Graphilbum. The most commonly encountered fungal associates of the examined insects were (a) Grosmannia cucullata, G. piceiperda, Grosmannia sp. 1, Ophiostoma macroclavatum and O. piceae with the spruce-infesting bark beetles; (b) O. pseudocatenulatum and Sporothrix sp. 1 with the larch-infesting bark beetles; and (c) O. piceae, Ophiostoma sp. 2 and Graphilbum sp. 2 with the fir-infesting bark beetles. The differences in fungal associates among the bark beetle species occurring on P. abies, L. decidua and A. alba could be linked to the different habitats that these beetles occupy.