Breeding performance and longevity of Tomicus destruens on Mediterranean and continental pine species

The pine shoot beetle Tomicus destruens (Wollaston) (Coleoptera: Curculionidae: Scolytinae) is one of the main pests of Mediterranean forests, where it is oligophagous on Mediterranean pines. However, possible global warming may make the insect move to higher latitudes and altitudes, allowing it to attack new pine species. In this respect, the aim of the present article was to assess both the acceptance and performance of T. destruens offered host and non‐host pine species. A no‐choice breeding experiment was set up under laboratory conditions, using logs of three Mediterranean (Pinus pinea L., Pinus pinaster Miller, and Pinus halepensis Aiton) and two continental (Pinus nigra Arnold and Pinus sylvestris L.) pine species. Log debarking at the end of adult emergence assessed parent fecundity, egg, and larval mortality. The quality of callow adults emerging from each tested pine was evaluated on the basis of their longevity on a semiartificial diet. Tomicus destruens colonised all tested pine species, but did not reproduce in Scots pine, taking about 79 days to complete development with no differences among pines tested. The best breeding performance, evaluated as female fecundity and adult production, was observed in P. halepensis, and the lowest in P. pinaster. On average, adults emerging from P. pinea survived longer (83 days) than from other pines, and adult longevity was the lowest in males emerging from P. nigra. Austrian pine, which under natural conditions is usually not a host of T. destruens, allowed insect development and adult production similar to P. pinea and P. pinaster.     

Morphological and genetic identification of the three pine pests of the genus Tomicus (Coleoptera, Scolytidae) in Europe

Abstract

• 1 Morphological characters were elaborated and part of the mitochondrial COI gene was sequenced in order to facilitate the determination of the three European pine bark beetles Tomicus piniperda, T. destruens and T. minor. The sequence information also provided the first information on the phylogenetic and phylogeographical relationships of these species.
• 2 Three hair rows were found on the antennal club of T. destruens between the second and third suture. Tomicus piniperda had only one row. Three different hair types were detected on the elytra – two hair types were found on T. piniperda, whereas the third hair type was only detected on the elytra of T. destruens.
• 3 The COI region (445 bp) revealed high sequence divergence among T. destruens, T. piniperda and T. minor. The three species proved to be monophyletic species with 16.98–19.23% sequence divergence. A phylogenetic approach placed T. minor and T. destruens as sister taxa, which contradicts the morphological findings.
• 4 European populations of T. piniperda shared two haplotypes, indicating a homogenous distribution of the genotypes. In the American populations only one of these European haplotypes was found. The Greek, Italian and Spanish T. destruens populations revealed three population‐specific haplotypes, indicating restricted gene flow.
• 5 Species‐specific primers were designed to allow a rapid and definitive determination of the two sibling Tomicus species by PCR.

     

Descriptive biogeography of Tomicus (Coleoptera: Scolytidae) species in Spain

Aim, location Tomicus (Coleoptera, Scolytidae) species are some of the principal pests of Eurasian forest and are represented by three coexisting species in Spain, Tomicus piniperda (Linnaeus, 1758), Tomicus destruens (Wollaston, 1865) and Tomicus minor (Harting, 1834). The distribution of two taxa are unknown as they have until recently been considered separate species. Therefore, we model the potential distribution centres and establish the potential distribution limits of Tomicus species in Iberia. We also assess the effectiveness of different models by comparing predicted results with observed data. These results will have application in forest pest management. Methods Molecular and morphological techniques were used to identify species from 254 specimens of 81 plots. For each plot, a Geographical Information System was used to extract a set of 14 environmental (one topographic, six climatic) and biotic variables (seven host tree distributions). General Additive Models and Ecological Niche Factor Analysis models are applied for modelling and predicting the potential distribution of the three especies of Tomicus. Results The results of both modelling methodologies are in agreement. Tomicus destruens is the predominant species in Spain, living in low and hot areas. Tomicus piniperda occurs in lower frequency and prefers wet and cold areas of north‐central Spain. We detected sympatric populations of T. destruens and T. piniperda in Northern coast of Spain, infesting mainly P. pinaster. Tomicus minor is the rarest species, and it occupies a fragmented distribution located in high and wet areas. The remarkable biotic variable is the distribution of P. sylvestris, incorporated into the models of T. destruens and T. piniperda. Main conclusions These results indicate that in wet areas of north‐central Spain where T. piniperda occurs (and possibly the high altitudes of the southern mountains), T. destruens has a climatic distribution limit. In the northern border of this area, both species overlap their distributions and some co‐occurrences were detected. Tomicus minor potentially occurs in high and wet fragmented areas.     

Predicting the distribution of the two bark beetles Tomicus destruens and Tomicus piniperda in Europe and the Mediterranean region

• 1 Various factors such as climate and resource availability influence the geographical distributions of organisms. Species sensitive to small temperature variations are known to experience rapid distribution shifts as a result of current global warming, sometimes leading to new threats to agriculture and forests. Tomicus piniperda and Tomicus destruens (Coleoptera, Curculionidae, Scolytinae) cause economic damage to pines in Europe and around the Mediterranean Basin. However, their respective potential distributions have not yet been studied at a large scale. The present study aimed to investigate the influence of climatic and host factors on the geographical distributions of both Tomicus species in Europe and around the Mediterranean Sea, and to establish maps of suitable areas.
• 2 Using 114 published localities where the presence or absence of both species was unambiguously recorded, we gathered WorldClim meteorological records to correlate the occurrence of insects with bioclimatic variables and to build potential distribution maps.
• 3 The two studied Tomicus species presented parapatric distributions and opposite climate demands, with T. destruens occurring in locations with warmer temperatures, whereas T. piniperda occurs under a colder climate. Amongst the investigated climate variables, temperature appeared to be most correlated with both species distributions.
• 4 The potential ranges of both species were further restricted by the availability of pine hosts. It appeared that setting new pine plantations in regions where T. destruens or T. piniperda are still absent could favour a rapid expansion of their distributions. Our data will be useful when aiming to apply management strategies adapted to each species, and to forecast their potential range expansions/contractions as a result of climate warming.

     

Shoot feeding ecology of Tomicus piniperda and T. minor (Col., Scolytidae) in southern China

Abstract: The pine shoot beetle, Tomicus piniperda, is causing substantial tree mortality to Pinus yunnanensis in south‐western China, whereas the same species in Europe seldom kills Pinus sylvestris. In order to understand this difference in aggressiveness, we studied the shoot feeding ecology of the pine shoot beetles in Yunnan, and compared it with European results. We found many similarities in the shoot feeding behaviour of T. piniperda, and also that of Tomicus minor, which was common locally. In contrast to Europe, however, the pine shoot beetles in Yunnan seem to be able to predispose the host trees for stem attack by intensive shoot‐feeding. It has also been observed that beetles aggregate in certain trees during shoot feeding, but we could not verify that in our experiment.     

Attack dynamics of the pine shoot beetle, Tomicus piniperda (Col.; Scolytinae) in Scots pine stands defoliated by Bupalus piniaria (Lep.; Geometridae)

Abstract 1 After a 1‐year, extensive pine looper (Bupalus piniaria) outbreak, plots were laid out to study tree susceptibility to attack, and performance of Tomicus piniperda in pine trees suffering from varying levels of defoliation. 2 Tomicus piniperda was the dominating stem‐attacking species among the primary stem colonizers, and 82% of all trees that died had been colonized by T. piniperda. 3 Beetle attacks primarily struck severely defoliated trees, i.e. trees that suffered from 90% to 100% defoliation. 4 Beetle attacks peaked in the second year after cessation of the outbreak, and suppressed trees were both more frequently attacked and more susceptible to beetle attack than intermediate and dominant trees. 5 Trees surviving beetle attacks carried more foliage than trees that did not survive the attacks. 6 A single year of severe defoliation is enough to render pine trees susceptible to secondary pests, such as T. piniperda.     

Impacts of Tomicus minor on distribution and reproduction of Tomicus piniperda (Col., Scolytidae) on the trunk of the living Pinus yunnanensis trees

This study investigated the impacts of Tomicus minor on Tomicus piniperda when the two Tomicus species coexist in the trunks of living Yunnan pine (Pinus yunnanensis L.) trees growing in the Kunming region, in south-western China. Tomicus piniperda mostly locates in the mid- and upper trunks of Yunnan pine tree; whereas T. minor mainly attacks the mid- and lower trunks. In the mid-trunk area from 1.0 to 5.0 m above ground, there are overlapping attack zones for the two Tomicus species, which accounts for an average of 80% of the entire zone occupied by T. piniperda and an average of 70% of the zone occupied by T. minor. In correspondence with their attack distributions, the average attack densities of the two species varied with height along the trunk, with 165.3 egg galleries per m² at a height of 6 m for T. piniperda, and 138.2 egg galleries per m² at the 1 m height for T. minor. It is suggested that T. minor adjusts its attack pattern with respect to T. piniperda, and thereby minimizes interspecific competition. No remarkable difference of average T. piniperda egg gallery length was found between the zone in which only T. piniperda occurred and the zone in which T. piniperda and T. minor coexisted; this is suggested to be due to low host quality in the upper trunk region where only T. piniperda was present. The number of T. piniperda larval galleries was highest when only T. piniperda was present, and decreased as T. piniperda and T. minor coexisted, particularly in the case when the density of T. piniperda was less than that of T. minor. Average larval density was 1649 larval galleries per m² where only T. piniperda occurred. However, when T. piniperda coexisted with T. minor, T. piniperda larval density averaged 1010 per m² when T. piniperda density was higher than T. minor, and averaged 442 per m² when T. piniperda density was less than T. minor, which led to the conclusion that T. minor makes a negative impact on T. piniperda reproduction when the two Tomicus species jointly colonize the same trunk of Yunnan pine tree.     

纵坑切梢小蠹对云南松蛀害研究

在昆明地区,纵坑切梢小蠹Tomicus piniperda L.表现出枝梢聚集、树干蛀害等重要的行为学特征,形成三种基本蛀害模式。横坑切梢小蠹、蓝色伴生真菌参与了纵坑切梢小蠹危害过程,并在其中发挥积极作用。上述因素的综合影响,加强了纵坑切梢小蠹对云南松Pinus yunnanensis寄主树木的危害能力。     

Hierarchical structure of mitochondrial lineages of Tomicus destruens (Coleoptera,Scolytidae) related to environmental variables

We investigated here the relation of environmental variables with the distribution of mitochondrial lineages using a bark beetle species of Mediterranean distribution as a model. We analysed a total number of 460 DNA sequences of Tomicus destruens provided by intensive and extensive collection and GenBank entries. We combined phylogeography and regression models to study the role of five environmental predictors at fine scale in the distribution of a local genealogy. The analysis revealed a high genetic diversity, with 52 haplotypes present in Sierra Espuña forest (SE Spain) and 21 haplotypes in the other 14 Spanish populations, all included in the western clade of the Mediterranean phylogeography of the species. We found a micro‐distribution of the species related to altitude and putative niche segregation between lineages associated with the micro‐environmental conditions of their host pine trees. We compared the phylogeographic hypothesis obtained here with the phylogeography obtained integrating our data with all data published elsewhere. Here, we demonstrate a relation between the environmental heterogeneity and the haplotypic diversity at Mediterranean Basin scale. This analysis allows us to support the evolutionary scenario where the phylogeography and current molecular diversity of T. destruens is a consequence of the recolonization from two principal refugia at both extremes of the Mediterranean Basin and, according to our data, we propose that the heterogeneity of habitats allows fixation of the mitochondrial lineages.     

Prolonged development time of the bark beetle predator Thanasimus formicarius (Col.: Cleridae) in relation to its prey species Tomicus piniperda (L.) and Ips typographus (L.) (Col.: Scolytidae)

1 The generation time of the bark beetle predator Thanasimus formicarius (L.) (Col.: Cleridae) was found to be predominantly two years both in the field and in rearing experiments conducted with two of its main prey species, the pine shoot beetle Tomicus piniperda (L.) and the spruce bark beetle Ips typographus (L.) (Col.: Scolytidae). 2 Emergence of T. formicarius adults in the first summer was only observed in one of the two rearing experiments, and these individuals represented only 6% of that generation. 3 All individuals not emerging as adults in the first summer remained as larvae in their pupal chambers until the second summer. Pupae were found starting around mid-June, and adults (in pupal chambers) were found from late July through to the end of August. 4 Newly emerged adults had to feed in order to survive hibernation. 5 The existence of T. formicarius races, specialized on certain bark beetle species and with phenologies matching their hosts, could not be demonstrated. After hibernation there was no difference in feeding activity, timing of egg-laying or proportion of egg-laying females between the T. formicarius adults reared as larvae on T. piniperda (flight period in April) and those reared as larvae on I. typographus (main flight period generally starting in late May or early June) when exposed to a temperature and day-length typical of the early spring conditions prevailing during the flight period of T. piniperda. 6 T. formicarius was parasitized by Enclisis vindex (Tschek) (Hym.: Ichneumonidae) in the pupal chamber. 7 The importance of these findings for the population dynamics of bark beetles is discussed.     

Attraction of the bark beetle Tomicus piniperda to Scots pine trees in relation to tree vigor and attack density

The attraction of Tomicus piniperda (L.) (Coleoptera: Scolytidae) to Scots pine trees of low and higher vigor with and without previous attacks was monitored with sticky traps. Somewhat higher numbers of beetles were caught on low vigor trees than on trees of higher vigor, indicating differences in olfactory stimuly between the two classes of trees.Many more beetles were caught on trees attacked by T. piniperda and on trees with simulated bark beetle galleries (manually drilled holes) than on control trees with no or only a few attacks. The strong attraction of beetles to attacked trees is attributed to the beetles responding to host volatiles released from the galleries.

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Zusammenfassung Der Anflug von Tomicus piniperda (L.) (Coleoptera: Scolytidae) an Kiefern in schlechtem oder besserem Gesundheitszustand, mit oder ohne vorherigen Befall, wurde mit Hilfe von Leimfallen untersucht. An Bäumen in schlechtem Zustand wurden etwas mehr Käfer gefangen als an Bäumen in besserem Gesundheitszustand. Das deutet auf Unterschiede in Geruchsreizen zwischen den beiden Klassen des Baumzustands hin.An Kiefern, die von T. piniperda befallen waren oder simulierte Einbohrungen (von Hand gebohrte Löcher) hatten, wurden viel mehr Käfer gefangen als an Kontrollbäumen ohne oder mit sehr geringem Befall. Der starke Anflug von Käfern an befallene Bäume wird gedeutet als Reaktion auf Duftstoffe des Wirts, die aus den Borkenkäfergängen abgegeben werden.

     

Development of an improved attractive lure for the pine shoot beetle, Tomicus piniperda (Coleoptera: Scolytidae)

Abstract 1 The pine shoot beetle, Tomicus piniperda (L.) (Coleoptera: Scolytidae), is an exotic pest of pine, Pinus spp., and was first discovered in North America in 1992. 2 Although primary attraction to host volatiles has been clearly demonstrated for T. piniperda, the existence and role of secondary attraction to insect‐produced pheromones have been widely debated. 3 Currently, commercial lures for T. piniperda include only the host volatiles α‐pinene in North America and α‐pinene, terpinolene and (+)‐3‐carene in Europe. Several potential pheromone candidates have been identified for T. piniperda. 4 We tested various combinations of host volatiles and pheromone candidates in Michigan, U.S.A., and Ontario, Canada, to determine an optimal blend. 5 Attraction of T. piniperda was significantly increased when trans‐verbenol (95% pure, 3.2%cis‐verbenol content) was added with or without myrtenol to α‐pinene or to blends of α‐pinene and other kairomones and pheromone candidates. 6 Our results, together with other research demonstrating that trans‐verbenol is produced by T. piniperda, support the designation of trans‐verbenol as a pheromone for T. piniperda. A simple operational lure consisting of α‐pinene and trans‐verbenol is recommended for optimal attraction of T. piniperda.     

Monoterpenes governing host selection in the bark beetles Hylurgops palliatus and Tomicus piniperda

Under field conditions, both bark beetles, Hylurgops palliatus Gyll. and Tomicus piniperda L. (Col., Scolytidae) preferred flight barrier traps baited with ethanol and oleoresin of their favoured host, Picea abies L., and Pinus sylvestris L., respectively, compared to traps baited with ethanol and various monoterpenes. However, replacing the host-specific oleoresin with -pinene and the pine-characteristic terpinolene enhanced trap catches of both species. Response of H. palliatus increased with myrtenol, but decreased with (+)-trans-verbenol, while T. piniperda was attracted by both terpene alcohols.

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Zusammenfassung Die Lockwirkung wirtsspezifischer Terpenmuster auf die sekundären Rindenbrüter Hylurgops palliatus und Tomicus piniperda wird im Freiland untersucht. Beide Arten fliegen bevorzugt Fallen an, die mit Ethanol und dem Rohharz ihrer Wirtsbäume, Picea abies bzw. Pinus sylvestris, beködert sind. Der Ersatz der Rohharze durch eine Mischung aus -Pinen und dem für Kiefernharz charakteristischen Terpinolen erhöht die Fangzahlen beider Arten deutlich. Die Präferenzen für Monoterpene deuten unterschiedlich enge Wirtsbindungen an: Bei breitem Wirtsspektrum bevorzugt H. palliatus wenig spezifische Terpene und hohe Ethanol-Konzentrationen, der zur Monophagie an Frischholz neigende Waldgärtner T. piniperda jedoch geringe Ethanol-Konzentrationen und spezifische Terpenkombinationen. H. palliatus wird von dem Terpenalkohol (-)-Myrtenol deutlich angelockt, während das von aggressiven Scolytiden bekannte Pheromon (+)-trans-Verbenol seinen Anflug im Gegensatz zu T. piniperda zu hemmen scheint.

     

Evolving Management Strategies for a Recently Discovered Exotic Forest Pest: The Pine Shoot Beetle, Tomicus Piniperda (Coleoptera)

Established populations of the Eurasian pine shoot beetle (Tomicus piniperda (L.); Coleoptera: Scolytidae) were first discovered in North America in Ohio in 1992. As of 31 December 2000, T. piniperda was found in 303 counties in 12 US states (Illinois, Indiana, Maine, Maryland, Michigan, New Hampshire, New York, Ohio, Pennsylvania, Vermont, West Virginia, and Wisconsin) and in 43 counties in 2 Canadian provinces (Ontario and Quebec). A federal quarantine imposed in November 1992 regulates movement of pine (Pinus) trees, logs, and certain pine products from infested to uninfested areas within US. The forest products, Christmas tree, and nursery industries are affected by the quarantine. This paper summarizes information on the discovery and spread of T. piniperda in North America, survey efforts, recent interception history, development and changes in the federal quarantine, development of a national compliance management program, and extension and research efforts.     

Interactions between the predators thanasimus formicarius (Col.: Cleridae) and rhizophagus depressus (Col: Rhizophagidae), and the bark beetle tomicus Piniperda (Col.: Scolytidae)

The occurrences of Thanasimus formicarius (L.) (Cleridae), Rhizophagus depressus (F.) (Rhizophagidae), and Epuraea marseuli Reitter (Nitidulidae) in cut Scots pines, Pinus sylvestris L., attacked by Tomicus piniperda (L.) (Col.: Scolytidae) were recorded in the field, and interactions between the species were studied in caged pine bolts attacked by T. piniperda. T. formicarius eggs and R. depressus adults were abundant in the T. piniperda attacked trees, whereas only a few individuals of E. marseuli were found. T. formicarius and R. depressus, but not E. marseuli, reproduced in the caged bolts. T. piniperda offspring production per unit area of bark was reduced by 41% when reared with R. depressus, by 81% when reared with T. formicarius, and by 89% when all three species were reared together, compared with T. piniperda alone. The interaction between T. formicarius and R. depressus was mutually antagonistic. When both species were present in the same bolt the total number of larvae was reduced by 49% for R. depressus and the number of large larvae (length > 10 mm) was reduced by 35% for T. formicarius compared with their respective production values when each species was present alone with the bark beetle. There was a positive relationship between T. piniperda egg gallery density and the production of R. depressus larvae per m². Larvae of both R. depressus and T. formicarius developed into new adults during the first summer.

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Résumé Interactions entre les prédateursThanasimus formicarius (Col.: Cleridae) etRhizophagus depressus (Col.: Rhizophagidae) et le scolyteTomicus piniperda (Col.: Scolytidae) La présence deThanasimus formicarius (L.) (Cleridae),Rhizophagus depressus (F.) (Rhizophagidae), etEpuraea marseuli Reitter (Nitidulidae) a été étudiée sur le terrain dans des pins sylvestres (Pinus sylvestiris L) abattus, attaqués partomicus piniperda (L.) (Col.: Scolytidae) et les relations entre les espèces ont été étudiées sur des rondins de pin attaqués parT. piniperda et placés sous cage. Le nombre d’∄ufs deT. formicarius et d’adultes deR. depressus était important dans les arbres attaqués parT. piniperda, alors que seuls quelques individus deE. marseuli ont pu être trouvés.T. formicarius etR. depressus, mais pasE. marseuli, se sont reproduits dans les rondins sous cage. La production de descendants deT. piniperda par unité de surface d’écorce était réduite de 41% quand le scolyte était élevé avecR. depressus, de 81% en élevage avecT. formicarius et de 89% quand les trois espèces sont élevées ensemble, par comparaison avecT. piniperda seul. Les interactions entreT. formicarius etR. depressus étaient mutuellement antagonistes. Quand les deux espèces étaient présentes dans le même rondin, le nombre total de larves était réduit de 49% pourR. depressus et le nombre de grosses larves (longueur > 10 mm) était réduit de 35% pourT. formicarius comparé aux valeurs respectives quand chaque espèce était présente isolément avec le scolyte. Il y a une corrélation positive entre la densité de galeries avec des ∄ufs deT. piniperda et la production de larves deR. depressus par m². Les larves deR. depressus et deT. formicarius se sont transformés en adultes l’été suivant.

     

Characterization of five microsatellite loci in the pine shoot beetle Tomicus piniperda (Coleoptera: Scolytidae)

Tomicus piniperda is a bark beetle that causes damage to various pine species across a wide geographical range. We developed five microsatellite polymorphic markers using an enrichment protocol. All loci could be successfully amplified with no evidence of null alleles and will be useful for population genetic studies. Cross‐species amplifications show that at least some of the markers could be useful in four other Tomicus species.     

Interactions between the phloem-feeding speciesTomicus piniperda (Col.: Scolytidae) andAcanthocinus aedilis (Col.: Cerambycidae), and the predatorThanasimus formicarius (Col.: Cleridae) with special reference to brood production

Interactions betweenTomicus piniperda (L.) (Col.: Scolytidae),Acanthocinus aedilis (L.) (Col.: Cerambycidae) andThanasimus formicarius (L.) (Col.: Cleridae) were investigated in caged pine bolts. The treatments wereT. piniperda alone,A. aedilis alone,T. piniperda together withA. aedilis, T. piniperda together withT. formicarius and all three species together. T. piniperda offspring production per m² was reduced by 92% when reared withT. formicarius, by 78% when reared withA. aedilis, and by 94% when all three species were reared together, compared withT. piniperda reared alone.A. aedilis had a negative influence on the offspring production ofT. formicarius and vice versa. When both species were present in the same bolt (together withT. piniperda) offspring production was reduced by 74% forA. aedilis and by 42% forT. formicarius compared with their respective production values when each species was present alone with the bark beetle. The new generation ofT. formicarius emerged as larvae from June to August while most of theA. aedilis offspring emerged as adults from September to October, leaving only a few larvae in the bolts to hibernate.A. aedilis only reproduced in a small part of one of the bolts without bark beetles.     

Species of Bursaphelenchus Fuchs, 1937 (Nematoda: Parasitaphelenchidae) and other nematode genera associated with insects from Pinus pinaster in Portugal

Insects associated with maritime pine, Pinus pinaster, in Portugal were collected and screened for the presence of Bursaphelenchus species. Nematodes were identified using Internal Transcribed Spacers‐Restriction Fragment Length Polymorphism (ITS‐RFLP) analysis of dauer juveniles and morphological identification of adults that developed from dauer juveniles on fungal cultures or on cultures in pine wood segments at 26°C. Several associations are described: Bursaphelenchus teratospicularis and Bursaphelenchus sexdentati are associated with Orthotomicus erosus; Bursaphelenchus tusciae, B. sexdentati and/or Bursaphelenchus pinophilus with Hylurgus ligniperda and Bursaphelenchus hellenicus with Tomicus piniperda, Ips sexdentatus and H. ligniperda. An unidentified Bursaphelenchus species is vectored by Hylobius sp. The previously reported association of Bursaphelenchus xylophilus with Monochamus galloprovincialis was confirmed. The association of Bursaphelenchus leoni with Pityogenes sp. is not definitively established and needs further studies for clarification. Other nematode genera besides Bursaphelenchus were found to be associated with the insects sampled, including two different species of Ektaphelenchus, Parasitorhabditis sp., Parasitaphelenchus sp., Contortylenchus sp. and other unidentified nematodes. The Ektaphelenchus species found in O. erosus is morphologically similar to B. teratospicularis found in the same insect; adults of both the species are found in cocoon‐like structures under the elytra of the insects.     

The influence of nematodes on the structure of genitalia of tomicus piniperda (Coleoptera: Scolytidae)

Larvae and adults of the greater pine shoot beetle, Tomicus piniperda, were collected from geographically distinct areas of Poland and examined for mortality causing agents. Endoparasitic nematodes were found in 1535 (62%) of the adults examined. Primary pathologies caused by the nematodes were reduction in size of fat body and structural damage to the genitalia of male adults. The latter, though not a major cause of mortality, may lead to reduced fertility of the beetle.     

Distribution of genetic diversity in Pinus pinaster Ait. as revealed by chloroplast microsatellites

 Using nine chloroplast simple sequence.repeats (cpSSRs) markers, we evaluated haplotypic variation within and among natural populations of Maritime pine (Pinus pinaster Ait.) in order to shed light on the history of this species. Seven out of the nine cpSSRs analysed were polymorphic, giving a total of 24 different variants. The 24 variants combined in 34 different haplotypes. The populations which generally showed the lowest level of haplotypic diversity are those located in Portugal. The Landes (France) and Pantelleria (Italy) populations represent the two main reservoirs of haplotypic diversity. The proportion of genetic differentiation among populations, estimated using R_st, which is a measure based upon a strict stepwise mutation model, was 0.235. The high level of differentiation was also confirmed by the AMOVA analysis (Φ_ST=0.254, P<0.001). Four main groups of populations were identified on the basis of Principal Component Analysis, with the differences being statistically significant (Φ_CT=0.299, P<0.001). Based on our results the presence of refugia located in the South of Portugal, previously proposed for this species, may be excluded, and a different possible recolonization process of Maritime pine in the post-glacial period has been proposed. Populations from North Africa and France might have represented a starting point of the recolonization process of Portugal and of the Italian part of the natural range, respectively. This hypothesis seems to be confirmed by the analysis of the distribution of the pairwise differences among individuals within populations: Landes and Pantelleria populations showed a bimodal distribution, as would be expected for ancient gene pools. Received: 5 November 1997 / Accepted: 5 January 1998