Seasonal Terpene Variation in Needles of Pinus radiata (Pinales: Pinaceae) Trees Attacked by Tomicus piniperda (Coleoptera: Scolytinae) and the Effect of Limonene on Beetle Aggregation.

Concentrations of four monoterpenes were determined in needles of Pinus radiata (D.Don) (Pinales: Pinaceae) trees that were attacked or nonattacked by Tomicus piniperda (L.) (Coleoptera: Scolytinae). Compounds were identified and quantified by gas chromatography-mass spectrometry. The mean ambient temperature was obtained using climate-recording data loggers. The effect of limonene on field aggregation was also evaluated at three limonene release rates using Lindgren attractant-baited traps and trap logs. Attacked trees produced less α-pinene in March, July, and November than nonattacked trees, less ß-pinene in July and November, and less limonene from May to November. Limonene reduced the attraction of T. piniperda to attractant-baited traps and trap logs. Results were linked to better responses to high temperatures, with respect to terpene contents, by the nonattacked trees after the spring attack.

Variation in virulence of Beauveria bassiana and B. pseudobassiana to the pine weevil Pissodes nemorensis in relation to mycelium characteristics and virulence genes.

Entomopathogenic fungi such as Beauveria spp. have potential applications in the biocontrol of insect pests but little is known regarding their infectivity to the pine weevil Pissodes nemorensis. In this study, five isolates of Beauveria pseudobassiana and five isolates of Beauveria bassiana were tested for characteristics correlating with virulence on P. nemorensis. Isolate UAMH301 had the lowest mean lethal concentration value whereas the highest value was obtained with isolate LRC137. Growth rate was negatively correlated with virulence in B. bassiana, because isolate LRC137, the least virulent isolate, grew much more rapidly than the other B. bassiana isolates on SDYA. In contrast, its growth on a hyperosmotic medium was the slowest. Sporulation rate and conidial area were not correlated with virulence. Mycelial cell density was positively correlated with virulence in both species, and the four tested genes appear to be one-copy genes. Bbchit1 and Bbhog1, genes respectively encoding a chitinase and a protein kinase, induced relative expression levels were positively correlated with virulence in B. pseudobassiana. We discuss in terms of previous morphological, physiological and genetic parameters related to virulence in Beauveria and the importance of testing the expression of putative virulence genes in comparison with their basal transcript levels.

Ophiostomatoid fungi including two new fungal species associated with pine root-feeding beetles in northern Spain.

Many bark beetles live in a symbiosis with ophiostomatoid fungi but very little is known regarding these fungi in Spain. In this study, we considered the fungi associated with nine bark beetle species and one weevil infesting two native tree species (Pinus sylvestris and Pinus nigra) and one non-native (Pinus radiata) in Cantabria (Northern Spain). This included examination of 239 bark beetles or their galleries. Isolations yielded a total of 110 cultures that included 11 fungal species (five species of Leptographium sensu lato including Leptographium absconditum sp. nov., five species of Ophiostoma sensu lato including Ophiostoma cantabriense sp. nov, and one species of Graphilbum). The most commonly encountered fungal associates of the bark beetles were Grosmannia olivacea, Leptographium procerum, and Ophiostoma canum. The aggressiveness of the collected fungal species was evaluated using inoculations on two-year-old P. radiata seedlings. Leptographium wingfieldii, Leptographium guttulatum, and Ophiostoma ips were the only species capable of causing significant lesions.

Multigene phylogenies of Ophiostomataceae associated with Monterey pine bark beetles in Spain reveal three new fungal species.

Ophiostoma species, some of which cause sapstain in timber and/or are mild pathogens, are common fungal associates of bark beetles (Coleoptera: Scolytinae). Three new Ophiostomataceae from Spain are recognized in the present study based on comparisons of sequence data for three gene regions as well as morphological characteristics. The new taxa are described as Ophiostoma nebulare sp. nov., Ophiostoma euskadiense sp. nov. and Graphilbum crescericum sp. nov.

Ophiostoma species (Ascomycetes: Ophiostomatales) associated with bark beetles (Coleoptera: Scolytinae) colonizing Pinus radiata in northern Spain.

Bark beetles (Coleoptera: Scolytinae) are known to be associated with fungi, especially species of Ophiostoma sensu lato and Ceratocystis. However, very little is known about these fungi in Spain. In this study, we examined the fungi associated with 13 bark beetle species and one weevil (Coleoptera: Entiminae) infesting Pinus radiata in the Basque Country of northern Spain. This study included an examination of 1323 bark beetles or their galleries in P. radiata. Isolations yielded a total of 920 cultures, which included 16 species of Ophiostoma sensu lato or their asexual states. These 16 species included 69 associations between fungi and bark beetles and weevils that have not previously been recorded. The most commonly encountered fungal associates of the bark beetles were Ophiostoma ips, Leptographium guttulatum, Ophiostoma stenoceras, and Ophiostoma piceae. In most cases, the niche of colonization had a significant effect on the abundance and composition of colonizing fungi. This confirms that resource overlap between species is reduced by partial spatial segregation. Interaction between niche and time seldom had a significant effect, which suggests that spatial colonization patterns are rarely flexible throughout timber degradation. The differences in common associates among the bark beetle species could be linked to the different niches that these beetles occupy.

Quantitative association of bark beetles with pitch canker fungus and effects of verbenone on their semiochemical communication in Monterey pine forests in Northern Spain.

The association between 11 species of bark beetles (Coleoptera: Scolytinae) and one weevil (Coleoptera: Entiminae) with the pitch canker fungus, Fusarium circinatum Nirenberg and O'Donnell, was determined by crushing beetles on selective medium and histone H3 gene sequencing. Pityophthorus pubescens (Marsham) (25.00%), Hylurgops palliatus (Gyllenhal) (11.96%), Ips sexdentatus (Börner) (8.57%), Hypothenemus eruditus Westwood (7.89%), Hylastes attenuatus Erichson (7.40%), and Orthotomicus erosus (Wollaston) (2.73%) were found to carry the inoculum. In addition, the root weevil Brachyderes incanus L. (14.28%) had the second highest frequency of occurrence of the fungus. The responses of the insects to a range of verbenone doses were tested in field bioassays using funnel traps. Catches of P. pubescens, a species colonizing branch tips of live trees, were significantly reduced in a log-linear dose-dependent relationship. Catches of I. sexdentatus, an opportunistic species normally attacking fresh dead host material, were also gradually reduced with increasing verbenone dose. Catches of Tomicus piniperda L., O. erosus, Dryocoetes autographus (Ratzeburg), H. eruditus, Xyleborus dryographus (Ratzeburg), Hylastes ater (Paykull), Hylurgus ligniperda (F.), H. attenuatus, and B. incanus were not significantly affected by verbenone. The effects of verbenone were consistent with differences in host-age preference. Semiochemical disruption by verbenone in P. pubescens and I. sexdentatus could represent an integrated pest management strategy for the prevention of the spread of pitch canker disease between different stands. However, several species associated with F. circinatum were unaffected by verbenone, not supporting this compound for prevention of the establishment of potential vectors in Northern Spain.