Ophiostoma breviusculum sp. nov. (Ophiostomatales, Ascomycota) is a new species in the Ophiostoma piceae complex associated with bark beetles infesting larch in Japan

A new species of the Ophiostoma piceae-complex was isolated from bark beetles and the insect galleries of Larix kaempferi in Japan. This species was described as Ophiostoma breviusculum. The fungus was morphologically similar to O. piceae and O. quercus. However the average length of the perithecial necks and synnemata were shorter than for O. piceae and O. quercus. Synnemata morphological characteristics also differentiated O. breviusculum from the other species of the Ophiostoma piceae-complex isolated from conifers. Mating tests demonstrated that this fungus did not produce perithecia with O. floccosum, O. piceae and O. quercus. In phylogenetic trees using rDNA ITS O. breviusculum was placed in a clade with O. canum, O. piceae and O. subalpinum, but when using beta-tubulin it was placed into a separate clade.     

Genetic characterization of Raffaelea quercivora isolates collected from areas of oak wilt in Japan

This study was undertaken to improve understanding of the phylogenetic position of pathogenic fungi implicated in the oak wilt in Japan. Sequences were obtained from three regions of partial nuclear ribosomal DNA of 25 isolates of Raffaelea quercivora including an ex-type strain, all of which were collected from seven areas of disease outbreak. All the isolates formed one clear clade with high bootstrap values, distinctly delimited from the closest species, R. montetyi. These results indicate that the R. quercivora is phylogenetically a well-defined taxon.     

<Emphasis Type="Italic">Ceratocystis ficicola</Emphasis> sp. nov., a causal fungus of fig canker in Japan

The causal fungus of Ceratocystis canker of fig in Japan is described as Ceratocystis ficicola sp. nov. This species is characterized by galeated ascospores and is similar to Ceratocystis fimbriata sensu lato. However, the perithecia of the new species are much larger than those of C. fimbriata. The fungus grows more rapidly than C. fimbriata sensu stricto at 25°C but more slowly at 30°C. Molecular analysis of the nucleotide sequences of rDNA ITS regions showed that C. ficicola is phylogenetically placed in the clade of C. fimbriata s. l. but is easily distinguishable from other species of C. fimbriata s. l.     

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.     

Regeneration and coexistence of two Abies species dominating subalpine forests in central Japan

Summary The mechanism of coexistence of the dominant firs Abies veitchii and A. mariesii is described in relation to regeneration patterns for climax subalpine forests of the northern Yatsugatake Mountains, central Honshu, Japan. Two mature stand types, pure conifer stands of Abies spp., and mixed stands of Abies spp. and hardwoods (mainly the birch Betula ermanii), are distinguished. Pure stands are likely to show simultaneous decay, followed by evenaged regeneration of stand-floor seedlings (<20 cm tall), Rapidly growing A. veitchii dominates over A. mariesii in this type of regeneration, which is occasionally invaded by light-demanding Betula. In constrast, mixed stands degenerate rather slowly, followed by the regeneration of Abies from the bank of suppressed saplings (>20 cm tall), which persist only in mixed stands. The more shade-tolerant A. mariesii is supeior in this type of regeneration, while Betula does not succeed, and mixed stands change to pure stands with time. The fact that two patterns of Abies regeneration occur in a certain ratio in the forest is what enables the two Abies species to coexist. A simple dynamical system model supports this conclusion.     

Four new Ophiostoma species associated with hardwood-infesting bark beetles in Norway and Poland

Ophiostoma spp. (Ophiostomatales, Ascomycota) are well-known fungi associated with bark and ambrosia beetles (Curculionidae: Scolytinae, Platypodinae). Fungi in the Ophiostomatales include serious tree pathogens as well as agents of timber blue-stain. Although these fungi have been extensively studied in the northern hemisphere, very little is known regarding their occurrence on hardwoods in Europe. The aims of the present study were to identify and characterize new Ophiostoma spp. associated with bark and ambrosia beetles infesting hardwoods in Norway and Poland, and to resolve phylogenetic relationships of Ophiostoma spp. related to the Norwegian and Polish isolates, using multigene phylogenetic analyses. Results obtained from five gene regions (ITS, LSU, β-tubulin, calmodulin, translation elongation factor 1-α) revealed four new Ophiostoma spp. These include Ophiostoma hylesinum sp. nov., O. signatum sp. nov., and O. villosum sp. nov. that phylogenetically are positioned within the Ophiostoma ulmi complex. The other new species, Ophiostoma pseudokarelicum sp. nov. reside along with Ophiostoma karelicum in a discrete, well-supported phylogenetic group in Ophiostoma s. stricto. The results of this study clearly show that the diversity and ecology of Ophiostoma spp. on hardwoods in Europe is poorly understood and that further studies are required to enrich our knowledge about these fungi.     

危害松树的小蠹虫与其伴生菌的相互关系

危害健康松属植物的小蠹虫经常与一些特殊的真菌相联系。在小蠹虫危害松属植物的过程中,这些真菌被小蠹虫的一些特殊结构或者体表携带到松属植物上。小蠹虫与其伴生菌的联系表明小蠹虫和其伴生菌之间是一种互惠互利的关系。伴生菌随小蠹虫扩散而被带到新的寄主树木。而伴生菌或作为小蠹虫的食物来源,但更重要的是,有些伴生菌能够通过其菌丝渗透寄主组织,释放毒素,致死寄主树木,以帮助小蠹虫降低寄主抗性。许多研究致力于探索小蠹虫/伴生菌联合体与寄主树木之间关系的特征和确定小蠹虫与其伴生菌相互关系在生态学上的意义。然而,不同小蠹虫和其伴生菌所组成的共生体系,不同小蠹虫的种群数量,和不同环境条件下同种小蠹虫与其伴生菌相互作用方式的差异使我们在研究小蠹虫和其伴生菌这个共生体系时,对它们各自在成功聚集寄主树木过程中所发挥的重要作用的概括变得非常困难。     

Isolation and characterization of Beauveria spp. associated with exotic bark beetles in New Zealand Pinus radiata plantation forests

A survey of entomopathogenic fungi associated with Hylastes ater and Hylurgus ligniperda was undertaken in six Pinus radiata forest sites in New Zealand. In each site soil, bark and frass associated with beetle galleries were sampled, as well as a selection of live beetles, larvae and mycosed cadavers. The density of Beauveria spp. (as measured by colony forming units) varied both within and between the sites and substrates sampled. The highest CFU density detected was 4 × 10⁶ CFU/g dry frass, which was collected from a stump at the Mahurangi site in the North Island. A number of isolates were selected and characterized using PCR amplification of a terminal region of the EF1-α fragment. This analysis revealed that two Beauveria species, Beauveria caledonica and Beauveria bassiana, were commonly associated with bark beetle habitat. B. caledonica was the only species isolated from mycosed beetles, and was also recovered from soils, bark and frass, though not live insects. B. bassiana was also found in soils, bark, frass and live insects; however, it was not isolated from mycosed cadavers in this study. It is speculated that B. caledonica is the common ‘natural’ pathogen of bark beetles in their breeding habitat. A third species, Beauveria malawiensis, was isolated once from soil. This is the first report of this species from New Zealand. Thirteen isolates (selected from all substrates sampled and live beetles) were selected for laboratory bioassays. Under laboratory conditions all isolates were pathogenic to H. ater and H. ligniperda adults; there was little difference among the isolates in terms of efficacy despite the paucity of B. bassiana infection observed in the field. The mechanisms behind the lack of field infection observed for B. bassiana are not known, but are likely to be linked to limiting environmental factors in the breeding sites. These results indicate potential risks associated with the selection of strains for use in biological control programmes solely on the basis of laboratory bioassays, especially if the mode of action and biology of the species used are not fully understood. The importance of selecting strains which are environmentally competent is evident. Overall, however, this study demonstrates that entomopathogenic fungi can play a significant role in the regulation of H. ater and H. ligniperda populations in pine plantations.     

Species diversity estimation of ambrosia and bark beetles in temperate mixed forests in Japan based on host phylogeny and specificity

Understandings of the effect of host plant phylogeny on the structure of herbivore assemblage is useful for estimating global species richness of herbivores. Here we test the relationship between host plant phylogeny and two assemblages including ambrosia beetle assemblage that have been considered to be the lowest host specificity among plant-dependent guilds. These results of local scale were used for estimating regional species richness by extrapolating to the number of plant order and species in Japan. The estimated numbers were compared with the numbers of described species in Japan. Tree trunks of 17 plant species representing 17 orders of all major lineages of Japanese tree flora were exposed for collecting wood boring beetle species. A total of 12 ambrosia and four bark beetle species were collected. Similarity of both ambrosia and bark beetle assemblages showed a significant negative trend with phylogenetic distance between focal host plant species. The regression model for this relationship was well fit by a linear model whereas previous studies used a semi-log model, which suggests a difference in mechanism of host utilization with host taxonomic levels. Our results showed a broader host range of ambrosia beetle assemblage in temperate forest than to a comparable study in tropical rainforests. Species richness estimated is lower than the described species in Japan, suggesting the need for more samples along the altitudinal gradients for accurate estimation for the Japanese fauna.     

产于中国西南高山温带森林的二个棒瑚菌新种

本文根据形态学特征和分子系统学分析报道了产于中国西南山地温带森林的棒瑚菌属新种2个。第一个新种金黄棒瑚菌Clavariadelphus aurantiacus具有亮橙黄色的子实体和不育的子实体顶部,第二个新种纤细棒瑚菌C. tenuis具有细长的子实体并且菌丝不具锁状联合。为两个新种提供了详细的描述、绘制了黑白线条图、拍摄了彩色照片,并且与类似的物种进行了比较。运用核糖体转录区间序列分析了棒瑚菌属的系统发育关系。提供了中国已知棒瑚菌的检索表。     

Pathogenicity to western larch (Larix occidentalis) of two fungi, Ophiostoma pseudotsugae and Leptographium abietinum, associated with the Douglas fir beetle (Coleoptera: Scolytidae)

1 Pole-sized, live western larch Larix occidentalis Nutt. were mass-inoculated with Ophiostoma pseudotsugae (Romb.) von Arx or Leptographium abietinum (Peck) Wingf., two blue-stain fungi associated with the Douglas fir beetle Dendroctonus pseudotsugae Hopkins, to assess their pathogenicity. 2 Inoculation with O. pseudotsugae resulted in significantly greater percentages of necrotic phloem compared with L. abietinum inoculations. 3 The percentage of occluded sapwood was significantly greater after L. abietinum inoculations compared with O. pseudotsugae inoculations. 4 Within the inoculation band, all trees had more than 60% functional sapwood 4 months after treatment. 5 The results suggest that western larch can successfully limit colonization by O. pseudotsugae and L. abietinum. 6 The inability of the fungi to thrive in live western larch may be a factor in the consistent failure of Douglas fir beetle broods in this host tree species.     

Two new species of <Emphasis Type="Italic">Pyxidiophora</Emphasis> associated with bark beetles in Europe

Two new species of Pyxidiophora (Ascomycota, Laboulbeniomycetes, Pyxidiophorales) associated with bark beetles (Coleoptera, Scolytidae) colonising the sapwood and secondary phloem of conifers (Larix decidua and Picea abies) in Europe are described. Each of both fungal species appeared to be connected with a Gabarnaudia-like anamorph. Pyxidiophora corallisetosa could only be cultivated in the presence of the hyphomycete Clonostachys rosea, and a connection of P. corallisetosa with its Gabarnaudia-like anamorph was proven by the development of perithecia after axenic transfer of conidia to cultures of C. rosea. Pyxidiophora cuniculicola was found in cultures originating from bark beetles placed onto agar media. Conidia developing directly from germinating ascospores were identical to those developing from Gabarnaudia-like conidiophores also present in these cultures, which indicates an anamorph-teleomorph connection. The probable interactions between both species of Pyxidiophora with associated fungi are discussed.     

Evolutionary dynamics of introns and their open reading frames in the U7 region of the mitochondrial rnl gene in species of Ceratocystis

The mtDNA rnl-U7 region has been examined for the presence of introns in selected species of the genus Ceratocystis. Comparative sequence analysis identified group I and group II introns encoding single and double motif LAGLIDADG open reading frames (ORFs) at the following positions L1671, L1787, and L1923. In addition downstream of the rnl-U7 region group I introns were detected at positions L1971 and L2231, and a group II intron at L2059. A GIY-YIG type ORF was located within one mL1923 LAGLIDADG type ORF and a degenerated GIY-YIG ORF fused to a nad2 gene fragment was found in association with the mL1971 group I intron. The diversity of composite elements that appear to be sporadically distributed among closely related species of Ceratocystis illustrates the potential for homing endonucleases and their associated introns to invade new sites. Phylogenetic analysis showed that single motif LADGLIDADG ORFs related to the mL1923 ORFs have invaded the L1787 group II intron and the L1671 group I intron. Phylogenetic analysis of intron encoded single and double motif LAGLIDADG ORFs also showed that these ORFs transferred four times from group I into group II B1 type introns.     

Conservation value of forests attacked by bark beetles: Highest number of indicator species is found in early successional stages

Heavy natural disturbance in large protected areas of former commercial forests increasingly evokes European parliaments to call for management intervention because a loss of habitats and species is feared. In contrast, natural early successional habitats have recently been recognised as important for conservation. Current knowledge in this field mostly results from studies dealing only with selected taxa. Here we analyse the success of species across 24 lineages of three kingdoms in the Bavarian Forest National Park (Germany) after 15 years of a European spruce bark beetle (Ips typographus L.) outbreak that led to rapid canopy opening. Using indicator species analysis, we found 257 species with a significant preference for open forests and 149 species with a preference for closed forests, but only 82 species with a preference for the stand conditions transitional between open and closed forests. The large number of species with a preference for open forests across lineages supports the role of this bark beetle as a keystone species for a broad array of species. The slowdown of the outbreak after 15 years in the core zone of the national park resulted in less than half of the area being affected, due to variability in stand ages and tree species mixtures. Our case study is representative of the tree species composition and size of many large protected montane areas in Central European countries and illustrates that (1) natural disturbances increase biodiversity in formerly managed forests and (2) a montane protected area spanning 10,000 ha of low range mountains is likely sufficient to allow natural disturbances without a biased loss of closed-forest species.