Sequence data reflect the introduction pathways of the Sirex woodwasp parasitoid,Ibalia leucospoides (Ibaliidae,Hymenoptera)

1. The parasitoid wasp Ibalia leucospoides is native to the northern hemisphere and has been introduced to the southern hemisphere as a biological control agent for the invasive woodwasp Sirex noctilio. Two subspecies of the parasitoid, Ibalia leucospoides leucospoides (Palearctic distribution) and Ibalia leucospoides ensiger (Nearctic distribution), were introduced and are reported to have hybridized.
2. Despite extensive records of the numbers and origins of the wasps imported into the southern hemisphere, nothing is known regarding their current population diversity. We investigated the genetic variation of I. leucospoides in its native and introduced ranges using mitochondrial (COI) and nuclear (ITS) markers.
3. Mitochondrial DNA diversity in the introduced range was limited, with only five haplotypes, although sequence divergence between these haplotypes was high. Similarly, the ITS rDNA sequences revealed multiple clades present in the introduced range.
4. These results reflect introductions from a wide geographical range but where genetic bottlenecks have possibly reduced the genetic diversity. The data further reflect the origin of the I. leucospoides populations in South America and South Africa from New Zealand or Australia. We found no evidence of hybridization between the two subspecies of the parasitoid in its introduced range, and no evidence that I. leucospoides ensiger has established outside its native range.

     

Phylogeny and historical biogeography of the cynipoid wasp family Ibaliidae (Hymenoptera)

The Ibaliidae are a small family of cynipoid wasps, the members of which parasitize woodboring siricid larvae in hardwoods and conifers. The 19 currently recognized extant species occur mainly in the Northern Hemisphere. No fossils are known despite the presumed old age of the family. We present a cladistic analysis of ibaliid relationships at the species-level, mainly based on external skeletal characters of adults. The results indicate that the three genera ( Eileenella , Heteribalia , Ibalia ) and two subgenera of Ibalia ( Ibalia s. str. and Tremibalia ) recognized in the current classification are monophyletic. Three different categories of characters were compared for their phylogenetic usefulness. Homoplasy was found to be lowest for main structures, higher for sculptural characters, and still higher for colour differences. The historical biogeography of the family was reconstructed using dispersal–vicariance analysis in combination with palaeogeographical data. The results suggest that the family primarily diversified within the eastern Palaearctic–northern Oriental region. The nominate subgenus of Ibalia dispersed early to the western Nearctic, where it radiated; two species later spread throughout the Holarctic. The other subgenus of Ibalia shows an early eastern Palaearctic–eastern Nearctic disjunction which presumably dates back to the Eocene–Oligocene transition.     

Evaluation of the insect parasitoids of Sirex noctilio (Hymenoptera: Siricidae) in Tasmania

Summary Sampling of Sirex noctilio F. and the insect parasitoids released in a forest of Pinus radiata D. Don near Hobart, Tasmania, has shown that the rhyssines, Megarhyssa nortoni (Cresson) and Rhyssa persuasoria (L.) were mainly responsible for reducing the level of the population of S. noctilio between 1965 and 1974. Parasitic nematodes were not present during this period. Key factor analysis indicates that in this forest the two species act as one delayed density-dependent factor, whereas Ibalia leucospoides Hochenw. is apparently density-independent, its ability to parasitize the host being regulated by conditions within the tree.     

Erysiphe kenjiana (Erysiphales), a new invasive fungus in Europe

An Asian powdery mildew fungus Erysiphe (Uncinula) kenjiana (Erysiphales, Ascomycota) has been found in Ukraine. This is the first record of this fungus in Europe. In 2007, E. kenjiana was collected on four Ulmus species in Kiev. All locations adjoined railways or an airport. Development of E. kenjiana was epiphytotic. This species was not found on elms surveyed at towns situated north-east, east or south of Kiev. The fungus may have been brought directly to Ukraine by rail or air transport. In 2008, the fungus was also collected in Chernihiv situated north-east of Kiev. It is likely that E. kenjiana will spread over all Ukraine and into countries of central and western Europe in 2009 or later. Molecular phylogenetic analyses using D1/D2 domains of the 28S rDNA and ITS sequences revealed that the Ulmaceae-Cannabaceae-parasitic powdery mildews, including E. kenjiana, form a clade with strong supports, suggesting that these species diverged from a single ancestor and expanded their host ranges within the Ulmaceae and allied Cannabaceae. This hypothesis is supported by these species sharing the unique morphology of enlarged apices on their chasmothecial appendages. These fungi formed part of a larger grouping with species on Fagaceae, Nothofagaceae, Rosaceae, and Sapindaceae with strong statistical supports. These results suggest that Uncinula-like powdery mildew fungi on these plant families exhibit close evolutionary relationships with their hosts.     

Digestive capacities of leaf-cutting ants and the contribution of their fungal cultivar to the degradation of plant material

Leaf-cutting ants (tribe Attini) are a unique group of ants that cultivate a fungus that serves as a main source of their food. The fungus is grown on fresh leaves that are harvested by workers. We examine the respective contribution of ants and their symbiotic fungus in the degradation of plant material by examining the digestive capacities of seven Attini species in the genera Atta and Acromyrmex. The results show that both, the ants and their mutualistic fungi, have complementary enzymatic activities. Ants are specialized in the degradation of low molecular weight substrates (oligosaccharides and heterosides) whereas the fungus displays high polysaccharidase activity. The two genera Atta and Acromyrmex are not distinguished by a specific enzymatic activity. The seven different mutualistic associations examined display a similar enzymatic profile but have quantitative differences in substrate degradation activities. The respective contribution of ants and the fungus garden in plant degradation are discussed.     

Identification of the ambrosia fungus of <Emphasis Type="Italic">Xyleborus monographus</Emphasis> and <Emphasis Type="Italic">X. dryographus</Emphasis> (Coleoptera: Curculionidae,Scolytinae)

The scolytid ambrosia beetles Xyleborus monographus and X. dryographus were investigated to identify their nutritional ambrosia fungi. The examination of the oral mycetangia of the beetles, the specialized organs for fungal transport, revealed the dominant occurrence of Raffaelea montetyi, a fungus that was also predominant in the beetle tunnels in the immediate vicinity of the feeding larvae. R. montetyi was previously known only as the ambrosia fungus of the platypodid ambrosia beetle, Platypus cylindrus. These beetle species inhabit the same habitat, mainly trunks of oaks in the Western Palaeartic. The possibility of an exchange of the symbiotic fungus between the ambrosia beetles within their common breeding place is discussed. Consequently, the previous hypothesis of a species-specific association of a single ambrosia fungus with a single beetle species is questioned. A phylogenetic analysis based on DNA sequences classified R. montetyi within the Ophiostomatales of the ascomycetes. The investigation of conidiogenesis of R. montetyi by SEM supported this taxonomic placement and showed the development of the conidia by annellidic percurrent proliferation, identical to the conidiogenesis reported for many anamorph states of the Ophiostomatales.     

Fungus Symbionts Colonizing the Galleries of the Ambrosia Beetle <Emphasis Type="Italic">Platypus quercivorus</Emphasis>

Isolations were made to determine the fungal symbionts colonizing Platypus quercivorus beetle galleries of dead or dying Quercus laurifolia, Castanopsis cuspidata, Quercus serrata, Quercus crispula, and Quercus robur. For these studies, logs from oak wilt-killed trees were collected from Kyoto Prefecture, Japan. Fungi were isolated from the: (1) entrances of beetle galleries, (2) vertical galleries, (3) lateral galleries, and (4) the larval cradle of P. quercivorus in each host tree. Among the fungus colonies which appeared on YM agar plates, 1,219 were isolated as the representative isolates for fungus species inhabiting in the galleries based on their cultural characteristics. The validity of the visual classification of the fungus colonies was checked and if necessary properly corrected using microsatellite-primed PCR fingerprints. The nucleotide sequence of the D1/D2 region of the large subunit nuclear rRNA gene detected 38 fungus species (104 strains) of which three species, i.e., Candida sp. 3, Candida kashinagacola (both yeasts), and the filamentous fungus Raffaelea quercivora were isolated from all the tree species. The two yeasts were most prevalent in the interior of galleries, regardless of host tree species, suggesting their close association with the beetle. A culture-independent method, terminal restriction fragment length polymorphism (T-RFLP) analysis was also used to characterize the fungus flora of beetle galleries. T-RFLP patterns showed that yeast species belonging to the genus Ambrosiozyma frequently occurred on the gallery walls along with the two Candida species. Ours is the first report showing the specific fungi inhabiting the galleries of a platypodid ambrosia beetle.     

Diversity of fungus‐growing termites (Macrotermes) and their fungal symbionts (Termitomyces) in the semiarid Tsavo Ecosystem,Kenya

Fungus‐growing termites of the subfamily Macrotermitinae together with their highly specialized fungal symbionts (Termitomyces) are primary decomposers of dead plant matter in many African savanna ecosystems. The termites provide crucial ecosystem services also by modifying soil properties, translocating nutrients, and as important drivers of plant succession. Despite their obvious ecological importance, many basic features in the biology of fungus‐growing termites and especially their fungal symbionts remain poorly known, and no studies have so far focused on possible habitat‐level differences in symbiont diversity across heterogeneous landscapes. We studied the species identities of Macrotermes termites and their Termitomyces symbionts by excavating 143 termite mounds at eight study sites in the semiarid Tsavo Ecosystem of southern Kenya. Reference specimens were identified by sequencing the COI region from termites and the ITS region from symbiotic fungi. The results demonstrate that the regional Macrotermes community in Tsavo includes two sympatric species (M. subhyalinus and M. michaelseni) which cultivate and largely share three species of Termitomyces symbionts. A single species of fungus is always found in each termite mound, but even closely adjacent colonies of the same termite species often house evolutionarily divergent fungi. The species identities of both partners vary markedly between sites, suggesting hitherto unknown differences in their ecological requirements. It is apparent that both habitat heterogeneity and disturbance history can influence the regional distribution patterns of both partners in symbiosis.     

Beneath the bark: associations among Sirex noctilio development,bluestain fungi,and pine host species in North America

1. Invasive species with global distributions encounter unique environmental and biotic variables that can greatly affect the magnitude of their impact. The European woodwasp, Sirex noctilio Fabricius, is a prime example that has invaded climatically and ecologically distinct ecosystems across the Southern Hemisphere and, more recently, North America. 2. Northeastern North America presents a unique set of conditions, including pine host species, native parasitoids, a diverse assemblage of native co‐colonising insects, and fungal associates of these co‐colonisers. In North America, S. noctilio attacks both a naturalised ancestral host (Pinus sylvestris L.) from Europe and a naïve native host (P. resinosa). A large assemblage of insects and their associated bluestaining ophiostomatoid fungi colonise these pines. Competition between S. noctilio and this group is a hypothesised mechanism of biotic resistance in the invaded region of North America, possibly via superior resource capture abilities or alteration of host tissue by bluestain fungi. 3. Investigating these ecological interactions is challenging because they manifest deep in the xylem tissue. To overcome this, 30 experimentally stressed trees were systematically dissected with an electric log splitter to investigate the effects of bluestain fungi and tree factors on S. noctilio development and parasitism by native hymenopterans. 4. Body size and colonisation density were affected by pine species, with S. noctilio being 25% larger and densities three‐fold greater in P. sylvestris than P. resinosa. Survivorship was slightly negatively correlated with the proportional volume of bluestain infection. Interestingly, rhyssine (Rhyssa and Megarhyssa spp.) parasitism responded positively to greater S. noctilio density, but there was no density relationship with Ibalia leucospoides ensiger parasitism. Pine host species appears to play a strong role in S. noctilio development, which is important considering uninvaded regions of North America have a diversity of pine species that likely vary in their susceptibility to this woodborer.     

Halosarpheia japonica sp. nov. (Halosphaeriales, Ascomycota) from marine habitats in Japan

A new Halosarpheia species, collected from driftwood from Hakkeijima beach, Yokohama, Japan, is described and illustrated and is compared with other species of the genus. The new fungus was growing together with its anamorph on a piece of decaying wood. SSU and LSU rDNA sequences for both morphs were 99% similar. Phylogenetic analyses of SSU and LSU rDNA sequences of the both morphs confirm their anamorph–teleomorph relationship and placed the new fungus in the Halosarpheia sensu stricto clade with high statistical support. Halosarpheia japonica is characterized by its polar appendage that is initially enclosed in a cellular sheath and dissolves in water, the appendage then swells to form a huge tree-like structure. The other three species currently included in Halosarpheia sensu stricto differ from H. japonica by having two polar appendages that uncoil in water to form long filaments.     

ORCHID MYCORRHIZA: VITAMIN PRODUCTION AND REQUIREMENTS BY THE SYMBIONTS

A Rhizoctonia species isolated from Cymbidium has been cultured successfully on a defined medium consisting of minerals, sugar, thiamine, and folic acid. Thiamine can be replaced by its thiazole component, which is probably produced by germinating orchids. The fungus apparently produces the pyrimidine moiety of thiamine, a compound which may enhance growth of certain orchid seedlings. Niacin is also provided by the fungus. Para-aminobenzoic acid, a constituent of folic acid, produced and released by orchid seeds, can satisfy the vitamin requirement of the fungus. These findings point to the possibility that orchids and their fungi may have coevolved with respect to vitamin requirements. The data also suggest that exchanges of vitamins or their components between orchids and endophytes are important aspects of the symbiotic relationship.     

A new species of Melampsora on Populus yunnanensis from China

Melampsora nujiangensis, a rust fungus found on Populus yunnanensis in China, is described as a new species. Light and scanning electron microscopy with herbarium specimens of the rust fungus show that the shape of its urediniospores differs from that of other known species of Melampsora, and its urediniospore walls are thinner than the other species. Furthermore, in phylogenetic trees based on the DNA sequences (28S and ITS) the rust fungus is phylogenetically separated by high bootstrap values. These results indicate that the fungus is an isolated species among the genus Melampsora. Contribution no. 202 from the Laboratory of Plant Parasitic Mycology, Graduate School of Life and Environmental Sciences, University of Tsukuba, Japan     

Host Range of Mycoleptodiscus terrestris, a Microbial Herbicide Candidate for Eurasian Watermilfoil, Myriophyllum spicatum

A native strain of the fungal plant pathogen Mycoleptodiscus terrestris is capable of causing under experimental conditions necrotic shoot lesions and a generalized decline and disintegration of Eurasian watermilfoil (Myriophyllum spicatum), an exotic submerged aquatic weed in North America. The potential of this fungus to cause disease on nontarget plants was evaluated on 33 species and cultivars in 11 families. The nontarget selection, intended as a Tier I group, was biased toward probable suscepts consisting of submerged, floating, and emergent aquatic species and terrestrial crop plants. The plants were exposed to the fungal mycelium formulated in alginate beads. Pathogenicity rather than phytotoxicity was the primary mode of attack by the fungus. Of the 16 nontarget aquatic species tested, the fungus was pathogenic to Hydrilla verticillata (hydrilla), Myriophyllum aquaticum (parrotfeather), and Ceratophyllum demersum (coontail), but only in hydrilla did it cause plant mortality comparable to that in Eurasian watermilfoil. The remaining 13 submerged, floating, and emergent aquatic species were unaffected. Of the 17 terrestrial species screened, none was significantly affected by the fungus in germination studies, but postemergent disease symptoms developed on seedlings of 10 species. Four of the 10, Medicago sativa (alfalfa), Lotus corniculatus (birdsfoot trefoil), Trifolium hybridum (alyce clover), and T. repens (white clover), developed disease affecting 26-50% of their tissues. Thus, this host range testing scheme, based on the concept of screening plants under the maximum hazard potential, helped to identify susceptible species. Nonetheless, it exaggerated the nontarget risk due to the small number of host species tested. Screening a taxonomically diverse and larger selection of plants as well as testing under conditions of less severe exposure to the fungal inoculum are necessary to obtain a more realistic view of the host range than presently indicated.     

Effect of IAA on symbiotic germination of an Australian orchid and its production by orchid-associated bacteria

Seven isolates of orchid-associated bacteria (OAB) belonging to five species were tested for their effect on mycorrhiza-assisted germination of the terrestrial orchid Pterostylis vittata. Hormone standards were also tested to evaluate their potential roles in the germination and development of the orchid. Strains of Pseudomonas putida, Xanthomonas maltophilia and Bacillus cereus promoted symbiotic germination, whereas certain strains of P. putida and an Arthrobacter species reduced it. Symbiotic germination was enhanced by IAA, inhibited by gibberellic acid and suppressed by kinetin. Each species of OAB produced IAA, although the conditions of growth affected the production of the auxin. IAA was not produced by the mycorrhizal fungus from P. vittata under the test conditions. Enhancement of symbiotic germination development may have resulted either from the production of IAA by the OAB and/or by the induction of endogenous hormones in the orchid by the metabolites of the bacteria and/or mycorrhizal fungus.     

Phylogeny and evolution of the cryptic fungus‐farming ant genus Myrmicocrypta F. Smith (Hymenoptera: Formicidae) inferred from multilocus data

Fungus‐farming ants (Hymenoptera: Formicidae) have become model systems for exploring questions regarding the evolution of symbiosis. However, robust phylogenetic studies of both the ant agriculturalists and their fungal cultivars are necessary for addressing whether or not observed ant–fungus associations are the result of coevolution and, if so, whether that coevolution has been strict or diffuse. Here we focus on the evolutionary relationships of the species within the ant genus Myrmicocrypta and of their fungal cultivars. The fungus‐farming ant genus Myrmicocrypta was created by Fr. Smith in 1860 based on a single alate queen. Since then, 31 species and subspecies have been described. Until now, the genus has not received any taxonomic treatment and the relationships of the species within the genus have not been tested. Our molecular analyses, using ～40 putative species and six protein‐coding (nuclear and mitochondrial) gene fragments, recover Myrmicocrypta as monophyletic and as the sister group of the genus Mycocepurus Forel. The species M. tuberculata Weber is recovered as the sister to the rest of Myrmicocrypta. The time‐calibrated phylogeny recovers the age of stem group Myrmicocrypta plus its sister group as 45 Ma, whereas the inferred age for the crown group Myrmicocrypta is recovered as 27 Ma. Ancestral character‐state analyses suggest that the ancestor of Myrmicocrypta had scale‐like or squamate hairs and that, although such hairs were once considered diagnostic for the genus, the alternative state of erect simple hairs has evolved at least seven independent times. Ancestral‐state analyses of observed fungal cultivar associations suggest that the most recent common ancestor of Myrmicocrypta cultivated clade 2 fungal species and that switches to clade 1 fungi have occurred at least five times. It is our hope that these results will encourage additional species‐level phylogenies of fungus‐farming ants and their fungal cultivars, which are necessary for understanding the evolutionary processes that gave rise to agriculture in ants and that produced the current diversity of mutualistic ant–fungus interactions.     

Monocentric fungi on species of rhizosolenia from saline habitats

Summary Eight monocentric fungi are reported in two provisionally identified species ofRhizosolenia from sea-water. Only two species are assigned specific names. These areRhizophydium planktonicum Canter andR. fragilariae Canter. A fungus with depressed-globose sporangia is included as a form ofR. fragilariae. Two unidentified species ofChytridium are reported. A fungus tentatively assigned toZygorhizidium and a form provisionally assigned toPetersenia are also reported and described insofar as their characteristics are known.     

A new acidophilic fungus Teratosphaeria acidotherma (Capnodiales, Ascomycota) from a hot spring

A novel acidophilic fungus was isolated by an acidic enrichment culture of microbial mats and biofilms collected at an extremely acidic and high temperature hot spring. In culture studies, this fungus was revealed to produce ascomycetous teleomorph structures. Molecular phylogenetic study and morphological observation showed this fungus is a new species of the genus Teratosphaeria (Capnodiales, Dothideomycetes) and is phylogenetically close to Acidomyces acidophilus and Bispora sp., which were previously reported as acidophilic anamorphic fungi. This new fungus is described here as a new species of Teratosphaeria, and its physiological properties adapting to its habitat are demonstrated. This is the first report of a teleomorphic fungus having highly acidophilic and thermophilic properties.     

First record and characterization of a powdery mildew on a member of the Juncaginaceae: <Emphasis Type="Italic">Leveillula taurica</Emphasis> on <Emphasis Type="Italic">Triglochin maritima</Emphasis>

The powdery mildew fungus Leveillula taurica (Erysiphales) is reported for the first time from the monocot Triglochin maritima (Juncaginaceae), a widespread salt marsh plant that causes economic losses because of its high toxicity to young livestock. This is the first report of an erysiphaceous fungus on a member of the Juncaginaceae. Morphological data, obtained by light and scanning electron microscopy, and ITS sequence data provided evidence that this fungus is referable to L. taurica. The ITS sequence for this fungus was identical with those reported for L. taurica hosted by Capsicum annuum in Australia and Elaeagnus angustifolia in Iran. This is the third host species and second monocot, in addition to Allium cepa and Solanum tuberosum, reported for L. taurica from Washington State, where the fungus was unreported before 2004.     

Interactions among unrelated species: Granivorous rodents,a parasitic fungus,and a shared prey species

Summary Granivorous rodents and a parasitic fungus in the Sonoran Desert utilize a common prey species, Erodium cicutarium, a desert annual plant. Experimental removal of rodents from field exclosures resulted in significantly higher densities of E. cicutarium. Fungal infection was significantly higher in the absence of rodents, suggesting that, while they do not interact directly, rodents and the fungus affect each other's densities by their use of a common prey species.