Horizontal transmission of a parasitic nematode from a non-native to a native woodwasp?

In eastern North America, the exotic invasive woodwasp, Sirex noctilio, attacks pines (Pinus spp.) and often shares larval habitat with the native woodwasp, Sirex nigricornis. The parasitic nematode, Deladenus siricidicola, has been used widely in the southern hemisphere as a biological control agent because it sterilizes female S. noctilio. This nematode was introduced accidentally to North America along with S. noctilio. Historical reports indicate nematode-woodwasp fidelity: the parasitic nematode, D. siricidicola, exclusively infects S. noctilio, and the native nematode, Deladenus proximus, exclusively infects S. nigricornis. From two sites in southern Ontario, separated by 225 km, we collected woodwasps from three Pinus sylvestris, and identified the nematode species present in the abdomens of infected wasps. Both wasp species co-occurred in all three trees. D. siricidicola was present in the haemocoel, but not inside the eggs, of infected S. noctilio and S. nigricornis. This evidence suggests horizontal transmission of D. siricidicola likely occurred from S. noctilio to S. nigricornis.     

Biotic resistance and the spatiotemporal distribution of an invading woodwasp, <Emphasis Type="Italic">Sirex noctilio</Emphasis>

Quantifying the strength of interactions among introduced and native species across space and time is critical in understanding biological invasions as they can attenuate or amplify the magnitude of impact. The European woodwasp, Sirex noctilio F., a global threat to pines, is a recent invader to North America. It attacks and kills primarily Pinus resinosa and Pinus sylvestris, and encounters a diverse assemblage of potential competitors for this resource. We quantified spatial colonization patterns of this woodwasp and resident competitors including scolytine bark beetles, woodboring cerambycid and buprestid beetles, and the fungal root rot diseases Armillaria and Heterobasidion across an 80 year old pine plantation over 4 years. All xylophages were spatially aggregated, but only on a localized scale of 15–20 m. Colonizers occurred non-randomly within trees, with S. noctilio negatively or neutrally associated with all other colonizing agents, whereas all other insect and root rot colonizers were mostly positively co-associated. An autologistic regression with spatially-weighted variables indicated the probability of a dead tree exhibiting symptoms of S. noctilio attack was positively associated with tree density, host species (P. sylvestris), and density of S. noctilio-attacked trees from the current and previous year. Interspecific stand patterns were weaker; probability of attack was negatively associated only with root rot pathogens. Across spatial scales, there were mixed (woodborers) and neutral (bark beetles) associations between S. noctilio and other co-colonizing insects. Our results suggest that competitive interactions with resident species may be contributing to the limited success of S. noctilio in North America, but are unlikely to be driving it by themselves.     

Influence of Nematode Parasitism,Body Size,Temperature, and Diel Period on the Flight Capacity of <Emphasis Type="Italic">Sirex noctilio</Emphasis> F. (Hymenoptera: Siricidae)

Sirex noctilio F. (Hymenoptera: Siricidae) is a woodwasp of pine trees that has recently invaded and established in North American forests. Although S. noctilio has had a limited impact in North America to date, there is some concern that it could have a significant impact on pine plantations, especially in the southeastern U.S.A. Moreover, there are few data on the flight capacity of male S. noctilio. We found no association between parasitism by D. siricidicola and whether or not S. noctilio initiated flight on the flight mill. Male wasps that were parasitized by nematodes were heavier than non-parasitized males, but there was no significant difference in mass between parasitized and non-parasitized females. We also examined the flight capacity of male and female S. noctilio in relation to nematode parasitism, body mass, temperature (for only males), and diel period. Body mass, temperature, and diel period affected flight in S. noctilio such that wasps were generally observed to fly faster, farther, and more frequently if they were heavier, flying at warmer temperatures, and flying during the photoperiod. The fact that nematode-parasitized male wasps were found to fly farther than the non-parasitized males is consistent with the hypothesis that nematode parasitism does not negatively affect the flight capacity of S. noctilio.     

The fungal matrices of <Emphasis Type="Italic">Ophiostoma ips</Emphasis> hinder movement of the biocontrol nematode agent, <Emphasis Type="Italic">Deladenus siricidicola</Emphasis>, disrupting management of the woodwasp, <Emphasis Type="Italic">Sirex noctilio</Emphasis>

Deladenus (=?Beddingia) siricidicola (Tylenchida: Neotylenchidae) is the most effective biocontrol agent used against the invasive wood wasp, Sirex noctilio (Fabricius) (Hymenoptera: Siricidae). The nematodes feed and reproduce on the wood-inhabiting fungus, Amylostereum areolatum (Chaillet ex Fr.) Boidin (Russulales: Amylostereaceae) and parasitise larvae of S. noctilio. In the nematode biocontrol program, the nematodes are inoculated into herbicide-weakened ‘trap trees’. Recent declines in nematode parasitism of S. noctilio in Australia have coincided with an increased incidence of an exotic bark beetle, Ips grandicollis (Eichhoff) (Coleoptera: Curculionidae), attacking trap trees and vectoring a wood-inhabiting fungus, Ophiostoma ips (Rumbold) Nannfelt (Ophiostomatales: Ophiostomataceae), which may inhibit migration of the nematode within the tree to the detriment of S. noctilio biocontrol. Several in vitro and in vivo experiments were conducted to investigate the effect of fungal interactions on the ability of D. siricidicola to locate and reproduce on A. areolatum. Deladenus siricidicola showed preference to A. areolatum in the presence and absence of O. ips, but the presence of O. ips negatively affected the choice response and the number of eggs laid by the nematodes. Deladenus siricidicola was unable to survive and reproduce on O. ips. Results give a clearer understanding of the choice response of D. siricidicola in I. grandicollis infested trees, explaining the disruptive impact of bark beetles on biocontrol of S. noctilio, an effect that could extend from Australia to other important pine growing countries.     

Genetic diversity and population structure identify the potential source of the invasive red clover casebearer moth, <Emphasis Type="Italic">Coleophora deauratella</Emphasis>, in North America

The red clover casebearer, Coleophora deauratella, is an invasive pest of red clover grown for seed in North America. In 2006, an outbreak in Alberta, Canada was discovered that resulted in significant seed losses, while further invasion threatens the world’s largest red clover forage seed production region in Oregon, USA. Prior to the recent outbreak, C. deauratella was thought to be restricted to eastern North America in its invasive range. We sequenced a 615-bp fragment of the mitochondrial cytochrome c oxidase subunit 1 gene, and developed three microsatellite markers to assess the genetic diversity and population structure of C. deauratella in North America and its native range in Europe. We observed signatures of a founder effect in North American populations and a further loss of genetic diversity within Alberta populations. Most genetic differentiation was found between continents, with no evidence of isolation-by-distance within each continent. From the limited number of European populations sampled, a single introduction from Switzerland is the most probable source of North American populations based on similar mitochondrial diversity and lack of population differentiation. Within North America, based on increased genetic diversity compared to the rest of the continent, the first North American record from Ithaca, NY, and the first documented outbreak in southern Ontario in 1989, the initial C. deauratella invasion most likely occurred in southern Ontario, Canada or adjacent states in the USA, followed by transport throughout the continent. This study provides insight into the phylogeographic history of C. deauratella in North America and Europe and may help to identify a regional source of future classical biological control agents.     

Trapping for Sirex Woodwasp in Brazilian Pine Plantations: Lure,Trap Type and Height of Deployment

The Sirex woodwasp, Sirex noctilio (Hymenoptera: Siricidae) is considered a secondary pest of pine in its native range but has caused considerable economic losses in pine plantation forests in the southern hemisphere. In Brazil, trap trees are the primary tool used for early detection purposes but these are costly, labor-intensive to install and require stressing trees by herbicide application. Flight intercept traps baited with synthetic blends of host volatiles are an attractive alternative but have performed poorly in some settings. This study was carried out to look for alternatives to trap trees for use in Brazilian pine plantations for early detection of S. noctilio. Four field experiments were conducted in two consecutive flight seasons (2015–16 and 2016–17), in planted loblolly pine (Pinus taeda) stands, to compare captures among flight intercept traps baited with different lures, deployed at different heights and among different intercept trap designs. Two experiments compared different host volatile lures and a significant treatment effect was observed in one. No effect of trap design or height was observed.     

Genetic divergence of sympatric charrs of the genus <Emphasis Type="Italic">Salvelinus</Emphasis> from Nachikinskoe Lake (Kamchatka)

We studied genetic differentiation of two charr species, Dolly Varden Salvelinus malma malma Walbaum and resident lacustrine charr Salvelinus sp., which sympatrically inhabit Nachikinskoe Lake (the Bol’shaya River basin) in southwestern Kamchatka Peninsula. Using restriction analysis (RFLP), three mitochondrial DNA fragments (ND1/ND2, ND5/ND6, and Cyt b/D-loop) amplified in polymerase chain reaction (PCR) were compared. The divergence of the mtDNA sequences between Salvelinus sp. and S. malma malma was 2.8%; Salvelinus sp. and S. taranetzi, 0.36%; Salvelinus sp. and S. krogiusae, 0.21%; Salvelinus sp. and S. alpinus, 3.0%. These results point to reproductive isolation of charrs in Nachikinskoe Lake and support the earlier suggestion on a close relationship between Salvelinus sp., S. taranetzi, and S. krogiusae.     

Evidence of interaction between Sirex noctilio and other species inhabiting the bole of Pinus

• 1 Sirex noctilio is a woodwasp native to Eurasia and Northern Africa and has recently been found infesting pines in eastern North America. Its pest status in this new range is not yet known, although it is an important pest in other areas where it has been introduced. Pinus spp. in North America are hosts to several native and alien species of subcortical insects. Interactions between the woodwasp and these species may influence its distribution or the characteristics of its life history, thus affecting its population dynamics over time.
• 2 Sixty S. noctilio‐infested Pinus spp. were felled in Ontario, Canada, and all phloem‐feeding and woodboring insects were collected and identified from each 1‐m section of the tree.
• 3 Sirex noctilio was in a tree alone 10% of the time but commonly shared the tree with subcortical beetles, such as Tomicus piniperda, Pissodes nemorensis, Ips grandicollis, Gnathotrichus materiarius and Monochamus carolinensis. The woodwasp was distributed throughout the tree stem and this distribution overlapped with that of the beetles.
• 4 Fewer but larger S. noctilio males emerged from trees with beetles compared with those without, although there was no statistical difference in females.
• 5 These findings suggest that co‐habiting beetles could negatively affect S. noctilio population dynamics over time. The potential mechanisms for this interaction are discussed.

     

Evaluation and comparison of the genetic structure of <Emphasis Type="Italic">Bunias orientalis</Emphasis> populations in their native range and two non-native ranges

We studied the invasive warty cabbage Bunias orientalis (Brassicaceae) in three geographically distinct areas. Using inter-simple sequence repeat fingerprinting, we analyzed warty cabbages, including non-native populations, from the eastern Baltic and western Siberian regions and native populations from southwestern Russia. The eastern Baltic region and western Siberia represent the two opposite directions of B. orientalis spread in climatically different zones. The genetic structures of the native and non-native B. orientalis populations were assessed through analysis of molecular variance (AMOVA) and the Bayesian clustering method and by determining the main measures of genetic diversity. AMOVA revealed considerable population differentiation in both the native and invasive ranges. Our results did not indicate a decrease in genetic diversity in the non-native populations of B. orientalis. Similar measures of genetic diversity and genetic structure were determined in the invasive populations in two geographically and ecologically distinct, non-native regions located in Europe and Asia. In both of these regions, higher genetic diversity was detected in the non-native populations than in the native region populations, which may be due to multiple introductions. However, Bayesian clustering analysis revealed slightly different sources of invasive populations in the two non-native regions. Genetic diversity patterns revealed the lack of isolation by distance between populations and confirmed the influence of anthropogenic factors on the spread of B. orientalis. The significance of native populations as germplasm resources for breeding is discussed.     

Characterization and phylogenetic analysis of the complete mitogenome of a rare cavefish, <Emphasis Type="Italic">Sinocyclocheilus multipunctatus</Emphasis> (Cypriniformes: Cyprinidae)

The genus Sinocyclocheilus is a representative group of cave creatures. However, genetic studies on Sinocyclocheilus are rare. The primary objective of this study was to explore the structure and feature of the complete mitochondrial genome of S. multipunctatus, and reconstruct the mitogenomic phylogeny of Sinocyclocheilus. The mitochondrial DNA of S. multipunctatus was amplified by overlapping PCR fragments. The mitogenome was assembled by the SeqMan and annotated using MitoAnnotator. The phylogenetic tree was established using the Bayesian inference and Maximum likelihood methods. The mitogenome of S. multipunctatus is a typical circular molecule of 16,586 bp with base composition A (31.25%), T (25.90%), G (16.35%), and C (26.50%), and consists of 13 protein-coding genes (PCGs), 22 transfer RNAs (tRNAs) genes, two ribosomal RNAs, and a 931 bp control region. Phylogenetic analysis reveals two clades in the Sinocyclocheilus with robust support. S. multipunctatus is close to a newly discovered cavefish, S. ronganensis. We obtained and described the complete mitogenome of S. multipunctatus, and investigated its phylogenetic status, which may provide a valuable resource for future phylogenetic analyses and population genetic studies in Sinocyclocheilus.     

Genetic analysis of Japanese and American specimens of <Emphasis Type="Italic">Scirpus hattorianus</Emphasis> suggests its introduction from North America

Scirpus hattorianus is a possible alien species in Japan, and a clarification of its unclear taxonomy is required to reveal its origin. It is not known whether the plants initially described from Japan represent the same species distributed in North America. To clarify the origin of the species, we attempted to sequence old specimens collected about 80 years ago using newly designed primer pairs specific for short sequences, including the variable sites. Chloroplast sequences of ndhF were compared among Japanese and North American S. hattorianus, and the closely related species, S. atrovirens, S. flaccidifolius, and S. georgianus. We succeeded in sequencing all samples, and two haplotypes were detected in S. hattorianus: one was unique to the species and the other, detected from specimens potentially collected from the same population as the types, was shared by both North American S. hattorianus and two closely related species, S. atrovirens and S. flaccidifolius. Our results suggest that Japanese S. hattorianus is an alien species that was introduced from North America at least twice.     

Pelagic larval dispersal habits influence the population genetic structure of clam <Emphasis Type="Italic">Gomphina aequilatera</Emphasis> in China

Pelagic larval dispersal habits influence the population genetic structure of marine mollusk organisms via gene flow. The genetic information of the clam Gomphina aequilatera (short larval stage, 10 days) which is ecologically and economically important in the China coast is unknown. To determine the influence of planktonic larval duration on the genetic structure of G. aequilatera. Mitochondrial markers, cytochrome oxidase subunit i (COI) and 12S ribosomal RNA (12S rRNA), were used to investigate the population structure of wild G. aequilatera specimens from four China Sea coastal locations (Zhoushan, Nanji Island, Zhangpu and Beihai). Partial COI (685 bp) and 12S rRNA (350 bp) sequences were determined. High level and significant F_ST values were obtained among the different localities, based on either COI (F_ST?=?0.100–0.444, P?<?0.05) or 12S rRNA (F_ST?=?0.193–0.742, P?<?0.05), indicating a high degree of genetic differentiation among the populations. The pairwise N_m between Beihai and Zhoushan for COI was 0.626 and the other four pairwise N_m values were >?1, indicating extensive gene flow among them. The 12S rRNA showed the same pattern. AMOVA test results for COI and 12S rRNA indicated major genetic variation within the populations: 77.96% within and 22.04% among the populations for COI, 55.73% within and 44.27% among the populations for 12S rRNA. A median-joining network suggested obvious genetic differentiation between the Zhoushan and Beihai populations. This study revealed the extant population genetic structure of G. aequilatera and showed a strong population structure in a species with a short planktonic larval stage.     

Molecular genetic characterization of the yeast <Emphasis Type="Italic">Lachancea kluyveri</Emphasis>

A comparative study of Lachancea kluyveri strains isolated in Europe, North America, Japan, and the Russian Far East was performed using restriction analysis, sequencing of non-coding rDNA regions, molecular karyotyping, and the phylogenetic analysis of the α-galactosidase MEL genes. This study showed a close genetic relatedness of these L. kluyveri strains. The chromosomal DNAs of the L. kluyveri strains were found to range in size from 980 to 3100 kb. The haploid number of chromosomes is equal to eight. The IGS2 restriction patterns and single nucleotide substitutions in the ITS1/ITS2 rDNA region correlate neither with geographic origin nor with the source of the strains. The L. kluyveri strains isolated from different sources have a high degree of homology (79–100%) of their MEL genes. The phylogenetic analysis of all of the known α-galactosidases in the “Saccharomyces” clade showed that the MEL genes of the yeasts L. kluyveri, L. cidri, Saccharomyces cerevisiae, S. paradoxus, S. bayanus, and S. mikatae are species specific.     

Finding loci associated to partial resistance to white pine blister rust in sugar pine (<Emphasis Type="Italic">Pinus lambertiana</Emphasis> Dougl.).

White pine blister rust (WPBR) is an exotic disease threatening five-needle pines in North America. In spite of its relatively recent introduction, some five-needle pines such as sugar pine (Pinus lambertiana) have developed both complete (major) gene resistance and partial (quantitative) resistance to WPBR. While significant effort has been dedicated to clone and locate the position of the major gene of WPBR resistance in sugar pine, the genetic basis of quantitative resistance remains largely unknown in all Strobus pines. In this work, we took a preliminary approach to identify potential genotype × phenotype associations using the results of long-term survival and symptoms of infection in both experimental and applied breeding populations. Our study found significant associations between several genes and WPBR disease symptoms such as normal active cankers and blights, important symptoms in the development of partial resistance. No significant associations were found with percentage of survival, probably due to the complex inheritance of the disease and long time to infection. With this study, we hope to lay the ground for further genome-wide association studies using large phenotypic data sets in sugar pine and other Strobus pines.     

Molecular systematics of the critically-endangered North American spinymussels (Unionidae: <Emphasis Type="Italic">Elliptio</Emphasis> and <Emphasis Type="Italic">Pleurobema</Emphasis>) and description of <Emphasis Type="Italic">Parvaspina</Emphasis> gen. nov.

Despite being common in numerous marine bivalve lineages, lateral spines are extremely rare among freshwater bivalves (Bivalvia: Unionidae), with only three known species characterized by the presence of spines: Elliptio spinosa, Elliptio steinstansana, and Pleurobema collina. All three taxa are endemic to the Atlantic Slope of southeastern North America, critically endangered, and protected by the US Endangered Species Act. Currently, these species are recognized in two genera and remain a source of considerable taxonomic confusion. Because spines are rare in freshwater mussels and restricted to a small region of North America, we hypothesized that spinymussels represent a monophyletic group. We sequenced two mtDNA gene fragments (COI and ND1) and a fragment of the nuclear ITS-1 locus from >70 specimens. Bayesian and maximum-likelihood phylogenetic reconstructions suggest that the spinymussels do not comprise a monophyletic group. Elliptio steinstansana is sister to P. collina, forming a monophyletic clade that was estimated to have diverged from its most recent ancestor in the late Miocene and is distinct from both Elliptio and Pleurobema; we describe a new genus (Parvaspina gen. nov.) to reflect this relationship. Additionally, E. spinosa forms a monophyletic clade that diverged from members of the core Elliptio lineage in the mid-Pliocene. Furthermore, E. spinosa is genetically divergent from the other spinymussel species, suggesting that spines, while extremely rare in freshwater mussels worldwide, may have evolved independently in two bivalve lineages. Recognizing the genetic distinctiveness and inter-generic relationships of the spinymussels is an important first step towards effectively managing these imperiled species and lays the groundwork for future conservation genetics studies.     

<Emphasis Type="Italic">Spartina versicolor</Emphasis> Fabre: Another case of <Emphasis Type="Italic">Spartina</Emphasis> trans-Atlantic introduction?

Intercontinental introductions are widespread in the genus Spartina, with important ecological and evolutionary consequences. The native or introduced status of Spartina species is then critical with regard to biodiversity assessment, especially for vulnerable Mediterranean coastline ecosystems. Spartina versicolor was first recorded in southern France in 1849, then successively in various places on the European and North-African Mediterranean and Atlantic coasts. This species is considered to be either a European native or an invasive species introduced from North America which has a high morphological similarity to the Atlantic American species Spartina patens. We performed extensive sampling of S. versicolor in Europe and North Africa (from natural populations and herbarium collections) and compared these samples to other European and American Spartina species (including S. patens). Chromosome counts were reported for the first time and revealed that S. versicolor is tetraploid (2n = 4x = 40). Phylogenetic analyses based on chloroplast and nuclear ribosomal DNA sequences did not reveal any molecular variation within S. versicolor. In this species, a single haplotype, that is identical to one haplotype of S. patens, was found in the four chloroplast and the nuclear ribosomal ITS regions investigated. In addition, simple sequence repeat markers were used and revealed a low level of genetic diversity within S. versicolor, suggesting that the introduction of S. versicolor occurred from a narrow genetic pool of S. patens from North America.     

Chromosome numbers of some North American Salix

Chromosome numbers are reported for 19 species of North AmericanSalix, one natural hybrid, and one introduced species. The following 17 species are here examined cytologically for the first time:Salix amygdaloides Anderss.,S. arbusculoides Anderss.,S. brachycarpa Nutt.,S. Candida Willd.,S. discolor Muhl.,S. exigua Nutt.,S. kumilis Marsh.,S. interior Rowlee,S. lutea Nutt.,S. maccalliana Rowlee,S. monticola Rydb.,S. myrtillifolia Anderss.,S. pellita Anderss.,S. petiolaris J. E. Sm,S. scouleriana Barr.,S. silicicola Raup, andS. subcoerulea Piper. Of the 19 native North American species 11 are diploid, four are tetraploid, one is triploid, one hexaploid, one dodecaploid, and one exhibits more than one ploidal level.     

Molecular phylogeny of pectinase genes <Emphasis Type="Italic">PGU</Emphasis> in the yeast genus <Emphasis Type="Italic">Saccharomyces</Emphasis>

Using yeast genome databases and literature data, phylogenetic analysis of pectinase PGU genes from 112 Saccharomyces strains assigned to the biological species S. arboricola, S. bayanus (var. uvarum), S. cariocanus, S. cerevisiae, S. kudriavzevii, S. mikatae, S. paradoxus, and the hybrid taxon S. pastorianus (syn. S. carlsbergensis) was carried out. A superfamily of divergent PGU genes was found. Natural interspecies transfer of the PGU gene both from S. cerevisiae to S. bayanus and from S. paradoxus to S. cerevisiae may, however, occur. Within the Saccharomyces species, identity of the PGU nucleotide sequences was 98.8–100% for S. cerevisiae, 86.1–95.7% for S. bayanus (var. uvarum), 94–98.3% for S. kudriavzevii, and 96.8–100% for S. paradoxus/S. cariocanus. For the first time, a family of polymeric PGU1b, PGU2b, PGU3b and PGU4b genes is documented for the yeast S. bayanus var. uvarum, a variety important for winemaking.     

Genetic structure of Sakhalin spruce (<Emphasis Type="Italic">Picea glehnii</Emphasis>) in northern Japan and adjacent regions revealed by nuclear microsatellites and mitochondrial gene sequences

The genetic structure of Sakhalin spruce (Picea glehnii) was studied across the natural range of the species, including two small isolated populations in south Sakhalin and Hayachine, by using six microsatellite loci and maternally inherited mitochondrial gene sequences. We also analyzed P. jezoensis, a sympatric spruce in the range. Genetic diversity of P. glehnii was higher in central Hokkaido and the lowest in the Hayachine. Bayesian clustering and principal coordinate analysis by using the microsatellites indicated that the Hayachine was clearly distinct from other populations, implying that it had undergone strong genetic drift since the last glacial period. P. glehnii harbored four mitochondrial haplotypes, two of which were shared with P. jezoensis. One of the two was observed without geographical concentration, suggesting its derivation from ancestral polymorphism. Another was observed in south Sakhalin and in P. jezoensis across Sakhalin. The Bayesian clustering—by using four microsatellite loci, including P. jezoensis populations—indicated unambiguous species delimitation, but with possible admixture of P. jezoensis genes into P. glehnii in south Sakhalin, where P. glehnii is abundantly overwhelmed by P. jezoensis; this might explain the occurrence of introgression of the haplotype of P. jezoensis into P. glehnii.