Genetic differentiation in sympatric wood ants, <Emphasis Type="Italic">Formica rufa</Emphasis> and <Emphasis Type="Italic">F. polyctena</Emphasis>

Summary Direct observations have suggested that the closely related wood ants Formica polyctena and F. rufa represent different social organizations, with high queen number in F. polyctena and a high frequency of monogynous nests in F. rufa. We examined social organization and genetic population structure in a setup where populations of the two species are sympatric and gene flow between the species is possible. Our aim was to compare social organization in the species, and study evolutionary relationships between them. The observed relatedness among colony workers suggested that the difference in the level of polygyny is quantitative rather than qualitative, with a higher queen number in F. polyctena. The observed difference in polygyny was not accompanied by a difference in spatial genetic differentiation which was weak in both species. The genetic distance between the species is consistent with limited interspecific gene flow. Identification of a few possible F. rufa migrants in F. polyctena populations suggests potential interspecific gene flow. Thus, reproductive isolation of the species may not be complete when they are sympatric.Received 14 March 2003; revised 10 October 2003; accepted 20 October 2003.     

Independent hybrid populations of <Emphasis Type="Italic">Formica polyctena X</Emphasis><Emphasis Type="Italic">rufa</Emphasis> wood ants (Hymenoptera: Formicidae) abound under conditions of forest fragmentation

Combined genetic and morphological data indicate frequent hybridisation between the wood ants Formica polyctena Förster 1850 and F. rufa Linnaeus 1761 in Central Europe. The genetic and morphological traits give a concordant picture of hybridisation with a strong correlation between the genotypic admixture proportions at 19 microsatellite loci and the first vectors of a principal component analysis (P < 0.001) and of a 3-class discriminant analysis (P < 0.001) of 15 quantitative morphological characters. This integrative approach enabled a grouping into F. polyctena, the hybrid and rufa. Genetic differentiation between the hybrid and F. rufa is significantly larger than between the hybrid and polyctena, indicating gene flow mainly between the latter entities. A suggested gene flow bias towards F. polyctena agrees with differential queen acceptance and mating behaviour. Both genetic and phenotypic colony parameters indicate predominance of monogyny in F. rufa but of polygyny in polyctena and the hybrid. Hybrids are intermediate between the parental species in body size, diagnostic morphological characters, monogyny frequency, size of nest population, nest diameter and infestation rate with epizootic fungi. The three entities respond differently to woodland fragmentation. Hybrids are significantly more abundant in forests with a coherent area <300 ha than in woodland above this size. Regions with high hybrid frequency in Germany—the Eastern Oberlausitz (23%) and the Baltic Sea islands Darss, Hiddensee and Rügen (28%)—are characterised by a fragmented woodland structure whereas regions with low hybrid frequency—Brandenburg and the lower Erzgebirge (3.4%)—have clearly larger and more coherent forest systems. Data from other European countries indicate habitat fragmentation to be a facilitating factor but no essential precondition for interspecific hybridisation in these ants. Hybrids are hypothesised to have selective advantage in fragmented systems because of combining the main reproductive and dispersal strategies of the parental species.     

Long-term impact of agriculture on the survival of wood ants of the <Emphasis Type="Italic">Formica rufa</Emphasis> group (<Emphasis Type="Italic">Formicidae</Emphasis>)

The impact of agriculture on wood ants of the Formica rufa group was investigated in a small-scale agricultural landscape with many woodland fragments in the east of the Netherlands. An inventory of nests was carried out in 1986, and repeated in 2014. The number of nests of F. rufa and F. polyctena had drastically decreased during the intervening 28 years, especially in small patches of woodland bordering a maize field. The species F. rufa, which mainly disperses by flight of the queen, seems to be better adapted to habitat fragmentation than F. polyctena, where dispersal is mainly by colony splitting. However, the flying strategy can only be successful in a landscape where most woodland fragments have a good quality habitat; in the study area, the habitat quality had deteriorated. This is ascribed to the development of a tall, nitrophilous vegetation, a result of considerable nitrogen deposition from the atmosphere and the spraying of manure. The regional survival of wood ants in the east of the Netherlands is at risk in the agricultural landscapes we studied. This is due to the increasing probability of colonies becoming extinct and the decreased probability of a queen colonizing habitat patches. However, the negative influence of agriculture on the edge vegetation of patches of woodland can be decreased by leaving a wide strip between the arable land and the woodland that receives no manure.     

Nest-building in ants <Emphasis Type="Italic">Formica exsecta</Emphasis> (Hymenoptera,Formicidae)

Representatives of the subgenus Coptoformica build composite aboveground nests in much the same manner as red wood ants do. However, despite the superficial similarity, Coptoformica nests show a number of substantial differences that impact on the way in which ants use the nest. The aboveground part of a Formica (Coptoformica) exsecta nest consists of the temporary layer, the outer crust layer, the mound, and the intermediate layer. The material of all the layers includes soil and fragments of plants (picked up or excised), the relative proportion of the two components differing among the layers. The thick, soil-rich crust layer acts as armature and allows a variety of anthill shapes. The intermediate layer and the upper part of the mound harbor quite a few large chambers. Building material is often transferred from nest to nest. On the whole, F. exsecta nests are plastic and movable, which facilitates the activity of these ants in ecotone biotopes.     

The dynamics of vegetation on and around <Emphasis Type="Italic">Formica rufa</Emphasis> nests

The species composition and structure of plant communities related to the activity of Formica rufa ants were studied in green moss pine forests located in Nerusso-Desnyanskoe Polesye (Bryansk Province). Four types of microsites were established and examined: (1) the mound of an active ant nest, (2) the soil bank around an active ant nest, (3) the mound of an abandoned nest, and (4) the soil bank around an abandoned nest. The background plant community was considered as the fifth type. According to MRPP method, all pairs of microsites except (3–4) were different both in plant species composition and species abundance. The microsites created by Formica rufa increase the habitat capacity and β diversity of the plant communities due to the appearance of meadow species requiring richer soils.     

<Emphasis Type="Italic">Mycobacterium avium</Emphasis> disease in wild red-legged partridges (<Emphasis Type="Italic">Alectoris rufa</Emphasis>)

Three wild red-legged partridges (Alectoris rufa) from intensively managed hunting areas in Spain were received for necropsy. They showed granulomatous lesions in different parts of the body, mainly in liver and spleen. Microscope examination of the granulomas showed central caseous necrosis and large amounts of acid-fast bacilli, surrounded by epitheloid cells, giant cells, and lymphocytes. Attempts to isolate and culture the bacillus in Colestsos medium were unsuccessful, but the polymerase chain reaction technique revealed the presence of microorganisms belonging to the Mycobacterium avium complex in one of the partridges. This is the first report of avian tuberculosis in free-living red-legged partridges.     

Preadaptation of <Emphasis Type="Italic">Formica aquilonia</Emphasis> and <Emphasis Type="Italic">Formica lugubris</Emphasis> ants (Hymenoptera,Formicidae) to low wintering temperatures in Northeast Asia

The ants Formica aquilonia and F. lugubris which inhabit the entire forest zone of the North Palaearctic and are absent from the basins of the Yana, Indigirka, and Kolyma rivers were found in the coastal area of the Sea of Okhotsk. A possible climatic conditionality of their occurrence in the Northeast is considered based on the data on the biotopic distribution of ants, the temperature causation of their overwintering, and cold hardiness. On the Sea of Okhotsk coast, these ants overwinter at a depth of 40–200 cm in the soil. During winter, the minimum soil temperature at a depth of 40 cm under the anthill was ?5°C. The supercooling temperature of F. aquilonia was not lower than ?20.2 ± 0.5°C, that of F. lugubris, not lower than ?19.6 ± 0.4°C. Half of F. aquilonia individuals did not survive the daily exposure at ?13°C, F. lugubris, at ?16°C. These two cold-resistant species could inhabit some biotopes of the Kolyma River basin, similar to F. exsecta, F. lemani, and F. sanguinea, but they are absent there for some reasons that are not related to the temperature. A similar cold hardiness is characteristic of F. aquilonia in Estonia (Maavara, 1971, 1985), where it represents a side effect of diapause, since excessive cold hardiness has no adaptive significance for insects overwintering in the non-freezing soils of Estonia. Colonization of Siberia by ant species turned out to be possible only due to the existing cold hardiness, i.e. preadaptation to low temperature. On the Sea of Okhotsk coast, cold hardiness of the ants is non-adaptive due to the relatively mild conditions during winter.     

Phenotypic and phylogenetic characterization of actinomycetes isolated from <Emphasis Type="Italic">Lasius niger</Emphasis> and <Emphasis Type="Italic">Formica cunicularia</Emphasis> ants

Multiple actinomycete strains were isolated from two ant species, Lasius niger and Formica cunicularia, and their phenotypic properties and phylogenetic position were studied. Partial sequencing of 16S rRNA assigned the greater part of them to the genus Streptomyces, but only one belonged to Nocardia. However, some isolates had significant color and morphological differences from their closest phylogenetic relatives. The abundance and biodiversity of actinomycete communities isolated from L. niger ants greatly exceeded those found for F. cunicularia. All of the actinomycetes associated with F. cunicularia ants demonstrated cellulolytic activity, but only one had such ability among the strains associated with black ants.     

New combinations and typifications in <Emphasis Type="Italic">Bistorta</Emphasis>, <Emphasis Type="Italic">Persicaria</Emphasis>, <Emphasis Type="Italic">Polygonum,</Emphasis> and <Emphasis Type="Italic">Rumex</Emphasis> (Polygonaceae)

New combinations are proposed in anticipation of the Polygonaceae treatment in the forthcoming volume of Intermountain Flora: Polygonum kelloggii var. esotericum, P. kelloggii var. watsonii , Rumex densiflorus var. pycnanthus , R. salicifolius var. utahensis, and R. occidentalis var. tomentellus. Typifications are proposed to facilitate ongoing studies in Polygonaceae and to maintain current usage.     

<Emphasis Type="Italic">Ty</Emphasis>1<Emphasis Type="Italic">/copia</Emphasis>- and <Emphasis Type="Italic">Ty</Emphasis>3<Emphasis Type="Italic">/gypsy</Emphasis>-like DNA sequences in <Emphasis Type="Italic">Helianthus </Emphasis>species

Two repeated DNA sequences isolated from a partial genomic DNA library of Helianthus annuus, p HaS13 and p HaS211, were shown to represent portions of the int gene of a Ty3 /gypsy retroelement and of the RNase-Hgene of a Ty1 /copia retroelement, respectively. Southern blotting patterns obtained by hybridizing the two probes to BglII- or DraI-digested genomic DNA from different Helianthus species showed p HaS13 and p HaS211 were parts of dispersed repeats at least 8 and 7 kb in length, respectively, that were conserved in all species studied. Comparable hybridization patterns were obtained in all species with p HaS13. By contrast, the patterns obtained by hybridizing p HaS211 clearly differentiated annual species from perennials. The frequencies of p HaS13- and p HaS211-related sequences in different species were 4.3x10(4)-1.3x10(5) copies and 9.9x10(2)-8.1x10(3) copies per picogram of DNA, respectively. The frequency of p HaS13-related sequences varied widely within annual species, while no significant difference was observed among perennial species. Conversely, the frequency variation of p HaS211-related sequences was as large within annual species as within perennials. Sequences of both families were found to be dispersed along the length of all chromosomes in all species studied. However, Ty3 /gypsy-like sequences were localized preferentially at the centromeric regions, whereas Ty1/ copia-like sequences were less represented or absent around the centromeres and plentiful at the chromosome ends. These findings suggest that the two sequence families played a role in Helianthusgenome evolution and species divergence, evolved independently in the same genomic backgrounds and in annual or perennial species, and acquired different possible functions in the host genomes.     

Pedogamy in <Emphasis Type="Italic">Neidium</Emphasis> (<Emphasis Type="Italic">Bacillariophyceae</Emphasis>)

Single (unpaired) vegetative cells of freshwater pennate diatom Neidium cf. ampliatum differentiated into gametangia and produced a single zygote (auxospore) via a pedogamic process. The gametic nuclei fused after auxospore expansion had begun. The auxospore expanded in parallel to the apical axis of the gametangium.     

<Emphasis Type="Italic">In silico</Emphasis> and <Emphasis Type="Italic">in situ</Emphasis> characterization of the zebrafish (<Emphasis Type="Italic">Danio rerio</Emphasis>) <Emphasis Type="Italic">gnrh3</Emphasis> (sGnRH) gene

Background

Gonadotropin releasing hormone (GnRH) is responsible for stimulation of gonadotropic hormone (GtH) in the hypothalamus-pituitary-gonadal axis (HPG). The regulatory mechanisms responsible for brain specificity make the promoter attractive for in silico analysis and reporter gene studies in zebrafish (Danio rerio).

Results

We have characterized a zebrafish [Trp⁷, Leu⁸] or salmon (s) GnRH variant, gnrh 3. The gene includes a 1.6 Kb upstream regulatory region and displays the conserved structure of 4 exons and 3 introns, as seen in other species. An in silico defined enhancer at -976 in the zebrafish promoter, containing adjacent binding sites for Oct-1, CREB and Sp1, was predicted in 2 mammalian and 5 teleost GnRH promoters. Reporter gene studies confirmed the importance of this enhancer for cell specific expression in zebrafish. Interestingly the promoter of human GnRH-I, known as mammalian GnRH (mGnRH), was shown capable of driving cell specific reporter gene expression in transgenic zebrafish.

Conclusions

The characterized zebrafish Gnrh3 decapeptide exhibits complete homology to the Atlantic salmon (Salmo salar) GnRH-III variant. In silico analysis of mammalian and teleost GnRH promoters revealed a conserved enhancer possessing binding sites for Oct-1, CREB and Sp1. Transgenic and transient reporter gene expression in zebrafish larvae, confirmed the importance of the in silico defined zebrafish enhancer at -976. The capability of the human GnRH-I promoter of directing cell specific reporter gene expression in zebrafish supports orthology between GnRH-I and GnRH-III.

     

<Emphasis Type="Italic">Quinua</Emphasis> and Relatives (<Emphasis Type="Italic">Chenopodium</Emphasis> sect.<Emphasis Type="Italic">Chenopodium</Emphasis> subsect.<Emphasis Type="Italic">Celluloid</Emphasis>)

Traditionally viewed as an Andean grain crop,Chenopodium quinoa Willd. includes domesticated populations that are not Andean, and Andean populations that are not domesticated. Comparative analysis of leaf morphology and allozyme frequencies have demonstrated that Andean populations, both domesticated(quinua) and free-living(ajara), represent an exceptionally homogeneous unit that is well differentiated from allied domesticates of coastal Chile(quingua) and freeliving populations of the Argentine lowlands(C. hircinum). This pattern of relationships indicates that Andean populations represent a monophyletic crop/weed system that has possibly developed through cyclic differentiation (natural vs. human selection) and introgressive hybridization. Relative levels of variation suggest that this complex originated in the southern Andes, possibly from wild types allied withC. hircinum, with subsequent dispersal north to Colombia and south to the Chilean coast. Coastal populations were apparently isolated from post-dispersal differentiation and homogenization that occurred in the Andes. Other data point toward a center of origin in the northern Andes with secondary centers of genetic diversity subsequently developing in the southern Andes and the plains of Argentina. Comparative linkage of South American taxa, all tetraploid, with North American tetraploids of the subsection will eventually clarify this problem. While the possibility of a direct phyletic connection betweenC. quinoa and the Mexican domesticate(C. berlandieri subsp. nuttalliae,) cannot be excluded, available evidence indicates that the latter represents an autonomous lineage that is associated with the basal tetraploid, C. b. subsp.berlandieri, through var.sinuatum, whereas South American taxa show possible affinities to either var. zschackei or var.berlandieri. An extinct domesticate of eastern North America,C. b. subsp.jonesianum, represents either another instance of independent domestication, possibly from subsp. b. var.zschackei, or a northeastern outlier of subsp.nuttalliae.     

The Madagascan genus <Emphasis Type="Italic">Vaughania</Emphasis> is reduced to synonymy under <Emphasis Type="Italic">Indigofera</Emphasis> (<Emphasis Type="Italic">Leguminosae-Papilionoideae-Indigofereae</Emphasis>)

Summary  Eleven species comprising the Madagascan genus Vaughania are subsumed within the large pantropical genus Indigofera. Six new combinations are made; the remaining species were originally described in Indigofera.     

Floral development and systematic position of <Emphasis Type="Italic">Arillastrum</Emphasis>, <Emphasis Type="Italic">Allosyncarpia</Emphasis>, <Emphasis Type="Italic">Stockwellia</Emphasis> and <Emphasis Type="Italic">Eucalyptopsis</Emphasis> (Myrtaceae)

We have investigated the floral ontogeny of Arillastrum, Allosyncarpia, Stockwellia and Eucalyptopsis (of the eucalypt group, Myrtaceae) using scanning electron microscopy and light microscopy. Several critical characters for establishing relationships between these genera and to the eucalypts have been determined. The absence of compound petaline primordia in Arillastrum, Allosyncarpia, Stockwellia and Eucalyptopsis excludes these taxa from the eucalypt clade. Post-anthesis circumscissile abscission of the hypanthium above the ovary in Stockwellia, Eucalyptopsis and Allosyncarpia is evidence that these three taxa form a monophyletic group; undifferentiated perianth parts and elongated fusiform buds are characters that unite Stockwellia and Eucalyptopsis as sister taxa. No floral characters clearly associate Arillastrum with either the eucalypt clade or the clade of Stockwellia, Eucalyptopsis and Allosyncarpia.We gratefully acknowledge Clyde Dunlop and Bob Harwood (Northern Territory Herbarium) for collecting specimens of Allosyncarpia, and Bruce Gray (Atherton) for collecting specimens of Stockwellia. The Australian National Herbarium (CANB) kindly lent herbarium specimens of Eucalyptopsis for examination. This research was supported by a University of Melbourne Research Development Grant to Andrew Drinnan.     

<Emphasis Type="Italic">Galleria</Emphasis><Emphasis Type="Italic">mellonella</Emphasis> are Resistant to <Emphasis Type="Italic">Pneumocystis</Emphasis><Emphasis Type="Italic">murina</Emphasis> Infection

Studying Pneumocystis has proven to be a challenge from the perspective of propagating a significant amount of the pathogen in a facile manner. The study of several fungal pathogens has been aided by the use of invertebrate model hosts. Our efforts to infect the invertebrate larvae Galleria mellonella with Pneumocystis proved futile since P. murina neither caused disease nor was able to proliferate within G. mellonella. It did, however, show that the pathogen could be rapidly cleared from the host.     

Revision of the <Emphasis Type="Italic">Serrulati</Emphasis> species of <Emphasis Type="Italic">Penstemon</Emphasis> section <Emphasis Type="Italic">Saccanthera</Emphasis> subsection <Emphasis Type="Italic">Serrulati</Emphasis>

A revision of Penstemon sect. Saccanthera subsect. Serrulati includes a new species (P. salmonensis), a new variety (P. triphyllus var. infernalis), and the elevation of a subspecies to species (P. curtiflorus), bringing the total number of species to eight, which are keyed and described, complete with nomenclature and type citations.     

<Emphasis Type="Italic">Agrobacterium</Emphasis><Emphasis Type="Italic">tumefaciens</Emphasis>-mediated transformation of callus cells of <Emphasis Type="Italic">Crataegus</Emphasis><Emphasis Type="Italic">aronia</Emphasis>

A genetic transformation system has been developed for callus cells of Crataegus aronia using Agrobacterium tumefaciens. Callus culture was established from internodal stem segments incubated on Murashige and Skoog (MS) medium supplemented with 5 mg l⁻¹ Indole-3-butyric acid (IBA) and 0.5 mg l⁻¹ 6-benzyladenine (BA). In order to optimize the callus culture system with respect to callus growth and coloration, different types and concentrations of plant growth regulators were tested. Results indicated that the best average fresh weight of red colored callus was obtained on MS medium supplemented with 2 mg l⁻¹ 2,4-dichlorophenoxyacetic acid (2,4-D) and 1.5 mg l⁻¹ kinetin (Kin) (callus maintenance medium). Callus cells were co-cultivated with Agrobacterium harboring the binary plasmid pCAMBIA1302 carrying the mgfp5 and hygromycin phosphotransferase (hptII) genes conferring green fluorescent protein (GFP) activity and hygromycin resistance, respectively. Putative transgenic calli were obtained 4 weeks after incubation of the co-cultivated explants onto maintenance medium supplemented with 50 mg l⁻¹ hygromycin. Molecular analysis confirmed the integration of the transgenes in transformed callus. To our knowledge, this is the first time to report an Agrobacterium-mediated transformation system in Crataegus aronia.