The pollen beetle (Meligethes aeneus) is one of the most important insect pests of oilseed rape (Brassica napus), and extensive use of insecticides is required to protect crop yields. To meet the challenges set by agricultural demands for more sustainable production and changing climate more information about pest biology and population genetics is needed. Using genomic Amplified Fragment Length Polymorphism (AFLP) analysis, DNA polymorphism was studied in 14 field populations of pollen beetles, collected during 2004 in six European countries (Denmark, France, Finland, Germany, Sweden, and UK). Using one primer combination 410 polymorphic DNA fragments were obtained based on analysis of single beetles. AFLP profiles were analysed with similarity measures using the Nei and Li coefficient and dendrograms generated. Dendrograms constructed from distance matrices revealed clustering by population origin and assignment analysis generally supported the genotype classification. Principal component analysis of the fourteen groups resulted in wide dispersion but also connections between some groups. Statistical analysis using AMOVA showed that the levels of genetic variation within populations explained most of the variation. Migrant analysis suggested a low level of gene flow between pollen beetle populations at different geographical locations indicating little long range dispersal of pollen beetles. However, a Mantel test found no correlation between genetic and geographical distance. Apparently genetic differentiation among populations has a complex background and may involve factors such as local adaptation and founder effects.
Calopteryx splendens is a widely distributed palaearctic damselfly with a remarkably uniform morphology. Variation in the size and shape of the pigmented spot on the wing is the main diagnostic character used to discriminate subspecies across its huge geographic range. Here, AFLP analysis was used to assess the genetic structure and diversity of nine populations representing 3 putative subspecies and evaluate the pigment spot as a taxonomic marker. Genetic diversity was high, with the number of polymorphic loci per population ranging from 141 to 280 out of a total of 333 variable sites (42.3-84.1%) and Nei's gene diversity from 0.160 to 0.283 (overall 0.299). Overall population genetic differentiation (FST = 0.2766) suggests limited gene flow and adaptation to local environments. Restricted gene flow and genetic differentiation among populations are supported by significant FST estimates. High levels of gene flow (Nm >1) were only recorded among three Asian populations (Russia - Kazakhstan - Turkey). The patterns of genotypic diversity suggest that a given wing spot size and shape may arise from the hybridization of a limited number, possibly not more than four, ancestral gene pools in different ways and at different times. Clearly, the sample analyzed was not sufficient to capture all of the complex history of C. splendens, but sufficient to indicate the taxa ancilla, waterstoni, and orientalis possibly represent three of the four ancestral gene pools, and originated in western Asia. The origin of the fourth, xanthostoma, is the western Mediterranean.
Genetic variability within and among fragmented populations of Artemisia pancicii was investigated in order to obtain a general understanding of the genetic structure related to the successful protection of this highly endangered species. Genetic variation within and among 15 populations of A. pancicii in Central Europe was analysed using amplified fragment length polymorphism (AFLP) and sequencing of two chloroplast DNA regions. The resulting polymorphism of AFLP loci was interpreted using basic population genetic indices and statistical visualisation. The total genetic variability within the populations was high (Ht = 0.248) and a highly differentiated population pattern (Fst = 0.241) was revealed. An analysis of molecular variance (AMOVA) revealed high variation among the populations (82%). There was no significant correlation between the genetic and geographic distance matrices. This indicates that population relatedness is not reflected in their geography. This was also confirmed by cpDNA sequencing. Highly restricted gene flow among the populations and genetic drift has resulted in reduced genetic variability in the smaller and highly differentiated A. pancicii populations, and very probably implies the presence of self-incompatibility and prevalence of clonal reproduction. The conservation of genetic variability in A. pancicii requires the persistence of large and also of small populations (because of population differentiation). The most important factor for the preservation of this species in the localities studied is the application of appropriate conservation management (such as mowing, grazing or fire management).
This paper focuses on the morphology, taxonomy and ecology of the widespread cyanobacteria of the genus Nodularia Mertens ex Bornet & Flahault. In this study the benthic strain of N. sphaerocarpa, isolated from a sand-pit near Olomouc (Czech Republic), is compared with brackish and seawater strains. Changes in morphology and growth parameters (biomass and chlorophyll a) recorded in varying salinity gradients were studied and a 16S rRNA sequencing and AFLP analysis conducted. Morphological and ecophysiological characteristics found were in congruence with molecular data. Three major subgroups of the benthic Nodularia (N. sphaerocarpa, N. moravica and N. harveyana) were found using the polyphasic approach. The results of both the molecular and morphological study clearly separated N. moravica and N. sphaerocarpa, as freshwater species preferring a low salinity and the N. harveyana strains originating from a marine environment preferring a high salinity.
Ptomascopus morio of both sexes are attracted to vertebrate carcasses, a necessary resource for reproduction. The stage during reproduction that resource defense was most intense and the hypothesis that large beetles were better competitors and sired a larger share of the offspring were supported and tested. Male-male aggression (pushing, biting and mounting) was commonly observed before and during oviposition, but rarely after the larvae hatched. Few female-female aggressive interactions were observed at any time. Parentage analysis of the offspring of six groups of two males and two females each reproducing on a separate carcass revealed that the large males sired more of the offspring than small males. Paternity analysis, using AFLP markers, revealed that larger males had higher paternity than smaller males, but the number of eggs produced by each female did not differ between large and small females. This suggests that competition among males is intense until the end of oviposition and that resident (large) males can acquire more mates and sire more offspring than smaller males; competition among females was not evident at any time.
The phylogeographic pattern of the temperate shrub Lonicera nigra (Caprifoliaceae) in Europe was inferred from molecular and fossil data. Population samples and pollen data from most of the contemporary natural distribution were analysed. While chloroplast DNA sequences revealed no intraspecific variation, AFLP data show a non-random geographic pattern. Two genetically different groups, distinguished by Bayesian clustering, divided the distribution area of L. nigra into south-western and north-eastern regions with a contact zone situated approximately in the upper part of the Danube Valley. Iberian populations constitute an additional distinct genetic group. Pollen evidence supports the genetic data, indicating that L. nigra might have survived in glacial refugia located in Central Europe. Nevertheless, this evidence should be considered only as indicative and supplementary, as an unambiguous determination of the species is not possible based on the information on pollen in the literature.
Cicerbita alpina was selected to elucidate the phylogeography of tall-herb species, an ecological group whose Quaternary history is rarely addressed. This species is a typical component of subalpine herbaceous communities in the mountains of Europe. Samples collected for this study comprised the entire range of species, with a focus on those in the Carpathians. The analysis based on AFLP fingerprinting revealed a lack of a strong phylogeographical structure implying that the different parts of the present-day range have not been isolated for a long period of time probably due to the biological characteristics of the species, such as its ability to disperse over great distances. However, the genetic structure indicates some phylogeographical trends, which may reflect traces of survival in local refugia and subsequent diversification into separate lineages during the last glacial period. Within the Carpathians, the division into the Western and South-Eastern Carpathian population groups is apparent. This division is maintained at a larger scale. In particular, the South-Eastern Carpathian group is similar to the Balkan populations, while the Western Carpathian populations are closely related to those in the Eastern Alps and Sudetes. The Scandinavian populations also have a genetic affinity with the latter group and originated from a source in the Eastern Alps or Western Carpathians, presumably via a stepping stone in a northern refugium.
Pollen viability was analysed causally between and within Central European Cirsium species and their hybrids to determine (i) how frequently hybrids are fertile and produce viable pollen; (ii) how the pollen viability of hybrids and their parents are related and how this is affected by the genetic distance between parents; (iii) how species promiscuity relates to species pollen viability; (iv) to what extent the pollen viability of a hybrid may predetermine its frequency in nature; (v) how the pollen viability of a hybrid and sympatricity of its parental species are related; and (vii) how the frequency of females in populations of gynodioecious species may affect the observed pollen viability. Altogether, the viability of 656,363 pollen grains was analysed using Alexander’s staining (1185 flowers from 301 plants from 67 field populations of 13 pure species and 1693 flowers from 345 plants from 96 field populations of 16 natural hybrids). The particular characters potentially related with pollen viability were estimated using following methods: natural hybrid frequency and species interfertility (by herbarium data), genetic distance (by AFLP), sympatricity (in local scale based on herbaria and literature data; on a global scale using the similarity between digitized maps of natural ranges). The strengths of pre- or postzygotic isolation were estimated for hybridizing species pairs using geographical data and pollen viability analyses. All hermaphrodite plants of the Cirsium hybrids had viable pollen, generally at lower levels than those found in pure species. The pollen viability of a hybrid generally decreased with increasing genetic distance between the parents and when the parental species had lower pollen viability. The pollen viability was decreased in frequently hybridizing species where occasionally individuals of pure species morphology may show decreased pollen viability. In some instances these might represent some unrecognized hybrid backcrosses. In populations of gynodioecious species where females co-occurred, pollen viability (in hermaphrodites)was also lower, indicating some degree of inbreeding depression. Hybrids between sympatric species exhibited higher post-pollination isolation (decrease of pollen viability), which suggests that the reproductive isolation had been increased by natural selection (effect similar to the Wallace effect). The strength of the postzygotic barrier (based on pollen viability) was generally stronger than that of the prezygotic barrier (based on distribution overlap) in studied hybridizing species pairs.
The recognition of hybrids of linear-leaved taxa of Potamogeton (sect. Graminifolii;) based on morphology is difficult and often debatable. As a consequence, currently only a few hybrid taxa are considered valid and many linear-leaved hybrids described in the past are not now recognized. On the other hand, the use of molecular tools has recently allowed more efficient tests of the origin of morphological forms and the tracking of hybridization events in Potamogeton systematics. In this paper, Potamogeton ×maëmetsiae Zalewska-Gałosz et M. Ronikier nothosp. nov. (Potamogetonaceae), a hybrid between two linear-leaved species, P. friesii and P. rutilus, is described and illustrated. Hybrid plants were collected from two Central-European populations growing in Lake Skaidrys (Lithuania) and Soitsjärv (Estonia). The hybrid origin of the new entity was identified based on a morphological survey and independently confirmed using nuclear (ITS, 5S-NTS) and chloroplast (rpl32-trnL intergenic spacer) DNA sequence data and AFLP analysis of genetic structure. Differences between P. ×maëmetsiae and similar taxa are outlined and other relevant details of the new hybrid discussed.
The pattern of postglacial re-colonization of Europe and the present population structure are known for various plant and animal species. The reed beetle Macroplea mutica (Fabricius, 1792) has characteristics that should influence both aspects in a peculiar way and therefore complement the currently known scenarios: It is fully aquatic but cannot swim or fly. Samples from 25 European populations of M. mutica and five specimens from China were investigated using amplified fragment length polymorphism (AFLP, 251 loci). Assessment of error rates associated with this method showed that the data set contains a strong population genetic signal. As hypothesized pronounced population differentiation and signs of inbreeding were found. Italian populations are clearly differentiated from northern populations (and from each other), which underlines the role of the Alps as a major barrier. Specimens from Lake Balaton (Hungary) show some affiliation with the populations in the Baltic Sea, which are all relatively similar. Populations from the eastern part of Northern Germany are similar to the Baltic populations, while those from the western part are allied to the British populations. The hypothesis is that the recolonization of Europe was from both the Southeast and a western refugium in the area of present-day southern England or Ireland, which resulted in a suture zone in Northern Germany. The effect of passive dispersal by drift attached to host plant material (especially in the Baltic Sea) and by zoochory (migrating waterfowl) is discussed.