Numerical analysis of morphology of natural hybrids between Carex hostiana and the members of Carex flava agg. (Cyperaceae)

Uni‐variate and multi‐variate statistical methods, based on data taken from dried specimens, were used to determine the morphological variance of Carex hostiana 3 Carex flava agg. hybrids and to establish their parents among members of the C. flava complex. The following hybrids were found: C. demissa 3 hostiana [C. 3 fulva], C. hostiana 3 lepidocarpa [C. ×leutzii] and C. flava 3 hostiana [C. 3 xanthocarpa]. The least variable traits, namely beak length, utricle length, ratio of beak length to the overall utricle length, female spike width, and width of the lowest bract, proved to be the most useful in delimiting the hybrids. Carex flava 3 hostiana specimens usually have long utricles and beaks, wide male and female spikes, as well as wide bracts and leaf blades. Carex hostiana 3 lepidocarpa specimens are characterized by relatively short beaks (with low ratio of beak length to the overall utricle length) and narrow bracts. Common features of C. demissa 3 hostiana specimens, on the other hand, are male spikes with long peduncles, usually longly parted female spikes and a long beak compared to the overall utricle length.     

Within-species variation among populations of the Carex flava complex as a function of habitat conditions

Background: Variable habitat conditions contribute to morphological variability that plays a substantial part in evolution of plants. Understanding the extent of phenotypic plasticity has important implication for assigning individuals to taxa.

Aims: I analysed morphological variability among populations within species of the Carex flava group and tested to what extent morphological variability was associated with habitat conditions.

Methods: A total of 571 specimens from 20 populations of four species from the C. flava complex were sampled in Poland and tested by Discriminant Function Analysis (DFA). The relationship between morphological characters and habitat factors was explored by means of the redundancy analysis (RDA).

Results: Variability of the generative and vegetative traits was fairly similar in Carex lepidocarpa, C. flava and Carex demissa, while it was somewhat higher in Carex viridula. The morphological traits were mostly related to soil organic matter content, calcium and carbonate, as well as to habitat fertility (C:N ratio), elevation and habitat disturbance. The results obtained do not support the separation of C. viridula var. pulchella from C. viridula var. viridula.

Conclusions: Phenotypic variability in the species of the C. flava complex is related to habitat conditions and this can lead to the differentiation of morphotypes within species.     

Intraspecific shape variation in horseshoe crabs: The importance of sexual and natural selection for local adaptation

A morphometric analysis of the body shape of three species of horseshoe crabs was undertaken in order to infer the importance of natural and sexual selection. It was expected that natural selection would be most intense, leading to highest regional differentiation, in the American species Limulus polyphemus, which has the largest climatic differences between different populations. Local adaptation driven by sexual selection was expected in males but not females because horseshoe crab mating behaviour leads to competition between males, but not between females. Three hundred fifty-nine horseshoe crabs from nine populations, representing three species, were analyzed using a digitizer to position sixty morphometric landmarks in a three-dimensional space. Discriminant analysis revealed strong regional structuring within a species, which suggests strong philopatry, and showed the existence of geographically-based intraspecific variation. An admixture analysis showed regional intraspecific differentiation for males and females of L. polyphemus and males of the Asian horseshoe crab Carcinoscorpius rotundicauda, but not for females of C. rotundicauda and another Asian horseshoe crab, Tachypleus gigas. Differences in shape variation between sexes were tested with F-tests, which showed lower intrapopulation morphometric variation in males than females. These results indicate a lower degree of local adaptation on body shape in C. rotundicauda and T. gigas than in L. polyphemus and a lower degree of local adaptation in females than in males.     

Niche width and variation within and between populations in colonizing species (Carex flava group)

Summary Taxa of the Carex flava group in Switzerland show a trend towards increased colonizing ability (r-selection). High colonizing ability is correlated with a large fundamental but a small and discontinuous realized niche. It is argued that r-selected species with wide niches should be monomorphic with generalists rather than polymorphic with specialists since they have small effective population sizes in which high genetic variability cannot be maintained. The most r-selected taxon of the group, C. viridula spp. viridula, has indeed the lowest genetic variability within populations but, in ecologically important characters, expresses the highest plasticity. The taxonomically important characters (inflorescences) have high heritability and differences between populations of C. viridula ssp. viridula are probably much affected by genetic drift.     

Leaf anatomy of Carex humilis does not correlate with orographic,geological and bioclimatic habitat conditions in C&SE Europe

This paper presents the results of a multivariate morphometric study of leaf anatomical characters in different, geographically very distant populations of taxon Carex humilis from Hungary, Romania, Serbia, Bosnia & Herzegovina, Montenegro and Albania with the aim to reveal the trends of population differentiation. Analyses were performed on the cross-section of 173 leaves collected from 12 populations. In order to establish the overall morphological variation and relationships between individuals from all populations, principal component analyses (PCA) and canonical discriminant analysis (CDA) have been done. The UPGMA clustering analyses based on leaf anatomical characters and habitat climatic characteristics were carried out to explore whether the observed anatomical differences are a result of adaptive responses. Regression analysis (linear regression) was performed to identify the level of correlation between leaf anatomical characters and basic orographic, geological, and bioclimatic habitat characteristics. Quite unexpectedly, most of observed groups are formed of geographically very distant populations which are living in extremely different climatic and geological conditions, indicating that general anatomical differentiation in Carex humilis in C&SE Europe cannot be explained by the environmental impacts, and basically do not represent an adaptive response to different climatic or geological condition.     

Selection by climatic regime and neutral evolutionary processes in holocentric chromosomes (Carex gr. laevigata: Cyperaceae): A microevolutionary approach

The holocentric structure promotes chromosome rearrangements by fission, fusion, translocation, and inversion, which have been thought to promote differentiation and speciation. The Carex laevigata group (Cyperaceae) comprises four species: two restricted endemics from the western Mediterranean (Carex camposii, 2n = 72, and Carex paulo-vargasii, 2n = 74–75), and two more widespread species, found mostly in Western Europe (C. laevigata, 2n = 69–84, and Carex binervis, 2n = 72–74). We tested the selection for chromosome number by climatic variables by controlling for the non-independence of the data using generalized linear mixed model (GLMM). We obtained chromosome counts as well as DNA sequences for the 5′ trnK intron and the trnV-ndhC intergenic spacer in the chloroplast genome from 181 individuals from 53 populations representing these four species. We also climatically characterized the sites where the 53 populations were found using the WorldClim database. Our results show that the best predictor of chromosome number variation is the climatic environment rather than neutral evolutionary processes like founder events and migration patterns. These results support the adaptive value of the holocentric chromosomes and their role in promoting differentiation and eventually speciation.     

A diploid drop in the tetraploid ocean: hybridization and long-term survival of a singular population of Centaurea weldeniana Rchb. (Asteraceae), a taxon new to Austria

A population of Centaurea weldeniana was discovered recently near Wien (Vienna). The species is reported from Austria for the first time. Determination is confirmed by morphometric analysis of Central European and additional Balkan material of the C. jacea agg. and karyological data. All sampled populations of C. weldeniana are diploid, while only tetraploids have been reported from Central Europe within the C. jacea agg. so far. Detailed morphometric analysis of the newly discovered C. weldeniana population revealed several hybrids with tetraploid C. jacea. The hybrids are tetraploid and originate from unreduced gametes of diploids. No triploids were found. Indeed, the incidence of hybridization is surprisingly low. The population was probably discovered already in 1886 (determined as C. bracteata) and was described as a new species C. argyrolepis in 1901, but these data were lost. However, this isolated diploid population has survived for at least 125?years, which provides a good example of the strength of inter-cytotype reproductive barriers in Centaurea sect. Jacea.     

Reproductive allocation of <Emphasis Type="Italic">Carex flava</Emphasis> reacts differently to competition and resources in a designed plant mixture of five species

In natural plant communities, reproductive allocation can be affected by complex interactions among abiotic resources, species competition and plant size. This topic was addressed using a variety of designed mixed stands of five species (Carex elata, Carex flava, Lycopus europaeus, Lysimachia vulgaris and Mentha aquatica) under four abiotic conditions to investigate how competition and abiotic resources influence the reproductive allocation of one of the five species, C. flava. The plant mixtures varied systematically in both the relative abundance of the five species and the absolute density, and were each established with two levels of water and nutrients. In total, 176 mixtures were maintained for two growing seasons in large pots in an experimental garden. Reproductive allocation of C. flava increased from 6.8% to 9.7% under high nutrient application; however, for both nutrient levels, reproductive allocation was independent of shoot mass (size-independent allocation). Under low competition, reproductive allocation of C. flava decreased as its shoot mass increased, indicating a relatively high investment in vegetative structures under higher light availability. However, under strong competition, the allocation pattern changed and a constant reproductive allocation for different plant sizes was observed. Different water levels did not influence the shoot mass, seed mass or reproductive allocation of C. flava, indicating that the species was not stressed under dryer conditions. When under competitive pressure, however, the species responded with reduced shoot and seed production under more favourable water conditions. This behaviour indicates a trade-off between the ability to tolerate stress and the competitive and reproductive response of C. flava. In conclusion, C. flava was adversely affected by competition with some of the species, and competition, mediated by plant size, indirectly affected reproductive allocation. C. flava was able to modify its allocation pattern depending on the available resources and retained its reproductive allocation even under unfavourable conditions for varying plant sizes, which is interpreted as an advantageous reaction to greater competition pressure. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Island biogeography of North European parthenogenetic Lumbricidae: I. Clone pool affinities and morphometric differentiation of Åland populations

Genetic and morphometnc differences between island (Åland) and mainland (Sweden, Finland and Estonia) populations of five parthenogenetic and ecologically different Lumbricidae species were studied by means of enzyme electrophoresis and morphometric analysis Affinities of clones to the mainland populations show that the postglacial rates of clone flow to Åland differ widely among species In Octolasion cyaneum, the only clone found on Åland is not related to any of the studied clones from the nearby mainlands The Åland population evidently originates from one long dispersal jump through human agency The Åland clone pool of the anthropochorous Aporrectodea rosea is rich As many as 98% of the worms and 82% of the clones belong to genotypes found in the mainland populations Åland is an intergradation zone of A rosea clones that have arrived from the west (Sweden) and from the east (Finland and Estonia) The more intense cultural connections to Sweden explain the higher clonal affinities that A rosea in Åland shows to the west than to the east There are not many Octolasion tyrtaeum clones on Åland The few that are present are mainly clones also found on the mainlands On Åland Eisemella tetraedra has extensive diversity but low clonal affinities to the mainland populations This also holds true for Dendrobaena octaedra but clonal affinities are on an average higher than in E tetraedra The Aland clone pool of D octaedra shows more affinities to Finland than to the other mainlands Probably the main factor that has facilitated D octaedra clone flow from Finland is a continuous archipelago In most species there are at least some clones that are more abundant on Åland than on the mainlands They may have a wider niche in the island environment The Åland populations of A rosea, O tyrtaeum and D octaedra differed particularly in somatic traits from the mainland populations The secondary reproductive traits of the parthenogens on Åland did not differ from the mainland populations We conclude that except for O cyaneumÅland has a role as a stepping stone island in the postglacial dispersal of parthenogenetic earthworms across the Baltic Sea Different rates of flow by immigrant clones from the surrounding mainland populations have resulted in very dissimilar clone pool structures even on the same island The Åland populations also show some morphometric differences to the mainland populations     

Seed image analysis provides evidence of taxonomical differentiation within the Lavatera triloba aggregate (Malvaceae)

Thirty-four seed morphometric and colorimetric features of the five taxa belonging to the Lavatera triloba aggregate were measured using an image analysis system. Data were statistically analysed using a Linear Discriminant Analysis (LDA), which provided evidence to reassess the taxonomic position of some neglected or doubtful taxa. The results confirm the differentiation of the insular endemics L. agrigentina, L. triloba ssp. pallescens and L. triloba ssp. minoricensis. Moreover, the Sardinian populations of L. triloba ssp. triloba are clearly differentiated from their Iberian counterparts, but on the other hand, the seed morpho-colorimetric variation between L. triloba ssp. triloba and L. flava is not easy to interpret.     

GENETIC DIFFERENTIATION AND HYBRIDIZATION BETWEEN CAREX GYNODYNAMA AND C. MENDOCINENSIS (CYPERACEAE) IN CALIFORNIA

Patterns of variation within and between Carex gynodynama and C. mendocinensis were investigated by studying allozyme and chromosome variation in natural populations and structural variation using herbarium specimens. Multivariate analyses of structural data demonstrated that C. gynodynama is clearly distinct from C. mendocinensis, and that sterile specimens similar to C. mendocinensis are intermediate between that species and C. gynodynama. The mean genetic distance between the two species, based on allozyme phenotypes at 17 enzyme-coding loci, was 0.22 ± 0.12. The sterile putative hybrids had the expected heterozygous pattern at three enzyme-coding loci at which the parental species were fixed for different alleles. Chromosome numbers are reported for the first time for both species and their putative hybrid. Carex mendocinensis had a different number in each of the three populations examined with n = 28, n = 29, or n = 30. Chromosome counts from one population of C. gynodynama revealed five plants with n = 25 and one with n = 26. Putative hybrids from this population exhibited irregular pairing at meiosis with 2n = ca. 55–57. Patterns of allozyme variation also suggest that C. mendocinensis has an outcrossing or mixed mating system but that C. gynodynama is an inbreeding species. Carex gynodynama exhibited very little variation in structure, habitat, or at the enzyme-coding loci examined, suggesting that it may have experienced a genetic bottleneck relatively recently. Carex mendocinensis had higher levels of variation both within and between populations at enzyme-coding loci and in structural features. This pattern of variation and a geographic distribution centered in serpentine areas of the Klamath–Siskiyou region, with disjunct smaller populations in serpentine areas farther south, suggest that C. mendocinensis once may have been a more widespread species.     

Asp-376-Glu substitution endows target-site resistance to AHAS inhibitors in Limnocharis flava,an invasive weed in tropical rice fields

Limnocharis flava (L.) Buchenau is a problematic weed in rice fields and water canals of Southeast Asia, and in Malaysia this invasive aquatic weed species has evolved multiple resistance to synthetic auxin herbicide and acetohydroxyacid synthase (AHAS) inhibitors. In this study, it was revealed that, a single nucleotide polymorphism (SNP) at amino acid position 376, where C was substituted to G at the third base of the same codon (GAC to GAG), resulting in Aspartate (Asp) substitution by Glutamate (Glu) was the contributing resistance mechanism in the L. flava population to AHAS inhibitors. In vitro assay further proved that, all the L. flava individuals carrying AHAS resistance mutation exhibited decreased-sensitivity to AHAS inhibitors at the enzyme level. In the bensulfuron-methyl whole-plant bioassay, high resistance indices (RI) of 328- and 437-fold were recorded in the absence and presence of malathion (the P450 inhibitor), respectively. Similarly, translocation and absorption of bensulfuron-methyl in both resistant and susceptible L. flava populations showed no remarkable differences, hence eliminated the possible co-existence of non-target-site resistance mechanism in the resistant L. flava. This study has confirmed another new case of a target-site resistant weed species to AHAS-inhibitors.     

Microbial Population Analysis of the Salivary Glands of Ticks; A Possible Strategy for the Surveillance of Bacterial Pathogens

Ticks are one of the most important blood-sucking vectors for infectious microorganisms in humans and animals. When feeding they inject saliva, containing microbes, into the host to facilitate the uptake of blood. An understanding of the microbial populations within their salivary glands would provide a valuable insight when evaluating the vectorial capacity of ticks. Three tick species (Ixodes ovatus, I. persulcatus and Haemaphysalis flava) were collected in Shizuoka Prefecture of Japan between 2008 and 2011. Each tick was dissected and the salivary glands removed. Bacterial communities in each salivary gland were characterized by 16S amplicon pyrosequencing using a 454 GS-Junior Next Generation Sequencer. The Ribosomal Database Project (RDP) Classifier was used to classify sequence reads at the genus level. The composition of the microbial populations of each tick species were assessed by principal component analysis (PCA) using the Metagenomics RAST (MG-RAST) metagenomic analysis tool. Rickettsia-specific PCR was used for the characterization of rickettsial species. Almost full length of 16S rDNA was amplified in order to characterize unclassified bacterial sequences obtained in I. persulcatus female samples. The numbers of bacterial genera identified for the tick species were 71 (I. ovatus), 127 (I. persulcatus) and 59 (H. flava). Eighteen bacterial genera were commonly detected in all tick species. The predominant bacterial genus observed in all tick species was Coxiella. Spiroplasma was detected in Ixodes, and not in H. flava. PCA revealed that microbial populations in tick salivary glands were different between tick species, indicating that host specificities may play an important role in determining the microbial complement. Four female I. persulcatus samples contained a high abundance of several sequences belonging to Alphaproteobacteria symbionts. This study revealed the microbial populations within the salivary glands of three species of ticks, and the results will contribute to the knowledge and prediction of emerging tick-borne diseases.     

Genetic variability in macaques: Correlations between biochemical and dermatoglyphic systems

The palmar dermatoglyphics of four groups of macaque, Macaca nemestrina, M. fuscata yakui, M. f. fuscata and M. fascicularis, were compared, and the genetic affinities of the four groups were assessed using a dermatoglyphic mean square distance measure. According to this measure the two subspecies of M. fuscata were dermatoglyphically more distant from one another than was M. fuscata fuscata from M. nemestrina, suggesting that the two insular populations of M. fuscata had evolved in genetic isolation. The distributions of specific dermatoglyphic patterns were then compared with the frequency distributions of alleles at different protein structural loci in other macaque populations. The results of the comparison were consistent in that insular populations showed reduction in variability for both kinds of systems. The implications of this correlation are discussed in terms of population size and natural selection.     

Genetic and morphological variation of three freshwater lake populations of Coilia ectenes (Engraulidae)

Population structure of the important commercial fish, Coilia ectenes was investigated in samples from three freshwater lakes in the Eastern China using a multivariate approach of morphometries and mitochondrial DNA control region sequencing. A total of eighteen morphological distances of truss method and eight morphometric variables were taken from each fish. Multivariate analyses of the morphometric data revealed significant morphological differences among the three lake populations, especially for those samples from Taihu Lake. Discriminant functions were used to compare sites, and these permitted an 83% success rate in distinguishing fish from the three sites. However, no obviously geographical differentiation was found among those populations of C. ectenes based on the genetic data. In the AMOVA analysis, only 2.2% genetic variability came from different populations, and most of them were present within the sub-populations. Experience a recent population expansion and some movement of fish among those areas, quite possibly enough to bring about relative genetic homogeneity, but there is insufficient to prevent the three populations from differing phenotypically. The diversified environmental factors may be playing an important role in shaping morphological variations among those populations.     

Inter- and Intraspecific Variation in Wild-type and Single Female-derived Populations of Xiphinema americanum-group Nematodes

Ten populations of Xiphinema americanum-group nematodes were reared from individual females to evaluate inter- and intraspecific variation under identical host and environmental conditions. Data indicated that morphometric variability of X. americanum was the result of genetic variation rather than phenotypic plasticity and that genetic heterogeneity was greater than previously thought. Morphometrics of single female derived (SFD) populations identified different genotypes present in the field populations. Stylet length was the least variable morphometric character of SFD populations, but collectively stylet measurements of all individuals formed an uninterrupted continuum ranging from 107-148 μm. Range and frequency of stylet measurements of field populations could be accounted for by the relative proportion of different genotypes in the population. Nine SFD populations were identified as X. americanum sensu stricto, and one SFD population was similar to X. californicum.     

Large scale SNP unearthing and genetic architecture analysis in sea-captured and cultured populations of Cynoglossus semilaevis

Knowledge on population structure and genetic diversity is a focal point for association mapping studies and genomic selection. Genotyping by sequencing (GBS) represents an innovative method for large scale SNP detection and genotyping of genetic resources. Here we used the GBS approach for the genome-wide identification of SNPs in a collection of Cynoglossus semilaevis and for the assessment of the level of genetic diversity in C. semilaevis genotypes. GBS analysis generated a total of 55.12 Gb high-quality sequence data, with an average of 0.63 Gb per sample. The total number of SNP markers was 563, 109. In order to explore the genetic diversity of C. semilaevis and to select a minimal core set representing most of the total genetic variation with minimum redundancy, C. semilaevis sequences were analyzed using high quality SNPs. Based on hierarchical clustering, it was possible to divide the collection into 2 clusters. The marine fishing populations were clustered and clearly separated from the cultured populations, and the cultured populations from Hebei was also distinct from the other two local populations. These analyses showed that genotypes were clustered based on species-related features. Differential significant SNPs were also captured and validated by GBS and SNaPshot, with linkage disequilibrium and haplotype analysis, seven SNPs have been confirmed to have obvious differentiation in two populations, which may be used as the characteristic evaluation sites of sea-captured and cultured Cynoglossus semilaevis populations. And SNP markers and information on population structure developed in this study will undoubtedly support genome-wide association mapping studies and marker-assisted selection programs. These differential SNPs could be also employed as the characteristic evaluation sites of sea-captured and cultured Cynoglossus semilaevis populations in future.     

Intercontinental effect on sexual shape dimorphism and allometric relationships in the beetle pest Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae)

Hind wing shape variation was examined in 686 adult Diabrotica virgifera virgifera collected from maize plants in Europe and the USA Corn Belt, using geometric morphometric techniques. Sexual dimorphism at an Intercontinental scale was assessed using canonical variates analysis, a multivariate statistical method used to find the shape characters that best distinguish among groups of specimens. Our results showed that each of the populations of D. v. virgifera investigated in this study showed high levels of sex based hind wing shape dimorphism. In particular a stronger and more obvious pattern of hind wing shape variation was found in the USA than in Europe. These results support previous studies on D. v. virgifera wing shape that show that female D. v. virgifera have more elongated wings than males. These differences raise the question of whether sexual dimorphism may be modulated by natural selection.     

The east‐west‐north colonization history of the Mediterranean and Europe by the coastal plant Carex extensa (Cyperaceae)

Coastal plants are ideal models for studying the colonization routes of species because of the simple linear distributions of these species. Carex extensa occurs mainly in salt marshes along the Mediterranean and European coasts. Variation in cpDNA sequences, amplified fragment length polymorphisms (AFLPs) and simple sequence repeats (SSRs) of 24 populations were analysed to reconstruct its colonization history. Phylogenetic relationships indicate that C. extensa together with the South American Carex vixdentata and the southern African Carex ecklonii form a monophyletic group of halophilic species. Analyses of divergence times suggest that early lineage diversification may have occurred between the late Miocene and the late Pliocene (Messinian crisis). Phylogenetic and network analyses of cpDNA variation revealed the monophyly of the species and an ancestral haplotype contained in populations of the eastern Mediterranean. The AFLP and SSR analyses support a pattern of variation compatible with these two lineages. These analyses also show higher levels of genetic diversity and differentiation in the eastern population group, which underwent an east‐to‐west Mediterranean colonization. Quaternary climatic oscillations appear to have been responsible for the split between these two lineages. Secondary contacts may have taken place in areas near the Ligurian Sea in agreement with the gene flow detected in Corsican populations. The AFLP and SSR data accord with the ‘tabula rasa’ hypothesis in which a recent and rapid colonization of northern Europe took place from the western Mediterranean after the Last Glacial Maximum. The unbalanced west‐east vs. west‐north colonization may be as a result of ‘high density blocking’ effect.