When sexual meets apomict: genome size,ploidy level and reproductive mode variation of Sorbus aria s.l. and S. austriaca (Rosaceae) in Bosnia and Herzegovina

Background and Aims Allopolyploidy and intraspecific heteroploid crosses are associated, in certain groups, with changes in the mating system. The genus Sorbus represents an appropriate model to study the relationships between ploidy and reproductive mode variations. Diploid S. aria and tetraploid apomictic S. austriaca were screened for ploidy and mating system variations within pure and sympatric populations in order to gain insights into their putative causalities.Methods Flow cytometry was used to assess genome size and ploidy level among 380 S. aria s.l. and S. austriaca individuals from Bosnia and Herzegovina, with 303 single-seed flow cytometric seed screenings being performed to identify their mating system. Pollen viability and seed set were also determined.Key Results Flow cytometry confirmed the presence of di-, tri- and tetraploid cytotype mixtures in mixed-ploidy populations of S. aria and S. austriaca. No ploidy variation was detected in single-species populations. Diploid S. aria mother plants always produced sexually originated seeds, whereas tetraploid S. austriaca as well as triploid S. aria were obligate apomicts. Tetraploid S. aria preserved sexuality in a low portion of plants. A tendency towards a balanced 2m : 1p parental genome contribution to the endosperm was shared by diploids and tetraploids, regardless of their sexual or asexual origin. In contrast, most triploids apparently tolerated endosperm imbalance.Conclusions Coexistence of apomictic tetraploids and sexual diploids drives the production of novel polyploid cytotypes with predominantly apomictic reproductive modes. The data suggest that processes governing cytotype diversity and mating system variation in Sorbus from Bosnia and Herzegovina are probably parallel to those in other diversity hotspots of this genus. The results represent a solid contribution to knowledge of the reproduction of Sorbus and will inform future investigations of the molecular and genetic mechanisms involved in triggering and regulating cytotype diversity and alteration of reproductive modes.     

The Structure of Morphological and Genetic Diversity in Natural Populations of Carica papaya (Caricaceae) in Costa Rica

In Costa Rica, dioecious Carica papaya has been observed growing in disturbed areas and within secondary lowland forests. Such populations can serve as a reservoir of genetic and morphological diversity for this tropical fruit crop. We quantify the levels and patterns of the diversity of naturally occurring populations of C. papaya and address the demographic history of these populations. We measured 29 vegetative and reproductive morphological traits in situ from 252 plants and found significant heterogeneity among regional populations in the majority of these traits. Significant variation was found among regional populations with respect to fruit size and shape, with plants in the Nicoya Peninsula possessing smaller, less fleshy fruit, a characteristic of previously described wild populations of papaya. We then assessed the levels and patterns of genetic diversity in 164 plants from natural populations and 20 cultivars. Natural populations exhibit a deficiency of heterozygotes; however, this is much more pronounced within the cultivars. Although there is little genetic differentiation among natural populations, we did find evidence of cryptic genetic population structure. Analyses of population demography indicate that these natural populations have undergone a recent genetic bottleneck, followed by recent population expansion, possibly promoted by the transformation of the Costa Rican landscape for agricultural use. Abstract in Spanish is available at http://www.blackwell‐synergy.com/loi/btp .     

WEEDY ADAPTATION IN SETARIA SPP. I. ISOZYME ANALYSIS OF GENETIC DIVERSITY AND POPULATION GENETIC STRUCTURE IN SETARIA VIRIDIS

Setaria viridis is an important self-pollinating, cosmopolitan weed of temperate regions worldwide. Allozyme markers were used to investigate genetic diversity and structure in 168 accessions (including four S. italica) collected mainly from North America and Eurasia. Genetic diversity in green foxtail, and its population genetic structure, provided important clues about this weed's evolutionary history. Genetic diversity was low, with marked population differentiation: the percentage of polymorphic loci was 25% (0.95 criterion); mean number of alleles per locus was 1.86; mean panmictic heterozygosity was 0.07; and the coefficient of population genetic differentiation was 0.65. A common genotype occurred in 25 accessions distributed in six countries from both the Old World and New World, in a wide variety of ecological situations. Relatively little genetic divergence occurred between Eurasia and North America, with Nei's unbiased genetic identity between the two regions equaling 1.0. Populations from these two continents also had equivalent genetic diversity. Within North America, regional differentiation was indicated by northern and southern groups separated at 43.5° N latitude. No geographic pattern in genetic diversity was found within Eurasia. The size of the geographic range from which populations were sampled was not an accurate indicator of the extent of genetic diversity found among populations from that region. These results suggest that present patterning among green foxtail populations in North America is the consequence of multiple introductions into the New World followed by local adaptation and regional differentiation. Finally, S. italica and several green foxtail varieties did not differ isozymatically from typical forms of green foxtail. This supports the view that S. italica and S. viridis are conspecific, that the former (foxtail millet) is a domesticated form of the latter, and also questions the taxonomic validity of formally recognizing morphological varieties within green foxtail.     

Genetic characterization of grey wolves (<Emphasis Type="Italic">Canis lupus</Emphasis> L. <Emphasis Type="Italic">1758</Emphasis>) from Bosnia and Herzegovina: implications for conservation

The grey wolves of Bosnia and Herzegovina form a subpopulation of the Dinaric-Balkan wolf population and represent one of Europe’s least studied wolf populations. Since the Dinaric-Balkan population is a valuable source of genetic diversity for neighboring populations, comprehensive assessments are warranted. We aimed to determine the genetic variability and structure of the grey wolf population from Bosnia and Herzegovina, as well as estimate levels of gene flow and inbreeding and evaluate genetic signals of a bottleneck. To do this, we analyzed the variability of eighteen microsatellite loci. We found moderately high genetic heterozygosity for wolves from Bosnia and Herzegovina, as described for other Dinaric-Balkan wolf populations. We reveal weak genetic structuring with two genetic clusters identified. Wolves from the eastern part of the region formed a relatively distinct cluster, whereas individuals in the second cluster overlapped quite considerably with admixed individuals. Despite the signal of genetic structure being weak, clustering of individuals from the eastern part of the country extended through all analyses. Thus, this cluster could be considered a separate management unit, perhaps requiring specific conservation attention.     

Genetic structure of Barbus spp. populations in the Marches Region of central Italy and its relevance to conservation actions

A genetic survey of Barbus spp. populations in the Marches Region (Adriatic River basins), central Italy, was carried out using mitochondrial and nuclear markers (partial D‐loop, cyt b sequences and microsatellite loci) in order to ascertain their systematic position and to address their genetic structure which is key to conservation action planning. Analyses were conducted on sequences obtained from 91 individuals collected from eight sampling sites in five different rivers, from two specimens provided by the Ichthyological Centre of Rome and mitochondrial sequences of Barbus spp. retrieved from GenBank. Presumptive classification based on external morphological characters was not confirmed by genetic analysis, by means of which all specimens collected in the Marches Region were ascribed to Barbus plebejus. Genetic diversity values (h and π) of sampling groups were all different from 0 except the one sample collected from the upper reaches of the River Tenna, above a hydroelectric dam. Population connectivity and colonization patterns of the studied area were inferred from an analysis of molecular variance distribution and evolutionary relationships among haplotypes. The results point to different levels of isolation among sampling groups due to ecological and anthropogenic factors and the effect of an artificial barrier on genetic variability and conservation status of the population. Finally, this study confirms the uncertainty associated with systematic classification of Barbus spp. based on morphological characters due to the phenotypic plasticity of the species.     

Morphological evolution and genetic differentiation in Daphnia species complexes

Despite many ecological and evolutionary studies, the history of several species complexes within the freshwater crustacean genus Daphnia (Branchiopoda, Anomopoda) is poorly understood. In particular, the Daphnia longispina group, comprising several large-lake species, is characterized by pronounced phenotypic plasticity, many hybridizing species and backcrossing. We studied clonal assemblages from lakes and ponds comprising daphnids from several species complexes. In order to reveal patterns of reticulate evolution and introgression among species, we analysed three data sets and compared nuclear, mtDNA and morphological divergence using animals from 158 newly established clonal cultures. By examining 15 nuclear and 11 mitochondrial (12S/16S rDNA) genetic characters (allozymes/restriction enzymes), and 48 morphological traits, we found high clonal diversity and discontinuities in genotypic and morphological space which allowed us to group clones by cytonuclear differentiation into seven units (outgroup D. pulex). In contrast to six groups emerging from nuclear divergence (related to three traditional species, D. cucullata, D. galeata, D. hyalina and three pairwise intermediate hybrids), a seventh group of clones was clearly resolved by morphological divergence: distinct mtDNA haplotypes within one nuclear defined cluster, ‘D. hyalina’, resembled traditional D. hyalina and D. rosea phenotypes, respectively. In other nuclear defined clusters, association between mtDNA haplotype and morphology was low, despite hybridization being bidirectional (reciprocal crosses). Morphological divergence was greatest between young sister species which are separated on the lake/pond level, suggesting a significant role for divergent selection during speciation along with habitat shifts. Phylogenetic analyses were restricted to four cytonuclear groups of clones related to species. mtDNA and nuclear phylogenies were consistent in low genetic divergence and monophyly of D. hyalina and D. rosea. Incongruent patterns of phylogenies and different levels of genetic differentiation between traditional species suggest reticulate evolutionary processes.     

Reticulate evolution: ancient introgression of the Adriatic brown trout mtDNA in softmouth trout Salmo obtusirostris (Teleostei: Salmonidae)

Two populations of softmouth trout ( Salmo obtusirostris ) from the rivers Neretva (Bosnia and Herzegovina) and Jadro (Croatia), along with two neighbouring populations of brown trout ( Salmo trutta ) were analysed with a suite of genetic markers (two mtDNA genes, two nuclear genes, and nine microsatellites) as well as morphological characters. The Jadro softmouth trout were fixed for a brown trout mtDNA haplotype of the Adriatic lineage, which is 1.7% divergent from a previously described haplotype characteristic for the Neretva softmouth trout. All other genetic markers, as well as morphological analysis, supported the clear distinction of softmouth trout from the rivers Neretva and Jadro from brown trout in neighbouring populations, and thus a mtDNA capture event is assumed. Population specific microsatellite allele profiles, as well as a high number of private alleles for both populations of softmouth trout, support the hybridization between brown trout and the Jadro softmouth trout most likely being of ancient origin, thus leading to a reticulate evolutionary pattern of mtDNA in this taxon.  © 2007 The Linnean Society of London, Biological Journal of the Linnean Society , 2007, 90 , 139–152.     

Genetic variation in Pueraria lobata (Fabaceae), an introduced, clonal, invasive plant of the southeastern United States

Pueraria lobata (kudzu), a clonal, leguminous vine, is invading the southeastern United States at a rate of 50 000 ha per year. Genetic variability and clonal diversity were measured in 20 southeastern U.S. populations using 14 allozyme loci. Within its U.S. range, 92.9% of the loci were polymorphic and overall genetic diversity was 0.290. Such high levels of genetic diversity are consistent with its history of multiple introductions over an extended period of time. The average proportions of polymorphic loci and genetic diversity within populations were 55.7% (range = 28.6–85.7%) and 0.213 (range = 0.114–0.317), respectively. The proportion of total genetic diversity found among populations was similar to species with equivalent life history characters (G_ST = 0.199). No regional patterns of variation were seen. The number of putative genotypes in each population ranged from 2 to 26. Mean genotypic diversity was 0.694, ranging from 0.223 to 0.955. Such high levels of genotypic diversity indicate that local sites are often colonized by several propagules (most likely seeds) and/or that sexual reproduction occurs within populations after establishment. An excess of heterozygosity was observed in populations with few unique genets, implying that selection for highly heterozygous individuals may occur in populations of P. lobata.     

Drainage-independent genetic structure and high genetic diversity of endangered freshwater pearl mussels (<Emphasis Type="Italic">Margaritifera margaritifera</Emphasis>) in northern Europe

Freshwater pearl mussels (Margaritifera margaritifera) are among the most critically threatened bivalve molluscs worldwide. An understanding of spatial patterns of genetic diversity is crucial for the development of integrative conservation strategies. We used microsatellites to study the genetic diversity and differentiation of 14 populations of M. margaritifera in central Sweden, an area which was described as a major secondary contact zone in postglacial colonisation for other species. Genetic diversity of Swedish pearl mussel populations was much greater than in central and southern Europe but similar to the genetic diversity observed in the northeastern portion of their European range. Genetic differentiation among populations was pronounced but to a large extent independent from present-day drainage systems. The complex patterns of genetic diversity and differentiation in pearl mussel seem to be strongly influenced by the species’ high degree of specialisation and extraordinary life history strategy which involves facultative hermaphrodism and an obligatory encystment stage on a host fish. Genetic drift effects and anthropogenic disturbances resulting in reduction of population size and loss of connectivity are less pronounced in northern pearl mussel populations compared to those in central and southern Europe.     

Molecular and morphological differences between two geographic populations of Salanx ariakensis (Salangidae) from Korea and Japan

Geographic variations of a salangid, Salanx ariakensis, from Korea and Japan, based on partial mtDNA cyt b gene sequences and morphometric data, are presented and compared with Salanx cuvieri from China. Analyses of molecular variance (AMOVA) suggested that the two populations of S. ariakensis differed significantly from each other in pairwise fixation index (F _ST = 0.43), although their reciprocal monophyly was not constructed in the phylogenetic analysis. The phylogenetic analysis, however, supported the reciprocal monophyly of S. ariakensis (Korean population + Japanese population) and S. cuvieri. Differences in genetic structure between the two populations of S. ariakensis were suggested by diversity indices, mismatch distribution shape, and minimum spanning network. In particular, diversity indices indicated that the Korean population was historically larger and more stable than the Japanese population. In addition, both the principal component analysis (PCA) and canonical discriminant analysis (CDA) for morphometric characters, and the Kruskal–Wallis test for some meristic characters, showed that the two populations of S. ariakensis differed from each other, although morphological differences between the two populations of S. ariakensis were smaller than those between S. ariakensis and S. cuvieri. Such morphological differences were consistent with the differences in mtDNA.     

Towards a better understanding of polyploid Sorbus (Rosaceae) from Bosnia and Herzegovina (Balkan Peninsula), including description of a novel,tetraploid apomictic species

Sorbus subgenus Soraria encompasses taxa originating from spontaneous hybridization between members of subgenera Aria and Sorbus disjunctly distributed across Europe and Asia. Using molecular data (amplified fragment length polymorphisms, plastid DNA sequences and nuclear microsatellites), flow cytometry (allowing for the determination of ploidy and mode of reproduction) and morphology, we disentangled the relationships among polyploid cytotypes and explored their relationships with their diploid ancestors. Among others, we focused on a large, geographically isolated hybrid population in Bosnia and Herzegovina. Molecular and morphological analyses confirmed the distinct position of this population in relation to its parental (S. aria and S. aucuparia) and other hybridogenous taxa originating independently from the same parents in different parts of Europe. After establishing its genetic and morphological divergence, we describe the isolated Bosnian population as a new tetraploid apomictic species, S orbus bosniaca , discuss its taxonomic status and propose conservation measures to protect the locus classicus of this new Balkan endemic. © 2015 The Linnean Society of London, Botanical Journal of the Linnean Society, 2015, 178 , 670–685.     

Overview of human population-genetic studies in Bosnia and Herzegovina during the last three centuries: history and prospective

Modern Bosnia and Herzegovina is a multi-ethnic and multi-religion country, with a very stormy history. Certain archaeological findings indicate continuous population of its territory since the Paleolithic. In time, vast number of different factors jointly influenced fascinating diversity of local human populations. A great number of small, more or less isolated, indigenous populations, make this area quite attractive for population-genetic surveys of different levels and approaches. Austro-Hungarian military physicians conducted the very first known bio-anthropological analyses of Bosnia-Herzegovina population at the end of the 19th century. Thus, the first step towards resolving the genetic structures of local B&H human populations was made. The studies that followed (conducted throughout most of the 20th century) were primarily based on the observation of various phenotypic traits. This stage was followed by the examination of various cytogenetic and fundamental DNA based molecular markers. The efforts undertaken over the last three centuries revealed "human genetic treasure" in Bosnia and Herzegovina. However, even now, after all the studies that were conducted, many interesting features remain to be discovered and described within the existing local human populations.     

Human-mediated processes affecting distribution and genetic structure of Squalidus multimaculatus,a freshwater cyprinid with small spatial range

Endemic species typically have a narrow niche breadth, and are likely more vulnerable to extinction than more widespread taxa. Squalidus multimaculatus is a small cyprinid endemic to the Korean Peninsula, and its reported geographical range was restricted to several small rivers located along the southeast coast. Several populations of S. multimaculatus have supposedly been subject to a variety of recent anthropogenic actions. Here, we analyzed the pattern of genetic diversity within and among populations of S. multimaculatus using nine microsatellite loci to quantify the relative contributions of human-mediated processes to the contemporary distribution and genetic structure. Overall, low levels of genetic diversity were exhibited in the populations of S. multimaculatus. Genetic differentiations among populations were not completely represented by their geographical proximity, likely resulting from the low intrapopulation genetic variability and anthropogenic transplants. The most conspicuous outcome of the anthropogenic activities was the introgression of alleles from a closely related species, S. gracilis majimae. Our study showed that anthropogenic transplanting, even with only a small number of individuals, can challenge our conservation goal to maintain the species integrity that has long been shaped in evolutionary processes.     

Effects of latitude and allopatry on body size variation in European water shrews

We studied the intra- and interspecific size variability of 271 water shrewsNeomys fodiens (Pennant, 1771) andN. anomalus Cabrera, 1907 from seven sample sites along a latitudinal transect from Bosnia and Herzegovina to Poland.Neomys anomalus was the only water shrew in three Dinaride karst fields, while it was sympatric with N.fodiens in remaining sites. The first principal component scores (PC1; 72.2% of variance explained), derived from principal components analysis of 13 cranial, mandibular and dental measurements, were used as the size factor. One-way ANOVA detected significant interpopulation variation in both species; intraspecific variation, however, was much more pronounced inN. anomalus. No latitudinal size pattern was found in N. fodiens (r = −0.42, p = 0.58), while mean PC1 scores correlated significantly and negatively with latitude inN. anomalus (r = −0.92, p = 0.004). Therefore, along a north to south transect,N. anomalus converged in size towards N. fodiens, which suggests that the former species occupies increasingly more aquatic habitats in the same direction. Individuals from allopatric populations ofN. anomalus from Slovenia and Bosnia and Herzegovina were, on average, larger than sympatric conspecific populations from the same latitudinal zone, which is consistent with the hypothesis of character displacement.     

Genetic diversify among Balkan trout populations based on RAPD analysis

Random amplified polymorphic DNA (RAPD) markers were used to estimate the population structure and phylogenetic relationships among samples of the Salmo trutta complex that inhabit the Balkan Peninsula. Five random oligodecamers were selected to amplify DNA from 140 fish from seven populations. Using these primers, 55 discernible DNA fragments were generated, of which 50 (90.91%) were polymorphic. The statistical results indicated that there was low genetic diversity within populations (with an average percentage of polymorphic bands (P) of 11.69% and a Nei’s genetic diversity index (h) of 0.035), but at the same time high genetic differentiation among populations (F _ST = 0.89). The distribution of genetic diversity among Balkan trout may result from their evolutionary history and reflects genetic drift coupled with bottleneck phenomena. Overall, RAPDs proved valuable tools for quick and reliable stock discrimination and provided information that might be useful regarding conservation and management of trout.     

Analysis of meristic and mitochondrial DNA variation in Syngnathus abaster (Teleostea: Syngnathidae) from two western Mediterranean lagoons

Syngnathus abaster is a euryhaline pipefish distributed in the Mediterranean, Black Sea, and the north-eastern Atlantic. Although its populations are characterised by high morphological plasticity, neither congruent information about the morphological differentiation of S. abaster populations from the Mediterranean lagoons is available, nor population genetic surveys have been so far performed. In this context, the aims of our study were as follow: i) to describe the variation at nine meristic characters of S. abaster from two western Mediterranean brackish-water areas: the Tunis north lagoon (Tunisia) and the Mauguio lagoon (France); ii) to analyse sequences of four mitochondrial DNA regions in order to evaluate the occurrence of genetic variation between the two areas, if any. The morphological survey revealed a subdivision into two distinct groups: the first included the Tunisian specimens, the second the French ones. Genetic analysis evidenced the occurrence of a sharp genetic structuring with high levels of genetic differentiation between Tunisian and French S. abaster populations. Results suggest that the evolutionary forces driven by the different biogeographical and ecological conditions of the two Mediterranean brackish-water areas have boosted the morphological and genetic divergence of the populations here analysed. The scarce potential of long-distance dispersal of S. abaster may have also enhanced the divergence retrieved.     

Morphology and mitochondrial DNA variation of the Siberian whitefish <Emphasis Type="Italic">Coregonus lavaretus pidschian</Emphasis> (Gmelin) in the upstream water bodies of the Ob and Yenisei Rivers

Combining morphological, ecological and genetic analyses, we compared patterns of diversification within and among populations of the southern Siberian whitefish species Coregonus lavaretus pidschian (Gmelin) to illuminate their evolutionary history. Using sequencing data from 1,930 bp of NADH dehydrogenase subunit 1 (ND1) mitochondrial DNA regions, we documented phylogeographic relationships among populations and developed a phylogeny of mtDNA haplotypes. We found significant differences in the perforated lateral-line scale numbers within and between some populations. Clear differences in the number of gill rakers on the first branchial arch were only exhibited between populations of C. l. pidschian and Coregonus lavaretus pravdinellus Dulkeit. Concordance between different morphological groups based on two meristic traits and mtDNA patterns was also tested.     

Genetic structure and differentiation of Plantago major reveals a pair of sympatric sister species

Seeds of the widespread weed Plantago major were collected from 10 European countries, as well as Trinidad and North America. The seed collections were from populations of two taxa which are ecologically rather than geographically separated and formally recognized as the subspecies Plantago major ssp. major and P.m. ssp. intermedia (also called P.m. ssp. pleiosperma). Eight polymorphic allozyme loci and 73 random-primed DNA fragments were scored, as well as 11 morphological characters. Complete concordance between morphological traits and genetic data provides evidence that these two taxa, although very similar, are distinct species. They are both widespread, they are broadly sympatric and capable of interbreeding. However, slight morphological and ecological differences coincide with genetic clustering of populations from widely separated locations. In addition, P. major and P. intermedia differ in their population structure: P. intermedia has greater genetic diversity among populations and less genetic variance within populations than P. major. We suggest that differences between the two species in their levels of selfing may explain the distinctive genetic structure of each species. We hypothesize a link between selfing rate and lifespan of the two taxa. P. major is characterized by lower genetic variation among populations, a higher rate of outcrossing, longer lifespan and production of fewer seeds per seed capsule. P. intermedia is more highly structured with much differentiation among populations, a higher rate of inbreeding and it often grows as an annual.     

油松天然群体的种实性状表型多样性研究

为了揭示油松天然种群在不同地理环境条件下表型变异的程度和规律,在油松整个天然分布范围内选择了12个具有代表性的居群作为研究对象,对其球果、种子、种翅等12个种实性状的变异程度及其与环境因子间关系进行了比较分析。结果显示:(1)各个性状在居群内和居群间均存在较大的变异(CV>12%)。其中千山(QS),曾家镇(ZJ)和互助(HZ)3个居群表现出了较高的变异(CV>20%),而球果干重(CDW)和种子长(CL)是所有表型性状中变异幅度最大的(CV分别为31%和21%),但种翅性状与其他性状相比具有较高的稳定性。(2)巢式设计方差分析表明,在居群内表型分化系数(Vst)变化在3.18%~89.86%之间,而群体间的Vst为38.97%;与其他针叶树种相比,油松拥有较高的表型分化系数,且居群内的变异程度远高于居群间的变异,尤其是千山(QS)、曾家镇(ZJ)和互助(HZ)3个居群,这说明油松具有较高的环境异质性适应能力或恶劣环境耐受能力。(3)相关性分析表明,该研究的各形态特征与潜在蒸发量均为负相关,且大部分形态指标间及它们与潜在蒸发量间存在显著相关性,表明潜在蒸发量是油松形态特征变化的最重要环境影响因子,预示油松最适宜生长于温暖潮湿的环境中;并表明因各形态特征间相互紧密关联,所以它们受环境条件影响而共变。