Poor genetic differentiation but clear cytoform divergence among cryptic species in Simulium damnosum complex (Diptera: Simuliidae)

A complex cryptic species taxonomy has been developed for the flies that vector Onchocerca volvulus L euckart in A frica. This taxonomy names cytoforms of Simulium damnosum T heobald s.l. based on chromosomal inversion polymorphisms. Researchers have linked variation in cytoforms to variation in geographical distributions and biological traits, but have failed to demonstrate that many of these cytoforms are differentiated genetically or morphologically. This may be explained by recent divergence and incomplete lineage sorting of ancestral polymorphisms. Alternatively, it may be the outcome of ongoing gene flow, with inversions maintaining sets of co‐adapted alleles and selection driving inversion frequency differences across habitats. Here, our goals were to test species delimitations in the S. damnosum complex, and determine if the poor differentiation between these species is better explained by the ancestral polymorphisms or gene‐flow hypotheses. Geometric morphometric analysis and more intensive genetic sampling of populations in N igeria only reinforced the lack of clear divergence among S. damnosum complex cytoforms. By contrast, the first statistical estimates of evolutionary relationships based on chromosomal inversion polymorphism data revealed clear signs of divergence, which lends support to the ancestral polymorphisms hypothesis. But the latter analyses may be biased by non‐neutral evolution of inversion polymorphisms; more rigorous tests of cytoform‐based species hypothesis will require deeper sampling across populations and neutral genetic markers along with explicit modelling of genetic isolation and migration.     

Genetic diversity and population differentiation of Capparis spinosa (Capparaceae) in Northwestern China

To explore the influence of vicariance on differentiation patterns of taxa in arid regions, we systematically assessed the genetic diversity and variability of Capparis spinosa, a typical xerophyte that is widely distributed in the Tianshan Mountains and adjacent areas. In total, 300 individuals from 25 populations were sampled, and 14 haplotypes were identified using two cpDNA sequences (rpS12-rpL20 and ndhF). A high level of total genetic diversity (H_T = 0.859) was detected, and this was attributed to the extensive distribution range, which included numerous large populations. The SAMOVA results suggested that the 25 populations were clustered into 4 major geographical groups; a similar divergence trend was found by constructing a BEAST phylogenetic tree and a network diagram. The AMOVA results revealed that significant genetic differentiation occurred among groups. Our results indicated a considerable correlation between genetic divergence and geographical distribution. Isolation due to complex mountain and desert geography might limit gene exchange between disjunct populations, resulting in high differentiation between geographical groups.     

Patterns and processes in the genetic differentiation of the Brachionus calyciflorus complex, a passively dispersing freshwater zooplankton

Elucidating the evolutionary patterns and processes of extant species is an important objective of any research program that seeks to understand population divergence and, ultimately, speciation. The island-like nature and temporal fluctuation of limnetic habitats create opportunities for genetic differentiation in rotifers through space and time. To gain further understanding of spatio-temporal patterns of genetic differentiation in rotifers other than the well-studied Brachionus plicatilis complex in brackish water, a total of 318 nrDNA ITS sequences from the B. calyciflorus complex in freshwater were analysed using phylogenetic and phylogeographic methods. DNA taxonomy conducted by both the sequence divergence and the GMYC model suggested the occurrence of six potential cryptic species, supported also by reproductive isolation among the tested lineages. The significant genetic differentiation and non-significant correlation between geographic and genetic distances existed in the most abundant cryptic species, BcI-W and Bc-SW. The large proportion of genetic variability for cryptic species Bc-SW was due to differences between sampling localities within seasons, rather than between different seasons. Nested Clade Analysis suggested allopatric or past fragmentation, contiguous range expansion and long-distance colonization possibly coupled with subsequent fragmentation as the probable main forces shaping the present-day phylogeographic structure of the B. calyciflorus species complex.     

中国南方双斑长跗萤叶甲地理种群遗传结构及Wolbachia感染

[目的]双斑长跗萤叶甲Monolepta hieroglyphica为多食性害虫,可取食为害多种农作物.本研究旨在探究中国南方地区分布的双斑长跗萤叶甲地理种群的遗传多样性、遗传结构及种群间的遗传分化程度与基因流水平,探究共生菌Wolbachia 在中国南方双斑长跗萤叶甲地理种群中的多样性和感染情况.[方法]以线粒体CO...     

Population genetics of the high elevation black fly Simulium (Nevermannia) feuerborni Edwards in Thailand

In this study, we examined the genetic structure and population history of the high elevation black fly Simulium feuerborni in Thailand at both cytogenetic and molecular genetic levels. Cytological examination revealed two cytoforms differentiated by fixed chromosome inversions. The distributions of the cytoforms were associated with geographic origins. Cytoform A was found in the lower north and northeast, and cytoform B was found in the upper northern region of Thailand. Molecular data based on the mitochondrial cytochrome oxidase subunit I (COI) barcoding sequence supports the separation of the cytoforms. The average sequence divergence between the two cytoforms was 3.75%, which is higher than the threshold value for the species level based on a COI barcoding sequence. Median joining network clearly differentiated the haplotypes of the cytoforms into different lineages. Population pairwise F_ST and amova analyses reveal significant genetic differentiation between cytoforms. This indicates that the low land areas separating these populations act as a gene flow barrier. No genetic differentiation was detected within cytoforms. This could be due to a recent sharing of population history. Mismatch distribution analysis revealed population expansion in the northern lineage of the cytoform B approximately 220 000 years ago. More recent expansion (32 000 years ago) was found in the lower north and northeast (cytoform A) lineage. The demographic history of S. feuerborni mirrored previous findings in black flies and other insect species in Thailand. This indicates the important role of Pleistocene climatic change on genetic structure and diversity of Southeast Asian mainland species.     

基于线粒体COI、Cytb和COII基因的中国草地螟不同地理种群遗传分化分析

【目的】揭示中国草地螟Loxostege sticticalis不同地理种群的遗传分化程度。【方法】采用PCR技术扩增中国西北和华北地区草地螟11个地理种群的线粒体 COI, Cytb和COII基因序列,基于其序列变异及单倍型贝叶斯系统发育树和单倍型网络图分析,探讨不同地理种群间的遗传距离、分子系统发生关系及遗传分化程度。【结果】草地螟11个地理种群的线粒体 COI, Cytb和COII基因序列分别有24, 12和69个变异位点(分别占总序列的3.6%, 2.7%和8.8%),检测到的单倍型分别为22, 14和16个,单倍型多样度(Hd)分别为0.7600, 0.5842和0.7341,核苷酸平均差异度(K)分别为1.704, 0.752和3.997,不同单倍型间的遗传距离平均值分别为0.004, 0.005和0.013。总种群的Tajima’s D和Fu’s Fs值皆不显著,表明草地螟不同地理种群间的遗传分化不明显,群体大小稳定。根据各地理种群的单倍型建立的系统发育树和单倍型网络图表明,各单倍型散布在不同的地理种群中,无明显的地理分布格局。【结论】草地螟各地理种群的遗传距离与地理距离间不具有显著的相关性,其遗传分化不明显。     

Multilevel studies on the two phenological forms of Large Blue (Maculinea arion) (Lepidoptera: Lycaenidae)

The main goal of our research was to study comprehensively the differences between the two phenological forms of the socially parasitic and globally threatened Large Blue (Maculinea arion) in the Carpathian Basin using four character sets (mitochondrial sequences, allozymes, male genitalia and wing morphometrics). Comparative analyses of distance matrices, phylogenetic trees and ordination patterns have been applied. The genetic and morphometric patterns revealed by our studies were discordant. While we experienced a significant differentiation between the ‘spring’ and ‘summer type’ of M. arion in both wing and genital traits, the two phenological forms did not show any genetic differentiation on two mitochondrial loci and in allozymes. At the same time, all individuals were infected by Wolbachia. Although certain wing traits may not represent reliable tracers of phylogeny because of the particular adaptive significance, the wing characteristics involved in our research are probably determined genetically. Additionally, the significant differentiation of male genitalia also indicates incipient prezygotic isolation arising from phenological differentiation between the ‘spring and summer arion’. It is possible that all extant differences between the two forms are attributable to (1) different host‐ant use, (2) incipient speciation, (3) cytoplasmatic incompatibility (CI) by Wolbachia or the combination of these factors. In addition, discordant results indicate that the combined use of different approaches and data sets is strictly necessary to clarify systematic and evolutionary relationships.     

DNA barcoding of tropical black flies (Diptera: Simuliidae) of Thailand

The ecological and medical importance of black flies drives the need for rapid and reliable identification of these minute, structurally uniform insects. We assessed the efficiency of DNA barcoding for species identification of tropical black flies. A total of 351 cytochrome c oxidase subunit 1 sequences were obtained from 41 species in six subgenera of the genus Simulium in Thailand. Despite high intraspecific genetic divergence (mean = 2.00%, maximum = 9.27%), DNA barcodes provided 96% correct identification. Barcodes also differentiated cytoforms of selected species complexes, albeit with varying levels of success. Perfect differentiation was achieved for two cytoforms of Simulium feuerborni, and 91% correct identification was obtained for the Simulium angulistylum complex. Low success (33%), however, was obtained for the Simulium siamense complex. The differential efficiency of DNA barcodes to discriminate cytoforms was attributed to different levels of genetic structure and demographic histories of the taxa. DNA barcode trees were largely congruent with phylogenies based on previous molecular, chromosomal and morphological analyses, but revealed inconsistencies that will require further evaluation.     

Calculation of Evolutionary Correlation between Individual Genes and Full-Length Genome: A Method Useful for Choosing Phylogenetic Markers for Molecular Epidemiology

Individual genes or regions are still commonly used to estimate the phylogenetic relationships among viral isolates. The genomic regions that can faithfully provide assessments consistent with those predicted with full-length genome sequences would be preferable to serve as good candidates of the phylogenetic markers for molecular epidemiological studies of many viruses. Here we employed a statistical method to evaluate the evolutionary relationships between individual viral genes and full-length genomes without tree construction as a way to determine which gene can match the genome well in phylogenetic analyses. This method was performed by calculation of linear correlations between the genetic distance matrices of aligned individual gene sequences and aligned genome sequences. We applied this method to the phylogenetic analyses of porcine circovirus 2 (PCV2), measles virus (MV), hepatitis E virus (HEV) and Japanese encephalitis virus (JEV). Phylogenetic trees were constructed for comparisons and the possible factors affecting the method accuracy were also discussed in the calculations. The results revealed that this method could produce results consistent with those of previous studies about the proper consensus sequences that could be successfully used as phylogenetic markers. And our results also suggested that these evolutionary correlations could provide useful information for identifying genes that could be used effectively to infer the genetic relationships.     

中国甜菜夜蛾地理种群的遗传分化与基因流

【目的】为了明确我国甜菜夜蛾Spodoptera exigua地理种群间的遗传分化及基因流,阐明该种害虫在我国的种群历史动态。【方法】本研究对采自我国20个地理种群的529头甜菜夜蛾样品进行线粒体COI基因序列测定与分析,利用DnaSP 5.0和Arlequin 3.11软件分析种群间遗传多样性、遗传分化、基因流水平及分子变异,构建了单倍型系统发育树与单倍型网络图。【结果】在所分析的所有529个序列样本中,共检测出10个单倍型,其中Hap_1为所有种群所共享。总群体遗传多样性较低（Hd=0.257±0.025,Pi=0.0007±0.0001,Kxy=0.323）,群体间遗传分化较小（F_ST=0.211）,基因流水平较高（N_m=1.870）。AMOVA分析表明,甜菜夜蛾遗传变异主要来自种群内,种群间变异水平较低。各种群间遗传分化程度与地理距离无显著相关性（R²=0.005,P>0.05）。各单倍型相互散布在不同种群中,未形成明显系统地理结构。中性检验（Tajima’s D=-2.177, P<0.05; Fu’s F_S=-8.629, P<0.05)与错配分布分析表明,我国甜菜夜蛾种群曾经历种群近期扩张。【结论】研究结果揭示,甜菜夜蛾各种群间的基因交流并未受到地理距离的影响,验证了甜菜夜蛾具有高度的迁飞能力。     

Evidence of Gene Conversion Events Between Paralogous Sequences Produced by Tetraploidization in Salmoninae Fish

We investigated the occurrence of gene conversions between paralogous sequences of Salmoninae derived from ancestral tetraploidization and their effect on the evolutionary history of DNA sequences. A microsatellite with long flanking regions (750 bp) including both coding and noncoding sequences was analyzed. Microsatellite size polymorphism was used to detect the alleles of both paralogous counterparts and infer linkage arrangement between loci. DNA sequencing of seven Salmoninae species revealed that paralogous sequences were highly differentiated within species, especially for noncoding regions. Ten gene conversion events between paralogous sequences were inferred. While these events appears to have homogenized regions of otherwise highly differential paralogous sequences, they amplified the differentiation among orthologous sequences. Their effects were larger on coding than on noncoding regions. As a consequence, noncoding sequences grouped by orthologous lineages in phylogenetic trees, whereas coding regions grouped by taxa. Based upon these results, we present a model showing how gene conversion events may also result in the PCR amplification of nonorthologous sequences in different taxa, with obvious complications for phylogenetic inferences, comparative mapping, and population genetic studies. Received: 11 October 2000 / Accepted: 18 September 2001     

High variation and very low differentiation in wide ranging plains zebra (Equus quagga): insights from mtDNA and microsatellites

Patterns of genetic differentiation in the plains zebra ( Equus quagga ) were analysed using mitochondrial DNA control region variation and seven microsatellites. The six morphologically defined subspecies of plains zebra lacked the population genetic structure indicative of distinct evolutionary units. Both marker sets showed high levels of genetic variation and very low levels of differentiation. There was no geographical structuring of mitochondrial DNA haplotypes in the phylogenetic tree, and the plains zebra showed the lowest overall differentiation recorded in any African ungulate studied so far. Arid-adapted African ungulates have shown significant regional genetic structuring in support of the Pleistocene refuge theory. This was not the case in the zebra, and the data are discussed in relation to the impact of Pleistocene climate change on a nonbovid member of the savannah ungulate community. The only other species showing a similar absence of genetic structuring is the African buffalo ( Syncerus caffer ), but this taxon lacks the high levels of morphological variation present in the plains zebra.     

Lack of genetic structure among Eurasian populations of the tick Ixodesricinus contrasts with marked divergence from north-African populations

Host-parasite interactions may select for significant novel mutations with major evolutionary consequences for both partners. In poor active dispersers such as ticks, their population structures are shaped by their host movements. Here, we use population genetics and phylogeography to investigate the evolutionary history of the most common tick in Europe, Ixodes ricinus, a vector of pathogenic agents causing diseases in humans and animals. Two mitochondrial and four nuclear genes were sequenced for 60 individuals collected on four geographical scales (local, regional, Eurasian and western Palearctic scales). The overall level of nucleotide diversity was low and the variability did not differ at the local, regional or Eurasian scales but increased two fold for the western Palearctic scale. Moreover, the phylogenetic trees indicated an absence of genetic structure among Eurasian ticks, contrasting with a strong differentiation of the north-African ticks which formed a divergent clade. The homogeneity in Eurasian ticks may be explained by gene flows due to passive dispersal of ticks by hosts within a continuous population and recent range expansion of I. ricinus as shown by the fit of the observed frequency distribution of numbers of mismatches between pairwise sequences with the demographic expansion model (Harpending raggedness index, P = 0.74). The genetic divergence of the north-African populations could be explained by genetic drift in these small populations that are geographically isolated and/or selection pressures due to different ecological conditions (seasonal activity, pathogenic agents and hosts communities). The consequences of these results on the epidemiology of vector-borne diseases are discussed.     

Cytotaxonomy,morphology and molecular systematics of the Bioko form of Simulium yahense (Diptera: Simuliidae)

Cytotaxonomic analysis of the polytene chromosomes from larvae of the Simulium damnosum Theobald complex from the island of Bioko in Equatorial Guinea is reported, and a new endemic cytoform is described. Chromosomally this cytoform is close to both S. squamosum (Enderlein) and S. yahense Vajime & Dunbar, but is not identical to either. However, it is morphologically and enzymatically identical to S. yahense. The Bioko form was also found to differ from other cytoforms of the S. damnosum complex in West Africa in the copy number or RFLP pattern of several different repetitive DNA sequences. It is clear that the Bioko form is genetically distinct from other populations of the S. damnosum complex, and whilst it is closest to S. yahense, it shows features that suggest a high degree of geographical and genetic isolation. Such isolation is an important consideration in the assessment of the potential for onchocerciasis vector eradication on Bioko.     

Genetic diversity and structure of a Mediterranean endemic plant in Corsica (<Emphasis Type="Italic">Mercurialis corsica</Emphasis>, Euphorbiaceae)

The island of Corsica is a Mediterranean hotspot of plant biodiversity characterized by a high rate of plant endemism, but also by a lack of studies combining genetic diversity and conservation. In Corsica, the dioecious and Corso-Sardinian endemic Mercurialis corsica Cosson (Euphorbiaceae) occurs across a wide ecological gradient, but the number of populations have decreased considerably over the last century. The main aim of this study was to examine the patterns of genetic diversity occurring in the Corsican populations of M. corsica, depending on their location and demographic structure. The rDNA sequences did not show the existence of any polymorphism, whereas the cpDNA sequences revealed the divergence of the western Corsican populations. By contrast, when the AFLP markers were examined, although significant levels of differentiation were detected between populations, no distinct geographical patterns were observed except for the pronounced isolation of the Cap Corse genotypes. No significant correlations were found to exist between population size and the genetic diversity indexes used. The results of this study suggest that M. corsica has undergone a complex gene flow history involving past population admixtures, followed by fragmentation processes resulting in population differentiation but no geographical patterns of isolation. These results support the existence of three evolutive conservation units which have to be monitored in priority to determine whether the current pattern of demographic structure is still declining or has stabilized.     

基于线粒体COI基因序列的黄胫小车蝗不同地理种群的遗传分化及基因流分析

黄胫小车蝗Oedaleus infernalis Saussure是一种在我国分布广泛、 对农牧业生产危害严重的经济害虫。本文对黄胫小车蝗10个地理种群的线粒体COI基因序列进行测序和分析, 利用DnaSP 5.0和Arlequin 3.5.1.2软件对该蝗虫种群间的遗传多样性、 遗传分化程度、 基因流水平及分子变异进行了分析, 建立了单倍型贝叶斯系统发育进化树和单倍型网络图。结果表明： 在所分析的144个序列样本中, 共检测到21种单倍型, 其中1种单倍型为10个地理种群所共享。总群体的单倍型多样性指数为0.653, 各地理种群单倍型多样度范围在0.423~0.790之间。总群体和各种群的Tajima’s D检验结果皆不显著, 说明该种害虫在较近的历史上未经历群体扩张。总群体的遗传分化系数Gst为0.04436, 固定系数Fst为0.05255, 基因流Nm为9.01。AMOVA分子方差分析结果表明, 黄胫小车蝗的遗传分化主要来自种群内部, 种群间的遗传变异水平较低。各地理种群的遗传距离的大小与其地理距离间没有显著的相关性。贝叶斯系统发育进化树与单倍型网络图显示, 黄胫小车蝗各地理种群中的单倍型散布在不同的分布群中, 分布格局较为混杂, 未形成明显的系统地理结构。研究结果揭示, 黄胫小车蝗各种群间的基因交流并未受到地理距离的影响。     

Genomic diversity and evolution of the lyssaviruses

Lyssaviruses are RNA viruses with single-strand, negative-sense genomes responsible for rabies-like diseases in mammals. To date, genomic and evolutionary studies have most often utilized partial genome sequences, particularly of the nucleoprotein and glycoprotein genes, with little consideration of genome-scale evolution. Herein, we report the first genomic and evolutionary analysis using complete genome sequences of all recognised lyssavirus genotypes, including 14 new complete genomes of field isolates from 6 genotypes and one genotype that is completely sequenced for the first time. In doing so we significantly increase the extent of genome sequence data available for these important viruses. Our analysis of these genome sequence data reveals that all lyssaviruses have the same genomic organization. A phylogenetic analysis reveals strong geographical structuring, with the greatest genetic diversity in Africa, and an independent origin for the two known genotypes that infect European bats. We also suggest that multiple genotypes may exist within the diversity of viruses currently classified as 'Lagos Bat'. In sum, we show that rigorous phylogenetic techniques based on full length genome sequence provide the best discriminatory power for genotype classification within the lyssaviruses.     

Hidden genetic diversity in the green alga Spirogyra (Zygnematophyceae, Streptophyta)

ABSTRACT: BACKGROUND: The unbranched filamentous green alga Spirogyra (Streptophyta, Zygnemataceae) is easily recognizable based on its vegetative morphology, which shows one to several spiral chloroplasts. This simple structure falsely points to a low genetic diversity: Spirogyra is commonly excluded from phylogenetic analyses because the genus is known as a long-branch taxon caused by a high evolutionary rate. RESULTS: We focused on this genetic diversity and sequenced 130 Spirogyra small subunit nuclear ribosomal DNA (SSU rDNA) strands of different origin. The resulting SSU rDNA sequences were used for phylogenetic analyses using complex evolutionary models (posterior probability, maximum likelihood, neighbor joining, and maximum parsimony methods). The sequences were between 1672 and 1779 nucleotides long. Sequence comparisons revealed 53 individual clones, but our results still support monophyly of the genus. Our data set did not contain a single slow-evolving taxon that would have been placed on a shorter branch compared to the remaining sequences. Out of 130 accessions analyzed, 72 showed a.     

Phylogeography of Simulium siamense Takaoka and Suzuki complex (Diptera: Simuliidae) in Thailand

The phylogeography of Simulium siamense complex was inferred from mitochondrial DNA sequences. A 586‐bp fragment of the cytochrome oxidase I was sequenced for 92 individuals from 13 populations throughout Thailand, representing five cytoforms (A, B, C, F and G). The cytoforms are not genetically different at the molecular level except for cytoform B, which is genetically distinct from the others. This might indicate that cytoform B is a distinct species. Further morphological and molecular work using other genes is needed to clarify this. Our results also argue for the need of integrated approach, both cytological and molecular studies to understanding biodiversity of black flies. The star‐like shape of the mtDNA genealogy is consistent with the sudden population expansion of the mismatch distribution analysis and large negative values of Fu's F_s and Tajima's D‐tests, indicating a population demographic expansion. The expansion time is estimated to be in the late Pleistocene (about 120 000 years ago). Therefore, the overall low level of genetic structure could be due to sharing a recent history. The ancestral haplotype was found in the mountainous area in northeastern Thailand, suggesting that this area could have been the refugium of the species complex during the Pleistocene glaciations. Our results are consistent with previous findings about population expansion in response to the Pleistocene climatic change, thus revealing the importance of this historical event in shaping the genetic structure and diversity of Southeast Asian mainland species.     

The green-veined white (Pieris napi L.), its Pierine relatives, and the systematics dilemmas of divergent character sets (Lepidoptera, Pieridae)

The butterfly Pieris napi (L.) and relatives exemplify recently evolving taxa, exhibiting variation that makes their evolutionary dynamics interesting, but their systematics difficult. Wing-pattern characters commonly used to distinguish these Holarctic insects display both genetic polymorphism and environmentally-cued polyphenism. Often, these causes of variation are confounded, impairing the characters' phylogenetic usefulness. DNA sequences of four mitochondrial genes offer an independent view of pierine phylogeny. Sampling diverse relatives within family Pieridae assists resolution of the P. napi complex, suggests previous underestimation of clade diversity in subfamily Pierinae, and shows that other genera near Pieris also display confusions of wing-pattern-based phylogenetic inference. The European P. napi is sister to all North American taxa, and is well diverged from them all in sequences. The North American taxa comprise a northern subclade including Pieris oleracea , and questionably distinct Pieris ' angelika ', and a southern subclade including distinct Pieris virginiensis , Pieris marginalis , and Pieris macdunnoughii , and other regional entities yet to be clarified. Weak bootstrap support for some nodes in this group arises from a closeness of sequence identity rather than character conflict; more sequence data and denser geographical sampling may resolve these nodes more clearly. Evidence of reproductive isolation, from other experimental hybridization studies, agrees with the DNA results where these conflict with other divergent character sets. The system offers much promise for a deeper understanding of character evolution in relation to phyletic differentiation.  © 2006 The Linnean Society of London, Biological Journal of the Linnean Society , 2006, 88 , 413–435.