Median-joining Haplotype Network Research Articles

Mitochondrial DNA Variation Reveals a Sharp Genetic Break within the Distribution of the Blue Land Crab Cardisoma guanhumi in the Western Central Atlantic.

The blue land crab Cardisoma guanhumi is widely distributed throughout tropical and subtropical estuarine regions in the Western Central Atlantic (WCA). Patterns of population genetic structure and historical demographics of the species were assessed by mtDNA control region sequence analysis to examine the connectivity among five populations (n = 97) within the region for future conservation strategies and decision-making of fishery management. A total of 234 polymorphic nucleotides were revealed within the sequence region, which have defined 93 distinct haplotypes. No dominant mtDNA haplotypes were found but instead a distribution of a few low-frequency recurrent haplotypes with a large number of singletons. A NJ-tree and a median-joining haplotype network revealed two distinct clusters, corresponding to individuals from estuaries located along the Caribbean Sea and Brazilian waters, respectively. AMOVA and FST statistics supported the hypothesis that two main geographic regions exists. Phylogeographical discontinuity was further demonstrated by the Bayesian assignment analysis and a significant pattern of isolation-by-distance. Additionally, tests of neutral evolution and analysis of mismatch distribution indicate a complex demographic history in the WCA, which corresponds to bottleneck and subsequent population growth. Overall, a sharp genetic break between Caribbean and Brazilian populations raised concerns over the conservation status of the blue land crab.

Cytogenetic and Molecular Evidence of Additional Cryptic Diversity in High Elevation Black fly Simulium feuerborni (Diptera: Simuliidae) Populations in Southeast Asia.

Simulium feuerborni Edwards is geographically widespread in Southeast Asia. Previous cytogenetic study in Thailand revealed that this species is a species complex composed of two cytoforms (A and B). In this study, we cytologically examined specimens obtained from the Cameron Highlands, Malaysia, and Puncak, Java, Indonesia. The results revealed two additional cytoforms (C and D) of S. feuerborni. Specimens from Malaysia represent cytoform C, differentiated from other cytoforms by a fixed chromosome inversion on the long arm of chromosome III (IIIL-5). High frequencies of the B chromosome (33-83%) were also observed in this cytoform. Specimens from Indonesia represent the cytoform D. This cytoform is differentiated from others by a fixed chromosome inversion difference on the long arm of chromosome II (IIL-4). Mitochondrial DNA sequences support genetic differentiation among cytoforms A, B, and C. The pairwise F(ST) values among these cytoforms were highly significantly consistent with the divergent lineages of the cytoforms in a median-joining haplotype network. However, a lack of the sympatric populations prevented us from testing the species status of the cytoforms.

Cytochrome Oxidase-I Sequence Based Studies of Commercially Available Pangasius Hypophthalmus in Italy

Pangasius hypophthalmus is one of the fish consumed in the Italian diet. It is farmed and imported from Mekong delta region of Vietnam. Among several types of Pangasius, Tra (Pangasius hypophthalmus) is permitted for sales by the European Union. Since these fish species are often allegedly substituted with other morphologically similar fish due to commercial benefits, authentication of the products in the international markets become often necessary to prevent fraud and safety issues. In addition, this fish is imported as fillets without skin and bone, thus leaving the consumer’s at the risk of buying a substandard nutritional food. In this article we present the molecular approach we developed to identify Pangasius hypophthalmus from other closely related species based on cytochrome oxidase-I (COI) mitochondrial barcoding gene and further described the variants in the studied population genetic of this species. Fifty-one samples of Pangasius hypophthalmus fillets labelled as Pangasio were obtained from various markets around Milan and their COI mitochondrial barcoding gene was sequenced and studied in our bioinformatics pipeline. All samples were successfully amplified and Basic Local Alignment Search Tool results of the amplified region confirmed that all sequences analysed belonged to Pangasius hypophthalmus. Based on the variations in their barcoding region single nucleotide polymorphisms were identified and delineative statistics was calculated on the sequences. Although Pangasius hypophthalmus is considered as a monophyly, seven polymorphisms were identified. The neighbour-joining tree and the Median-joining network of haplotypes showed for all the identified haplotypes a unique cluster, with the exception of one sample.

Effects of habitat specialization on population genetic structure of black fly Simulium weji Takaoka (Diptera: Simuliidae)

Ecologically specialized species often show disjunctive distribution because suitable habitats have patchy occurrence. Consequently, a high level of genetic differentiation is expected because gene flow is limited. In this study, we examined the population genetic structure of the habitat-specialized black fly Simulium weji using mitochondrial cytochrome c oxidase subunit II sequences. A high level of genetic structure was observed with all FST values showing highly significant differences. The median joining haplotype network revealed two genetically divergent lineages that associated with the different microhabitats. Therefore, our results indicate the important role of the ecological conditions in the habitats on black fly evolution. As the genetic differentiation has not been observed in previous cytogenetic studies, our results support an integrated approach as being necessary for black fly biodiversity assessments.

Support for a ‘Center of Origin’ in the Coral Triangle: Cryptic diversity, recent speciation, and local endemism in a diverse lineage of reef fishes (Gobiidae: Eviota)

The Coral Triangle is widely regarded as the richest marine biodiversity hot-spot in the world. One factor that has been proposed to explain elevated species-richness within the Coral Triangle is a high rate of in situ speciation within the region itself. Dwarfgobies (Gobiidae: Eviota) are a diverse genus of diminutive cryptobenthic reef fishes with limited dispersal ability, and life histories and ecologies that increase potential for speciation. We use molecular phylogenetic and biogeographic data from two clades of Eviota species to examine patterns, processes and timing associated with species origination within the Coral Triangle. Sequence data from mitochondrial and nuclear DNA were used to generate molecular phylogenies and median-joining haplotype networks for the genus Eviota, with emphasis on the E. nigriventris and E. bifasciata complexes – two species groups with distributions centered in the Coral Triangle. The E. nigriventris and E. bifasciata complexes both contain multiple genetically distinct, geographically restricted color morphs indicative of recently-diverged species originating within the Coral Triangle. Relaxed molecular-clock dating estimates indicate that most speciation events occurred within the Pleistocene, and the geographic pattern of genetic breaks between species corresponds well with similar breaks in other marine fishes and sessile invertebrates. Regional isolation due to sea-level fluctuations may explain some speciation events in these species groups, yet other species formed with no evidence of physical isolation. The timing of diversification events and present day distributions of Eviota species within the Coral Triangle suggest that both allopatric speciation (driven by ephemeral and/or ‘soft’ physical barriers to gene flow) and sympatric speciation (driven by niche partitioning and assortative mating) may be driving diversification at local scales within the Coral Triangle. The presence of multiple young, highly-endemic cryptic species of Eviota within the Coral Triangle suggests that (i) the Coral Triangle is indeed a “cradle” of reef fish biodiversity and that (ii) our current approximations of reef fish diversity in the region may be significantly underestimated.

Two kinorhynch species (Cyclorhagida, Echinoderidae, Echinoderes) show different distribution patterns across Tsugaru Strait, Northern Japan.

We investigated the geographic population structures of two intertidal kinorhynch species, Echinoderes sensibilis and Echinoderes sp. A, in the vicinity of Tsugaru Strait between Honshu and Hokkaido Islands, Japan, to examine whether the distribution or connectivity of populations of either species has been constrained by the strait. For each species, we examined the geographic distribution of COI haplotypes, constructed a median-joining haplotype network, and calculated statistics of genetic variation and connectivity. Tsugaru Strait is the northern range limit for E. sensibilis, which comprises a large, evolutionarily stable metapopulation that appears to have undergone a reduction in size followed by expansion; connectivity is low among most local populations, including across Tsugaru Strait. A divergent haplotype lineage showing no variation occurred only at Horozuki, suggesting recent immigration there from outside the study area. Echinoderes sp. A underwent a severe population bottleneck followed by rapid expansion. It occurred at all sampling sites on both sides of the strait, with high connectivity between populations across the strait. There is a zone of secondary contact between moderately divergent, presumably previously allopatric lineages in eastern Hokkaido. Present-day conditions in the strait have existed only for the past 8000 years, and differences in these species' distributions and apparent connectivity across the strait may relate to conditions existing in the strait when the species underwent population expansions or shifts in range; these historical events were not necessarily concurrent between the species, and occurred more than 8000 years ago. We discuss dispersal mechanisms for kinorhynchs, which could include suspension transport or rafting.

Haplotype diversity in mitochondrial DNA hypervariable region in a population of southeastern Brazil

Brazilian population derives from Native Amerindians, Europeans, and Africans. Southeastern Brazil is the most populous region of the country. The present study intended to characterize the maternal genetic ancestry of 290 individuals from southeastern (Brazil) population. Thus, we made the sequencing of the three hypervariable regions (HV1, HV2, and HV3) of the mitochondrial DNA (mtDNA). The statistical analyses were made using Arlequin software, and the median-joining haplotype networks were generated using Network software. The analysis of three hypervariable regios showed 230 (79.3 %) unique haplotypes and the most common haplotype was "263G" carried by 12 (4.1 %) individuals. The strikingly high variability generated by intense gene flow is mirrored in a high sequence diversity (0.9966 ± 0.0010), and the probability of two random individuals showing identical mtDNA haplotypes were 0.0068. The analysis of haplogroup distribution revealed that 36.9 % (n = 107) presented Amerindian haplogroups, 35.2 % (n = 102) presented African haplogroups, 27.6 % (n = 80) presented European haplogroups, and one (0.3 %) individual presented East Asian haplogroup, evidencing that the southeastern population is extremely heterogeneous and the coexistence of matrilineal lineages with three different phylogeographic origins. The genetic diversity found in the mtDNA control region in the southeastern Brazilian population reinforces the importance of increased national database in order to be important and informative in forensic cases.

Genetic Structure and Demographic History of Endemic Micronesian Blue Riffle Goby,Stiphodon caeruleus(Gobiidae) Inferred from Mitochondrial DNA Sequence Analysis

The Blue Riffle Goby, Stiphodon caeruleus (Gobiidae: Sicydiinae), is an amphidromous species endemic to the volcanic islands of Kosrae and Pohnpei, Federated States of Micronesia, in the eastern Caroline Islands. This study evaluated the population genetic structure and demographic history of S. caeruleus using mitochondrial NADH subunits 4 (ND4) and 4L (ND4L) DNA sequences. A total of 876 base pairs were successfully sequenced for 208 specimens from 15 localities. Twenty-three haplotypes were recovered with diversity indices of h = 0.489±0.043 and π = 0.097±0.012% that are lower than many other marine species. The median-joining network of haplotypes is star-like in formation and displayed no genetic structure. AMOVA results show that 99% of the genetic variation is found within populations, rather than between streams or islands. Mismatch distribution analysis and associated statistical tests indicate that a very recent demographic expansion has occurred for the species. Bayesian skyline plot confirms a gradual demographic expansion over the past 5000 years. This expansion is coincident with a reduction in sea level that produced coastal plains and shallow coastal waters, and human settlement of the islands. Our analyses suggest that populations of S. caeruleus among streams are historically unstructured for ND4 and ND4L haplotypes, that larvae may be presently recruited among streams of both islands, and that further nuclear DNA investigations are warranted to resolve potential genetic divergence of populations.

Mitochondrial phylogeography of a Beringian relict: the endemic freshwater genus of blackfish Dallia (Esociformes)

Mitochondrial genetic variability among populations of the blackfish genus Dallia (Esociformes) across Beringia was examined. Levels of divergence and patterns of geographic distribution of mitochondrial DNA lineages were characterized using phylogenetic inference, median-joining haplotype networks, Bayesian skyline plots, mismatch analysis and spatial analysis of molecular variance (SAMOVA) to infer genealogical relationships and to assess patterns of phylogeography among extant mitochondrial lineages in populations of species of Dallia. The observed variation includes extensive standing mitochondrial genetic diversity and patterns of distinct spatial segregation corresponding to historical and contemporary barriers with minimal or no mixing of mitochondrial haplotypes between geographic areas. Mitochondrial diversity is highest in the common delta formed by the Yukon and Kuskokwim Rivers where they meet the Bering Sea. Other regions sampled in this study host comparatively low levels of mitochondrial diversity. The observed levels of mitochondrial diversity and the spatial distribution of that diversity are consistent with persistence of mitochondrial lineages in multiple refugia through the last glacial maximum.

Extinction and recolonization of maritime Antarctica in the limpet Nacella concinna (Strebel, 1908) during the last glacial cycle: toward a model of Quaternary biogeography in shallow Antarctic invertebrates

Quaternary glaciations in Antarctica drastically modified geographical ranges and population sizes of marine benthic invertebrates and thus affected the amount and distribution of intraspecific genetic variation. Here, we present new genetic information in the Antarctic limpet Nacella concinna, a dominant Antarctic benthic species along shallow ice-free rocky ecosystems. We examined the patterns of genetic diversity and structure in this broadcast spawner along maritime Antarctica and from the peri-Antarctic island of South Georgia. Genetic analyses showed that N.concinna represents a single panmictic unit in maritime Antarctic. Low levels of genetic diversity characterized this population; its median-joining haplotype network revealed a typical star-like topology with a short genealogy and a dominant haplotype broadly distributed. As previously reported with nuclear markers, we detected significant genetic differentiation between South Georgia Island and maritime Antarctica populations. Higher levels of genetic diversity, a more expanded genealogy and the presence of more private haplotypes support the hypothesis of glacial persistence in this peri-Antarctic island. Bayesian Skyline plot and mismatch distribution analyses recognized an older demographic history in South Georgia. Approximate Bayesian computations did not support the persistence of N.concinna along maritime Antarctica during the last glacial period, but indicated the resilience of the species in peri-Antarctic refugia (South Georgia Island). We proposed a model of Quaternary Biogeography for Antarctic marine benthic invertebrates with shallow and narrow bathymetric ranges including (i) extinction of maritime Antarctic populations during glacial periods; (ii) persistence of populations in peri-Antarctic refugia; and (iii) recolonization of maritime Antarctica following the deglaciation process.

Phylogenetic Relationships among Populations of the Vineyard SnailCernuella virgata(Da Costa, 1778)

Cernuella virgata(Da Costa, 1778) (Mollusca: Hygromiidae), commonly known as the “vineyard snail,” is endemic species in Mediterranean and Western Europe including the British Isles, but in the Eastern USA and Australia it represents an introduced invasive species. The present work examines the genetic variability and phylogenetic relationships among the four populations of this land snail sampled along the east Adriatic region of Croatia using mitochondrial markers (partial 16S rDNA and COI gene) in addition to traditional methods of shell’s shape analysis. All the three molecular-phylogenetic approaches (median joining haplotype network analysis and Bayesian analysis, as well as maximum likelihood analysis) revealed two-three major subnetworks for both 16S rDNA and COI, with a clear distinction between south Adriatic haplotypes (Pisak) and north Adriatic haplotypes (Krk and Cres). The population from Karlobag was comprised of both north and south haplotypes, thus representing a putative contact zone between these two groups. The morphometric analysis showed that individuals from Cres island population were statistically significantly wider and higher than individuals from Pisak population. Analysis of the SW/SH ratio and the relationship between shell width and shell height showed no differences in shell growth between the two examined populations, indicating equal shell growth and shape, which gives the possibility that differences in size of individuals between those two populations could be influenced by biotic (physiological) or abiotic (environmental) factors. This study represents the first analysis of genetic variability and relatedness among native populations ofC. virgata.

Natural selection and population genetic structure of domain-I of Plasmodium falciparum apical membrane antigen-1 in India

Development of a vaccine against Plasmodium falciparum infection is an urgent priority particularly because of widespread resistance to most traditionally used drugs. Multiple evidences point to apical membrane antigen-1(AMA-1) as a prime vaccine candidate directed against P. falciparum asexual blood-stages. To gain understanding of the genetic and demographic forces shaping the parasite sequence diversity in Kolkata, a part of Pfama-1 gene covering domain-I was sequenced from 100 blood samples of malaria patients. Statistical and phylogenetic analyses of the sequences were performed using DnaSP and MEGA. Very high haplotype diversity was detected both at nucleotide (0.998±0.002) and amino-acid (0.996±0.001) levels. An abundance of low frequency polymorphisms (Tajima’s D=−1.190, Fu & Li’s D∗ and F∗=−3.068 and −2.722), unimodal mismatch distribution and a star-like median-joining network of ama-1 haplotypes indicated a recent population expansion among Kolkata parasites. The high minimum number of recombination events (Rm=26) and a significantly high dN/dS of 3.705 (P<0.0001) in Kolkata suggested recombination and positive selection as major forces in the generation and maintenance of ama-1 allelic diversity. To evaluate the impact of observed non-synonymous substitutions in the context of AMA-1 functionality, PatchDock and FireDock protein–protein interaction solutions were mapped between PfAMA-1-PfRON2 and PfAMA-1-host IgNAR. Alterations in the desolvation and global energies of PfAMA-1-PfRON2 interaction complexes at the hotspot contact residues were observed together with redistribution of surface electrostatic potentials at the variant alleles with respect to referent Pf3D7 sequence. Finally, a comparison of P. falciparum subpopulations in five Indian regional isolates retrieved from GenBank revealed a significant level of genetic differentiation (FST=0.084–0.129) with respect to Kolkata sequences. Collectively, our results indicated a very high allelic and haplotype diversity, a high recombination rate and a signature of natural selection favoring accumulation of non-synonymous substitutions that facilitated PfAMA-1-PfRON2 interaction and hence parasite growth in Kolkata clinical isolates.

Phylogenetic relationship and species delimitation of matsutake and allied species based on multilocus phylogeny and haplotype analyses

Tricholoma matsutake (S. Ito & S. Imai) Singer and its allied species are referred to as matsutake worldwide and are the most economically important edible mushrooms in Japan. They are widely distributed in the northern hemisphere and established an ectomycorrhizal relationship with conifer and broadleaf trees. To clarify relationships among T. matsutake and its allies, and to delimit phylogenetic species, we analyzed multilocus datasets (ITS, megB1, tef, gpd) with samples that were correctly identified based on morphological characteristics. Phylogenetic analyses clearly identified four major groups: matsutake, T. bakamatsutake, T. fulvocastaneum and T. caligatum; the latter three species were outside the matsutake group. The haplotype analyses and median-joining haplotype network analyses showed that the matsutake group included four closely related but clearly distinct taxa (T. matsutake, T. anatolicum, Tricholoma sp. from Mexico and T. magnivelare) from different geographical regions; these were considered to be distinct phylogenetic species.

Genetic population structure of the malaria vector Anopheles baimaii in north-east India using mitochondrial DNA

BackgroundAnopheles baimaii is a primary vector of human malaria in the forest settings of Southeast Asia including the north-eastern region of India. Here, the genetic population structure and the basic population genetic parameters of An. baimaii in north-east India were estimated using DNA sequences of the mitochondrial cytochrome oxidase sub unit II (COII) gene.MethodsAnopheles baimaii were collected from 26 geo-referenced locations across the seven north-east Indian states and the COII gene was sequenced from 176 individuals across these sites. Fifty-seven COII sequences of An. baimaii from six locations in Bangladesh, Myanmar and Thailand from a previous study were added to this dataset. Altogether, 233 sequences were grouped into eight population groups, to facilitate analyses of genetic diversity, population structure and population history.ResultsA star-shaped median joining haplotype network, unimodal mismatch distribution and significantly negative neutrality tests indicated population expansion in An. baimaii with the start of expansion estimated to be ~0.243 million years before present (MYBP) in north-east India. The populations of An. baimaii from north-east India had the highest haplotype and nucleotide diversity with all other populations having a subset of this diversity, likely as the result of range expansion from north-east India. The north-east Indian populations were genetically distinct from those in Bangladesh, Myanmar and Thailand, indicating that mountains, such as the Arakan mountain range between north-east India and Myanmar, are a significant barrier to gene flow. Within north-east India, there was no genetic differentiation among populations with the exception of the Central 2 population in the Barail hills area that was significantly differentiated from other populations.ConclusionsThe high genetic distinctiveness of the Central 2 population in the Barail hills area of the north-east India should be confirmed and its epidemiological significance further investigated. The lack of genetic population structure in the other north-east Indian populations likely reflects large population sizes of An. baimaii that, historically, were able to disperse through continuous forest habitats in the north-east India. Additional markers and analytical approaches are required to determine if recent deforestation is now preventing ongoing gene flow. Until such information is acquired, An. baimaii in north-east India should be treated as a single unit for the implementation of vector control measures.

Genetic structure of Pallas’s squirrel ( Callosciurus erythraeus) populations from artificial forests in Hongya County, Sichuan, China

To understand levels of population differentiation in Pallas’s squirrel ( Callosciurus erythraeus) in fragmented habitats, we collected 83 samples from three patches of artificial forest in Hongya County, Sichuan Province, China. Sample numbers from each patch were as follows: 16 from Hanwang (HW), 27 from Muchansi (MCS) and 40 from Yanyandong (YYD). The mitochondrial DNA control region was sequenced and 18 haplotypes were observed. Our results showed that haplotype diversities of the three C. erythraeus populations were similar (0.771, 0.791 and 0.733). Fixation indices ( F s t) of pairwise populations were between 0.21 and 0.31, and the estimated gene flow ( Nm) was between 1 and 2. Analysis of molecular variance (AMOVA) showed that most molecular variation occurred within populations (74.82%); variances among populations were small but there was significant genetic differentiation. In addition, the neighbour-joining (NJ) tree showed three clades in the phylogenetic tree for population genetic structure. This was confirmed by the median-joining haplotype network. Furthermore, analysis of isolation by distance (IBD) showed that genetic differentiation among the three populations was positively related to geographical distance. However, tests of neutrality and the observed mismatch distribution of pairwise differences between sequences indicated that C. erythraeus populations were relatively stable in the past.

Genome-Wide Polymorphism and Comparative Analyses in the White-Tailed Deer (Odocoileus virginianus): A Model for Conservation Genomics

The white-tailed deer (Odocoileus virginianus) represents one of the most successful and widely distributed large mammal species within North America, yet very little nucleotide sequence information is available. We utilized massively parallel pyrosequencing of a reduced representation library (RRL) and a random shotgun library (RSL) to generate a complete mitochondrial genome sequence and identify a large number of putative single nucleotide polymorphisms (SNPs) distributed throughout the white-tailed deer nuclear and mitochondrial genomes. A SNP validation study designed to test specific classes of putative SNPs provides evidence for as many as 10,476 genome-wide SNPs in the current dataset. Based on cytogenetic evidence for homology between cow (Bos taurus) and white-tailed deer chromosomes, we demonstrate that a divergent genome may be used for estimating the relative distribution and density of de novo sequence contigs as well as putative SNPs for species without draft genome assemblies. Our approach demonstrates that bioinformatic tools developed for model or agriculturally important species may be leveraged to support next-generation research programs for species of biological, ecological and evolutionary importance. We also provide a functional annotation analysis for the de novo sequence contigs assembled from white-tailed deer pyrosequencing reads, a mitochondrial phylogeny involving 13,722 nucleotide positions for 10 unique species of Cervidae, and a median joining haplotype network as a putative representation of mitochondrial evolution in O. virginianus. The results of this study are expected to provide a detailed template enabling genome-wide sequence-based studies of threatened, endangered or conservationally important non-model organisms.

Demographic history and incomplete lineage sorting obscure population genetic structure of the Texas mouse (Peromyscus attwateri)

Abstract The Texas mouse, Peromyscus attwateri, is a relatively habitat-specific species that exhibits a discontinuous distribution across the south-central United States. To examine the evolutionary history and contemporary population genetic structure of P. attwateri we sequenced an 1,100-base pair fragment of the mitochondrial cytochrome b gene for 210 individuals from 22 localities, resulting in 89 unique haplotypes. Low nucleotide diversity (π = 0.0043) and a median-joining haplotype network indicated low levels of divergence among haplotypes with little geographic structure. Demographic analyses indicated the presence of 2 significant range expansions: the 1st coinciding with the end of the last glacial maximum of the Pleistocene approximately 14 thousand years ago (kya) and the 2nd more recent expansion during the hypsithermal (9–5 kya) of the Holocene. Coalescent simulations under a model of no gene flow indicated that the lack of divergence among populations most likely is attributable to incom...

Population genetic structure of the caddisfly Rhyacophila pubescens, Pictet 1834, north of the Alps

In this study we analyse the genetic population structure of the caddisfly Rhyacophila pubescens, a montane aquatic insect with a Central European distribution range. The species exhibits an insular distribution pattern due to its strong binding to unimpaired tufa streams and thus to mountain ranges with calcareous geology. We examined sequence data (mtCOI) of 197 individuals from 33 sites of the northern part of the distribution range to elucidate whether genetic population structure reflects this insular distribution. 28 haplotypes were identified, one of which was central in the median-joining haplotype network, occurred all across the study area, and seemed to be ancestral to others. In almost every mountain we also found closely related, private haplotypes indicating recent differentiation processes. Exact tests of population differentiation and pairwise F ST values showed that most mountain ranges that were studied are significantly differentiated from one another, indicating limited gene flow between mountain ranges. Analysis of Molecular Variance (AMOVA) revealed that most variation was within (68.54 %) and among mountains (30.48 %, for both p < 0.0001), but not among major mountain ranges (0.98 %, p = 0.31). Analysis of demographic history inferred recent demographic expansion in every region of the study area and the entire data set. Together these results provide evidence that limited extant gene flow and isolation between populations are currently structuring the populations north of the Alps. We present different hypotheses how the present-day genetic pattern of R. pubescens could have developed from its contrasting pattern in the past.

Genetic evidence from Indian red jungle fowl corroborates multiple domestication of modern day chicken

BackgroundDomestication of chicken is believed to have occurred in Southeast Asia, especially in Indus valley. However, non-inclusion of Indian red jungle fowl (RJF), Gallus gallus murghi in previous studies has left a big gap in understanding the relationship of this major group of birds. In the present study, we addressed this issue by analyzing 76 Indian birds that included 56 G. g. murghi (RJF), 16 G. g. domesticus (domestic chicken) and 4 G. sonneratii (Grey JF) using both microsatellite markers and mitochondrial D-loop sequences. We also compared the D-loop sequences of Indian birds with those of 779 birds obtained from GenBank.ResultsMicrosatellite marker analyses of Indian birds indicated an average FST of 0.126 within G. g. murghi, and 0.154 within G. g. domesticus while it was more than 0.2 between the two groups. The microsatellite-based phylogenetic trees showed a clear separation of G. g. domesticus from G. g. murghi, and G. sonneratii. Mitochondrial DNA based mismatch distribution analyses showed a lower Harpending's raggedness index in both G. g. murghi (0.001515) and in Indian G. g. domesticus (0.0149) birds indicating population expansion. When meta analysis of global populations of 855 birds was carried out using median joining haplotype network, 43 Indian birds of G. g. domesticus (19 haplotypes) were distributed throughout the network sharing haplotypes with the RJFs of different origins.ConclusionOur results suggest that the domestication of chicken has occurred independently in different locations of Asia including India. We found evidence for domestication of Indian birds from G. g. spadiceus and G. g. gallus as well as from G. g. murghi, corroborating multiple domestication of Indian and other domestic chicken. In contrast to the commonly held view that RJF and domestic birds hybridize in nature, the present study shows that G. g. murghi is relatively pure. Further, the study also suggested that the chicken populations have undergone population expansion, especially in the Indus valley.