Genetic Divergence

The ability of a population to evolve in a changing environment may be compromised by human-imposed barriers to gene flow. We investigated the population structure and the possible occurrence of a genetic bottleneck in two isolated populations of the black-faced lion tamarin (Leontopithecus caissara), a species with very reduced numbers (less than 400) in a very restricted range in the Atlantic Forest of southeast Brazil. We determined the genotypes of 52 individuals across 9 microsatellite loci. We found genetic divergence between the populations, each exhibiting low genetic diversity. Analysis revealed broad-and fine-scale population structuring. Both populations have evidently experienced population reduction and a genetic bottleneck without presenting any apparent detrimental effect. Anyway, measures should be taken to effectively protect the forests where L. caissara occurs in order to allow its populations to increase and counteract the eventual effects of genetic impoverishment.

Genes involved in the reproductive isolation are particularly useful as molecular markers in speciation studies. Lutzomyia longipalpis (Diptera: Psychodidae: Phlebotominae), a putative species complex, is a vector of visceral leishmaniasis in Latin America. We isolated from this species a fragment homologous to cacophony, a Drosophila gene that encodes features of the lovesong, an acoustic signal that is important in the sexual isolation of closely related species and known to vary considerably among L. longipalpis putative siblings species. Using an intron of the sandfly cacophony as a marker, we analyzed the molecular variation and sequence divergence among five populations of L. longipalpis from Brazil, three allopatric (Jacobina, Lapinha and Natal) and two putative sympatric sibling species from the locality of Sobral. A high level of polymorphism was found and analysis of the data indicates that very little gene flow is occurring among the populations of Jacobina, Lapinha, and Natal. A high level of differentiation was also observed between the two putative sympatric species of Sobral, one of which seems to be the same sibling species found in Natal, while the other is somewhat more related to Jacobina and Lapinha. However, the amount of estimated gene flow among the Sobral siblings is about seven times higher than the previously estimated for period, another lovesong gene, perhaps indicating that introgression might be affecting cacophony more than period. The results suggest that L. longipalpis is not a single species in Brazil, but it is yet not clear whether the different populations studied deserve species status rather than representing an incipient speciation process.

Several nuclear RFLP loci have been discovered recently that exhibit extensive allele frequency variation among Norwegian coastal and north-east Arctic populations of Atlantic cod Gadus morhua. One of these polymorphisms was detected by hybridizing an anonymous cDNA ...

Twenty two mutants of Urdbean variety T9 were evaluated to assess genetic divergence for nine quantitative traits. Mutants were grouped in four clusters based on D values. Clusters II, II and IV were 2 divergent from the cluster, I, which contained the parent variety, T9. Cluster IV having a single mutant culture AAI-UB8 exhibited higher value for yield/plant, pod length, seeds/pod and 100-seed weight. Correlations coefficient measure the mutual relationship between various plant characters on which selection can be based for yield improvement. Path analysis revealed that pod length, pod cluster/plant and 100-seed weight had positive direct effects on seed yield/plant.

Restriction fragment length polymorphisms (RFLP) were used to assess genetic diversity of mitochondrial DNA (mtDNA) among 184 isolates of Fusarium proliferatum recovered from maize, asparagus, palms and reed. All strains were cross-fertile with standard mating type tester strains of Gibberella intermedia. Sixteen mitochondrial haplotypes were identified following digestion of DNAs with HaeIII, with seven, seven, five and six different haplotypes from maize, asparagus, palms and reed, respectively. Four haplotypes (I, III, IV and VII) were found on more than one host. Of these four, haplotype I was dominant on maize, representing 71% of the isolates. The banding patterns for haplotypes III and IV were >90% similar to the banding pattern of haplotype I. Haplotypes I, III and IV accounted for 87% of the isolates from maize, but were less common on the other hosts, accounting for 70%, 52% and 33% of the isolates from asparagus, palms and reed, respectively. Thirteen of the 16 haplotypes were recovered from only a single host plant species. When comparing the banding patterns and frequencies of these haplotypes, at least five were recovered at a higher frequency from one host relative to the others. Our results suggest that mtDNA RFLP analysis is a useful indicator of genetic divergence in Fusarium proliferatum.

Penaeoidea is a diverse group of economically important marine shrimps. Attention to the evolutionary history of the penaeoids has been raised since studies using mitochondrial DNA markers and sperm ultrastructure contradict classification of the penaeoid families based on morphology and hence challenge the long standing taxonomy of this superfamily. In this study, DNA sequences of two nuclear protein-coding genes, phosphoenolpyruvate carboxykinase and sodium-potassium ATPase alpha-subunit, were determined from 37 penaeoid genera to reconstruct the evolutionary relationships and to estimate divergence ages of the penaeoid shrimps. Phylogenetic analyses using maximum likelihood and Bayesian approaches strongly support the monophyly of Solenoceridae, Aristeidae and Benthesicymidae, but find Sicyoniidae nested within Penaeidae, making this family paraphyletic. Penaeoidea comprises two lineages: the former three families in one while the latter two in another. The diversification of the...

Genetic divergence of 40 lime accessions was estimated using D2 and principal component analysis. The genotypes under study fell into 6 clusters. The distribution pattern indicated that the maximum number of genotypes were included in cluster III and the minimum number in cluster VI. The inter cluster distance in most of the cases were higher than the intra cluster distance indicating wider genetic diversity among the accessions of different groups. The highest inter cluster distance was observed between cluster II and III (1568.51) followed by cluster II and VI (1223.72) showing wide diversity among the groups. The highest intra-cluster distance was observed for the cluster III and the lowest for the cluster VI. Leaf area and number of stamen showed maximum contribution towards total divergence among the genotypes.

The objective of this study was to evaluate the genetic divergence among the common bean group Carioca by the Tocher method (based on Mahalanobis distance) and graphic dispersion of canonic variables, aiming to identify populations with wide genetic variability. Eighteen genotypes were evaluated in four seasons using a randomized block design with four replications. The mean weight of 100 seeds, in three experiments, and the mean number of pods per plant, in one experiment, were the most important characteristics for the genetic divergence, representing more than 46% of the total variation in the first canonic variable. The first two canonic variables were sufficient to explain about 88.23% of the total variation observed in the average of the four environments. The results showed that CNFC 8008 and CNFC 8009 genotypes presented the best yield averages in all the experiments. While Pérola, Princesa and CNFC 8005 cultivars were the most dissimilar for morphoagronomic traits. Therefore, the combinations of Pérola)/CNFC 8008, CNFC 8005)/CNFC 8009, Pérola)/CNFC 8009, Princesa)/CNFC 8008 and Princesa)/CNFC 8009 were indicated for interpopulational breeding.

There has been a recent appreciation of the ecological impacts of zooplanktonic species invasions. The North American brine shrimp Artemia franciscana is one such alien invader in hyper-saline water ecosystems at a global scale. It has been shown to outcompete native Artemia species, leading to their local extinction. We used partial sequences of the mitochondrial Cytochrome c Oxidase Subunit 1 (COI or cox1) gene to investigate the genetic diversity and phylogeography of A. salina, an extreme halophilic sexual brine shrimp, over its known distribution range (Mediterranean Basin and South Africa) and to assess the extent of local endemism, the degree of population structure and the potential impact of traditional human saltpan management on this species. We also examined the phylogenetic relationships in the genus Artemia using COI sequences. Our results show extensive regional endemism and indicate an early Pleistocene expansion of A. salina in the Mediterranean Basin. Subsequent population isolation in a mosaic of Pleistocene refugia is suggested, with two or three refugia located in the Iberian Peninsula. Two instances of longdistance colonization were also observed. Surprisingly, given its strong phylogeographical structure, A. salina showed a signature of correlation between geographical and genetic distance. Owing to strong 'priority effects', extensive population differentiation is retained, despite dispersal via migrant birds and human management of saltpans. The foreseeable expansion of A. franciscana is likely to be followed by substantial loss of genetic diversity in Mediterranean A. salina. Large genetic divergences between Mediterranean and South African A. salina suggest that the latter deserves species status. Fig. 3 Statistical parsimony network showing the nested design used in NCPA. Only second (continuous thin line), third (continuous thick line) and statistically informative 1stlevel clades (discontinuous thin line) are shown. Sizes of ellipses are proportional to the number of individuals included (ellipses with numbers bottom left). The populations where each haplotype was found are shown next to the haplotype code. Lines linking haplotypes indicate single substitutions. Small black circles indicate missing haplotypes.

The southern Rocky Mountains and adjacent Intermontane Plateau Highlands region of western North America is a geographically diverse area with an active geologic history. Given the topological complexity and extensive geologic activity, organisms inhabiting this region are expected to show some degree of morphological and genetic divergence, especially populations found on the southern montane 'sky islands' of this region. Here we examine the phylogeographic history and diversification of a montane forest inhabiting harvestmen, Sclerobunus robustus, using a combination of genetic and morphological data. Divergence time estimates indicate that much of the diversification within and between major groups S. robustus predate the Pleistocene glacial cycles. The most widespread subspecies, Sclerobunus robustus robustus, is recovered as six genetically distinct, geographically cohesive mitochondrial phylogroups. Gene tree data for a single nuclear gene reveals congruent, albeit slightly more conservative, patterns of genetic divergence. Despite high levels of genetic divergence throughout their distribution, phylogroups show extreme conservation in somatic and reproductive morphology. This uncoupling of morphological and genetic differentiation may be due to morphological conservatism associated with a conserved microhabitat preference. Based on these data, it is obvious that S. robustus has undergone some level of cryptic diversification.

... 1998). In India 5000 ha are cultivated in Rajasthan, Madhya Pradesh, Andhra Pradesh and Uttar Pradesh (Kothari et al. 2003). ... KA Suri Natural Products Chemistry Division, Regional Research Laboratory (CSIR), Jammu Tawi 180001, India 123 ...

The Antarctic region is renowned for its isolated, unusual, diverse, and disharmonic marine fauna. Holothuroids are especially diverse, with 187 species (including 51 that are undescribed) recorded south of the Antarctic Convergence. This represents 4% of the documented Antarctic marine biota, and 10% of the world's holothuroid diversity. We present evidence that both inter-regional speciation with southern cold-temperate regions and intra-regional diversification has contributed to species richness. The Antarctic fauna is isolated, with few shallow-water Antarctic species known from north of the Convergence, yet several species show recent transgression of this boundary followed by genetic divergence. Interchange at longer time scales is evidenced by the scarcity of endemic genera (10 of 55) and occurrence of all six holothuroid orders within the region. While most Antarctic holothuroid morphospecies have circum-polar distributions, mtDNA sequence data demonstrate substantial geographic differentiation in many of these. Thus, most of the 37 holothuroid species recorded from shelf/slope depths in the Weddell Sea have also been found in collections from Prydz Bay and the Ross Sea. Yet 17 of 28 morphospecies and complexes studied show allopatric differentiation around the continent, on average into three divergent lineages each, suggesting that morphological data fails to reflect the level of differentiation. Interchange and local radiation of colonizers appear to have rapidly built diversity in the Antarctic, despite the potential of cold temperatures (and associated long generation times) to slow the rate of evolution.

Chinook salmon, Oncorhynchus tshawytscha, from the Sacramento River, California, USA were introduced to New Zealand between 1901 and 1907, and colonized most of their present-day range within about 10 years. The New Zealand populations now vary in phenotypic traits typically used to differentiate salmon populations within their natural range: growth in freshwater and at sea, age at maturity, dates of return to fresh water and reproduction, morphology, and reproductive allocation. This paper reviews a large research program designed to determine the relative contributions of phenotypic plasticity and genetic adaptation to this variation, in an effort to understand the processes underlying the natural evolution of new populations. We found strong evidence of trait divergence between populations within at most 30 generations, particularly in freshwater growth rate, date of return, and reproductive output, with plausible adaptive bases for these differences. Importantly, we also demonstrated not only a genetic basis for post-release survival but higher survival, and hence fitness, of a population released from its established site compared to another population released from the same site. We conclude that divergence of salmon in different rivers probably resulted initially from phenotypic plasticity (e.g., habitat-specific growth rates, and effects of upriver migration on ovarian investment). Philopatry (homing to natal streams) combined with rapid evolution of distinct breeding periods to restrict gene flow, facilitating divergence in other traits. We also suggest that in addition to genetic divergence resulting from random founder effects, divergence may also arise during the very early stages of colonization when the original colonists are a non-random, pre-adapted subset of the source population. This 'favored founders effect' immediately improves the fitness of the new population. Overall, this research reveals the complex interplay of environmental and genetic controls over behavior, physiology and life history that characterize the early stages of population differentiation, a process that has taken place repeatedly during the history of salmon populations.

ORIGINAL ARTICLE Carlos Flores Æ Nicole Maca-Meyer Æ Jose M. Larruga Vicente M. Cabrera Æ Naif Karadsheh Æ Ana M. Gonzalez Isolates in a corridor of migrations: a high-resolution analysis ... (2002); (18) Nasidze et al. (2005) cReferences before the slash were used for ...

Amphibians are in decline worldwide. However, their patterns of diversity, especially in the tropics, are not well understood, mainly because of incomplete information on taxonomy and distribution. We assess morphological, bioacoustic, and genetic variation of Madagascar's amphibians, one of the first near-complete taxon samplings from a biodiversity hotspot. Based on DNA sequences of 2,850 specimens sampled from over 170 localities, our analyses reveal an extreme proportion of amphibian diversity, projecting an almost 2-fold increase in species numbers from the currently described 244 species to a minimum of 373 and up to 465. This diversity is widespread geographically and across most major phylogenetic lineages except in a few previously well-studied genera, and is not restricted to morphologically cryptic clades. We classify the genealogical lineages in confirmed and unconfirmed candidate species or deeply divergent conspecific lineages based on concordance of genetic divergences with other characters. This integrative approach may be widely applicable to improve estimates of organismal diversity. Our results suggest that in Madagascar the spatial pattern of amphibian richness and endemism must be revisited, and current habitat destruction may be affecting more species than previously thought, in amphibians as well as in other animal groups. This case study suggests that worldwide tropical amphibian diversity is probably underestimated at an unprecedented level and stresses the need for integrated taxonomic surveys as a basis for prioritizing conservation efforts within biodiversity hotspots.

Xylopia emarginata Mart. é uma espécie de ocorrência nas matas de galerias, caracterizada pela sua tolerância às saturações hídricas potencializando-a para a recuperação dos ecossistemas ciliares. Com o objetivo de caracterizar geneticamente a espécie, foram estudadas duas populações naturais localizadas em campo rupestre, divididas naturalmente em cinco subpopulações, com altitude entre 940 e 1010 m, no município de Itumirim, MG (S 21º21'42,8"; W 44º46'0,52"). O processamento das amostras e as análises genéticas foram realizados no Departamento de Ciências Florestais da Universidade Federal de Lavras. A eqüidade gênica observada na subpopulação 1 foi de 36%, na 2 de 9%, na 3 de 27% e de 72% nas subpopulações 4 e 5, indicando que as subpopulações 1, 2 e 3 apresentam menor variabilidade genética. As heterozigosidades observadas (Ĥ o ) foram 0,484 e 0,722 e as esperadas (Ĥ e ) 0,395 e 0,425 para as populações A e B, respectivamente. Estimativas significativas do índice de fixação (f ) ocorreram nas subpopulações 1 (-0,389), 4 (-0,45) e 5 (-0,848), ao contrário daquelas encontradas nas subpopulações 2 (-0,105) e 3 (-0,209). Os índices de diversidade genética demonstram a existência de excesso de heterozigotos entre as subpopulações (F = -0,29). Somente as subpopulações 1 x 4, 2 x 3, 4 x 5 não apresentaram divergência genética. Os baixos níveis de fluxo alélico detectados entre algumas subpopulações possivelmente são devidos à presença do divisor d'águas, que pode estar atuando como barreira física. Elevada identidade genética foi observada entre as subpopulações 1 e 4, confirmando os resultados obtidos de fluxo alélico. As subpopulações 1, 4 e 5 são mais próximas geneticamente, mesmo estando mais distantes geograficamente, o que pode ser explicado pelo fluxo alélico. Dados dessa natureza são importantes para programas de coleta de sementes visando a conservação genética da espécie.

Endemic Ponto-Caspian gobies include a flock of 24 ‘‘neogobiin” species (containing the nominal genera
and subgenera Apollonia, Babka, Neogobius, Mesogobius, Ponticola, and Proterorhinus; Teleostei: Gobiidae),
of which a large proportion (5 species; 21%) recently escaped to invade other freshwater Eurasian systems
and the North American Great Lakes. We provide its first comprehensive phylogenetic and biogeographic
analysis based on 4709 bp sequences from two mitochondrial and two nuclear genes with
maximum parsimony, likelihood, and Bayesian approaches. We additionally compare its relationships
with the tadpole gobies (Benthophilus and Caspiosoma), which comprise a related endemic Ponto-Caspian
gobiid group; along with a variety of postulated relatives and outgroups. Results of all phylogenetic
approaches are highly congruent and provide very strong support for recognizing the subfamily Benthophilinae;
which encompasses both the ‘‘neogobiins” and tadpole gobies, and genetically diverges from
other Gobiidae subfamilies—including (non-monophyletic) Gobiinae and Gobinellinae. Benthophilinae
contains three tribes: Neogobiini (Neogobius, which is synonymized here with Apollonia; containing the
type species N. fluviatilis, along with N. melanostomus and N. caspius), Ponticolini (containing the genera
Mesogobius, Proterorhinus, Babka, and Ponticola—elevating the latter two from subgenera and removing
them from the formerly paraphyletic Neogobius), and Benthophilini (tadpole gobies). Within Ponticolini,
Proterorhinus and Mesogobius comprise the sister clade of the Ponticola and Babka clade. Further work is
needed to clarify the interrelationships of the tadpole gobies. Invasiveness is widespread in freshwater
and euryhaline taxa of Neogobius, Proterorhinus, Babka, and Ponticola; but not in marine species, Mesogobius,
or tadpole gobies.

The first molecular phylogenetic analysis of Asaccus reveals high sequence divergence among the sampled species. Asaccus montanus is recovered as the sister to all other species. Two Arabian species (A. gallagheri, A. platyrhynchus) form a monophyletic group as suggested earlier by morphological analyses. Our data show that A. gallagheri from Nizwa, Oman likely represents a new species. Samples of two species (A. elisae, A. griseonotus) from north of the Persian Gulf form a well-supported clade. The traditional concept of A. elisae likely represents more than one species, as the samples included here are paraphyletic with respect to A. griseonotus. The taxonomy of A. elisae awaits comparison with samples from the type locality in Iraq. Our inability to resolve the phylogenetic position of two species (A. caudivolvulus and A. nasrullahi) prevents testing biogeographic hypotheses involving the occurrence of Asaccus on either side of the Persian Gulf.

Background: Coral reefs are hotspots of biodiversity, yet processes of diversification in these ecosystems are poorly understood. The environmental heterogeneity of coral reef environments could be an important contributor to diversification, however, evidence supporting ecological speciation in corals is sparse. Here, we present data from a widespread coral species that reveals a strong association of host and symbiont lineages with specific habitats, consistent with distinct, sympatric gene pools that are maintained through ecologically-based selection.

The present study was conducted to assess the magnitude of genetic variability, heritability in broad sense and genetic advance among thirteen mung bean accessions for growth and grain yield characters. The study was conducted at the Haramaya University Rare research site and Hirna research site during the 2012/2013 cropping season. Randomized Complete Block Design with three replications was used for both locations. Each genotype was planted on a 1.2 X 2m plot consisting of four rows, which accommodates ten plants per row and thus 40 plants per plot. The result showed that number of primary branches, pods per plant, seeds per plant and harvest index had high genotypic and phenotypic coefficients of variation at both locations. For all characters, phenotypic coefficient of variation was higher than genotypic coefficient of variation indicating that there was environmental influence on these traits. The combined results for heritability showed that the high estimates of heritability and genetic advance were scored for seeds per plant and seed yield indicating that these characters were under the control of additive genetic effects. High genetic advance expected as percent of mean coupled with high heritability was observed for number of primary branches at Hirna, number of seeds per plant at Rare and number of secondary branches, pods per plant and 100seed weight for combined analysis. The present findings could be useful for establishing selection criteria for high seed yield in the mung bean breeding.

The tetraploid Allium altyncolicum (2n = 4x = 32) is considered to be of hybrid origin, because most of its morphologie al characters are intermediate between those of its putative parents, A. schoenoprasum and A. ledebourianum. In the present work an attempt has beeil made to ascertain its parentage by several methods: Giemsa C-banding, genomic in situ hybridization (GISH), PCR-RFLP of cpDNA, restriction enzyme mapping of the rDNA, and RAPDs. C-banding and GISH indicates c1early that A. altyncolicum is a segmental allopolyploid. Allium schoenoprasum and A. ledebourianum are the most likely the parental species and the larger part of the genome of A. altyncolicum (26 chromosömes) is derived from A. schoenoprasum. The low genetic divergence between these three species was confirmed by the lack of sequence variation in the ITS sequences of nuc1ear rRNA genes and of the plastid rbcL-atpB intergenie spacer. Both parental species and A. altyncolicum could be distinguished by RFLP of the rDNA repeats. The geographie origin of the putative parental species was investigated using RAPDs.

Phylogenetic analysis using up to 1325 base pairs of mitochondrial DNA from 179 specimens and 30 species of Chalcides, Sphenops, Eumeces, Scincopus and Scincus indicates that Sphenops arose twice independently within Chalcides. It is consequently synonymized with that genus. Chalcides in this broader sense originated in Morocco, diversifying into four main clades about 10 Ma, after which some of its lineages dispersed widely to cover an area 40 times as large. Two separate lineages invaded the Canary Islands and at least five main lineages colonized southern Europe. At least five more spread across northern Africa, one extending into southwest Asia. Elongate bodies with reduced limbs have evolved at least four times in Chalcides, mesic 'grass-swimmers' being produced in one case and extensive adaptation to life in loose desert sand in two others.

South African Homopholis Boulenger, 1885 and Madagascan Blaesodactylus Boettger, 1893 are gekkonid genera originally distinguished by minor differences in scalation. Subsequent morphological studies have argued either for synonymy of the genera, or retention of the original 19 th century taxonomy. Moreover, the validity and boundaries of putative species within each genus, as well as the relationships of each genus to other gekkonid genera, remains unresolved and contentious. We sequenced 3433 aligned bp of mitochondrial (ND2 and ND4) and nuclear (RAG-1 and PDC) DNA sequence data from 14 samples of Homopholis and Blaesodactylus representing all six recognized species in these two genera, and based on a larger, unpublished study of all gekkotan relationships, eight samples of Geckolepis and two species of Afroedura for outgroups. Results indicated a strongly supported, monophyletic clade including Geckolepis as the sister group to Blaesodactylus + Homopholis, with the latter two genera reciprocally monophyletic. Given the clear morphological and biogeographic distinctness of Homopholis sensu stricto from Blaesodactylus, as well as deep genetic divergence between the two groups, we favor the retention of Blaesodactylus as the valid generic name for the Madagascan species. Homopholis mulleri is confirmed as a valid species, and evidence of cryptic speciation is suggested among divergent samples of H. walbergii (often misspelled in the literature), and among samples of Blaesodactylus and Geckolepis. The pattern observed among South African samples of H. walbergii suggests an important role for the Soutpansberg as a barrier to gene flow.

Acanthochromis polyacanthus is an unusual tropical marine damselfish that uniquely lacks pelagic larvae and has lost the capacity for broad-scale dispersal among coral reefs. On the modern Great Barrier Reef (GBR), three color morphs meet and hydridize at two zones of secondary contact. Allozyme electrophoreses revealed strong differences between morphs from the southern zone but few differences between morphs from the northern counterpart, thus suggesting different contact histories. We explore the phylogeography of Acanthochromis polyacanthus with mitochondrial cytochrome b region sequences (alignment of 565 positions) obtained from 126 individuals representing seven to 12 fish from 13 sites distributed over 12 reefs of the GBR and the Coral Sea. The samples revealed three major clades: (1) black fish collected from the southern GBR; (2) bicolored fish collected from the GBR and one reef (Osprey) from the northern Coral Sea; (3) black and white monomorphs collected from six reefs in the Coral Sea. All three clades were well supported (72-100%) by bootstrap analyses. Sequence divergences were very high between the major clades (mean ϭ 7.6%) as well as within them (2.0-3.6%). Within clades, most reefs segregated as monophyletic assemblages. This was revealed both by phylogenetic analyses and AMOVAs that showed that 72-90% of the variance originated from differences among groups, whereas only 5-13% originated within populations. These patterns are discussed in relation to the known geological history of coral reefs of the GBR and the Coral Sea. Finally, we ask whether the monospecific status of Acanthochromis should be revisited because the sequence divergences found among our samples is substantially greater than those recorded among well-recognized species in other reef fishes.

The aim of this research was to study the mating system of a natural population of Theobroma grandiflorum (cupuassu) from Nova Ipixuna, Pará state, using microsatellite markers. Eight polymorphic microsatellite loci were analyzed in eight families, each represented by 10 six-month old seedlings derived from open-pollinated pods. The estimation for the multilocus outcrossing rate ( $ t m = 1.0) and individual outcrossing rate ( $ t = 1.0) for this population suggests that T. grandiflorum may be a perfect outbreeding (allogamous) species. Likewise, for the studied population the estimate for single locus outcrossing rate ( $ t s ) was elevated (0.946), but lower than $ t m , confirming the likely outcrossing character of the species and suggesting the occurrence of 5.4% biparental inbreeding rate ( $ t m -$ t s ). The estimation of genetic divergence ( $ F ST ) between allelic frequencies in ovules and pollen revealed a deviation from random mating in 75% of the evaluated loci. Likewise, the estimate of correlation of paternity ( $ r p = 0.930) and the mean coefficient of co-ancestrality within families ( $ θ xy = 0.501) indicated that the outcrossings were predominantly correlated, and the offspring were full-sibs. These results suggested that for this particular population of T. grandiflorum, the sampling strategy for genetic conservation and breeding should adopt specific models for families derived from correlated outcrossing (full-sibs) and not the ones usually adopted in classic outcrossing species breeding programs (half-sibs).

Porcine reproductive and respiratory syndrome (PRRS) is an immunosuppressive chronic respiratory viral disease of pigs that is responsible for major economic losses to the swine industry worldwide. The efficacy of parenteral administration of widely used modified live virus PRRS vaccine (PRRS-MLV) against genetically divergent PRRSV strains remains questionable. Therefore, we evaluated an alternate and proven mucosal immunization approach by intranasal delivery of PRRS-MLV (strain VR2332) with a potent adjuvant to elicit cross-protective immunity against a heterologous PRRSV (strain MN184). Mycobacterium tuberculosis whole cell lysate (Mtb WCL) was chosen as a potent mucosal adjuvant due to its Th1 biased immune response to PRRS-MLV. Unvaccinated pigs challenged with MN184 had clinical PRRS with severe lung pathology; however, vaccinated (PRRS-MLV+ Mtb WCL) pigs challenged with MN184 were apparently healthy. There was a significant increase in the body weight gain in vaccinated compared to unvaccinated PRRSV challenged pigs. Vaccinated compared to unvaccinated, virus-challenged pigs had reduced lung pathology associated with enhanced PRRSV neutralizing antibody titers and reduced viremia. Immunologically, an increased frequency of Th cells, Th/memory cells, ␥␦ T cells, dendritic cells, and activated Th cells and a reduced frequency of T-regulatory cells were detected at both mucosal and systemic sites. Further, reduced secretion of immunosuppressive cytokines (IL-10 and TGF-␤) and upregulation of the Th1 cytokine IFN-␥ in blood and lungs were detected in mucosally vaccinated, PRRSVchallenged pigs. In conclusion, intranasal immunization of pigs with PRRS-MLV administered with Mtb WCL generated effective cross-protective immunity against PRRSV.

Bats use various roost types with a wide spectrum of ecological features. The greater mouse-eared bat Myotis myotis (Borkhausen, 1797), creates nurseries in attics and caves in Central Europe. The stable low temperature and high humidity cave microclimate contrasts that of attics, which may alter species adaptations and life strategies. We analysed population characteristics (composition, body condition, parasite load, and immune response) and genetic relatedness of two proximal M. myotis populations. Age, sexual and parasite species composition were similar between the cave and attic sites. However, a significantly higher parasite load and body condition was detected in the post-partum females and juveniles of the cave colony (n = 263 bats from the cave, 231 from the attic), with the cave colony females having a significantly stronger immune response (n = 2 caves and 2 attics, 20 females per site). There was no evidence for genetic divergence between cave and attic populations (n = 3 caves and 3 attics, 24 females per site), indicating that different population characteristics are not genetically based and that M. myotis is an example of a species with rather unique phenotypic plasticity.

Mesoamerica, defined as the broad linguistic and cultural area from middle southern Mexico to Costa Rica, might have played a pivotal role during the colonization of the American continent. The Mesoamerican isthmus has constituted an important geographic barrier that has severely restricted gene flow between North and South America in pre-historical times. Although the Native American component has been already described in admixed Mexican populations, few studies have been carried out in native Mexican populations. In this study, we present mitochondrial DNA (mtDNA) sequence data for the first hypervariable region (HVR-I) in 477 unrelated individuals belonging to 11 different native populations from Mexico. Almost all of the Native Mexican mtDNAs could be classified into the four pan-Amerindian haplogroups (A2, B2, C1, and D1); only two of them could be allocated to the rare Native American lineage D4h3. Their haplogroup phylogenies are clearly star-like, as expected from relatively young populations that have experienced diverse episodes of genetic drift (e.g., extensive isolation, genetic drift, and founder effects) and posterior population expansions. In agreement with this observation, Native Mexican populations show a high degree of heterogeneity in their patterns of haplogroup frequencies. Haplogroup X2a was absent in our samples, supporting previous observations where this clade was only detected in the American northernmost areas. The search for identical sequences in the American continent shows that, although Native Mexican populations seem to show a closer relationship to North American populations, they cannot be related to a single geographical region within the continent. Finally, we did not find significant population structure in the maternal lineages when considering the four main and distinct linguistic groups represented in our Mexican samples (Oto-Manguean, Uto-Aztecan, Tarascan, and Mayan), suggesting that genetic divergence predates linguistic diversification in Mexico.

We investigated genetic and morphological diversity and population structure of 14 traditional sheep populations originating from four ecological zones in Ethiopia (sub-alpine, wet highland, sub-humid lowland and arid lowland). All animals (n ¼ 672) were genotyped for 17 microsatellite markers and scored for 12 morphological characters. The sheep were initially classified as fat-tailed (11 populations), thin-tailed (one population) and fat-rumped sheep (two populations). These classifications are thought to correspond to three consecutive introduction events of sheep from the Near-East into East Africa. For the 14 populations, allelic richness ranged from 5.87 to 7.51 and expected heterozygosity (H E ) from 0.66 to 0.75. Genetic differentiations (F ST values) between all pairs of populations, except between sub-alpine populations, were significantly different from zero (P < 0.001). Cluster analysis of morphological characters and a dendrogram constructed from genetic distances were broadly consistent with the classification into fat-tailed, thin-tailed and fat-rumped sheep. Bayesian cluster analysis using microsatellite markers indicated that there has been further genetic differentiation after the initial introduction of sheep into Ethiopia. Investigation of factors associated with genetic variation showed that an isolationby-distance model, independently of other factors, explained most of the observed genetic variation. We also obtained a strong indication of adaptive divergence in morphological characters, patterns of morphological variation being highly associated with ecology even when the effect of neutral genetic divergence (F ST ) was parcelled out in partial Mantel tests. Using a combination of F ST values, Bayesian clustering analysis and morphological divergence, we propose a classification of Ethiopian sheep into six breed groups and nine breeds.

Electrophoretic analysis of 17 enzyme loci was carried out to assess the genetic differentiation and isolation of the Sardinian Polyommatus coridon gennargenti from R c. apennina of peninsular Italy and the related taxon P caelestissimus from central Spain. I? c. gennargenti is represented by a small, strongly inbred population, restricted to the central mountains of Sardinia, and probably derived from mainland populations as indicated by the reduction of genetic variation (P 17.6 per cent, H 0.024) with respect to the continental populations (P>52 per cent, H 0.170). Absence of gene flow is indicated by the presence of alternative fixed alleles at the Aat, Gpi and Pgm loci and significant differences in allele frequencies at other loci, which distinguish the Sardinian population from 1? C. apennina and P caelestissimus. The genetic differentiation of P c. gennargenti, as measured using Wright's FST values and Nei's genetic distances, suggests the evolution of the Sardinian taxon along an independent lineage, facilitated by isolation and the strict dependence of the butterflies on specific biotopes, thus confirming its taxonomic status.

The genetic divergence time between two species varies substantially across the genome, conveying important information about the timing and process of speciation. Here we develop a framework for studying this variation and apply it to about 20 million base pairs of aligned sequence from humans, chimpanzees, gorillas and more distantly related primates. Human-chimpanzee genetic divergence varies from less than 84% to more than 147% of the average, a range of more than 4 million years. Our analysis also shows that human-chimpanzee speciation occurred less than 6.3 million years ago and probably more recently, conflicting with some interpretations of ancient fossils. Most strikingly, chromosome X shows an extremely young genetic divergence time, close to the genome minimum along nearly its entire length. These unexpected features would be explained if the human and chimpanzee lineages initially diverged, then later exchanged genes before separating permanently.

We completed a taxonomic revision for the New Zealand Copper Skink (Cyclodina aenea) species complex using morphological and molecular data. Two new species are described on the basis of several morphological characteristics, with the specific status of each species supported by mitochondrial sequence data (ND2). We also redescribe C. aenea. One of the new species is restricted to the Poor Knights Islands, whereas the distribution of the other new species is limited to northernmost region of Northland. Both new species exhibit significant genetic divergence from C. aenea (,13.5% sequence divergence), indicating that each species has evolved in isolation from C. aenea for a substantial period of time.

Ceuthophilus spp. (Orthoptera: Rhaphidophoridae) in two subgenera (Ceuthophilus and Geotettix) were examined phylogenetically, morphologically, and biogeographically in order to improve our understanding of these species that carry nutrients into caves and are thus closely tied to rare and endangered troglobites in central Texas. Crickets were collected from 43 caves across central Texas, and outgroup taxa were collected in west Texas, Kentucky, Missouri, New Mexico, and Mexico. We analyzed 1263 base pairs of mitochondrial DNA from the COI and ND5 genes and complemented this data with morphometric analyses of 19 morphological characters. Phylogenetic trees from the molecular analysis allowed us to select representatives from 25 clades for morphological work.

The Afrotropical leaf-nosed bat Hipposideros caffer has been traditionally regarded as a complex of populations, currently pertaining to two recognized cryptic species, H. caffer and H. ruber. Extent of distribution and morphological variation of these bats has raised concerns over whether the current perception of the complex reflects true phylogenetic relationships and taxonomic diversity. Our phylogenetic analysis of nucleotide sequences of the mitochondrial cytochrome b gene challenged the hypothesis of two cryptic species. Instead of the two reciprocally monophyletic lineages expected, corresponding to the two species, we recovered four distinct lineages with deep internal divergences. Two sister clades within a lineage of bats of H. caffer represent respectively the nominotypical form H. c. caffer, restricted to Southern Africa, and H. c. tephrus, inhabiting the Maghreb, West Africa and the Arabian Peninsula. Geographical isolation and deep genetic divergence suggest species status of both the forms. Another lineage comprises specimens of both morphotypes from West and East Africa. It probably represents a distinct species but its taxonomic assignation remains obscure. A Central African lineage of H. ruber comprises two sister clades, which become sympatric in Cameroon. Their status has to be clarified with additional evidence, since nuclear gene flow might be taking place. A further divergent lineage with H. ruber morphotype, most probably representing another distinct species, is restricted to West Africa. Although all three genetic forms of H. ruber may correspond to named taxa, their proper taxonomic assignation has to be assessed by comparison with type material.

New molecular (allozyme and 16S mtDNA sequence) and morphological data on Euscorpius italicus (Herbst, 1800) (Scorpiones: Euscorpiidae) reveal two groups of populations. One including E. italicus from Italy, Switzerland, and Pindos Mts (Greece), and another, Greek populations from the Zakynthos Island (formerly described as E. i. zakynthi and the Peloponnese (formerly described as Scorpius naupliensis C. L. Koch, 1837). The genetic divergence between these two groups was similar or higher to that observed between other congeneric species, i.e. fixed for private alleles at eight out of 18 allozyme loci and ~5% mtDNA sequence divergence. A morphological study of the material covering the entire range of E. italicus (Italy, ) is consistent with genetic data. Euscorpius naupliensis (C. L. Koch, 1837) (=E. i. zakynthi Caporiacco, 1950, syn. n.) from Greece is restored from synonymy and elevated to the rank of species. It is diagnosed by a number of morphological features, i.e. the absence of the esb a external trichobothrial series on the pedipalp patella, position of trichobothria on the pedipalp fixed finger, and morphometric ratios.

We investigated phylogeography of Philippine slender toads (genus Ansonia) and used a temporal framework for diversification, statistical tests of alternate topologies, and Bayesian approaches to test previous hypotheses concerning dispersal to, and colonization routes within, the southern Philippine island of Mindanao. Two species of Ansonia previously have been documented, with ranges separated by an east-west split corresponding to the approximate boundaries of Mindanao's paleoisland precursors. We present new mtDNA sequence data (1946 bp from genes encoding ND1, 16S rRNA and tRNA Leu ) for 105 Ansonia specimens sampled from 20 localities on Mindanao Island. Our data suggest that Philippine Ansonia is composed of at least eight, well-supported population lineages, structured into a minimum of four highly divergent mtDNA clades. One clade corresponds to Ansonia mcgregori, a range-restricted species apparently limited to the distal portion of the Zamboanga Peninsula of western Mindanao. Two morphologically indistinguishable, but genetically divergent, lineages possibly are undescribed cryptic species from western Mindanao. We recognize the five remaining lineages as Ansonia muelleri pending data from morphology or bioacoustics that might diagnose separate species among these lineages. Regardless of their species status, the five allopatric lineages of A. muelleri should be viewed as important genetic units for future genetic conservation planning.

We examine the effects of isolation over both ancient and contemporary timescales on evolutionary diversification and speciation patterns of springtail species in circum-Antarctica, with special focus on members of the genus Cryptopygus (Collembola, Isotomidae).

Reintroductions are a common approach for preserving intraspecific biodiversity in fragmented landscapes. However, they may exacerbate the reduction in genetic diversity initially caused by population fragmentation because the effective population size of reintroduced populations is often smaller and reintroduced populations also tend to be more geographically isolated than native populations. Mixing genetically divergent sources for reintroduction purposes is a practice intended to increase genetic diversity. We documented the outcome of reintroductions from three mixed sources on the ancestral composition and genetic variation of a North American fish, the slimy sculpin (Cottus cognatus). We used microsatellite markers to evaluate allelic richness and heterozygosity in the reintroduced populations relative to computer simulated expectations. Sculpins in reintroduced populations exhibited higher levels of heterozygosity and allelic richness than any single source, but only slightly higher than the single most genetically diverse source population.