Determining Management Units in Southeastern Brazil: The Case of Astyanax bimaculatus (Linnaeus, 1758) (Teleostei: Ostariophysi: Characidae)

Characterization, management and protection of biodiversity are the most prominent challenges in conservation biology. Analyses on molecular similarity (Random Amplified Polymorphic DNA-Polymerase Chain Reaction, RAPD-PCR) and morphologic patterns (morphometric and meristic characters) were performed to evaluate the existence of population structuring in the Doce river basin, in a widespread small-sized characin, Astyanax bimaculatus. Three hundred (300) individuals were collected from six locations isolated by waterfalls or dams, in the Doce river basin, Minas Gerais State, Brazil. Genetic differentiation among tributaries was significant (p<0.00001) and accounted for 21% of total variance (p<0.00001). Only the Grande waterfall of the Casca river was an effective factor for differentiation of populations (p<0.00001), and meristic characters were also consistent with molecular data. We concluded that genetic and morphologic variation of this species was not necessarily associated with waterfalls and that molecular and meristic data are effective predictors of population divergence in this basin. The consequences of these findings for the protection and management of aquatic biodiversity were discussed.     

Classical and molecular cytogenetics of Atherinella brasiliensis (Teleostei, Atheriniformes) from South coast of Brazil

Cytogenetic studies were performed on specimens of Atherinella brasiliensis from Laranjeiras Bay (Paraná State, Brazil). All specimens had a diploid number of 48 chromosomes, with a karyotype constituted by 4m+14sm+18st+12a and fundamental number of 84. The C-positive heterochromatin was distributed over the nucleolar organizer regions (NORs) in the centromeric regions and on short arms of metacentric and submetacentric chromosomes. Most of this heterochromatin was AT-rich, except in the NORs, which were rich in GC, as detected by double staining with chromomycin A₃/4'-6-diamin-2-phenylindole. Single NORs were located at terminal positions of a submetacentric pair, as confirmed by fluorescent in situ hybridization with 18S rDNA probes. Both techniques showed a size heteromorphism between the homologous chromosomes. The 5S rDNA clusters were located in terminal positions on two chromosomal pairs and also displayed a size heteromorphism. Despite the conserved diploid number, the data on the karyotype microstructure help characterize the cytogenetic profile of this group.     

Life on the margin: genetic isolation and diversity loss in a peripheral marine ecosystem, the Baltic Sea

Marginal populations are often isolated and under extreme selection pressures resulting in anomalous genetics. Consequently, ecosystems that are geographically and ecologically marginal might have a large share of genetically atypical populations, in need of particular concern in management of these ecosystems. To test this prediction, we analysed genetic data from 29 species inhabiting the low saline Baltic Sea, a geographically and ecologically marginal ecosystem. On average Baltic populations had lost genetic diversity compared to Atlantic populations: a pattern unrelated to dispersal capacity, generation time of species and taxonomic group of organism, but strongly related to type of genetic marker (mitochondrial DNA loci had lost c. 50% diversity, and nuclear loci 10%). Analyses of genetic isolation by geographic distance revealed clinal patterns of differentiation between Baltic and Atlantic regions. For a majority of species, clines were sigmoid with a sharp slope around the Baltic Sea entrance, indicating impeded gene flows between Baltic and Atlantic populations. Some species showed signs of allele frequencies being perturbed at the edge of their distribution inside the Baltic Sea. Despite the short geological history of the Baltic Sea (8000 years), populations inhabiting the Baltic have evolved substantially different from Atlantic populations, probably as a consequence of isolation and bottlenecks, as well as selection on adaptive traits. In addition, the Baltic Sea also acts a refuge for unique evolutionary lineages. This marginal ecosystem is thus vulnerable but also exceedingly valuable, housing unique genes, genotypes and populations that constitute an important genetic resource for management and conservation.     

Population Genetic Structure of Astyanax scabripinnis (Teleostei, Characidae) from an Urban Stream

Despite the great anthropogenic interference on urban streams, information is still scarce about the genetic variability and structure of native fish populations inhabiting such streams. In the present study, random amplified polymorphic DNA (RAPD) markers were used to analyze genetic variability and structure of populations assigned to the Neotropical fish species Astyanax scabripinnis from an urban stream located in Londrina, Paraná State, southern Brazil. Thirty individuals of this species were collected from three sites throughout the upper Cambé stream. A total of 10 primers amplified 159 loci, of which 128 (80.5%) were polymorphic. Each of the three populations showed very similar proportions of polymorphic loci, which ranged from 63.5 to 64.8%. Unbiased genetic distances varied from 0.0612 to 0.0646. Theta_p-test values indicated moderate to high genetic differentiation among individuals from different localities. The number of migrants varied from 1.34 to 1.46, suggesting a low gene flow between populations. The genetic similarity among all individuals studied ranged from 0.424 to 0.848. The results suggest that populations of A. scabripinnis in Cambé stream are undergoing genetic differentiation.     

Seed bank influences on genetic diversity in the rare annual Clarkia springvillensis (Onagraceae)

The potential for seed banks to significantly affect the genetic structure of populations was recognized nearly two decades ago. However, there has been little empirical work that examines the problem. In this study, we explore the possibility that the seed bank of a rare annual, Clarkia springvillensis, could act as a buffer against the genetic consequences of small population size. We examined the adult and seed bank cohort in three natural populations. The seed bank was surveyed by collecting soil cores twice during the growing season: postgermination and post seed set. The genetic constitution of the adults and seed bank cohort was determined by examining eight polymorphic isozyme loci via starch gel electrophoresis. The total genetic diversity in the seed bank (Ht = 0.355) was significantly higher than in the adults (Ht = 0.260). Additionally, Fst estimates of genetic differentiation among populations showed significantly less differentiation among population seed banks (Fst = 0.008) than among adults (Fst = 0.045). These results are in agreement with the expectation that seed banks would act to maintain genetic diversity in populations as well as have the effect of slowing differentiation of populations.     

Ex situ cultivation affects genetic structure and diversity in arable plants

Worldwide, botanical gardens cultivate around 80,000 taxa, corresponding to approximately one‐quarter of all vascular plants. Most cultivated taxa are, however, held in a small number of collections, and mostly only in small populations. Lack of genetic exchange and stochastic processes in small populations make them susceptible to detrimental genetic effects, which should be most severe in annual species, as sowing cycles are often short. In order to assess whether ex situ cultivation affects genetic diversity of annuals, five annual arable species with similar breeding systems were assessed with 42 in situ populations being compared to 20 ex situ populations using a random amplified polymorphic DNA (RAPD) analysis approach. Population sizes tended to be lower under ex situ cultivation and levels of genetic diversity also tended to be lower in four of the five species, with differences being significant in only two. Ex situ populations showed incomplete representation of alleles found in the wild. The duration of cultivation did not indicate any effect on genetic diversity. This implies that cultivation strategies resulted in different genetic structures in the garden populations. Although not unequivocally pronounced, differences nonetheless imply that conservation strategies in the involved gardens may need improvement. One option is cold storage of seeds, a practice that is not currently followed in the studied ex situ collections. This may reflect that the respective gardens focus on displaying living plant populations.     

Influence of genetic drift on patterns of genetic variation: The footprint of aquaculture practices in Sparus aurata (Teleostei: Sparidae)

Aquaculture finfish production based on floating cage technology has raised increasing concerns regarding the genetic integrity of natural populations. Accidental mass escapes can induce the loss of genetic diversity in wild populations by increasing genetic drift and inbreeding. Farm escapes probably represent an important issue in the gilthead sea bream (Sparus aurata), which accounted for 76.4% of total escapees recorded in Europe during a 3‐year survey. Here, we investigated patterns of genetic variation in farmed and wild populations of gilthead sea bream from the Western Mediterranean, a region of long gilthead sea bream farming. We focused on the role that genetic drift may play in shaping these patterns. Results based on microsatellite markers matched those observed in previous studies. Farmed populations showed lower levels of genetic diversity than wild populations and were genetically divergent from their wild counterparts. Overall, farmed populations showed the smallest effective population size and increased levels of relatedness compared to wild populations. The small broodstock size coupled with breeding practices that may favour the variance in individual reproductive success probably boosted genetic drift. This factor appeared to be a major driver of the genetic patterns observed in the gilthead sea bream populations analysed in the present study. These results further stress the importance of recommendations aimed at maintaining broodstock sizes as large as possible and equal sex‐ratios among breeders, as well as avoiding unequal contributions among parents.     

Atherinella blackburni (Schultz, 1949) at Itamambuca Beach, Ubatuba, SP: ecological characterization and distribution on the Brazilian coast (Teleostei: Atheriniformes: Atherinopsidae)

Atherinella blackburni is a silverside species whose occurrence on the Brazilian coast was not properly registered until recently. So far, records of its distribution along the Brazilian shore were limited to Itaparica Island, Bahia State, and Porto Inhaúma, Rio de Janeiro State. In a recent survey of the ichthyofauna of Itamambuca Beach, Ubatuba, S?o Paulo State, 100 specimens of this species were collected, yielding a considerable source of information regarding its ecology and a new southern limit of its distribution. A detailed survey of an ichthyological collection revealed lots of this species from regions of the Brazilian Northeast, resulting in a northward expansion of the occurrence of A. blackburni in Brazilian waters. Besides the populations found on the Brazilian coast, the species also occurs discontinuously in the Atlantic coasts of Costa Rica, Panama and Venezuela. Meristic and morphometric analysis showed overlapped values between Brazilian populations and the ones closer to the Caribbean. An ecological study of A. blackburni at Itamambuca Beach revealed that it is a predominantly diurnal species with preference for warm and salty waters, often occurring in sandy-bottom environments. It was more abundant in Summer and Winter, when individuals with smaller values of length and weight also occurred.     

Chromosomes, RAPDs and evolutionary trends of the Neotropical fish Mimagoniates microlepis (Teleostei: Characidae: Glandulocaudinae) from coastal and continental regions of the Atlantic forest, Southern Brazil

Chromosome and random amplified polymorphic DNA (RAPD) markers of samples of Mimagoniates microlepis were studied to test the hypothesis that a vicariant event occurred as the result of the orogeny of the coastal mountain range (Serra do Mar; southeastern and southern Brazil). Conventional karyotypes and nucleolar organizer region (Ag-NOR) phenotypes of two samples of M. microlepis from the headwaters of the Iguaçu River (southern Brazil) were compared both with each other and with other local populations of the species in the coastal drainage of southeastern Brazil. Additional molecular data (RAPD markers and genetic diversity) were obtained from specimens from coastal and continental regions of southern Brazil. The same diploid number (52 chromosomes), karyotypic formula and Ag-NOR phenotype were found for both analysed samples from the Iguaçu River. A genetic discontinuity was discovered in the comparison of the karyotypical formula of the Iguaçu samples with those from coastal drainages of the region. Polymerase chain reaction-RAPD markers revealed strikingly different molecular profiles between coastal and continental samples and indications of a high degree of genetic variation. Based on these results, we provide some comments on the biogeographical patterns and evolutionary trends for M. microlepis from coastal and continental regions of southeastern/southern Brazil.     

四川西部濒危植物桃儿七遗传多样性的RAPD分析

桃儿七是一种具有重要药用价值的珍稀濒危植物。采用RAPD分子标记技术,对在四川西部地区的桃儿七7个自然种群的遗传多样性水平和遗传结构进行了分析。用12个RAPD引物对7个种群共140个样品进行了扩增,共得到111条清晰带,其中32条具有多态性,在物种水平上多态位点百分率(PPB)为28.83%,在种群内的多态位点百分率变动幅度为4.50%至16.22%。同其它一些濒危植物相比,桃儿七种群具有较低的遗传多样性(He=0.0622,Ho=0.0987)。7个自然种群间出现了很强的遗传分化,分化指数接近70%。种群间的基因流低(Nm=0.2240)。造成上述结果的可能原因是与桃儿七的繁育方式和有限的基因流等因素有关。应将遗传多样性相对较高的松潘县牟尼沟种群作为原位保护的核心种群进行保护,尽量保护所有现有种群。     

Phylogeography, genetic structure and diversity in the endangered bearded vulture (Gypaetus barbatus, L) as revealed by mitochondrial DNA

Bearded vulture populations in the Western Palearctic have experienced a severe decline during the last two centuries that has led to the near extinction of the species in Europe. In this study we analyse the sequence variation at the mitochondrial control region throughout the species range to infer its recent evolutionary history and to evaluate the current genetic status of the species. This study became possible through the extensive use of museum specimens to study populations now extinct. Phylogenetic analysis revealed the existence of two divergent mitochondrial lineages, lineage A occurring mainly in Western European populations and lineage B in African, Eastern European and Central Asian populations. The relative frequencies of haplotypes belonging to each lineage in the different populations show a steep East-West clinal distribution with maximal mixture of the two lineages in the Alps and Greece populations. A genealogical signature for population growth was found for lineage B, but not for lineage A; futhermore the Clade B haplotypes in western populations and clade A haplo-types in eastern populations are recently derived, as revealed by their peripheral location in median-joining haplotype networks. This phylogeographical pattern suggests allopatric differentiation of the two lineages in separate Mediterranean and African or Asian glacial refugia, followed by range expansion from the latter leading to two secondary contact suture zones in Central Europe and North Africa. High levels of among-population differentiation were observed, although these were not correlated with geographical distance. Due to the marked genetic structure, extinction of Central European populations in the last century re-sulted in the loss of a major portion of the genetic diversity of the species. We also found direct evidence for the effect of drift altering the genetic composition of the remnant Pyrenean population after the demographic bottleneck of the last century. Our results argue for the management of the species as a single population, given the apparent ecological exchangeability of extant stocks, and support the ongoing reintroduction of mixed ancestry birds in the Alps and planned reintroductions in Southern Spain.     

Phylogeography, genetic structure, and diversity in the dhole (Cuon alpinus)

The Asiatic wild dog or dhole was once very widely distributed across Asia but now has a very fragmented range. In this first genetic study of this little-known species, we obtained information on genetic diversity, phylogeography, and social structure using both mitochondrial control region sequencing and microsatellite genotyping of noninvasive faecal samples from wild populations, as well as from museum and captive samples. A pattern largely consistent with isolation by distance across the Asian mainland was observed, with no clear subspecies distinctions. However, two major phylogeographical groupings were found across the mainland, one extending from South, Central, and North India (south of the Ganges) into Myanmar, and the other extending from India north of the Ganges into northeastern India, Myanmar, Thailand and the Malaysian Peninsula. We propose a scenario involving glaciation events that could explain this pattern. The origin of the dhole populations in Sumatra and Java is enigmatic and requires further study. Very low levels of genetic diversity were observed among wild dholes from Baluran National Park in Java, Indonesia, but in contrast, high levels were observed in Mudumalai Wildlife Sanctuary in South India.     

巴西橡胶树SSR遗传图谱的构建

以热研88-13×IAN873的94个F1群体为试材, 利用简单序列重复(Simple sequence repeat, SSR)标记, 采用FsLinkageMAP 1.0软件, 构建了巴西橡胶树热研88-13×IAN873的遗传连锁图谱。从441对SSR引物中筛选出160对具有多态信息的引物, 在分离群体中共检测到206个多态性位点, 176个位点用于遗传图谱的构建; χ²检验结果显示, 有147个位点符合1:1分离比例, 有12个符合1:2:1分离比例, 有17个符合1:1:1:1的分离比例, 共有13个偏分离位点, 偏分离率低(7.38%); 91个SSR位点被分为18个连锁群, 覆盖橡胶树基因组1 937.06 cM, 每个连锁群包含2～16个位点, 标记间的平均距离为21.29 cM。     

Genetic characterization of Bhutanese native chickens based on an analysis of Red Junglefowl (Gallus gallus gallus and Gallus gallus spadecieus), domestic Southeast Asian and commercial chicken lines (Gallus gallus domesticus)

The genetic diversity of Bhutanese chickens needs to be understood in order to develop a suitable conservation strategy for these birds in Bhutan. In this, work, we used microsatellite markers to examine the genetic diversity of Bhutanese chickens. Four Bhutanese chicken varieties (Black plumage, Frizzle, Naked neck and Red Junglefowl-like, corresponding to Yuebjha Narp, Phulom, Khuilay and Seim, respectively), two subspecies of Red Junglefowl (Gallus gallus gallus and Gallus gallus spadecieus), two varieties of Thai native chickens (Pradhu Hang Dam and Chee; Gallus gallus domesticus) representing the Southeast Asian domestic chicken, and two commercial lines (Broiler and Single Comb White Leghorn) were genotyped with 18 microsatellites that included 16 loci recommended by the FAO/ISAG for investigations of genetic variability in chickens. All loci were polymorphic, with the number of alleles ranging from six (MCW0111) to 23 (MCW0183). Substantial genetic variation was observed in all populations, with the Bhutanese native chicken Yuebjha Narp (Black plumage chicken) showing the lowest genetic variability. Despite extensive intrapopulation variation, the genetic differentiation among 10 populations was moderate. A neighbor-joining tree revealed the genetic relationships involved while principal component analysis showed that Bhutanese native chickens should be given priority in conservation efforts because of their genetic distinctiveness. Chee chickens are especially valuable as a reservoir of predomestic diversity, as indicated by their greater genetic variation and their position in the phylogenetic tree.     

High genetic differentiation among remnant populations of the endangered Caesalpinia echinata Lam. (Leguminosae–Caesalpinioideae)

Forest fragments along the Atlantic coastland of Brazil have been highly impacted by extensive human activities for the last 400 years. Caesalpinia echinata (Leguminosae– Caesalpinioideae), brazilwood, was overexploited during this period due to its economical importance as a dye. As a result, the species has become endangered and today its total population size is very restricted. We have assessed the distribution of genetic variation between five natural populations of brazilwood by means of RAPD (random amplified polymorphic DNA) markers. Of the total genetic variability, 28.5% was attributable to differences between two geographical groups, 29.6% to population differences within groups and 42.0% to individual differences within populations. The high level of population differentiation observed is in contrast to that expected for a primarily outcrossed woody perennial plant, and suggests that there may be a degree of inbreeding. Our results are in agreement with previous studies which postulated that C. echinata has always occurred in clumps, being common in some places but rare in between. From a conservation point of view, different populations representing different regions should be protected and, yet, plants with different origins should not be synthesized into populations in a recovery process at the risk of loss and dilution of genetic information. This study demonstrates that RAPD markers were effective in establishing a clear correlation between genetic and geographical distance and in identifying areas of maximum diversity, and may be used as an initial approach to assess the partitioning of genetic variation in this endangered species.     

Landscape models for nuclear genetic diversity and genetic structure in white-footed mice (Peromyscus leucopus)

Dramatic changes in the North American landscape over the last 12 000 years haveshaped the genomes of the small mammals, such as the white-footed mouse (Peromyscusleucopus), which currently inhabit the region. However, very recent interactions ofpopulations with each other and the environment are expected to leave the most pronouncedsignature on rapidly evolving nuclear microsatellite loci. We analyzed landscapecharacteristics and microsatellite markers of P. leucopus populations along atransect from southern Ohio to northern Michigan, in order to evaluate hypotheses aboutthe spatial distribution of genetic heterogeneity. Genetic diversity increased to thenorth and was best approximated by a single-variable model based on habitat availabilitywithin a 0.5-km radius of trapping sites. Interpopulation differentiation measured byclustering analysis was highly variable and not significantly related to latitude orhabitat availability. Interpopulation differentiation measured as F_STvalues and chord distance was correlated with the proportion of habitat intervening, butwas best explained by agricultural distance and by latitude. The observed gradients indiversity and interpopulation differentiation were consistent with recent habitatavailability being the major constraint on effective population size in this system, andcontradicted the predictions of both the postglacial expansion and core-peripheryhypotheses.     

水杉孑遗居群AFLP遗传变异的空间分布

本研究采用空间自相关分析方法对水杉 (Metasequoiaglyptostroboides)孑遗居群AFLP遗传变异的空间结构进行了研究 ,以探讨水杉孑遗居群遗传变异的分布特征及其形成机制。根据 6对AFLP选择性引物扩增的 46个多态性位点 ,选择了其表型频率在 2 5 %～ 75 %的 2 7个AFLP标记 ,运用等样本频率方法和等地理距离间隔方法分别对 3 9株和 3 7株原生母树进行了空间自相关系数Moran’sI值计算。结果表明 :水杉孑遗居群缺乏空间结构 ,绝大多数AFLP位点变异为随机分布的空间模式 ,但也有少数位点存在显著性随机相关 ,在 4～ 8km地理距离间隔显示负相关 ,说明该间隔可能是水杉孑遗居群的部分基因交流的有效屏障。水杉原生母树分布存在 12～ 2 8km的明显距离间隔空挡 ,说明人类从迁入该区域起就影响着水杉孑遗居群的原始生境 ,导致其生境片断化、景观破碎 ,进而形成岛屿状分布格局 ,并引起了水杉残留居群的随机遗传漂变。根据本研究结果 ,结合水杉孑遗居群较低的遗传多样性 ,分析探讨了水杉孑遗居群濒危的机理 ,并提出了相应的保育策略 ,为水杉的有效保育提供了科学依据     

Isozymes in macroalgae (seaweeds): genetic differentiation,genetic variability and applications in systematics

Use of isozymes (including allozymes) in studies of population genetics and systematics of seaweeds has increased sufficiently in the last decade to allow some generalization. Only a single locus has been observed for about half the enzymes analysed in seaweeds, compared with 29% in vascular plants. Compared with higher plants, macroalgal species generally have low amounts of electrophoretically detectable genetic variation; the lowest levels of genetic variation found in natural populations are those reported for seaweeds. Nonetheless, seaweeds show an association between levels of genetic diversity as revealed by isozymes and species-specific attributes, such as mating system and predominance of asexual versus sexual reproduction. In systematic studies, isozymes have revealed cryptic species and identified pairs of sibling taxa. The quaternary structure of enzymes appears to be conserved at the phylum level. With the current availability of improved techniques for enzyme electrophoresis and for data interpretation, we expect future studies utilizing isozyme electrophoresis to provide further insight into population and evolutionary processes in seaweeds.     

孑遗植物桫椤种群遗传变异的RAPD分析

应用 RAPD标记分析了桫椤分布在海南尖峰岭及霸王岭 ,广东塘朗山、黑石顶及大西山孑遗种群的遗传变异。 2 8个引物共检测到 1 1 8个位点 ,其中多态位点 33个 ,多态位点比率 2 7.97%。Shannon多样性、Nei遗传分化和分子方差分析 ( AMOVA)结果一致显示尖峰岭种群遗传多样性最大 ,霸王岭种群次之、塘郎山种群第三 ,黑石顶种群第四 ,而大西山种群最小。种群的遗传变异主要发生在种群间 ;Shannon指数测出的种群间遗传多样性所占比率为 79.1 0 % (基于表型频率 )和 77.85 % (基于基因频率 ) ,Nei基因分化系数 ( Gst)达 80 .69% ,分子方差分析也表明种群分化极为显著 ( Φst=0 .82 86,P<0 .0 0 1 )。桫椤的种内遗传多样性水平很低 ,种内 Shannon多样性仅为 0 .0 641 (基于表型频率 )和 0 .1 1 2 4 (基于基因频率 ) ,总基因多样性只有 0 .0 770。另外 ,Jaccard相似性系数的 UPGMA分析结果显示 5个种群分为两个分支 :尖峰岭种群和霸王岭种群组成一分支 ;黑石顶、塘郎山和大西山种群组成另一分支 ,并且三者中前两者的遗传关系更为密切。聚类分析结果还得到主成分分析的支持。根据桫椤种群的遗传变异特点 ,初步探讨了保护策略。