Genetic diversity and genetic structure of Puya raimondii (Bromeliaceae) for its conservation in the Peruvian Andes

Resumen Puya raimondii es una especie endémica de los altos Andes de Perú y Bolivia. En el Perú se distribuye desde 8.068501°S, 16.170280°W hasta 16.180580° S, 70.658873° W, entre los 3600 y 4800 m de altitud, viviendo en condiciones climáticas extremas propias de la Puna, donde juega un papel ecológico importante. Pese a la amplia distribución de las poblaciones de P. raimondii en el Perú, aparentemente son bastante uniformes morfológicamente; por lo que surgen las siguientes preguntas: ¿Podrán las actuales herramientas moleculares mostrar diferencias entre las numerosas poblaciones? ¿Son suficientes las áreas de conservación establecidas para P. raimondii ya que albergan la variabilidad existente? Para responder a estas interrogantes, este trabajo tuvo como objetivo evaluar la diversidad genética y estructura genética en una población del norte del país, Pachapaqui (departamento de Ancash), una población del centro, Yanacancha (Junín), y una población del sur, Lampa - sector Choconchaca (Puno), utilizando marcadores microsatélites (SSR) específicos para la especie. Los parámetros de diversidad genética utilizados incluyeron número de alelos (A), alelos exclusivos (RA), heterocigosidad observada (Ho), heterocigosidad esperada (He) e índice de contenido polimórfico (PIC). Los resultados mostraron que el número total de A varió de 2 ‒ 13, los valores de He fueron 0 ‒ 0.723 y Ho 0 ‒ 0.929, con un He promedio de 0.217, indicando una diversidad genética moderada a alta, siendo la población de Lampa-sector Choconchaca, la que presentó mayor diversidad alélica y mayor diversidad genética. La prueba de Hardy-Weinberg mostró que las poblaciones se encuentran en desequilibrio HW, el análisis estadístico indica un 65% de variación genética a nivel poblacional y valores de FST (0.426) y RST (0.650) que indican alta diferenciación genética entre poblaciones, con dos grupos genéticos (K=2) que corresponden a las poblaciones del centro-norte y sur del Perú. Los resultados brindan información útil para establecer estrategias de conservación para P. raimondii, que conduzcan a la creación de una área de conservación adicional para proteger a las poblaciones del sur del Perú.

Abstract Puya raimondii is an endemic species from the high Andes of Peru and Bolivia. In Peru it is distributed from 8.068501°S, 16.170280°W to 16.180580°S, 70.658873°W, between 3600 and 4800 m, living in extreme climatic conditions typical of the Puna, where it plays an important ecological role. Despite the wide distribution of P. raimondii populations in Peru, they appear to be fairly uniform morphologically. The following questions arise: Will the current molecular tools be able to show differences between the numerous populations? Are the conservation areas established for P. raimondii sufficient since they harbor the existing variability? To answer these questions, this work aimed to evaluate the genetic diversity and genetic structure in a northern population, Pachapaqui (Ancash department), a central population, Yanacancha (Junin), and a southern population, Lampa - Choconchaca sector (Puno), using microsatellite markers (SSR) specific for the species. The genetic diversity parameters used included number of alleles (A), exclusive alleles (RA), observed heterozygosity (Ho), expected heterozygosity (He), and polymorphic content index (PIC). The results showed that the total number of A varied from 2 - 13, the He values were 0 ‒ 0.723 and Ho 0 ‒ 0.929, with an average He of 0.217, indicating a moderate to high genetic diversity, being the population of Lampa-Choconchaca sector, the one that presented the greatest allelic diversity and the greatest genetic diversity. The Hardy-Weinberg test showed that the populations are in HW disequilibrium, the statistical analysis indicates 65% of the genetic variation at the population level and values of FST (0.426) and RST (0.650) that indicate high genetic differentiation among populations, with two genetic groups (K=2) that correspond to the populations of northern-central and southern Peru. The results provide useful information to establish conservation strategies for P. raimondii, which lead to the creation of an additional conservation area to protect the populations in southern Peru.

DIVERSIDAD GENÉTICA DE POBLACIONES DE GUANÁBANA (Annona muricata L.) EN NAYARIT, MÉXICO MEDIANTE MARCADORES SSR Y SRAP / Genetic diversity of soursop populations (Annona muricata L.) in Nayarit, Mexico using SSR and SRAP markers

RESUMEN La guanábana (Annona muricata L.) es un cultivo de importancia económica para Nayarit, México. Los frutos han tenido una excelente aceptación en el mercado regional, dificultando su comercialización a lugares lejanos porque la producción es altamente perecedera, aunado a que los árboles de los huertos de guanábana son en su mayoría ecotipos o fenotipos sin ningún plan de mejoramiento genético. Debido a la falta de variedades comerciales y de un banco de germoplasma, es importante conocer la diversidad genética para identificar y seleccionar genotipos; una de las herramientas para este propósito es el uso de marcadores moleculares. El objetivo de esta investigación fue analizar la diversidad genética de guanábana de las principales zonas productoras de Nayarit. Se extrajo ADN genómico de hojas de guanábana, las cuales fueron recolectadas de 11 huertos (poblaciones) de las siguientes zonas: Compostela (cinco poblaciones), Tepic (tres poblaciones) y San Blas (tres poblaciones). Posteriormente, se realizó un análisis mediante marcadores moleculares SSR y SRAP. Los resultados indicaron que los SSR no mostraron polimorfismo entre las poblaciones. Por otro lado, en los marcadores SRAP se obtuvieron 116 loci polimórficos con un promedio de porcentaje de loci polimórfico (P) entre las zonas productoras de 29,55 %. Asimismo, se realizó un AMOVA, el cual mostró que el mayor porcentaje de varianza se encuentra dentro de las poblaciones. Además, los análisis de agrupamiento demostraron la formación de tres grupos independientes. Por tanto, se obtuvo una alta homocigocidad y baja diversidad genética de guanábana entre las zonas y poblaciones estudiadas.

ABSTRACT Soursop (Annona muricata L.) is a crop of economic importance for Nayarit, Mexico. Soursop fruits have had an excellent acceptance in the regional market, making it difficult its commercialization to distant places because the production is highly perishable, in addition to the fact that the trees in the soursop orchards are mostly ecotypes or phenotypes without any genetic improvement plan. Due to the lack of commercial varieties and a germplasm bank, it is important to know the genetic diversity to identify and select genotypes; one of the tools for this purpose is the use of molecular markers. The objective of this research was to analyze the genetic diversity of soursop in the main producing areas of Nayarit. Genomic DNA was extracted from soursop leaves from 11 orchards (populations) in the following areas: Compostela (five populations), Tepic (three populations) and San Blas (three populations). Subsequently, we performed molecular analysis using SSR and SRAP molecular markers. The results indicated that the SSRs showed no polymorphism between the populations. On the other hand, we found 116 polymorphic loci in the SRAP markers with an average percentage of polymorphic loci (P) among the producing areas of 29.55 %. Likewise, an AMOVA was performed, showing that the highest percentage of variance is found within the populations. Furthermore, cluster analyzes demonstrated the formation of three independent groups. Therefore, a high homozygosity and low genetic diversity of soursop were obtained between the areas and populations studied.

Population genetic structure of two herds of Aberdeen Angus cattle breed in Colombia / Estructura genética poblacional de dos hatos de ganado bovino Aberdeen Angus en Colombia / Estrutura genética da população de dois rebanhos da raça Aberdeen Angus na Colômbia

Abstract Background: Two biotypes of Aberdeen Angus cattle breed, known as Old Type and New Type, that differ in their origin and beef production are formally recognized. In Colombia, this breed has been commercialized for approximately 80 years. Studies on the origin, kinship and levels of genetic diversity of this breed in Colombian herds are scarce, yet important for planning crossing and management strategies. Objective: To measure the genetic diversity and structure of two Colombian herds of Old Type and New Type biotypes of Aberdeen Angus from Huila and Cundinamarca provinces and assess mitochondrial introgression with other breeds. Methods: A set of ten microsatellites and sequences of the Mitochondrial Control Region were characterized. Estimators of genetic diversity and population differentiation along with tests of population assignment were applied. Results: Nuclear loci were highly polymorphic as shown by the Polymorphic Information Content (0.599) and the Probability of Identity (1.896 10-08). Both populations were highly diverse and clearly differentiated into two groups corresponding to the Old Type and New Type phenotypes. In contrast, mitochondrial data failed to distinguish these two groups and showed extensive admixture. Conclusions: This study optimized a set of ten highly polymorphic nuclear markers that may be used for parentage and population genetic studies of Aberdeen Angus. Genetic differentiation in these loci agreed with phenotypic differences of the Old and New Types. However, mitochondrial data indicated ancestry of multiple European breeds in the origin of Colombian Aberdeen Angus.

Resumen Antecedentes: Dentro de la raza Aberdeen Angus existen dos biotipos conocidos como Old Type y New Type, las cuales difieren en su origen y producción de carne. En Colombia, esta raza se ha venido comercializando desde hace aproximadamente 80 años. No obstante, aún no se han realizado estudios sobre su origen, parentesco y niveles de diversidad genética de esta raza en hatos colombianos, lo cual es importante para planear estrategias de cruce y manejo. Objetivo: Medir la diversidad y estructura genética de dos hatos colombianos de Aberdeen Angus Old Type y New Type de Huila y Cundinamarca y evaluar la introgresión mitocondrial con otras razas. Métodos: Se caracterizó un grupo de diez loci microsatélite y se secuenció la Región Control Mitocondrial. Se aplicaron estimadores de diversidad genética y diferenciación poblacional, junto con pruebas de asignación poblacional. Resultados: Los loci microsatélite fueron altamente polimórficos, tal como lo indicaron el Contenido de Información Polimórfica (0,599) y la Probabilidad de Identidad (1,896 10-08). Las poblaciones evaluadas de Aberdeen Angus en Colombia fueron altamente diversas y se diferenciaron claramente en dos grupos correspondientes a los fenotipos Old Type y New Type. En contraste, los datos mitocondriales no recobraron estos dos grupos y mostraron una amplia mezcla genética. Conclusiones: Este estudio optimizó un grupo de diez marcadores altamente polimórficos que pueden ser usados para estudios de parentesco y genética poblacional de Aberdeen Angus. La diferenciación genética en loci nucleares concordó con las diferencias fenotípicas entre Old y New Types, pero los datos mitocondriales indicaron ancestría de múltiples razas europeas en el origen del Aberdeen Angus colombiano.

Resumo Antecedentes: Dentro da raça Aberdeen Angus há dois biótipos conhecidos como Old Type e New Type, que diferem em sua origem e produção de carne. Na Colômbia, esta raça é comercializada há aproximadamente 80 anos. Entretanto, estudos sobre a origem, o parentesco e os níveis de diversidade genética desta raça em rebanhos colombianos ainda não foram realizados, o que é importante para o planejamento de cruzamentos e estratégias de manejo. Objetivo: Medir a diversidade genética e a estrutura de dois rebanhos colombianos de biótipos de Old Type e New Type de Aberdeen Angus de Huila e Cundinamarca e avaliar a introgressão mitocondrial com outras raças. Métodos: Um grupo de dez loci de microssatélites foi caracterizado e a Região de Controle Mitocondrial foi sequenciada. As estimativas de diversidade genética e diferenciação populacional foram aplicadas, juntamente com testes de designação populacional. Resultados: Os locus microssatélites foram altamente polimórficos, conforme indicado pelo Conteúdo de Infomação Polimórfica (0,599) e Probabilidade de Identidade (1,896 10-08). As populações avaliadas de Aberden Angus na Colômbia eram altamente diversificadas e claramente diferenciadas em dois grupos correspondentes aos fenótipos do Old Type e New Type. Em contraste, os dados mitocondriais não recuperaram esses dois grupos e mostraram um amplo mix genético. Conclusões: Este estudo otimizou um grupo de dez marcadores altamente polimórficos que podem ser usados para estudos genéticos de parentesco e população de Aberdeen Angus. A diferenciação genética nos loci nucleares concordou com as diferenças fenotípicas entre os Old e New Types, mas os dados mitocondriais indicam ancestralidade de várias raças européias na origem do Aberdeen Angus colombiano.

Population genetics of three threatened catfish species in heterogeneous environments of the Cauca River, Colombia

Neotropical catfishes Ageneiosus pardalis, Pimelodus grosskopfii and Sorubim cuspicaudus are migratory fishes of commercial importance that exhibit decreasing populations due to overfishing and other anthropic interventions. This study used species-specific microsatellite loci to test the hypothesis that threatened fish populations show genetic vulnerability signs and are genetically structured in the middle and lower sections of the Cauca River. The studied species exhibit genetic diversity levels higher than the average values reported for Neotropical Siluriformes; however, they seem to have suffered recent bottlenecks and they present significant endogamy levels that are higher for the critically endangered catfish P. grosskopfii. Furthermore, both Ageneiosus pardalis and S. cuspicaudus are each formed by one genetic group, while Pimelodus grosskopfii comprises two coexisting genetic groups. The information obtained in this study is useful for the decision making in management plans that are appropriate for the sustainability of these three species populations within the proposal for the expansion of the hydroelectric development and other anthropic activities.(AU)

Los bagres Neotropicales Ageneiosus pardalis, Pimelodus grosskopfii y Sorubim cuspicaudus, son peces migratorios de importancia comercial cuyas poblaciones han disminuido debido a la sobrepesca y otras intervenciones antrópicas. En este trabajo, se utilizaron loci microsatélites especie-específicos para contrastar la hipótesis de que las poblaciones de peces amenazadas muestran señales de vulnerabilidad genética y están genéticamente estructuradas en los sectores medio y bajo del río Cauca. Las especies estudiadas exhiben niveles de diversidad genética superiores a los promedios reportados para Siluriformes Neotropicales; sin embargo, parecen haber sufrido cuellos de botella recientes y presentan niveles significativos de endogamia que son más altos para el bagre en peligro crítico, P. grosskopfii. Además, Ageneiosus pardalis y S. cuspicaudus están conformados cada uno por un solo grupo genético, mientras que Pimelodus grosskopfii comprende dos grupos genéticos que coexisten. La información obtenida en este estudio es útil para la toma de decisiones en planes de manejo que sean adecuados para la sostenibilidad de las poblaciones de estas tres especies de bagre dentro de las propuestas para la expansión de desarrollo hidroeléctrico y otras actividades antrópicas.(AU)

Metapopulation dynamics of the migratory fish Prochilodus lineatus (Characiformes: Prochilodontidae) in a lotic remnant of the Grande River, Southeastern Brazil

River impoundments for electricity generation lead to environmental changes which severely affect fish migration and species richness. However, little is known about their effect on the genetic structure and population dynamics downstream from the reservoir. Here, we analyzed a set of ten microsatellite loci of Prochilodus lineatus, an important South American migratory fish. Specimens (n = 150) were sampled from five sites in a remnant lotic system that includes sections of the Grande, Pardo and Mogi Guaçu rivers, southeastern Brazil. The data showed that all microsatellites were polymorphic with the allele number per locus ranging from 5 to 32, and genetic diversity (H e ) varied from 0.74 to 0.80. Indices of genetic differentiation and Bayesian analysis showed a significant genetic structure and three genetic clusters inhabiting this river system. An asymmetric gene flow suggests source-sink metapopulation dynamics from tributaries (genetic source) to the main river (genetic sink). A genetic cluster that was not detected in the upper Mogi and Pardo rivers tributaries may indicate there is a "trapped gene pool" downstream from the Porto Colômbia dam. Thus, here we provide new insights into the genetic structure and population dynamics of a migratory fish species in a highly dammed river basin.(AU)

Represamento de rios para geração de eletricidade levam a mudanças ambientais que afetam severamente a migração de peixes e riqueza de espécies. No entanto, pouco se sabe sobre seu efeito na estrutura genética e dinâmica populacional a jusante de reservatórios. Aqui, analisamos um conjunto de dez loci de microssatélites de Prochilodus lineatus, um importante peixe migratório sul-americano. Os espécimes (n = 150) foram amostrados em cinco locais de um sistema lótico remanescente que inclui seções dos rios Grande, Pardo e Mogi Guaçu, sudeste do Brasil. Os dados mostraram que todos microssatélites eram polimórficos com o número de alelos por locus variando de 5 a 32 e diversidade genética (H e ) variou de 0,74 a 0,80. Índices de diferenciação genética e análise de agrupamento baseada em modelo bayesiano indicou a presença de três agrupamentos genéticos habitando este sistema fluvial. Um fluxo gênico assimétrico sugere dinâmica metapopulacional de fonte-sumidouro dos tributários (fonte genética) para o rio principal (sumidouro genético). Um agrupamento genético que não foi detectado nos tributários rio Mogi e rio Pardo parecem indicar que há um "trapped gene pool" a jusante da represa de Porto Colômbia. Assim, nós provemos aqui novos conhecimentos sobre a estrutura genética e dinâmica populacional de uma espécie de peixe migratório em um rio altamente fragmentado por barramentos.(AU)

Population structure and ancestry prediction of Aedes aegypti (Diptera: Culicidae) supports a single African origin of Colombian populations

BACKGROUND A previous phylogeographic study revealed two Aedes aegypti African-related mitochondrial lineages distributed in Colombian's cities with different eco-epidemiologic characteristics with regard to dengue virus (DENV). It has been proposed these lineages might indicate independent invasion sources. OBJECTIVES Assessing to Colombian population structure and to support evidence of its probable source origin. METHODS We analysed a total of 267 individuals from cities of Bello, Riohacha and Villavicencio, which 241 were related to the West and East African mitochondrial lineages (termed here as WAL and EAL, respectively). Eight polymorphic microsatellite loci were analysed aiming population structure. FINDINGS Results indicate substantial gene flow among distant and low-connected cities composing a panmictic population with incipient local differentiation of Ae. aegypti is placed in Colombia. Likewise, genetic evidence indicates no significant differences among individuals related to WAL and EAL is placed. MAIN CONCLUSIONS Minimal genetic differentiation in low-connected Ae. aegypti populations of Colombia, and lack concordance between mitochondrial and nuclear genealogies suggest that Colombian Ae. aegypti shared a common demographic history. Under this scenario, we suggest current Ae. aegypti population structure reflects a single origin instead of contemporary migration, which founding populations have a single source from a mitochondrial polymorphic African ancient.

Isolation and characterization of 30 STRs in Temminck’s ground pangolin (Smutsia temminckii) and potential for cross amplification in other African species

Temminck’s ground pangolin (Smutsia temminckii) is one of four species of pangolin, endemic to Africa. Two of the African pangolin species are listed as vulnerable and two are listed as endangered on the International Union for Conservation of Nature Red List of Threatened Species due to their ongoing exploitation for traditional medicine and bushmeat. In this study, we developed 30 species-specific short-tandem repeats (STRs) in Temminck’s ground pangolin using next-generation sequencing. The markers were also optimized for crossamplification in other African species. All the markers amplified successfully in Temminck’s ground pangolin with allelic polymorphisms observed in 87% of the markers in giant pangolin (S. gigantea) whereas 60% of the markers were amplified polymorphic loci in both whitebellied pangolin (Phataginus tricuspis) and black-bellied pangolin (P. tetradactyla). Analysis of diversity estimates showed moderate levels of variability in Temminck’s ground pangolin (Na = 5; Ho = 0.559), giant pangolin (Na = 4.909; Ho = 0.514) and white-bellied pangolin (Na = 2.686; Ho = 0.541) with lower values being observed in black-bellied pangolin (Na = 3; Ho = 0.242). This study provides data of the first available STR markers which was amplified in all four African pangolin species that can now be used in conservation genetic and evolutionary aspects of population histories.

Genome-wide development of polymorphic microsatellite markers and their application in peanut breeding program

BACKGROUND: Cultivated peanut (Arachis hypogaea. L) represents one of the most important oil crops in the world. Although much effort has been expended to characterize microsatellites or Simple Sequence Repeats (SSRs) in peanut, the quantity and quality of the markers in breeding applications remain limited. Here, genome-wide SSR characterization and marker development were performed using the recently assembled genome of the cultivar Tifrunner. RESULTS: In total, 512,900 microsatellites were identified from 2556.9-Mb genomic sequences. Based on the flanking sequences of the identified microsatellites, 7757 primer pairs (markers) were designed, and further evaluated in the assembled genomic sequences of the tetraploid Arachis cultivars, Tifrunner and Shitouqi, and the diploid ancestral species, A. duranensis and A. ipaensis. In silico PCR analysis showed that the SSR markers had high amplification efficiency and polymorphism in four Arachis genotypes. Notably, nearly 60% of these markers were single-locus SSRs in tetraploid Arachis species, indicating they are more specific in distinguishing the alleles of the A and B sub-genomes of peanut. In addition, two markers closely related with purple testa color and 27 markers near to FAD2 genes were identified, which could be used for breeding varieties with purple testa and high-oleic acid content, respectively. Moreover, the potential application of these SSR markers in tracking introgressions from Arachis wild relatives was discussed. CONCLUSIONS: This study reported the development of genomic SSRs from assembled genomic sequences of the tetraploid Arachis Tifrunner, which will be useful for diversity analysis, genetic mapping and functional genomics studies in peanut

Complete chloroplast genome of Salvia plebeia: organization, specific barcode and phylogenetic analysis / 中国天然药物

Salvia plebeia has been in use as traditional Chinese medicine (TCM) for more than 500 years. In this study, the complete chloroplast (cp) genome of S. plebeia was sequenced, assembled and compared to those of other five published Salvia cp genomes. It was found that the cp genome structure of S. plebeia was well conserved and had a total size of 151 062 bp. Four parameters were used to display the usage conditions of the codons of the amino acids in Salvia genus. Although the number of protein-coding genes in each species was the same, the total number of codons was different. Except for amino acids Trp and Met whose Relative Synonymous Codon Usage (RSCU) value of one condon was equal to 1, the remaining 19 amino acids had 1-3 preferred codons. The preferred codon names of each amino acid were coincident. The period size for the tandem repeats of six species ranged from 9 to 410 bp. Salvia cp genomes mainly possessed tandem repeats with a copy number less than or equal to 3. The sequence length of tandem repeats of the six species ranged from 25 to 824 bp. Highly viarable regions including four intergenic spacers and six partial genes were discovered as potential specific barcodes for Salvia species through cp genome-wide comparison. Finally, we performed phylogenetic analyses based on the complete cp genome and coding sequences respectively. These results provide information to help construct the cp genome library for Salvia, which may support studies of phylogenetics, DNA barcoding, population and transplastomics.

Genetic evidence supports polygamous mating system in a wild population of Prochilodus lineatus (Characiformes: Prochilodontidae), a Neotropical shoal spawner fish

Behavioral observations made on fish have revealed remarkably diverse reproductive strategies, including polygamy by both sexes. Still, to date, most Neotropical species remain unstudied as to whether the observed reproductive behavior in natural populations correlates with their genetic mating systems. Here, we investigated the genetic mating system of a wild population of Prochilodus lineatus settled in the Middle Uruguay River basin. By using sibship reconstruction and parental inference methods based on microsatellites' genotypes, we inferred 45 females and 47 males as potential parents of the 87 larvae analyzed. We found evidence supporting polygamous mating in both sexes: while a high percentage of males (44.7%) fertilized the eggs of one female, 55.3% of the inferred males fertilized eggs of up to four females. Likewise, while 44.5% of the inferred females had their eggs fertilized by one only male, 55.5% of females were fertilized by multiple males. The estimated proxy of the effective population size (Nb) was 126, exhibiting moderate to high levels of genetic diversity. The genetic evidence contributed in this study complements earlier behavioral observations of formation of spawning nuclei of aggregating breeders, which may be promoting a polygamous mating strategy in this long-distance migratory fish.(AU)

Observações do comportamento de peixes neotropicais têm revelado estratégias reprodutivas marcadamente variáveis, incluindo poligamia nos dois sexos. Ainda assim, até então, a correlação entre comportamento reprodutivo observado em populações naturais e sistemas de acasalamento genético permanece pouco explorada para maioria de espécies Neotropicais. Neste estudo investigamos o sistema genético de acasalamento de Prochilodus lineatus em uma população natural estabelecida no Médio rio Uruguai. Utilizando métodos de reconstrução de grupos familiares e inferências parentais baseados em genótipos de microssatélites, inferimos 45 fêmeas e 47 machos como os possíveis parentais das 87 larvas amostradas. Encontramos evidência que permite apoiar a ocorrência de acasalamento poligâmico em ambos os sexos: enquanto uma percentagem alta de machos (44,7%) fertilizou somente uma fêmea, 55,3% dos machos inferidos fertilizaram mais de uma fêmea (até quatro por macho). Da mesma forma, enquanto que 44,5% das fêmeas inferidas tiveram seus ovos fertilizados por apenas um único macho, 55,5% das fêmeas tiveram ovos fertilizados por múltiplos machos. A estimativa do tamanho populacional efetivo (Nb) foi 126, exibindo níveis entre moderados e altos de diversidade genética. A evidência genética que apresentamos nesse estudo complementa observações iniciais da formação de núcleos de desova que podem promover estratégias de acasalamento poligâmico nessa espécie migratória de longa distância.(AU)

Caracterización morfológica y molecular de Oxalis tuberosa Mol. en el departamento de Boyacá / Morphological and molecular characterization of Oxalis tuberosa Mol. in the department of Boyacá

RESUMEN Dentro de los tubérculos andinos de mayor importancia, después de la papa, se encuentra la ibia (Oxalis tuberosa Mol.) ya que constituye un alimento básico para las comunidades campesinas. Boyacá es uno de los departamentos de Colombia, en donde todavía existe la tradición de cultivar y consumir tubérculos como los cubios, ullucus y rubas o ibias, sin embargo están amenazados por la erosión genética. No existen estudios sobre estos recursos fitogenéticos en Boyacá, por lo cual el objetivo de esta investigación fue colectar y caracterizar morfológica y molecularmente materiales de ibias en este departamento. El análisis morfológico mostró que las características más discriminantes fueron: color de los tallos aéreos, pigmentación de las axilas, color del follaje, color de la flor, color del pedúnculo y pedicelo, color predominante de la superficie del tubérculo, color secundario de la superficie del tubérculo, color predominante y secundario de la pulpa, distribución del color secundario de la pulpa y forma de los tubérculos. El análisis de similitud diferenció a los materiales en dos grandes grupos, de acuerdo principalmente a características morfológicas como el color y hábito de floración. El valor de heterocigosidad promedio para la población total fue de 0.39 el cual se considera alto al compararse con otros estudios de diversidad genética en ibias. El análisis de varianza molecular y el Fst (coeficiente de diferenciación genética) muestran que existe una alta variabilidad genética entre los materiales de ibias evaluados la cual debe ser conservada y aprovechada dentro de futuros programas de mejoramiento.

ABSTRACT Among the most important Andean tubers, after potato, is the ibia (Oxalis tuberosa Mol.) because it is a staple food for peasant communities. Boyacá in one of the departments of Colombia, where there is still tradition to grow and consume tubers like cubios, ullucus and rubas or ibias; however, they are threatened by genetic erosion. There are no studies on these plant genetic resources in Boyacá, therefore the goal of this research was to collect and morphological and molecularly characterize ibias in this department. Morphological analysis showed that the most discriminate characteristic were color aerial stems, pigmentation armpits, foliage color, flower color, color peduncle and pedicel, predominant color of the tuber surface, secondary color of the surface tuber, dominant and secondary color of the pulp, distribution of secondary pulp color and shape of tubers. The similarity analysis discriminate the materials in two groups according mainly to morphological characteristics such as color and flowering habit. The value of average heterozygosity for the total population was 0.39, which is considered high when compared with other studies of genetic diversity in ibias. The analysis of molecular variance and Fst (coefficient of genetic differentiation) show that existed a high genetic variability among the ibias evaluated which it should be maintained and exploited in future breeding programs of Andean tubers.

Genetic diversity of Piracanjuba populations in fish stocking programs in the Tietê River, Brazil / Diversidad genética de poblaciones de Piracanjuba del programa de repoblamiento del Río Tietê, Brasil / Diversidade genética de populações de Piracanjuba do programa de repovoamento do Rio Tietê, Brasil

Abstract Background: Piracanjuba (Brycon orbignyanus) is a fish species highly affected by anthropogenic actions such as overfishing, water pollution, and hydroelectric developments. This species is currently considered in danger of extinction. Objective: To analyze the genetic diversity of a natural population (NP) and two captive broodstocks (SA and SB) of B. orbignyanus. Methods: Samples of caudal fins (NP: 24, SA: 30, and SB: 30) were collected. DNA was extracted and amplified for six RAPD primers and four microsatellite loci. Results: Sixty polymorphic fragments and 17 microsatellite alleles were detected. High intrapopulation heterozygosity (NP: 0.692, SA: 0.724, and SB: 0.686) was observed. Thirty-eight fragments and six alleles were shared among NP, SA, and SB. The FIS and Shannon's Index of diversity revealed a lack of inbreeding within groups. AMOVA analyses and FST indicated very high (NP vs SA and SB) and small (SA vs SB) genetic differentiation, confirmed by genetic distance and identity, number of migrants and a dendrogram, which revealed the formation of two genetic groups. Conclusions: The two marker types showed similar variability. The groups have adequate genetic variability, with high differentiation between NP and SA-SB, and similarity between broodstocks.

Resumen Antecedentes: Piracanjuba (Brycon orbignyanus) es una especie de pez fuertemente impactada por acciones antrópicas como sobrepesca, contaminación del agua y proyectos hidroeléctricos. Esta especie está considerada en peligro de extinción. Objetivo: Analizar la diversidad genética de una población natural (NP) y de dos lotes de reproductores (SA y SB) de B. orbignyanus en cautiverio. Métodos: Se colectaron 84 muestras de aleta caudal (NP: 24, SA: 30 y SB: 30). El ADN fue extraído y amplificado para seis cebadores RAPD y cuatro loci microsatélites. Resultados: Se obtuvieron 60 fragmentos polimórficos y 17 alelos microsatélites. Se observó alta heterocigosidad intra-poblacional (NP: 0,692; SA: 0,724 y SB: 0,686). Treinta y ocho fragmentos y seis alelos fueron compartidos entre NP, SA y SB. Los valores de FIS e índice de Shannon mostraron ausencia de endogamia entre los grupos. Los análisis de ANOVA y FST indicaron alta (NP vs SA y SB) y pequeña (SA vs SB) diferenciación genética; resultados confirmados por la distancia e identidad genética, número de migrantes y dendograma, evidenciando la formación de dos grupos genéticos. Conclusiones: Los grupos poseen adecuada variabilidad genética, con alta diferenciación entre NP vs SA-SB y similitud entre los lotes de reproductores.

Resumo Antecedentes: Piracanjuba (Brycon orbignyanus) é uma espécie peixe fortemente impactada por ações antrópicas como sobrepesca, poluição e construção de hidrelétricas. Atualmente, essa espécie engloba a lista de peixes que correm perigo de extinção. Objetivo: Analisar a diversidade genética de uma população natural (NP) e de dois estoques de reprodutores em cativeiro (SA e SB) de B. orbignyanus. Métodos: Foram coletadas amostras de nadadeira caudal de 84 indivíduos (NP: 24, SA: 30 e SB: 30). O DNA foi extraido e amplificado para seis primers RAPD e quatro loci microssatélites. Resultados: Foram obtidos 60 fragmentos polimórficos e 17 alelos microssatélites. Foi observada uma alta heterozigosidade intra-populacional (NP: 0,692; SA: 0,724 e SB: 0,686). Trinta e oito fragmentos e seis alelos foram compartilhados entre NP, SA e SB. Os valores de FIS e índice de Shannon demonstraram ausência de endogamia entre os grupos. As análises de AMOVA e FST indicaram alta (NP vs SA e SB) e pequena (SA vs SB) diferenciação genética, resultados confirmados pela distância e identidade genética, número de migrantes e dendrograma, que evidenciaram a formação de dois grupamentos genéticos. Conclusões: Os grupos possuem adequada variabilidade genética, com alta diferenciação entre NP e SA-SB e similaridade entre os estoques de reprodutores.

Identification of microsatellite markers and their application in yellow catfish (Pseudobagrus fulvidraco Richardson, 1846) population genetics of Korea

Microsatellite markers from a fresh water yellow catfish, Pseudobagrus fulvidraco, were developed by whole-genome sequencing in the Ion S5 system. Of the 40 chosen sets of microsatellite markers, with tetra-repeat and penta-repeat motifs, from a total 19,743 sequence, only 13 markers were successfully applied in 78 individual fish sampled to detect genomic variability from four natural populations of Korea. On an average, the number of alleles per marker was 6.7. The observed heterozygosity varied from 0.048 to 0.810. Twelve microsatellite markers conformed to Hardy–Weinberg equilibrium and none exhibited significant linkage disequilibrium. In yellow catfish, genetic differentiation among four natural populations was further supported by FST (P < 0.05) and STRUCTURE analysis. The microsatellite markers identified could facilitate genetic diversity and population structure studies and thus aid in conservation of the yellow catfish.

Genetic variability in a population of Astyanax scabripinnis: recent bottleneck and the possible influence of individuals with B chromosomes

Access the genetic variability of endangered and isolated populations has become an important conservation tool. Astyanax scabripinnis is a well-known fish model for genetic studies, forming very isolated populations in headwaters. Besides that, this species frequently presents supernumerary chromosomes, which elevates the interest on genetic studies. Genetic diversity of an Astyanax scabripinnispopulation from the Atlantic Forest (Serra da Mantiqueira region, Brazil) was assessed with microsatellite markers for the first time. Since microsatellite markers are not described for this species, we tested markers described for a related species for transferability to A. scabripinnis. Six polymorphic loci were sufficiently reliable for population genetic analysis. We found that this population passed through a recent bottleneck because of the presence of an excess of heterozygotes, low allelic diversity, heterozygosity excess, and small effective population size. Individuals with and without B chromosomes were previously identified in this population and our study found private alleles in the individuals without B chromosomes. Furthermore, when individuals without B chromosomes were removed from the analysis, the population did not present heterozygosity excess, suggesting that the bottleneck event was driven by individuals with B chromosomes. Our results provide an insight into the value of microsatellite markers as molecular tools and is the first genetic study using molecular data of A. scabripinnis from this area.

Simulation of pattern of gene flow in Canjerana fragments in the Brazilian Atlantic Rainforest for evaluating genetic conservation strategies / Simulação do padrão de fluxo gênico em fragmentos de Canjerana no bioma Mata Atlântica para definição de estratégias de conservação genética

ABSTRACT: Gene flow is important for the conservation of genetic resources to allow connectivity of geographically isolated populations and which genetic variability is reduced. Gene movement is a function of flow rate and model. Understanding how gene flow occurs can contribute to the conservation and selection of priority populations that could benefit from an eventual intervention. Simulation softwares allow making inferences about past events based on current datasets or predict future phenomena under real genetic scenarios. Adverse phenomena can be predicted and actions can be taken to avoid them. The aim of this study was to identify a model and the gene flow rates that could explain genetic structure of eight forest fragments of Cabralea canjerana in development in the Brazilian Atlantic Rainforest. To do this, simulations were performed with the EASYPOP software using a microsatellite marker dataset obtained for the species by Melo and collaborators, in 2012, 2014 and 2016. We tested five models and nine migration rates and we selected the model that produced values closer to those previously obtained for them. Criteria used for selection were the observed and expected heterozygosity and the Wright's F Statistics obtained in the simulations. The gene flow model selected was the isolation by distance model that used a rate of 0.1. We observed high levels of genetic differentiation among the fragments as result of their reproductive isolation. To allow homogenization of the allelic frequencies through gene flow, the solution would be to create ecological corridors with the aim of connecting distant fragments.

RESUMO: O fluxo gênico, cuja efetividade é função do modelo e da taxa, assume especial importância na conservação de recursos genéticos por permitir a conectividade de populações isoladas geograficamente, sujeitas à redução da variabilidade genética. O entendimento de como o fluxo gênico ocorre pode contribuir no planejamento de ações para a conservação e na seleção de populações prioritárias para uma eventual intervenção. Programas de simulação permitem inferir sobre eventos passados, a partir de dados atuais ou prever fenômenos futuros sob cenários genéticos reais. Fenômenos adversos podem ser previstos e medidas podem ser tomadas para contorná-los. O objetivo deste estudo foi identificar o modelo e a taxa de fluxo gênico que melhor explicam a estrutura genética de oito fragmentos da espécie arbórea florestal Cabralea canjerana, em desenvolvimento na região brasileira do bioma Mata Atlântica. Foram realizadas simulações com o programa EASYPOP usando dados de marcadores microssatélites obtidos por Melo e colaboradores, em 2012, 2014 e 2016, sendo testados cinco modelos e nove taxas de migração, selecionando-se o modelo que apresentou os valores mais próximos daqueles que foram publicados. Os critérios usados para a seleção do modelo foram a heterozigosidade observada e esperada e as estatísticas F de Wright obtidas nas simulações. O modelo de fluxo gênico entre os fragmentos foi o de isolamento por distância a uma taxa de 0.1. Foram observados elevados índices de diferenciação genética entre os fragmentos em decorrência do seu isolamento reprodutivo. Desse modo, sugere-se a construção de corredores ecológicos com vistas a conectar fragmentos distantes e, desta forma, permitir a homogeneização das frequências alélicas por meio do fluxo gênico.

Repetitive DNAs and shrink genomes: A chromosomal analysis in nine Columbidae species (Aves, Columbiformes)

Abstract An extensive karyotype variation is found among species belonging to the Columbidae family of birds (Columbiformes), both in diploid number and chromosomal morphology. Although clusters of repetitive DNA sequences play an important role in chromosomal instability, and therefore in chromosomal rearrangements, little is known about their distribution and amount in avian genomes. The aim of this study was to analyze the distribution of 11 distinct microsatellite sequences, as well as clusters of 18S rDNA, in nine different Columbidae species, correlating their distribution with the occurrence of chromosomal rearrangements. We found 2n values ranging from 76 to 86 and nine out of 11 microsatellite sequences showed distinct hybridization signals among the analyzed species. The accumulation of microsatellite repeats was found preferentially in the centromeric region of macro and microchromosomes, and in the W chromosome. Additionally, pair 2 showed the accumulation of several microsatellites in different combinations and locations in the distinct species, suggesting the occurrence of intrachromosomal rearrangements, as well as a possible fission of this pair in Geotrygon species. Therefore, although birds have a smaller amount of repetitive sequences when compared to other Tetrapoda, these seem to play an important role in the karyotype evolution of these species.

Genetic tracing of farmed shrimp (Decapoda: Penaeidae) in wild populations from a main aquaculture region in Mexico / Rastreo genético de camarón de cultivo (Decapoda: Penaeidae) en poblaciones silvestres de la principal región de cultivo de México

Abstract Release or escapes of aquaculture organisms may impact the genetic composition and variability of wild populations, leading to diverse issues that may compromise long-term wild stock fitness. Therefore, it is relevant to determine if farmed stocks are currently interacting with wild populations. Shrimp farming is an aquaculture activity taking place along the tropical Pacific coast of the Americas, and represents the most important culture business of Northwestern Mexico. In this study, wild and farmed whiteleg shrimp Litopenaeus vannamei from the State of Sinaloa were genetically evaluated to determine admixture levels. A newly developed set of 14 microsatellite markers (mean number of alleles per locus 11.8, and 0.836 expected heterozygosity) was obtained by Next Generation Sequencing to characterize samples. Sampling consisted of 32 wild shrimps collected during three years (2002, 2012, and 2013) and three different sites, and two hatchery stocks from 2007. No significant differences were observed among years in the wild samples, but cluster analyses showed that hatchery-produced individuals were different from wild specimens. Deviations from Hardy-Weinberg Equilibrium and genotype assignment tests indicated that a fraction from each sample could contain individuals from hatchery origin. Even though the estimated fraction of escaped farmed individuals in the most recent samples (2012-2013; mean = 7.1 %) is considered of low genetic risk, management recommendations for hatcheries and farms were provided. Besides, the reasons that explain the intended and unintended farmed shrimp release into the wild were discussed. Rev. Biol. Trop. 66(1): 381-393. Epub 2018 March 01.

Resumen La liberación o escape de lotes de cultivo pueden impactar la composición y variabilidad genética de las poblaciones silvestres, dando lugar a diversos problemas que pueden comprometer la eficacia biológica a largo plazo. Por lo tanto, es relevante determinar si las poblaciones de cultivo se encuentran actualmente interactuando con las poblaciones silvestres. El cultivo de camarón es una actividad de acuicultura que tiene lugar a lo largo de la costa del Pacífico tropical de América, y es la más importante en el noroeste de México. En este estudio, el camarón blanco Litopenaeus vannameisilvestre y de cultivo proveniente del Estado de Sinaloa, México, fueron evaluados genéticamente para determinar los niveles de mezcla. Se desarrolló un lote de 14 marcadores microsatélites nuevos (número de alelos promedio por locus de 11.8 y heterocigosidad esperada promedio de 0.836), mediante secuenciación de nueva generación, para la caracterización de las muestras. El muestreo consistió en camarón silvestre recolectado durante tres años (2002, 2012 y 2013) y dos lotes de unidades productoras de larva del 2007. No se observaron diferencias significativas entre años en las muestras silvestres, pero el análisis de agrupamiento indicó que los lotes de las unidades productoras de larva fueron distintos a los ejemplares silvestres. Desviaciones del equilibrio de Hardy-Weinberg y los análisis de asignación de genotipos indicaron que una fracción de cada una de las muestras silvestres podría contener individuos originados del larvicultivo. Se discuten las razones que explican la liberación de camarón de cultivo intencional y no intencional al medio silvestre. Aun cuando la fracción estimada de individuos de origen de cultivo en las muestras silvestres más recientes (2012-2013; promedio = 7.1 %) se considera de bajo riesgo, se dan recomendaciones de manejo para unidades de larvicultura y granjas de cultivo.

Effective population size and the genetic consequences of commercial whaling on the humpback whales (Megaptera novaeangliae) from Southwestern Atlantic Ocean

Abstract Genotypes of 10 microsatellite loci of 420 humpback whales from the Southwestern Atlantic Ocean population were used to estimate for the first time its contemporary effective (Ne) and census (Nc) population sizes and to test the genetic effect of commercial whaling. The results are in agreement with our previous studies that found high genetic diversity for this breeding population. Using an approximate Bayesian computation approach, the scenario of constant Ne was significantly supported over scenarios with moderate to strong size changes during the commercial whaling period. The previous generation Nc (Ne multiplied by 3.6), which should corresponds to the years between around 1980 and 1990, was estimated between ~2,600 and 6,800 whales (point estimate ~4,000), and is broadly compatible with the recent abundance surveys extrapolated to the past using a growth rate of 7.4% per annum. The long-term Nc in the constant scenario (point estimate ~15,000) was broadly compatible (considering the confidence interval) with pre-whaling catch records estimates (point estimate ~25,000). Overall, our results shown that the Southwestern Atlantic Ocean humpback whale population is genetically very diverse and resisted well to the strong population reduction during commercial whaling.

Caracterización genética de cuatro poblaciones de ovinos criollos de Argentina / Genetic characterization of four populations of Argentinian creole sheep

Los ovinos criollos son los fundadores de la ganadería ovina en la Argentina y han contribuido de manera sostenida al desarrollo económico, social y cultural de algunas regiones del país. A pesar de ello, es un recurso zoogenético escasamente valorizado y por ende poco estudiado. En orden de caracterizar genéticamente a los ovinos criollos argentinos, se tomaron muestras de ADN de cuatro poblaciones representativas localizadas en las provincias de Buenos Aires, Corrientes, Santiago del Estero y Salta. Estas majadas se seleccionaron por ser grupos conservados, que presentan las características fenotípicas de la raza y no registran la introducción de animales de otras razas en el sistema de reproducción. Un total de 30 marcadores microsatélites y la región D-loop del ADN mitocondrial fueron analizados. El análisis de los microsatélites permitió evidenciar una alta diversidad genética intrapoblacional (Ho= 0,676; He= 0,685; PIC= 0,713). Dicha variabilidad es explicada por diferencias entre los patrones moleculares de los individuos estudiados que pueden clasificarse en 3 grupos de poblaciones significativamente diferentes: BA, SA, SE+CO. Dado que dichas poblaciones explican muy poco de la variabilidad total (7,6%), ellas deberían considerarse perteneciente a una misma raza. El análisis del D-loop mitocondrial demostró que los individuos analizados están relacionados con el haplogrupo asiático, el cual está ampliamente distribuido en las razas españolas que son las antecesoras de la raza criolla argentina. Los resultados obtenidos en este trabajo proveerán información para establecer criterios de manejo de este recurso genético de Argentina con el fin de implementar planes de conservación, recuperación y/o mejora de los programas.

Creole sheep are the founders of sheep farming in Argentina and have contributed in a sustained way to the economic, social and cultural development of some regions of this country. However, it is a scarcely valorised and poorly studied genetic resource. In order to genetically characterize the Argentinian Creole sheep, DNA samples were taken from four representative populations located in the provinces of Buenos Aires, Corrientes, Santiago del Estero and Salta. These flocks were selected because they are considered to be conserved groups, they have the phenotypic characteristics of the creole breed and there are no records about the introduction of animals of other breeds into those systems. A total of 30 microsatellites and the D-loop region of mitochondrial DNA were analysed. Microsatellite analysis showed high level of genetic diversity within populations (Ho= 0.676; He= 0.685; PIC= 0.713). This variability is explained by differences between molecular patterns of the studied individuals, which can be classified into three significantly different population groups: BA, SA, SE+CO. Since these populations explain very little of the total variability (7.6%), it can be considered that they belong to a same race. The analysis of the mitochondrial D-loop showed that Argentinian Creole sheep have haplotypes belonging to the Asian haplogroup, which is widely distributed in the Spanish breeds, which are considered to be their ancestors. The results obtained in the present study will provide information to develop management criteria for this genetic resource in Argentina, in order to implement their conservation, recovery and/or to develop breeding programs.

Variation and genetic structure of the endangered Lepus flavigularis (Lagomorpha: Leporidae) / Variación y estructura genética de la liebre en peligro de extinción, Lepus flavigularis (Lagomorpha: Leporidae)

Abstract Lepus flavigularis, is an endemic and endangered species, with only four populations inhabiting Oaxaca, México: Montecillo Santa Cruz, Aguachil, San Francisco del Mar Viejo and Santa María del Mar. Nevertheless, human activities like poaching and land use changes, and the low genetic diversity detected with mitochondrial DNA and allozymes in previous studies, have supported the urgent need of management strategies for this species, and suggest the definition of management units. For this, it is necessary to study the genetic structure with nuclear genes, due to their inheritance and high polymorphism, therefore, the objective of this study was to examine the variation and genetic structure of L. flavigularis using nuclear microsatellites. We sampled four populations of L. flavigularis and a total of 67 jackrabbits were captured by night sampling during the period of 2001 to 2006. We obtained the genomic DNA by the phenol-chloroform-isoamyl alcohol method. To obtain the diversity and genetic structure, seven microsatellites were amplified using the Polymerase Chain Reaction (PCR); the amplifications were visualized through electrophoresis with 10 % polyacrylamide gels, dyed with ethidium bromide. Genetic diversity was determined using the software GenAlEx v. 6.4, and genetic structure was obtained with ARLEQUIN v. 3.1; null alleles were evaluated using the program Micro-Checker v.2.2.2. Additionally, a Bayesian analysis was performed with software STRUCTURE v. 2.2.3., and the isolation by distance (IBD) was studied using the program PASSAGE v.2.0.11.6. Our results showed that the genetic variation found was low ( HO = 0.30, HE = 0.24) when compared to other jackrabbit species. Fixed alleles and moderate levels of genetic differentiation (F ST = 0.18, P = 0.001) were detected among populations, indicating the effect of the genetic drift and limited gene flow. Bayesian clustering analysis revealed two groups: (1) jackrabbits from Montecillo Santa Cruz, and (2) individuals living in Aguachil, San Francisco del Mar Viejo and Santa María del Mar. No evidence was found of isolation by distance. It is possible that the geographic barriers present between populations (e.g. lagoons, human settlements), rather than the geographical distance between them, may explain the observed genetic structure. The inbreeding coefficient was negative ( FIS =-0.27, P = 0.03), indicating genetic sub-structure in populations. We suggest two management units based on the genetically closer populations, which will help define precise conservation actions in L. flavigularis. This research is the basis for defining translocation of individuals between populations, nevertheless, a more extensive future study, with specific molecular markers for L. flavigularis, is required. In addition, it is necessary to analyze the barriers that limit the gene flow, since it is urgent to reduce the genetic differentiation between populations and increase the genetic diversity of this species.

Resumen Lepus flavigularis es una especie endémica y en peligro, con solo cuatro poblaciones ubicadas en Oaxaca, México: Montecillo Santa Cruz, Aguachil, San Francisco del Mar Viejo y Santa María del Mar. Las actividades humanas (e.g. cacería, cambios de uso de suelo) y la baja diversidad genética detectada con ADN mitocondrial y aloenzimas muestran la urgencia de desarrollar estrategias de manejo para esta especie. Para definir unidades de manejo es necesario estudiar la estructura genética con genes nucleares debido a su herencia y alto polimorfismo, por lo tanto, el objetivo de este estudio fue examinar la variación y estructura genética de L. flavigularis con microsatélites nucleares. Se obtuvo el ADN genómico de 67 liebres de las cuatro poblaciones de L. flavigularis, capturadas mediante muestreo nocturno de 2001 a 2006, mediante el método fenol-cloroformo-alcohol isoamílico. Para obtener la diversidad y estructura genética se amplificaron siete microsatélites con la Reacción en Cadena de la Polimerasa (PCR). Las amplificaciones se visualizaron mediante electroforesis con geles de poliacrilamida al 10 %, teñidas con bromuro de etidio. La diversidad genética se determinó con el programa GenAlEx v.6.4, y la estructura genética se obtuvo con el ARLEQUIN v.3.1. Se evaluaron los alelos nulos con el programa Micro-Checker v.2.2.2. Adicionalmente, se realizó un análisis bayesiano con el software STRUCTURE v.2.2.3, y se estudió el aislamiento por distancia (IBD) mediante el programa PASSAGE v.2.0.11.6. La variación genética encontrada fue baja ( HO = 0.30, HE = 0.24) en comparación con otras especies de liebres. Se detectaron alelos fijos y diferenciación genética moderada (F ST = 0.18, P < 0.001) entre las poblaciones, lo que indica el efecto de la deriva genética y flujo genético limitado. El análisis Bayesiano reveló dos grupos: (1) liebres de Montecillo Santa Cruz, e (2) individuos de Aguachil, San Francisco del Mar Viejo y Santa María del Mar. No se detectó evidencia de aislamiento por distancia. Es posible que las barreras geográficas presentes entre las poblaciones (e.g. lagunas, asentamientos humanos), más que la distancia geográfica entre ellas, expliquen la estructura genética observada. El coeficiente de endogamia fue negativo ( FIS =-0.27, P = 0.03), indicando sub-estructura genética en las poblaciones. Sugerimos dos unidades de manejo con base en las poblaciones más cercanas genéticamente, lo que ayudará a definir acciones precisas de conservación en L. flavigularis. Esta investigación es la base para definir la translocación de individuos entre las poblaciones, sin embargo, se requiere un estudio futuro más amplio que incorpore marcadores moleculares específicos para L. flavigularis. Asimismo, es necesario analizar las barreras que limitan el flujo genético, ya que es urgente reducir la diferenciación genética entre poblaciones e incrementar la diversidad genética de esta especie.