Spatial Scale Patterns of Genetic Diversity and Gene Flow in Populations of Sweet Detar (Detarium microcarpum Guill. & Perr.; Fabaceae)

The main objective of this study is to investigate the patterns of genetic diversity and phylogenetic relationships within populations of Detarium microcarpum (Fabaceae) relative to different spatial conditions. Seventy-eight (78) accessions of D. microcarpum belonging to six populations (Phytogeographic districts) were sampled. In order to have very good quality DNA for molecular analysis, an optimization of the DNA isolation protocol was made. The molecular analysis of the accessions was carried out using 7 chloroplast microsatellite markers. The polymorphism rate (P) is 85.71% and the Polymorphism Information Content (PIC) was in the range of 0.43 (Ntcp_9) to 0.73 (Ccmp_2) with an average of 0.59. Allelic richness (A) ranged from 1.41 to 2.85 with an average of 2.04. The observed heterozygosity (Ho) ranged from 0.23 to 0.60 with an average of 0.39. The expected heterozygosity (He) ranged from 0.43 to 0.60 with a mean of 0.50. Wright's fixation index (FIS) ranged from -0.17 to 0.47. The effective allele (Ae) is between 1.77 and 2.53 with an average of 2.02. Wright differentiation index (FST) was 0.024. Phylogenetic analysis revealed that the NST value was significantly higher than the GST value (NST = 0.452; GST = 0.190; p <0.05). A relatively low hd haplotype diversity is obtained (Hd = 0.320). AMOVA analysis showed that 17.35% of the variation existed within populations but 45.80% among populations within the species. Neighbor-Joining phylogenetic tree of D. microcarpum revealed three non-distinct clusters haplotypes showing the existence of gene flow between populations of the species. Our findings of genetic structure and gene flow of D. microcarpum populations based on different spatial conditions is caused by evolutionary forces such as scattering and pollination.

The spawning method affects the reproductive efficiency of piracanjuba fish (Brycon orbignyanus) in restocking programs / El método de desove afecta la eficiencia reproductiva de piracanjuba (Brycon orbignyanus) en programas de repoblación / O método de desova afeta a eficiência reprodutiva de piracanjuba (Brycon orbignyanus) em programas de repovoamento

Abstract Background: Preserving the genetic diversity of wild fish is an important consideration for restocking programs, as inbreeding can compromise progeny survival as well as impact the resilience of natural populations. Objective: To evaluate the influence of spawning method: semi-natural (SN) or strip-spawning (ST), and the number of breeders (1♀:3♂ and 2♀:6♂) on the reproductive efficiency and genetic diversity of B. orbignyanus progeny destined for restoration of wild stocks. Methods: Rates of fertilization, hatching and broodfish mortality were recorded. For genetic evaluations (allele frequency, observed and expected heterozygosity, Shannon index, inbreeding coefficient, molecular variance analysis, and genetic differentiation), breeders (n=24), and their progenies (90 larvae/treatment) were sampled and analyzed using eight microsatellite markers. Results: Higher fertilization and hatching rates, and lower broodfish mortality were observed for the SN method (p<0.05), whereas the number of breeders did not affect these parameters (p>0.05). Interaction between spawning method and number of breeders was not significant (p>0.05). The amplified microsatellite loci produced a total of 30 alleles, with sizes between 80 and 225 bp and their frequencies indicated an increase (p<0.05) of genetic diversity in the progenies, but low genetic differentiation between treatments (p>0.05). Conclusion: The spawning methods and number of breeders tested increased equally the genetic diversity of the progeny, with low genetic differentiation between treatments. In contrast, rates of fertilization, hatching and brood fish mortality revealed that the SN method resulted in the best reproductive efficiency due to the handling stress and injuries caused by ST. Thus, SN proves to be the most suitable spawning-method for B. orbignyanus in restocking programs.

Resumen Antecedentes: Mantener la diversidad genética de los peces salvajes es una consideración importante para los programas de repoblación, ya que la endogamia puede comprometer la supervivencia de la progenie y afectar la supervivencia de las poblaciones naturales. Objetivo: Evaluar la influencia del método de desove (seminatural - SN o en franjas - ST) y el número de reproductores (1♀:3♂ y 2♀:6♂) sobre la eficiencia reproductiva y la diversidad genética de progenies de B. orbignyanus destinados a la restauración de poblaciones silvestres. Métodos: Se monitorearon las tasas de fertilización, eclosión y mortalidad de reproductores. Para las evaluaciones genéticas (frecuencias alélicas, heterocigosidad observada y esperada, índice de Shannon, coeficiente de endogamia, análisis de varianza molecular y diferenciación genética) los reproductores (n=24) y su progenie (90 larvas/tratamiento) se muestrearon y analizaron utilizando ocho marcadores microsatélites. Resultados: La interacción entre el método de desove y el número de reproductores no fue significativa (p>0,05). Se obtuvieron mejores tasas de fecundación y eclosión (p<0,05), y una menor mortalidad de reproductores (p<0,05) con el método SN, mientras que el número de reproductores no tuvo efecto (p>0,05). Los loci de microsatélites amplificados produjeron un total de 30 alelos con tamaños entre 80 y 225 pb, y sus frecuencias indicaron un aumento (p<0,05) en la diversidad genética de las progenies, pero una baja diferenciación genética entre tratamientos (p>0,05). Conclusión: Los métodos de desove y el número de reproductores evaluados aumentaron de la misma manera la diversidad genética de las progenies, con baja diferenciación genética entre tratamientos. En contraste, las tasas de fecundación, eclosión y mortalidad de peces reproductores revelaron que el SN tuvo la mejor eficiencia reproductiva, un hecho relacionado con el estrés del manejo y las lesiones causadas por el ST. Por lo tanto, el SN demuestra ser el método de desove más adecuado para B. orbignyanus en los programas de repoblación.

Resumo Antecedentes: A manutenção da diversidade genética dos peixes selvagens é uma importante consideração para programas de repovoamento, já que a endogamia pode comprometer a sobrevivência da progênie, além de impactar na resiliência das populações naturais. Objetivo: Avaliar a influência do método de desova (semi-natural - SN ou por extrusão - ST) e número de reprodutores (1♀:3♂ e 2♀:6♂) na eficiência reprodutiva e na diversidade genética de progênie de B. orbignyanus destinados à recuperação de estoques selvagens. Métodos: Taxas de fertilização, eclosão e mortalidade de reprodutores foram monitorados. Para avaliações genéticas (frequências alélicas, heterozigosidade observada e esperada, índice de Shannon, coeficiente de endogamia, análise de variância molecular e diferenciação genética), reprodutores (n=24) e sua progênie (90 larvas/tratamento) foram amostrados e analisados utilizando oito marcadores microssatélites. Resultados: Interação entre método de desova e número de reprodutores não foi significante (p>0,05). Melhores taxas de fertilização e eclosão (p<0,05), e menor (p<0,05) mortalidade de reprodutores foram observados para o método SN, enquanto que o número de reprodutores não afetou esses parâmetros (p>0,05). Os loci microssatélites amplificados produziram um total de 30 alelos, com tamanhos entre 80 e 225 pb e suas frequências indicaram aumento (p<0,05) da diversidade genética nas progênies, mas baixa diferenciação genética entre os tratamentos (p>0,05). Conclusão: Os métodos de desova e números de reprodutores avaliados aumentaram igualmente a diversidade genética das progênies, com baixa diferenciação genética entre tratamentos. Em contraste, as taxas de fecundação, eclosão e mortalidade de reprodutores revelaram que SN obteve a melhor eficiência reprodutiva, fato relacionado com o estresse de manipulação e injurias causadas por ST. Por isso, SN se mostrou como o método de desova mais adequado para B. orbignyanus em programas de repovoamento.

Molecular characterization and core collection evaluation of Manihot esculenta Crantz / Caracterização molecular e avaliação de coleção nuclear de Manihot esculenta Crantz

Cassava is one of the most important subsistence crops in tropical regions. It is necessary to preserve and to know the genetic diversity existent for the adequate use of genetic resources. The evaluation of genetic diversity among genotypes results in information about potential parents in breeding programs, allows duplicates identification, and facilitates germplasm exchange between research institutions. The objective of this study was to characterize the genetic diversity of cassava accessions of North Brazil region. A total of 106 accessions were analyzed using ten microsatellite markers. The genetic parameters estimated were: expected heterozygosity (HE), observed heterozygosity (HO) and polymorphic information content (PIC). Clustering was performed using the UPGMA and Neighbor-Joining (NJ) method. Bayesian analysis, analysis of principal coordinates and identification of a core collection were also used. The ten loci amplified 8,40 alleles on average. The average heterozygosity estimates were: HE = 0.71, HO = 0.58 and PIC = 0.72. Genetic distances ranged from 0.158 to 0.908. Six (5,66%) accesses were redundant. Clustering and dispersion analysis didn't differentiate bitter from sweet cassava, and there wasn't correlation between groups and collect origin. The core collection consisted of 22 individuals that represented 94% of total allelic diversity and 20,75% of the base collection. The results indicate high dissimilarity between the accessions and allowed the detection of redundant genotypes, showing the use of genetic markers as informative tools for the management of collections. (AU)

A mandioca é uma das mais importantes culturas de subsistência em países tropicais. É preciso conservar e conhecer a diversidade genética para o uso adequado dos recursos genéticos. A avaliação da diversidade genética entre os genótipos resulta em informações sobre potenciais genitores em programas de melhoramento, possibilita a identificação de duplicatas, além disso, facilita o intercâmbio de germoplasma entre instituições de pesquisa. O objetivo deste trabalho foi caracterizar a diversidade genética dos acessos de mandioca da Região Norte do Brasil. Foram analisados 106 acessos por meio de dez marcadores microssatélites. Os parâmetros de diversidade genética estimados foram: heterozigosidade esperada (HE), heterozigosidade observada (HO) e conteúdo de informação polimórfica (PIC). Agrupamentos foram realizados pelo método UPGMA e Neighbor-Joining (NJ). Utilizou-se também análises bayesianas, dispersão por coordenadas principais e a identificação de uma coleção nuclear. Os dez locos amplificaram 8,40 alelos em média. A média das estimativas de diversidade foram altas: HE = 0,71, HO = 0,58 e PIC = 0,72. As distâncias genéticas variaram de 0,158 a 0,908. Seis (5,66%) acessos estão redundantes. Os agrupamentos e análises de dispersão não evidenciaram distinção entre variedades bravas e mansas e não foi identificada estrutura genética correspondente a origem dos acessos. A coleção nuclear foi formada por 22 indivíduos, que representaram 94% da diversidade alélica total e 20,75% da coleção base. Os resultados indicam alta dissimilaridade entre os acessos e permitiram a detecção de genótipos redundantes, mostrando o uso de marcadores genéticos como ferramentas informativas para o manejo de coleções. (AU)

Genetic diversity and differentiation of nine populations of the hard clam (Meretrix petechialis) assessed by EST-derived microsatellites

BACKGROUND: Meretrix petechialis is one of the commercially important marine bivalves. In this study, we selected six highly polymorphic EST-derived microsatellite markers to assess the genetic diversity and population differentiation on nine wild populations of Meretrix petechialis. RESULTS: The number of alleles detected per-locus ranged from 4 to 30 (mean NA = 27.5) with a total of 165 alleles. The mean value of observed and expected heterozygosities varied from 0.717 to 0.861 and from 0.797 to 0.856, respectively. Meanwhile, the result of Neighbor-joining and overall FST = 0.214 (P < 0.01) reveled that M. petechialis populations from GX are the farthest populations, a certain degree of genetic variation among individuals in each population and the genetic differentiation is significant. CONCLUSIONS: GX population has high genetic diversity among individual, and there are certain differences in genetic characteristics among different populations. This study will provide a basis for the domestication and cultivation of genetic diversity of M. petechialis population and the protection of clam germplasm resources.

Variation of grain quality characters and marker-trait association in rice (Oryza sativa L.)

A set of 24 genotypes were studied for 17 grain quality characters and validated with the reported associated rice microsatellite markers with grain quality characters. Using 23 polymorphic markers distributed across 11 chromosomes marker-trait associations were studied. The percentage of polymorphism information content (PIC) of the markers ranged between 54.0 and 86.7. Eight markers with [80% and seven markers with[70% of PIC were found to be efficient in differentiating the studied grain quality characters. A total of 37 significant marker-trait associations (P B 0.09) were found with R2 ranging from 4.70% to 43.80%. Eight markers a (RM246, RM11, RM241, RM16427, RM421, RM3, RM234 and RM257) showed association with more than one character suggesting their utility for the selection for grain quality characters which can be deployed in the rice crop improvement programme

Genetic diversity in oocyte donors used in in vitro bovine embryo production programs in Brazil / Diversidad genética en donadoras de ovocitos utilizadas en programas de producción de embriones bovinos in vitro en Brasil / Diversidade genética em doadoras de oócitos utilizadas em programas de produção de embriões bovinos in vitro no Brasil

Abstract Background: Current reproductive management of bovine elite populations involves the use of assisted reproductive technologies (ARTs), aiming to obtain the greatest genetic gain. However, inadequate use of ARTs may lead to loss of genetic diversity in the offspring. Objective: To assess the genetic diversity in elite female cattle populations used in commercial in vitro embryo production. Methods: Using genetic and ecological approaches for the study of populations based on microsatellite markers, we assessed the genetic diversity between and within populations of cows used in commercial in vitro embryo production programs in Brazil. Results: Endogamy within populations varied from zero to 9.1%, while heterozygosity between populations (FST) was <0.05 in the different population interactions. AMOVA showed 1% variation between populations, 8% between individuals and 91% within individuals. The dimensionality reduction method utilized indicated a lack of structure in the populations analyzed, identifying two main clusters in the three populations. Conclusions: Low genetic diversity between cow populations associated with commercial programs of in vitro embryo production in Brazil was evidenced. Variable levels of endogamy within the populations were observed. Approaches of population genetics as well as ecological diversity can be implemented to more thoroughly estimate genetic diversity in livestock populations.

Resumen Antecedentes: El actual manejo reproductivo en poblaciones de bovinos de élite incluye la utilización de tecnologías de reproducción asistida (ARTs) con el fin de obtener mayor ganancia genética. Sin embargo, el uso inadecuado de las ART puede llevar a la pérdida de diversidad genética en los descendientes. Objetivo: Evaluar la diversidad genética en poblaciones de vacas de élite utilizadas en la producción comercial de embriones bovinos in vitro. Métodos: Utilizando abordajes de la genética y ecología de poblaciones basados en marcadores microsatélites, evaluamos la diversidad genética entre y dentro de poblaciones de vacas participantes de programas comerciales de producción de embriones in vitro en Brasil. Resultados: La endogamia dentro de las poblaciones varió de cero a 9,1%, mientras que la heterocigosidad entre poblaciones (FST) fue <0,05 en las diferentes interacciones de la población. El AMOVA mostró variación del 1% entre poblaciones, 8% entre individuos y 91% dentro de individuos. El método de reducción de dimensionalidad utilizado indicó una falta de estructura en las poblaciones analizadas, identificando dos grupos principales en las tres poblaciones. Conclusiones: Se evidenció una baja diversidad genética entre las poblaciones de vacas asociadas a programas comerciales de producción de embriones in vitro en Brasil. Se evidenciaron niveles variables de endogamia entre las poblaciones. Abordajes de la genética poblacional, así como de diversidad ecológica pueden ser implementados para estimar de manera más amplia la diversidad genética en poblaciones animales de interés pecuario.

Resumo Antecedentes: O atual manejo reprodutivo das populações de elite em bovinos envolve o uso de tecnologias de reprodução assistida (ARTs), visando obter o maior ganho genético. No entanto, o uso inadequado de ARTs pode levar à perda de diversidade genética na prole. Objetivo: Avaliar a diversidade genética em populações de vacas de elite utilizadas na produção comercial de embriões bovinos in vitro. Métodos: Utilizando abordagens da genética e ecologia de populações baseadas em marcadores microssatélites, foi avaliada a diversidade genética entre e dentro das populações de vacas participantes de programas comercias de produção in vitro de embriões. Resultados: A endogamia dentro das populações variou de zero a 9,1%, enquanto a heterozigosidade entre populações (FST) foi <0,05 nas diferentes interações populacionais. AMOVA mostrou variação de 1% entre populações, 8% entre indivíduos e 91% dentro de indivíduos. O método de redução de dimensionalidade utilizado indicou uma falta de estrutura nas populações analisadas, identificando dois clusters principais nas três populações. Conclusões: Baixa diversidade genética entre populações de vacas associadas a programas de produção in vitro de embriões foi evidenciada. Níveis de endogamia variáveis dentro das populações foram observados. Abordagens da genética populacional assim como de diversidade ecológica podem ser implementadas na tentativa de estimar de maneira mais abrangente a diversidade genética em populações animais de interesse pecuário.

The degree of major histocompatibility complex matching between purebred Maltese and mongrel dogs using microsatellite markers

Long-term maintenance of transplanted organs is one of the major factors that increases survival time of recipients. Although obtaining a major histocompatibility complex (MHC)-matched donor with the recipient is essential for successful organ transplantation, there have been limited reports on MHC matching between dogs. In this study, we analyzed the canine MHC matching rates using Maltese, one of the most popular purebred dogs, and mongrel dogs in Korea. Genomic DNA was extracted from blood leukocytes and DNA was amplified by polymerase chain reaction with primers specific to MHC microsatellite markers. The MHC matching degree was confirmed by the microsatellite markers using polyacrylamide gel electrophoresis. The MHC matching rates of each donor-recipient groups including Maltese-Maltese, mongrel-mongrel and Maltese-mongrel were 4.76%, 5.13% and 6.67%, respectively. There were no significant differences in the MHC matching degree between each group. These results demonstrate that MHC-matched donors could be selected from other breeds as much as from the same breed for transplantation. Knowledge of the MHC matching degree of purebred and mongrel dogs would offer valuable information not only for improving the success rate of organ transplantation surgery in canine patients but also for transplantation research using experimental canine models.

Molecular characterization and genetic diversity of different genotypes of Oryza sativa and Oryza glaberrima

Background: Availability of related rice species is critical for rice breeding and improvement. Two distinct species of domesticated rice exist in the genus Oryza: Oryza sativa (Asian rice) and Oryza glaberrima (African rice). New rice for Africa (NERICA) is derived from interspecific crosses between these two species. Molecular profiling of these germplasms is important for both genetics and breeding studies. We used 30 polymorphic SSR markers to assess the genetic diversity and molecular fingerprints of 53 rice genotypes of O. sativa, O. glaberrima, and NERICA. Results: In total, 180 alleles were detected. Average polymorphism information content and Shannon's information index were 0.638 and 1.390, respectively. Population structure and neighbor-joining phylogenetic tree revealed that 53 genotypes grouped into three distinct subpopulations conforming to the original three groups, except three varieties (IR66417, WAB450-4, MZCD74), and that NERICA showed a smaller genetic distance from O. sativa genotypes (0.774) than from O. glaberrima genotypes (0.889). A molecular fingerprint map of the 53 accessions was constructed with a novel encoding method based on the SSR polymorphic alleles. Ten specific SSR markers displayed different allelic profiles between the O. glaberrima and O. sativa genotypes. Conclusions: Genetic diversity studies revealed that 50 rice types were clustered into different subpopulations whereas three genotypes were admixtures. Molecular fingerprinting and 10 specific markers were obtained to identify the 53 rice genotypes. These results can facilitate the potential utilization of sibling species in rice breeding and molecular classification of O. sativa and O. glaberrima germplasms.

Combined association of Presenilin-1 and Apolipoprotein E polymorphisms with maternal meiosis II error in Down syndrome births

Abstract Alzheimer's disease and Down syndrome often exhibit close association and predictively share common genetic risk-factors. Presenilin-1 (PSEN-1) and Apolipoprotein E (APOE) genes are associated with early and late onset of Alzheimer's disease, respectively. Presenilin −1 is involved in faithful chromosomal segregation. A higher frequency of the APOE ε4 allele has been reported among young mothers giving birth to Down syndrome children. In this study, 170 Down syndrome patients, grouped according to maternal meiotic stage of nondisjunction and maternal age at conception, and their parents were genotyped for PSEN-1 intron-8 and APOE polymorphisms. The control group consisted of 186 mothers of karyotypically normal children. The frequencies of the PSEN-1 T allele and TT genotype, in the presence of the APOE ε4 allele, were significantly higher among young mothers (< 35 years) with meiosis II nondisjunction than in young control mothers (96.43% vs. 65.91% P = 0.0002 and 92.86% vs. 45.45% P < 0.0001 respectively) but not among mothers with meiosis I nondisjunction. We infer that the co-occurrence of the PSEN-1 T allele and the APOE ε4 allele associatively increases the risk of meiotic segregation error II among young women.

Comparative genome-wide polymorphic microsatellite markers in Antarctic penguins through next generation sequencing

Abstract Microsatellites are valuable molecular markers for evolutionary and ecological studies. Next generation sequencing is responsible for the increasing number of microsatellites for non-model species. Penguins of the Pygoscelis genus are comprised of three species: Adélie (P. adeliae), Chinstrap (P. antarcticus) and Gentoo penguin (P. papua), all distributed around Antarctica and the sub-Antarctic. The species have been affected differently by climate change, and the use of microsatellite markers will be crucial to monitor population dynamics. We characterized a large set of genome-wide microsatellites and evaluated polymorphisms in all three species. SOLiD reads were generated from the libraries of each species, identifying a large amount of microsatellite loci: 33,677, 35,265 and 42,057 for P. adeliae, P. antarcticus and P. papua, respectively. A large number of dinucleotide (66,139), trinucleotide (29,490) and tetranucleotide (11,849) microsatellites are described. Microsatellite abundance, diversity and orthology were characterized in penguin genomes. We evaluated polymorphisms in 170 tetranucleotide loci, obtaining 34 polymorphic loci in at least one species and 15 polymorphic loci in all three species, which allow to perform comparative studies. Polymorphic markers presented here enable a number of ecological, population, individual identification, parentage and evolutionary studies of Pygoscelis, with potential use in other penguin species.

Fatal toxoplasmosis in an immunosuppressed domestic cat from Brazil caused by Toxoplasma gondii clonal type I / Toxoplasmose fatal em um gato doméstico imunossuprimido do Brasil causada por Toxoplama gondii clonal tipo I

Abstract The objective of the study was to report on a fatal case of feline toxoplasmosis with coinfection with the feline leukemia virus (FeLV). A domestic cat (Felis silvestris catus) presented intense dyspnea and died three days later. In the necropsy, the lungs were firm, without collapse and with many white areas; moderate lymphadenomegaly and splenomegaly were also observed. The histopathological examination showed severe necrotic interstitial bronchopneumonia and mild necrotic hepatitis, associated with intralesional cysts and tachyzoites of Toxoplasma gondii that were positive by anti-T. gondii immunohistochemical (IHC) evaluation. The bone marrow showed chronic myeloid leukemia and the neoplastic cells were positive by anti-FeLV IHC evaluation. DNA extracted from lungs was positive for T. gondii by PCR targeting REP-529. T. gondii was characterized by PCR-RFLP and by the microsatellites technique. ToxoDB-PCR-RFLP #10, i.e. the archetypal type I, was identified. Microsatellite analysis showed that the strain was a variant of type I with two atypical alleles. This was the first time that a T. gondii clonal type I genotype was correlated with a case of acute toxoplasmosis in a host in Brazil.

Resumo O objetivo deste estudo foi relatar um caso de toxoplasmose felina fatal com coinfecção com o vírus da leucemia felina (FeLV). Um gato doméstico (Felis silvestris catus) apresentou intensa dispneia e morreu três dias depois. Na necropsia, observaram-se pulmões firmes, não colabados e com múltiplas áreas brancas, além de linfoadenomegalia e esplenomegalia moderadas. No exame histopatológico, evidenciaram-se broncopneumonia intersticial necrótica acentuada e hepatite necrótica discreta associada a cistos e taquizoítas de T. gondii intralesionais positivos na imuno-histoquímica (IHC) anti-T. gondii. Evidenciou-se ainda, na medula óssea, leucemia mieloide crônica com IHC anti-FeLV positiva nas células neoplásicas. O DNA extraído dos pulmões foi positivo para T. gondii por meio da PCR-REP-529. T. gondii foi caracterizado por PCR-RFLP e pela técnica de microssatélites. Foi identificado o genótipo ToxoDB-PCR-RFLP #10, i.e., o arquétipo tipo I. A análise por microssatélites mostrou que a cepa era uma variante do tipo I, com dois alelos atípicos. Esta é a primeira vez que T. gondii clonal tipo I foi relacionado com um caso agudo de toxoplasmosis em um hospedeiro no Brasil.

Analysis of the genetic relationships and diversity among 11 populations of Xanthoceras sorbifolia using phenotypic and microsatellite marker data

Background: Assessments of genetic diversity are essential for germplasm characterization and exploitation. Molecular markers are valuable tools for exploring genetic variation and identifying germplasm. They play key roles in a Xanthoceras sorbifolia breeding program. Results: We analyzed the genetic diversity of populations of this species using 23 simple sequence repeat (SSR) loci and data on kernel oil content. The 11 populations included in the study were distributed across a large geographic range in China. The kernel oil content differed significantly among populations. The SSR marker analysis detected high genetic diversity among the populations. All SSRs were polymorphic, and we identified 80 alleles across the populations. The number of alleles at each locus ranged from two to six, averaging 3.48 per primer pair. The polymorphism information content values ranged from 0.35 to 0.70, averaging 0.51. Expected heterozygosity, observed heterozygosity, and Shannon's information index calculations detected large genetic variations among populations of different provenance. The high average number of alleles per locus and the allelic diversity observed in the set of genotypes analyzed indicated that the genetic base of this species was relatively wide. The statistically significant positive correlation between genetic and geographic distances suggested adaptations to local conditions. Conclusions: Microsatellite markers can be used to efficiently distinguish X. sorbifolia populations and assess their genetic diversity. The information we have provided will contribute to the conservation and management of this important plant genetic resource.

SNP discovery and characterisation in White Rhino (Ceratotherium simum) with application to parentage assignment

Abstract The white rhino is one of the great success stories of modern wildlife conservation, growing from as few as 50-100 animals in the 1880s, to approximately 20,000 white rhinoceros remaining today. However, illegal trade in conservational rhinoceros horns is adding constant pressure on remaining populations. Captive management of ex situ populations of endangered species using molecular methods can contribute to improving the management of the species. Here we compare for the first time the utility of 33 Single Nucleotide Polymorphisms (SNPs) and nine microsatellites (MS) in isolation and in combination for assigning parentage in captive White Rhinoceros. We found that a combined dataset of SNPs and microsatellites was most informative with the highest confidence level. This study thus provided us with a useful set of SNP and MS markers for parentage and relatedness testing. Further assessment of the utility of these markers over multiple (> three) generations and the incorporation of a larger variety of relationships among individuals (e.g. half-siblings or cousins) is strongly suggested.

Screening of microsatellite loci with polymorphism among individuals and strains of guinea pigs / 中国比较医学杂志

Objective To screen the polymorphism microsatellite markers of guinea pigs, and provide basic information for identification of genetic structure and gene mapping in guinea pigs. Methods The genomic DNA of guinea pigs was found out on foreign websites,and microsatellite sequences characterized by AC/GT were selected. A total of 400 pairs of primers were designed using software according to their flanking sequences. After two rounds of PCR and electrophoresis,with the electrophoresis bands serving as genetic markers, the polymorphic primers were screened by the evaluation of microsatellite structure of guinea pigs. Results At the first step,10 DNA samples from 5 strains of guinea pigs were taken for PCR, and 110 pairs of polymorphic microsatellite markers with 1 -3 electrophoretic bands were screened from the designed 400 pairs of primers. At the second step,60 DNA samples from 5 strains of guinea pigs were assayed by PCR and 51 pairs of polymorphic primers with high resolution electrophoresis bands were screened from the above 110 pairs of primers. Among these screened 51 primers, more than 10 pairs of primers showed accordant or mostly accordant differences among strains. Conclusions The 51 pairs of primers are polymorphic among individuals and strains, and have not been found in other reports so far.They can be used for conventional genetic detection and related studies,and some of them can be used for gene mapping of the traits in guinea pigs.

Monitoramento e conservação genética de populações naturais de Prochilodus lineatus dos rios Pardo, Mogi-Guaçu e Tietê, São Paulo / Monitoring and conservation genetics of Prochilodus lineatus wild populations of Pardo, Mogi Guaçu and Tiete rivers, São Paulo

O objetivo do presente estudo foi determinar a diversidade e a estrutura genética de seis populações naturais de Prochilodus lineatus em usinas hidrelétricas (UHE) dos rios Pardo (UHE Limoeiro - LMO), Mogi-Guaçu (UHE Mogi-Guaçu - MOG) e Tietê (UHE Promissão - PRO, UHE Barra Bonita - BAB, UHE Nova Avanhandava - NAV e UHE Bariri - BAR). Foi encontrado um total de 47 alelos, com tamanhos entre 118pb e 330pb. Os resultados de heterozigosidade média observada (0,490 a 0,625) refletiram uma alta variabilidade genética intrapopulacional. Os valores de distância genética (0,149 a 0,773), Fst (0,006 a 0,218) e Nm (1,2 a 4,2) mostraram a presença de similaridade genética entre as populações. De acordo com a AMOVA, houve maior variação dentro das populações do que entre elas. O dendograma mostrou a formação de dois agrupamentos (LMO-PRO-MOG e BAR-BAB-NAV). Concluiu-se que as populações naturais apresentaram alta variabilidade genética, com similaridade genética entre elas, possivelmente causada pelo programa de repovoamento realizado nesses rios.(AU)

The aim of this study was to determine the genetic diversity and structure of six wild populations of Prochilodus lineatus in Hydroelectric Power Plants (HPP) of the Pardo (HPP Limoeiro - LMO), Mogi Guacu (HPP Mogi-Guaçu - MOG), and Tiete (HPP Promissão - PRO, HPP Barra Bonita - BAB, HPP Nova Avanhandava - NAV and HPP Bariri - BAR) rivers. A total of 47 alleles, ranging in size from 118bp to 330bp were found. The results of observed heterozygosity average (0.490 to 0.625) reflected a high intra-population genetic variability. The values of genetic distance (0.149 to 0.773), Fst (0.006 to 0.218), and Nm (1.2 to 4.2) showed that between the populations there is genetic similarity. According to AMOVA there was higher variation within populations than between them. The dendrogram demonstrated the formation of two groups (LMO-PRO-MOG and BAR-BAB-NAV). It was concluded that wild populations had high genetic variability with genetic similarity between them, possibly caused by the restocking program performed in these rivers.(AU)

Diversity and spatial genetic structure of a natural population of Theobroma speciosum (Malvaceae) in the Brazilian Amazon / Diversidad y estructura genética espacial de una población natural de Theobroma speciosum (Malvaceae) en la Amazonía brasileña

Abstract:The quantification of genetic diversity and intrapopulation spatial genetic structure (SGS) of tree species are important aspects for in and ex situ conservation practices. In this study we seek to understand the importance of conservation areas by quantifying the genetic diversity and the spatial genetic structure of a natural population of Theobroma speciosum. Within this population, 49 adults and 51 subadults were genotyped for five microsatellite loci. The results showed that adults and subadults have similar levels of genetic diversity and inbreeding (adults: A= 10.4, Ae = 10.3, F= 0.68, subadults: A= 10.6, Ae= 10.6, F= 0.57). Genetic diversity was spatially structured within the population, and the results suggest that near-neighbor trees up to a distance of 70 m are likely related. SGS is likely the result of short-distance seed dispersal, the short-distance range of pollinators, and infrequent breaches of the self-incompatible mating system. Considering the high demographic density of the species and size of the study area, as well as the high average number of alleles per locus and the presence of rare alleles, we believe that the study population is an excellent resource for in situ genetic conservation of T. speciosum. The study area is also a useful resource for collecting germplasm for ex situ conservation and seed collection, either for breeding programs used in the restoration of degraded areas or forest improvement. Rev. Biol. Trop. 64 (3): 1091-1099. Epub 2016 September 01.

ResumenLa cuantificación de la estructura de la diversidad genética e genética intrapoblacional espacial (SGS) de especies de árboles son importantes en las prácticas de conservación ex situ. En este estudio se busca determinar la importancia de las unidades de conservación mediante la cuantificación de la diversidad genética y la estructura espacial de una población natural de Theobroma speciosum. Dentro de esta población, a 49 adultos y 51 subadultos se les analizó el genotipo para cinco microsatélites loci. Los resultados mostraron que los adultos y subadultos tienen niveles similares de diversidad genética y endogamia (adultos: A= 10.4, Ae = 10.3, F= 0.68; subadultos: A= 10.6, Ae = 10.6, F= 0.57). La diversidad genética detectada fue estructurada espacialmente dentro de la población y los árboles vecinos hasta una distancia de 70 m estaban probablemente relacionados. Esta SGS es probablemente el resultado de la dispersión de semillas de corta distancia. El establecimiento de unidades de conservación permanentes como el Parque Nacional Juruena mostró que las áreas de conservación son herramientas valiosas en la preservación de la diversidad genética en las poblaciones naturales. Debido a la alta densidad demográfica de la especie y el tamaño del área de estudio, así como el elevado número promedio de alelos por locus y la presencia de alelos raros, creemos que la población de estudio es un excelente recurso para la conservación genética in situ de T. speciosum y también es un recurso útil de germoplasma para la recolección de conservación ex situ y la semilla, ya sea para programas de mejoramiento utilizados en la restauración de áreas degradadas o mejora de los bosques.

Development, characterization and use of genomic SSR markers for assessment of genetic diversity in some Saudi date palm (Phoenix dactylifera L. ) cultivars

Background: The present study was undertaken towards the development of SSR markers and assessing genetic relationships among 32 date palm (Phoenix dactylifera L.) representing common cultivars grown in different geographical regions in Saudi Arabia. Results: Ninety-three novel simple sequence repeat markers were developed and screened for their ability to detect polymorphism in date palm. Around 71% of genomic SSRs were dinucleotide, 25% tri, 3% tetra and 1% penta nucleotide motives. Twenty-two primers generated a total of 91 alleles with a mean of 4.14 alleles per locus and 100% polymorphism percentage. A 0.595 average polymorphic information content and 0.662 primer discrimination power values were recorded. The expected and observed heterozygosities were 0.676 and 0.763 respectively. Pair-wise similarity values ranged from 0.06 to 0.89 and the overall cultivars averaged 0.41. The UPGMA cluster analysis recovered by principal coordinate analysis illustrated that cultivars tend to group according to their class of maturity, region of cultivation, and fruit color. Analysis of molecular variations (AMOVA) revealed that genetic variation among and within cultivars were 27% and 73%, respectively according to geographical distribution of cultivars. Conclusions: The developed microsatellite markers are additional values to date palm characterization tools that can be used by researchers in population genetics, cultivar identification as well as genetic resource exploration and management. The tested cultivars exhibited a significant amount of genetic diversity and could be suitable for successful breeding program. Genomic sequences generated from this study are available at the National Center for Biotechnology Information (NCBI), Sequence Read Archive (Accession numbers. LIBGSS_039019).

ESTUDIO DE LA DIVERSIDAD GENÉTICA DEL CERDO DOMÉSTICO EN EL DEPARTAMENTO DE CÓRDOBA (COLOMBIA) UTILIZANDO MARCADORES MICROSATÉLITES / STUDY OF GENETIC DIVERSITY OF THE DOMESTIC PIG IN THE DEPARTMENT OF CÓRDOBA (COLOMBIA) USING MICROSATELLITE MARKERS

En este estudio se analizaron los niveles de diversidad y estructura genética de 161 cerdos domésticos pertenecientes a tres poblaciones del departamento de Córdoba, mediante 20 marcadores microsatélites. Todos los microsatélites utilizados resultaron polimórficos. Para todos los loci, el valor promedio de la heterocigosidad esperada fue mayor al valor promedio de la heterocigosidad observada, lo cual puede sugerir una posible endogamia en el sistema de apareamiento. El índice (0,12 ± 0,08) mostró un 88% de la varianza en las frecuencias alélicas reportadas dentro de cada población y solo el 12% de la varianza atribuible a diferencias entre poblaciones. Los valores de F IS (0,079) y F IT (0,13), indican deficiencia de heterocigotos dentro de cada población y a nivel global. Desviaciones significativas del equilibrio Hardy-Weinberg (p < 0,05) fueron observadas en ocho de los marcadores utilizados. El árbol Neighbor-Joining obtenido reveló que Momil estuvo más estrechamente relacionada con Cereté, mientras Tierralta se mostró más alejada. El análisis de componentes principales (ACoP) genera la individualización geográfica de cada población, siendo distante la población de Tierralta de las poblaciones Momil y Cereté, resultados similares a los obtenidos con la metodología Neighbor-Joining. El programa Structure con un K = 3, confirma la existencia de tres grupos o poblaciones distintas, generándose un patrón filogeográfico observado en la relación entre Momil, Cereté y Tierralta. Es importante señalar que son 3 grupos raciales diferentes, valiosos y deben conservarse.

Diversity and genetic structure of 161 domestic pigs from three populations of the department of Cordoba, were analyzed by means 20 microsatellite markers; all of them were polymorphic microsatellites. The expected average value of heterozygosity was higher than the observed average value for all loci, which may suggest a possible inbreeding mating system. The F ST index (0.12 ± 0.08) showed 88% of variance in allele frequencies reported within each population and only 12% of the variance was attributable to differences between populations. F K values (0.079) and F IT (0.13) indicate heterozygote deficiency within each population and globally. Significant deviations from Hardy-Weinberg balance (p < 0.05) were observed in eight of the markers used. The Neighbor-Joining tree showed than Momil was more closely related with Cerete while Tierralta was further. The main components analysis (ACoP) generates the geographical identification of each population, being more far Tierralta from Momil and Cerete populations, similar results were obtained with the Neighbor-Joining method. The Structure program with K = 3, confirms the existence of three groups or different populations, which generates a phylogeographic pattern observed in the relationship between Momil, Cerete and Tierralta. It is important to note that there are 3 different racial groups, valuable and must be preserved.

Diversidad y estructura genética de accesiones de palma de aceite (Elaeis guineesis Jacq.)provenientes de Camerún / Diversity and genetic structure of oil palm accession (Elaeis guineesis Jacq.) from Cameroon

La palma de aceite Elaeis guineesis Jacq. posee gran importancia debido al aceite que se extrae de sus frutos, del cual se obtienen derivados refinados de gran valor comercial como el biodiesel, entre otros. Esta investigación buscó determinar la estructura y diversidad genética de 311 muestras de palma de aceite proveniente de la República de Camerún mediante 10 marcadores microsatélite. Los resultados mostraron valores promedio para el número medio de alelos por locus de Na=8.433 y un número efectivo de alelos por locus de Ne=4.756; las diferencias entre estos valores permiten inferir que los 106 alelos encontrados para estas poblaciones son considerados alelos raros. Adicionalmente, el valor de diversidad genética fue alto (valor medio de He= 0.781) respecto a reportes de varios autores. La varianza molecular obtenida evidenció que el mayor porcentaje (80 %) se encuentra dentro de los individuos. Los análisis mostraron que no se definió ningún tipo de estructura poblacional, lo que permitió inferir un alto flujo genético entre las zonas geográficas, esto corroborado por los altos valores de diversidad genética obtenidos. Los 311 genotipos evaluados fueron definidos como una población natural heterogénea heterocigota, apta para favorecer el aumento de la base genética de las poblaciones cultivadas de palma de aceite.

Oil palm Elaeis guineesis Jacq. is of great importance because of the oil extracted from its fruits, whose refined derivatives are commercially valuable as biodiesel, among others uses. This study sought to determine the structure and genetic diversity of 311 oil palm samples from the Republic of Cameroon with 10 microsatellite markers. The results showed values for the average number of alleles per locus of Na= 8.433 and effective number of alleles per locus of Ne= 4.756; from the differences between these values, it can be inferred that the 106 alleles found for these populations could be considered rare alleles. Additionally the value of genetic diversity was high (mean value of He= 0.781) compared to reports of several authors. The obtained molecular variance showed that the highest percentage (80 %) was found within the individuals. The analysis did not show any defined population structure, which allowed us to infer a high gene flow among the geographic zones, corroborated it by the high genetic diversity values obtained. The 311 genotypes were defined as a heterogeneous heterozygous natural population suitable to increase the genetic base of oil palm cultivated populations.

Linkage study of DFNB3 responsible for hearing loss in human.

BACKGROUND: Hearing disorders represent a significant health problem worldwide. Recessive inherited cases of the deafness are more prevalent in Pakistan due to consanguineous marriages. Deafness caused by DFNB3 is due to mutation in the gene MYO XVA and its prevalence among Pakistani population is about 5%. MATERIALS AND METHODS: Families with at least two or more individual affected with deafness were selected from different areas of District Okara of Pakistan. Six consanguineous families of different ethnic groups having deaf individuals were studied. All these families had three or more deaf individuals in either two or more sib ships. Family history was taken to minimize the chances of other abnormalities. Pedigrees drawn by using Cyrillic software (version 2.1) showed that all the marriages were consanguineous and the families have recessive mode of inheritance. Three STR markers were selected and amplified on all the samples of six families through PCR. The PCR products were then genotyped on non denaturing polyacrylamide gel electrophoresis (PAGE). Haplotypes were constructed to determine the pattern of inheritance and also to determine whether a family was linked or unlinked with known DFNB3 locus. RESULTS: One out of six families showed linkage to the DFNB3 while rest of the families remained unlinked. Carriers of deafness genes were identified and information was provided to the families on request. CONCLUSION: Knowledge about the genetic causes of deafness provide insight into the variable expression of genes involved in this hereditary problem and may allow the prediction and prevention of associated health problems.