Genomewide association study of C-peptide surfaces key regulatory genes in IndiansC

Insulin is a commonly used measure of pancreatic β-cell function but exhibits a short half-life in the human body. During biosynthesis, insulin release is accompanied by C-peptide at an equimolar concentration which has a much higher plasma half-life and is therefore projected as a precise measure of β-cell activity than insulin. Despite this, genetic studies of metabolic traits haveneglected the regulatory potential of C-peptide for therapeutic intervention of type-2 diabetes. The present study is aimed to search genomewide variants governing C-peptide levels in genetically diverse and high risk population for metabolic diseases—Indians. We performed whole genome genotyping in 877 healthy Indians of Indo-European origin followed by replication of variants with P ≤ 1 × 10−3 in an independent sample-set of 1829 Indians. Lead-associated signals were also tested in-silico in 773 Hispanics. To secure biological rationale for observed association, we further carried out DNA methylation quantitative trait loci analysis in 233 Indians and publicly available regulatory data was mined. We discovered novel lncRNA gene AC073333.8 with the strongest association with C-peptide levels in Indians that however missed genomewide significance. Also, noncoding genes, RP1-209A6.1 and RPS3AP5; protein gene regulators, ZNF831 and ETS2; and solute carrier protein gene SLC15A5 retained robust association with C-peptide after meta-analysis. Integration of methylation data revealed ETS2 and ZNF831 single-nucleotide polymorphisms as significant meth-QTLs in Indians. All genes showed reasonable expression in the human lung, signifying alternate important organs for C-peptide biology. Our findings mirror polygenic nature of C-peptide where multiple small-effect size variants in the regulatory genome principally govern the trait biology.

Identifying signatures of recent selection in Holstein cattle in the Tropic / Identificación de señales de selección reciente en ganado Holstein en el trópico / Identificação de sinais de seleção recente no gado Holandês no trópico

Abstract Background: Holstein cattle have undergone strong selection processes in the world. These selection signatures can be recognized and utilized to identify regions of the genome that are important for milk yield. Objective: To identify recent selection signatures in Holstein from the Province of Antioquia (Colombia), using the integrated haplotype score (iHS) methodology. Methods: Blood or semen was extracted from 150 animals with a commercial kit. The animals were genotyped with the BovineLD chip (6909 SNPs). The editing process was carried out while preserving the loci whose minor allele frequency (MAF) was greater than 0.05. In addition, genotypes with Mendelian errors were discarded using R and PLINK v1.07 software programs. Furthermore, the extended haplotype homozygosity (EHH), iHS and the p-value were determined with the "rehh" package of R language. Results: The minor allele frequencies showed a tendency toward intermediate frequency alleles. In total, 144 focal markers were significant (p<0.001) for selection signatures. Some chromosomes showed a greater number of signatures than others. Many of the variants were found inside genes, although they were in intronic regions. Some important regions were associated with genes TRAPPC12, PANK3, ZNF16, OPLA and DPYSL4, which are related with cellular transport, excretion or metabolism. Conclusion: Identifying signatures of selection using the iHS method made it possible to determine some important regions for selection in Holstein cattle in the high tropics, some of which had been previously reported to be associated with quantitative traits loci (QTLs).

Resumen Antecedentes: El ganado Holstein ha sido sometido a procesos fuertes de selección en el mundo. Estas señales de selección pueden ser reconocidas y utilizadas para identificar regiones del genoma importantes para la producción de leche. Objetivo: Identificar señales de selección recientes en ganado Holstein de la Provincia de Antioquia (Colombia), mediante la metodología de puntaje haplotípico integrado (iHS). Métodos: A 150 animales se les extrajo DNA de sangre o semen mediante un kit comercial y posteriormente se genotiparon los animales con el chip BovineLD (6909 SNPs). Se realizó edición conservando los loci con frecuencia del alelo menor (MAF) superior a 0,05. Además, se descartaron los genotipos con errores mendelianos, usando el software R y PLINK v1.07. La determinación de la homocigosidad haplotípica extendida (EHH), iHS y el valor p se realizó utilizando el paquete "rehh" de R. Resultados: Las frecuencias del alelo menor mostraron una tendencia hacia alelos de frecuencias intermedias. En total, 144 marcadores focales fueron significativos (p<0,001) para las señales de selección. Algunos cromosomas presentaron mayor número de señales de selección que otros. Muchas de las variantes focales se encontraron al interior de genes, aunque comúnmente en regiones intrónicas. Algunas de las regiones importantes estuvieron asociadas con genes como TRAPPC12, PANK3, ZNF16, OPLA y DPYSL4 que en general se encuentran asociados con funciones relacionadas con el transporte, excreción o metabolismo celular. Conclusión: La identificación de señales de selección usando el método iHS permitió determinar algunas regiones importantes para la selección en ganado Holstein del trópico alto, algunas de las cuales han sido previamente reportadas por su asociación a loci de características cuantitativas (QTLs).

Resumo Antecedentes: O gado holandês tem sido objeto de processos de seleção fortes no mundo. Estes sinais de seleção podem ser reconhecidos e utilizados para identificar regiões do genoma importantes para a produção de leite. Objetivo: Identificar sinais de seleção recente em gado Holandês de la Província de Antioquia (Colômbia), através da metodologia de pontuação haplotípica integrada (iHS). Métodos: Foram usados 150 animais para a extração de DNA a partir de sangre ou sêmen usando kit comercial, os animais foram posteriormente genotipados com o chip BovineLD (6909 SNPs). A edição foi feita mantendo os loci com frequência do alelo menor (MAF) de 0,05; além disso, genótipos com erros mendelianos foram descartados usando o programa R e PLINK v1.07. A determinação da homozigosidade haplotípica estendida (EHH), iHS e valor p foi realizada utilizando o pacote estatístico R "reeh". Resultados: As frequências do alelo menor mostraram uma tendência inclinada a frequências intermédias. No total, 144 marcadores focais foram significativos (p<0,001) para os sinais de seleção. Alguns cromossomos apresentaram mais numero de sinais de seleção que outros. Muitas dos variantes focais foram encontradas dentro dos genes, embora comumente em regiões intrônicas. Algumas das regiões importantes foram associadas com genes como TRAPPC12, PANK3, ZNF16, OPLA e DPYSL4 que geralmente estão associadas a funções relacionadas com o transporte, a excreção ou metabolismo celular. Conclusão: A identificação de sinais de seleção usando o método iHS permitiu determinar algumas regiões importantes para a seleção no gado holandês do tropico alto, algumas destas regiões foram previamente relatados por sua associação com loci de características quantitativas (QTLs).

IDENTIFICACIÓN DE QTLs ASOCIADOS A CARACTERES DE ARQUITECTURA VEGETAL EN YUCA (Manihot esculenta) / Identification of QTLs Associated to Plant Architecture in Cassava (Manihot esculenta)

La yuca (Manihot esculenta) es el cuarto cultivo en importancia a nivel mundial como fuente de calorías para la población humana después del arroz, el azúcar y el maíz, posicionándose por esta razón como un cultivo primordial para la seguridad alimentaria. Su arquitectura ha sido considerada como un factor clave que subyace a la fisiología del rendimiento, relacionando características morfológicas con productividad. En este trabajo se evaluaron diferentes características de arquitectura vegetal en yuca. Los caracteres fueron evaluados en una población F1 compuesta por 133 hermanos completos (familia K) sembrados en dos lugares biogeográficamente diferentes: La Vega (Cundinamarca) y Arauca (Arauca) en Colombia. Las características evaluadas relacionadas con la arquitectura vegetal fueron altura de la planta (AT), número de brotes (NB), longitud entrenudos (LE), número de raíces (NR), peso de raíces (PR), pigmentación del peciolo (PP), área de la hoja (AH) y tipo de hoja (TH). A partir de los datos obtenidos y empleando un mapa genético de alta densidad basado en SNPs (Single Nucleotide Polymorphisms) se llevó a cabo un análisis de QTLs (Quantitative Trait Loci). Se lograron identificar tres QTLs para La Vega asociados con los caracteres altura total, número de brotes y área de la hoja. Para Arauca se detectaron tres QTLs asociados con altura total, longitud de entrenudos y número de brotes. Los QTLs se distribuyeron en cuatro grupos de ligamiento y explicaron entre 18,93 y 41,92 % de la variación genética.

Cassava (Manihot esculenta) is the fourth most important crop worldwide as a source of calories for the human population after rice, sugar and corn and therefore it is considered as a staple crop. Cassava's architecture has been considered as a key factor underlying the physiology of yield, relating morphological traits with productivity. In this work different characteristics of plant architecture were evaluated in a cassava F1 population composed by 133 complete siblings (family K) planted in two biogeographically different zones: La Vega (Cundinamarca) and Arauca (Arauca) in Colombia. The characteristics evaluated related to the vegetal architecture were plant height (AT), number of shoots (NB), internodes length (LE), number of roots (NR), root weight (PR), petiole pigmentation (PP), leaf area (AH) and leaf type (TH). From the data obtained and using a SNP- (Single Nucleotide Polymorphism) high-density genetic map a QTLs analysis (Quantitative Trait Loci) was carried out. It was possible to identify three QTLs for La Vega associated with characters plant height, internodes length and leaf area. From the Arauca's dataset, three QTLs were detected associated with plant height, number of shoots and internodes length. The QTLs were distributed into four linkage groups and explained between 18.93 and 41.92 % of genetic variation.

La biotecnología como herramienta para la propagación, conservación y el mejoramiento genético del guayabo / Biotechnology as a tool for propagation, con-servation and genetic breeding in guava

Las técnicas biotecnológicas contribuyen positiva y significativamente en los programas de propagación, conservación y mejoramiento de las especies vegetales. Dentro de éstas, el cultivo de tejidos, el desarrollo de mapas de ligamientos genéticos y de QTLs y la detección de genes de interés han demostrado ser de gran utilidad para los mencionados propósitos. En este sentido, se estandarizó una técnica para la multiplicación in vitro de la forma silvestre de guayabo en tres fases de cultivo: establecimiento, multiplicación de propágulos y enraizamiento. La misma constituye una vía de utilidad para la propagación, la conservación de germoplasma y el mejoramiento genético en la especie. Además, se estandarizó un método de conservación a corto-mediano plazo. Por otra parte, se construyó un mapa de ligamiento genético para la especie empleando marcadores AFLP y SSR. Los 11 grupos del mapa de ligamiento genético y los 50 QTLs relacionados con caracteres vegetativos y de calidad interna y externa del fruto, constituyen el punto de partida para el clonaje de genes de interés agrícola y la implementación futura de la selección asistida por marcadores en el guayabo. De igual forma, las 176 secuencias candidatas a genes de resistencia (RGL) y del desarrollo de la planta (MADS-box y HOMEO-box) detectadas pueden ser de gran utilidad en la saturación del mapa de ligamiento referido, el estudio de la variabilidad presente en el cultivo, así como en la solución de problemas relacionados con el rendimiento, la producción y la resistencia a estrés biótico y abiótico

Biotechnologies contribute positively and signifi¬cantly in the propagation, conservation and breeding programs of many plant species. From them, tissue culture, linkage maps and QTLs detection for interesting genes have been proved to be of great utility for these purposes. In this sense, a technique for in vitro multiplication of wild guava was standardized in three culture phases: establishment, multiplication and rooting. This technique constituted a useful way for propagation, germplasm conservation and genetic breeding in the specie. A method for short-medium term conservation was also standardized. On the other hand, a genetic linkage map was constructed for the specie using AFLP and SSR markers. The 11 groups of the genetic linkage map and the 50 QTLs related with vegetative and internal/external fruit characters constitute the starting point for genes cloning of agricultural interest and the future imple¬mentation of markers assisted selection in guava. Also, the 176 candidate sequences for resistance-gene-like (RGL) and plant development (MADS-box and HOMEO-box) genes detected can be of great utility in linkage map saturation, variability studies in this crop, as well as in the solution of problems rela¬ted with yielding and resistance to biotic and abiotic stresses.

Perspectivas en genómica poblacional: genes, poblaciones y arquitectura genética / Perspectives in population genomics: genes, populations and genetic architecture

La Arquitectura Genética (AG) se refiere a los patrones de los efectos genéticos que construyen y controlan un carácter fe no-típico dado y sus propiedades variacionales. Una descripción de AG puede incluir afirmaciones acerca del gen y número de alelos, la distribución de los efectos alélicos, mutacionales, y los patrones de pleiotropía, dominancia y epistasis. La Genética de poblaciones clásica tiende a tratar la AG como un grupo de parámetros invariantes y no como variables evolutivas. El paradigma Neo-Darviniano más o menos define la evolución como un cambio en las frecuencias alélicas y deja poco margen para la evolución de los efectos alélicos. Conceptos como la canalización genética, la evolución de la variabilidad genética reducida, y la asimilación genética-respuestas evolutivas basadas en la variación medioambiental inducidaeran difíciles de enmarcar en esta línea de trabajo y fue seguida por unos pocos investigadores, siendo tratada de manera empírica. La canalización ha tenido finalmente, una sóidal interpretación de genética poblacional en términos de evolución de los efectos reducidos de un gen, a través de las interacciones epis-táticas con un trasfondo genético evolutivo, y es actualmente el foco de considerable interés empírico y teórico. El rápido desarrollo de las tecnologías moleculares ha permitido la generación de un número casi ilimitado de marcadores que especifican la estructura y organización del genoma de cualquier organismo. La integración de la estadística con la genética molecular permitió tener la primera herramienta para disgregar un rasgo cuantitativo en sus componentes genéticos individuales (qTLs). Posterior a esto se han desarrollado múltiples métodos estadísticos para localizar rasgos complejos y su aplicación posterior a la genética vegetal, animal y humana.

Genetic Architecture (GA) refers to the patterns of genetic effects that build and control a given phenotypic character and their variation properties. A description of GA can include declarations about the gene and number of alleles, the distri-bution of the allelic and mutational effects, and the patterns of pleiotropy, dominance and epistasis. The classical population genetics tends to treat the GA as a group of invariable para-meters and not as an evolutionary variable. The Neon-Dar - winian paradigm more or less defines the evolution as a chan ge in the allelic frequencies and gives small scope for the evolution of the allelic effects. Concepts as the genetic canalization, the evolution of the reduced genetic variation, and the genetic assimilation -evolutionary answers based on the induced environmental variation- were difficult to frame in this line of work and were followed by a few investigators, being treated in an empirical way. The canalization has had finally; a solid interpretation in terms of population genetics and evolution of reduced effects of a gene, through epistatic interactions with an evolutionary genetic background, and actually is the focus of considerable theoretical and empirical interest. The fast development of the molecular technologies has permitted the generation of an almost unlimited number of markers that specify the structure and organization of the genome of any organism. The integration of the statistics with the molecular genetics permitted to have the first tool to disin-tegrate a quantitative characteristic in its individual genetic components (quantitative Trait Loci, qTL). After that, has been developed multiple statistical methods to locate com-plex characteristics and their applications to the animal, vegetal, and human genetics.

Linkage disequilibrium, genetic association mapping and gene localization in crop plants

DNA-based molecular markers have been extensively utilized for a variety of studies in both plant and animal systems. One of the major uses of these markers is the construction of genome-wide molecular maps and the genetic analysis of simple and complex traits. However, these studies are generally based on linkage analysis in mapping populations, thus placing serious limitations in using molecular markers for genetic analysis in a variety of plant populations. Therefore, alternative approach has been suggested, linkage disequilibrium-based association analysis which detects and locates quantitative trait loci (QTL) by the strength of the correlation between a trait and a marker. Although association analysis has already been used for studies on genetics of complex traits in humans, its use in plants has newly started. In the present review, we describe what is known about variation in linkage disequilibrium (LD) and summarize published results on association studies in crop plant species. We give a list of different factors affecting LD, and discuss the current issues of LD research in plants. Later, we also describe the various uses of LD in crop plants research and summarize the present status of LD researches in different plant genomes. Finally, future key issues about the application of these studies on the localization of genes in these crop plants have been also discussed.

Identificação de QTLs em feijoeiro por meio de marcadores SSR influenciados pela seleção natural / QTLs identification in common bean through SSR markers affected by natural selection

Para identificar QTLs para produtividade de grãos e peso de 100 sementes em feijoeiro (Phaseolus vulgaris L.), foram usados microssatélites influenciados pela seleção natural, identificados na população derivada do cruzamento 'Carioca MG' x 'ESAL 686', conduzida pelo método da população até a geração F24. Foram avaliadas 107 linhagens da geração F8 e 107 da F24 em três épocas distintas: inverno de 2001 (F8:9 e F24:25) em Ijaci; águas de 2001 (F8:10 e F24:26) e secas de 2002 (F8:11 e F24:27) ambos em Lavras. Utilizou-se o delineamento látice simples 18 x 18 em Ijaci, e triplo nas duas outras épocas. Entre os 105 pares de primers utilizados, 30 foram polimórficos nos genitores e no bulk de DNA das linhagens F24 e utilizados, juntamente com as avaliações experimentais, na análise de regressão linear múltipla - Stepwise. Foram identificados sete QTLs para a produtividade de grãos em F8 e seis QTLs em F24, sendo o marcador derivado do 'ESAL686' (BM156) o que exibiu maior efeito para aumentar a produtividade. Para peso de 100 sementes foram identificados cinco marcadores em F8 e dois em F24, todos provenientes do genitor 'ESAL686' e o que contribuiu com o maior peso foi o X61293. A maioria dos QTLs se expressou em um só ambiente.

Aiming to identify QTLs for grain yield and for 100 seed weight of common bean (Phaseolus vulgaris L.), microsatellite markers (SSR) affected by natural selection were selected in a population derived from the cross 'Carioca MG' x 'ESAL 686', advanced by the bulk breeding until F24. One hundred and seven F8 lines, and 107 F24, were evaluated in three environments: Winter of 2001 (F8:9 and F24:25) at Ijaci county; spring/summer of 2001 (F8:10 and F24:26) and summer/fall of 2002 (F8:11 and F24:27), both at Lavras county. Simple lattice 18 x 18 experimental design was used at Ijaci, and triple lattice in the others environments. Thirty polymorphic pair of primers were selected among 105, through the parents and a bulk of the F24 lines. The molecular and the experimental data were used in the multiple linear regression analysis (stepwise). Seven QTLs for grain yield were identified in F8 and six in F24, and the BM156 marker (derived from 'ESAL 686') showed the major effect. For 100 seed weight five QTLs were identified in F8 and two in F24, all of them derived from 'ESAL 686', and the major effect was exhibited by X61293 marker. Most of the QTLs expressed in only one environment.