High-throughput targeted genotyping using next-generation sequencing applied in Coffea canephora breeding.

The use of molecular markers to detect polymorphism at DNA level is one of the most significant developments in molecular biology techniques. With the development of new next-generation sequencing technologies, the discovery of SNP became easier and faster, and the costs of data point were reduced. The development and use of SNP markers for coffee have provided new perspectives for the evaluation of genetic diversity and population structure via different statistical approaches. In this study, 72 Coffea canephora genotypes were analyzed to identify the SNP markers and apply them to genetic studies and selection of parents/hybrids in genetic breeding. As many as 117,450 SNP were identified using the RAPiD Genomics platform. After quality analyses, 33,485 SNP were validated for analyses of genetic diversity and population structure. Genotypes were separated based on their varietal groups, and Hybrids were differentiated using the clustering and Bayesian approach. Coffee accessions mistakenly identified in the germplasm and breeding program were detected. The Conilon varietal group presented the lowest genetic dissimilarity values, suggesting the introduction of new accessions in the germplasm bank. The highest genetic distances values were observed among genotypes of the heterotic groups (Conilon and Robusta). The markers were efficient in evaluating the genetic diversity and population structure of C. canephora. Promising crosses were selected within and between the varietal groups. Hybrids with greater genetic distances were selected, which were important for C. canephora breeding programs

Seleção assistida por marcadores moleculares para resistência à ferrugem do cafeeiro.

Marcadores moleculares intimamente ligados a dois genes maiores (SH3 e SH?) que conferem resistência à ferrugem do cafeeiro foram previamente identificados e têm potencial para serem usados em programas de melhoramento visando identificar e monitorar genótipos contendo resistência a esse patógeno. Dessa forma, o objetivo do trabalho foi incorporar a estratégia de seleção assistida por marcadores moleculares (SAM) associados a esses genes no programa de melhoramento do cafeeiro. Inicialmente os marcadores moleculares disponíveis na literatura foram validados e então utilizados na seleção assistida de 160 genótipos. Esses cafeeiros fazem parte do germoplasma da UFV/Epamig e foram introduzidos do CIFC por serem potencialmente portadores de resistência à ferrugem. Por meio dos marcadores validados foi possível identificar nove indivíduos resistentes homozigotos para o gene SH3, UFV311-48 (F2), UFV311-56 (F2), UFV313-133 (F2), UFV329-78 (F2), UFV334-65 (F2), UFV335-12 (F2), UFV335-77 (F2), UFV399-45 (F2) e UFV409-08 (F2). O gene SH3 confere resistência a diferentes raças do patógeno e a sua incorporação em variedades de interesse tem sido sugerida como importante estratégia para obter resistência a essa doença. Também foram identificados cafeeiros que além se serem portadores do gene SH3 em homozigose, apresentaram o gene SH? proveniente do Híbrido de Timor (HT). Tais genótipos são o UFV311-48 (F2), UFV 311-56 (F2), UFV313-133 (F2), UFV334-65 (F2), UFV399-45 (F2) e UFV409-08 (F2). Esse gene originado do HT confere resistência a outras raças do patógeno. Logo, por meio da SAM, foram identificados cafeeiros portadores de dois diferentes genes de resistência que juntos são capazes de conferir resistência a um amplo espectro de raças de H. vastatrix, fungo causador da ferrugem, permitindo obter resistência durável. Os cafeeiros identificados poderão ser utilizados no avanço de geração ou em cruzamentos para a introdução desses genes em outros materiais genéticos de interesse nos programas de melhoramento. A utilização dos marcadores moleculares validados, nos programas de melhoramento, serão imprescindíveis para monitoramento dos genes nas diferentes gerações e cruzamentos

Caracterização molecular de cultivares de café resistentes à ferrugem.

Os ganhos alcançados com o melhoramento genético do cafeeiro no Brasil resultaram na obtenção de cultivares com potencial produtivo muito superior ao das cultivares tradicionais. Atualmente, 123 cultivares de C. arabica estão registradas, destas oito são protegidas. Para que uma cultivar possa ser registrada e ou protegida é obrigatório que ela seja distinta, homogênea e estável (DHE). Na comprovação de DHE de uma cultivar são usados descritores mínimos, sendo que os marcadores morfológicos tem sido os mais utilizados. No entanto, as cultivares comerciais são cada vez mais próximas fenotipicamente, o que dificulta a discriminação precisa desses materiais por meio desses descritores. Assim sendo, uma alternativa que venha auxiliar nos testes de DHE é de extrema necessidade e proporcionará grandes benefícios. Nesse sentido, o uso de marcadores moleculares pode discriminar com maior precisão e segurança as cultivares as quais se deseja registrar e ou a proteger. Nesse contexto, o presente trabalho objetivou estabelecer um conjunto de marcadores microssatélite para a caracterização molecular (fingerprinting) de cultivares de café portadores de resistência à ferrugem (Hemileia vastatrix Berk. et Br.). Foram analisadas 34 cultivares/progênies de C. arabica portadoras de resistência à ferrugem. Trinta e um primers microssatélites foram testados, 27 amplificaram, sendo 20 polimórficos. Foi construído um dendrogroma e estabelecido o perfil molecular das cultivares. Com os resultados obtidos, demonstrou-se que os marcadores microssatélites são capazes de diferenciar genótipos aparentados e próximos fenotipicamente, podendo ser utilizados como ferramenta auxiliar nos testes de DHE das cultivares. Além disso, foi estabelecido um conjunto de marcadores microssatélites sendo discriminadas 29 das 34 cultivares de C. arabica

Early selection enabled by the implementation of genomic selection in Coffea arabica breeding.

Genomic Selection (GS) has allowed the maximization of genetic gains per unit time in several annual and perennial plant species. However, no GS studies have addressed Coffea arabica, the most economically important species of the genus Coffea. Therefore, this study aimed (i) to evaluate the applicability and accuracy of GS in the prediction of the genomic estimated breeding value (GEBV); (ii) to estimate the genetic parameters; and (iii) to evaluate the time reduction of the selection cycle by GS in Arabica coffee breeding. A total of 195 Arabica coffee individuals, belonging to 13 families in generation of F2, susceptible backcross and resistant backcross, were phenotyped for 18 agronomic traits, and genotyped with 21,211 SNP molecular markers. Phenotypic data, measured in 2014, 2015, and 2016, were analyzed by mixed models. GS analyses were performed by the G-BLUPmethod, using the RKHS (Reproducing Kernel Hilbert Spaces) procedure, with a Bayesian algorithm. Heritabilities and selective accuracies were estimated, revealing moderate to high magnitude for most of the traits evaluated. Results of GS analyses showed the possibility of reducing the cycle time by 50%, maximizing selection gains per unit time. The effect of marker density on GS analyses was evaluated. Genomic selection proved to be promising for C. arabica breeding. The agronomic traits presented high complexity for they are controlled by several QTL and showed low genomic heritabilities, evidencing the need to incorporate genomic selection methodologies to the breeding programs of this species

Population structure and genetic diversity of coffee progenies derived from Catuaí and Híbrido de Timor revealed by genome-wide SNP marker.

The use of single nucleotide polymorphism (SNP) molecular markers has provided advances in selection methodologies used in breeding programs of different crops, reducing cost and time of cultivar release. Despite the great economic and social importance of Coffea arabica, studies with SNP markers are scarce and a small number of SNP are available for this species, when compared with other crops of agronomic mportance. Thus, the objective of this study was to identify and validate SNP molecular markers for the species Coffea arabica and to introduce these markers to genetic breeding by means of an accurate analysis of the diversity and genetic structure of breeding populations of this species. After quality filtering, 11,187 SNP markers were selected from the coffee population obtained from crosses between the genotypes Catuaí and Híbrido de Timor. A great number of markers were distributed in the 11 chromosomes, within transcribed regions, and were used to estimate the genetic dissimilarity among the individuals of the breeding population. Dendrogram analysis and a Bayesian approach demonstrated the formation of two groups and the discrimination of all genotypes evaluated. The expressive number of SNP molecular markers distributed throughout C. arabica genome was efficient to discriminate all the accessions evaluated in the experiment, clustering them according to their genealogies. This work identified mixtures within the progenies. The genotyping data also provided detailed information about the parental genotypes and led to the identification of new candidate parents to be introduced to the breeding program. The study discussed population structure and its consequence in obtaining improved varieties of C. arabica

Aceleração do melhoramento do cafeeiro via seleção genômica: agilidade e eficácia no lançamento de novas cultivares.

Neste documento, foram demonstrados os usos da seleção assistida por marcadores moleculares e da seleção genômica nas espécies Coffea arabica e Coffea canephora. O uso dessas técnicas seletivas possibilitou o aumento na eficiência nos programas de melhoramento dessas espécies. Além disso, evidencia a importância da utilização de diferentes metodologias de seleção, sendo explorado os benefícios de cada técnica seletiva, tornando os programas de melhoramento mais eficientes e competitivos.Projeto SI04 - Genotipagem e seleção genômica em populações de melhoramento de Coffea arabica

Tamanho amostral e detecção de genes via GWAS em características quantitativas do cafeeiro.

O presente documento refere-se à avaliação prática do potencial da GWAS no melhoramento dos cafeeiros das espécies Coffea arabica e Coffea canephora. O objetivo geral é orientar a aplicação eficiente da GWAS em características produtivas, agronômicas e de resistência a doenças e pragas do cafeeiro. Para isso, um novo método para o estudo de associações genômicas foi desenvolvido e avaliado via simulação, em termos de poder de detecção de genes. O objetivo específico é possibilitar a identificação de marcadores genéticos úteis ao melhoramento do cafeeiro

Violence and local development in Fortaleza, Brazil: A spatial regression analysis

Fortaleza is the fifth largest city of Brazil, and has become the most violent state capital in the last years. In this paper, we investigate whether violent crime rates are associated with the local development of the city. Using an unexplored data source about georeferenced murders and deaths due to bodily injury and theft, we show that violent crime rates exhibit a positive spatial dependence across clusters of census tracts. In other words, small urban areas with high (low) violent crime rates have neighbors, on average, with similar pattern of violent crime rates. Investigating the relationship between violent crime rates and variables associated with local development, spatial regressions suggest that high violent crime rates are related with low-income neighborhood, with high spatial isolation of poor households, low access to urban infrastructure, and high prevalence of illiterate adolescents and young black males. The study also provides important evidence about spillover effects that helps to understand how the absence of local development can expose neighbors to violence

Marker-assisted selection provides arabica coffee with genes from other Coffea species targeting on multiple resistance to rust and coffee berry disease.

Selecting superior genotypes is facilitated by marker-assisted selection (MAS), which is particularly suitable for transferring disease resistance alleles because it nullifies environmental effects and allows selection of resistant individuals in the absence of the pathogen or race, enabling preventive breeding. Molecular markers linked to two major genes (SH3 and SH?), conferring resistance to coffee rust, and those linked to the Ck-1 gene, conferring resistance to coffee berry disease (CBD), have previously been identified. These markers were validated and used in a progeny of crosses between Indian selections with Coffea arabica cultivars. Eleven resistant individuals homozygous for SH3 were identified by MAS. Of these, seven carry SH? from Híbrido de Timor and the gene introduced from Coffea liberica (SH3). SH? was characterized as derived from Coffea canephora. Thus, it was possible to identify C. arabica genotypes carrying important genes for rust resistance introgressed from other coffee species. MAS also allowed identification of sources of CBD resistance for use in preventive breeding for resistance to this serious disease. Using two validated molecular markers, two coffee plants carrying Ck-1 were identified: the UFV 328-60 genotype (F2) was resistant and homozygous based on both molecular markers but exhibited no markers related to SH3 and SH?, and the UFV 317- 12 genotype (F1) was resistant and homozygous but resistant and heterozygous based on CBD-Sat207 and CBD-Sat235, respectively. Along with possessing Ck- 1, the latter carries SH?. Overall, plants carrying different genes for resistance to rust and CBD were identified. These plants are important sources for gene pyramiding in breeding programs aimed at multiple and durable resistance.201