Analysis of genomic and functional RFLP derived markers associated with sucrose content, fiber and yield QTLs in a sugarcane (Saccharum spp.) commercial cross

Expressed sequence tags derived markers have a great potential to be used in functional map construction and QTL tagging. In the present work, sugarcane genomic probes and expressed sequence tags having homology to genes, mostly involved in carbohydrate metabolism were used in RFLP assays to identify putative QTLs as well as their epistatic interactions for fiber content, cane yield, pol and tones of sugar per hectare, at two crop cycles in a progeny derived from a bi-parental cross of sugarcane elite materials. A hundred and twenty marker trait associations were found, of which 26 at both crop cycle and 32 only at first ratoon cane. A sucrose synthase derived marker was associated with a putative QTL having a high negative effect on cane yield and also with a QTL having a positive effect on Pol at both crop cycles. Fifty digenic epistatic marker interactions were identified for the four traits evaluated. Of these, only two were observed at both crop cycles.

Identificació de QTLs implicats en l'adaptació a canvis de temperatura en la planta model Arabidopsis thaliana

Estudi realitzat a partir d’una estada al Max Planck Institute for Plant Breeding Research, a Alemanya, entre 2006 i 2008. En aquest treball s´ha identificat una nove interacció epistàtica entre l'ecotip Landsberg, originari d'Europa del nord, i Kashmir-2 o Kondara, ambdós originaris d'Àsia Central. Els anàlisi de QTLs en poblacions recombinants Ler x Kas-2 i Ler x Kond indica el requeriment de 3 loci en Ler x Kas-2 i 2 loci en Ler x Kond. Els híbrids incompatibles crescuts a temperatures baixes (16°C) mostren seriosos defectes en el desenvolupament, mort cel.lular espontània i resistència a Hyaloperonospora parasitica. Aquests fenotips es suprimeixen a elevades temperatures o per mutació d'EDS1, o depleció dels nivells d'àcid sal.licílic per transformació amb salicil.lat hidroxilasa (NahG). El grau de severitat en els fenotips observats correlaciona amb els nivells d'àcid sal.licílic, indicant que aquesta molècula és essencial per a la senyalització d'incompatibilitats genètiques en Arabidopsis.

Multiple QTLs influencing triglyceride and HDL and total cholesterol levels identified in families with atherogenic dyslipidemia.

We conducted a genome-wide scan using variance components linkage analysis to localize quantitative-trait loci (QTLs) influencing triglyceride (TG), high density lipoprotein-cholesterol (HDL-C), low density lipoprotein-cholesterol, and total cholesterol (TC) levels in 3,071 subjects from 459 families with atherogenic dyslipidemia. The most significant evidence for linkage to TG levels was found in a subset of Turkish families at 11q22 [logarithm of the odds ratio (LOD)=3.34] and at 17q12 (LOD=3.44). We performed sequential oligogenic linkage analysis to examine whether multiple QTLs jointly influence TG levels in the Turkish families. These analyses revealed loci at 20q13 that showed strong epistatic effects with 11q22 (conditional LOD=3.15) and at 7q36 that showed strong epistatic effects with 17q12 (conditional LOD=3.21). We also found linkage on the 8p21 region for TG in the entire group of families (LOD=3.08). For HDL-C levels, evidence of linkage was identified on chromosome 15 in the Turkish families (LOD=3.05) and on chromosome 5 in the entire group of families (LOD=2.83). Linkage to QTLs for TC was found at 8p23 in the entire group of families (LOD=4.05) and at 5q13 in a subset of Turkish and Mediterranean families (LOD=3.72). These QTLs provide important clues for the further investigation of genes responsible for these complex lipid phenotypes. These data also indicate that a large proportion of the variance of TG levels in the Turkish population is explained by the interaction of multiple genetic loci.

Mapeamento de QTLs para reação ao oídio e mancha-angular do feijoeiro-comum em diferentes locais

Os objetivos deste trabalho foram mapear, em diferentes locais, marcadores RAPD ligados a QTLs (Quantitative Trait Loci) de reação do feijoeiro-comum ao oídio e à mancha-angular, avaliar a interação de QTLs por locais e comparar os métodos de mapeamento e regressão múltipla no processo de detecção de QTLs. Foram avaliadas 196 linhagens recombinantes, oriundas do cruzamento entre as cultivares Carioca e Flor de Mayo, em duas épocas de cultivo: época da seca, para mancha-angular, e inverno para oídio, nos anos de 1996, 1997 e 1998, em dois locais de Minas Gerais. Foram conduzidos sete experimentos de avaliação fenotípica utilizando o delineamento experimental em látice quadrado simples. Os resultados mostraram que a interação QTLs por locais foi significativa, mas foram identificados alguns QTLs com maior estabilidade. O método da regressão múltipla detectou maior número de marcadores ligados a QTLs do que o processo de mapeamento por intervalo composto; não houve concordância entre os resultados apresentados pelos dois métodos. Os marcadores que se mostraram mais estáveis e promissores para serem utilizados em programas de seleção assistida foram OPR02-832, OPD08-759 e OPN10-851 para reação a oídio; OPN02-436, e OPN07-1072 para reação à mancha-angular.

QTLs for resistance to soybean cyst nematode, races 3, 9, and 14 in cultivar Hartwig

The objective of this work was to identify major and minor-effect quantitative trait loci (QTL) for resistance to races 3, 9, and 14 of soybean cyst nematode (SCN) in Hartwig cultivar; to map new resistance QTLs for these races; and to check for the existence of epistatic interactions between QTLs. Cultivar Hartwig is an important resistance source to SCN. Recombinant inbred lines (RIL) obtained from a cross between 'Hartwig' (resistant) and Y23 (susceptible) were evaluated regarding resistance to the three races. New genomic regions for resistance to SCN were identified by microsatellites. Four QTLs, which explained between 12 and 34% of phenotypic variance, were detected for resistance to race 3 in linkage groups (LG) A2, G, J, and M. The QTL in LG G is also important for resistance to race 9. Epistatic interactions were detected between loci, which indicate resistance to races 9 and 14. There are high and low-effect resistance QTLs to SCN.

Genetic associations in the detection of QTLs for wheat spike-related traits

The objective of this work was to assess the genetic diversity and population structure of wheat genotypes, to detect significant and stable genetic associations, as well as to evaluate the efficiency of statistical models to identify chromosome regions responsible for the expression of spike-related traits. Eight important spike characteristics were measured during five growing seasons in Serbia. A set of 30 microsatellite markers positioned near important agronomic loci was used to evaluate genetic diversity, resulting in a total of 349 alleles. The marker-trait associations were analyzed using the general linear and mixed linear models. The results obtained for number of allelic variants per locus (11.5), average polymorphic information content value (0.68), and average gene diversity (0.722) showed that the exceptional level of polymorphism in the genotypes is the main requirement for association studies. The population structure estimated by model-based clustering distributed the genotypes into six subpopulations according to log probability of data. Significant and stable associations were detected on chromosomes 1B, 2A, 2B, 2D, and 6D, which explained from 4.7 to 40.7% of total phenotypic variations. The general linear model identified a significantly larger number of marker-trait associations (192) than the mixed linear model (76). The mixed linear model identified nine markers associated to six traits.

Divergência em QTLs e variância genética para teores de proteína e óleo em soja

Resumo: O objetivo deste trabalho foi avaliar a relação entre os parâmetros de divergência em regiões de QTLs e a variância genética em genótipos de soja, quanto aos teores de proteína e óleo nos grãos. Dois grupos de genótipos foram avaliados, em diferentes ambientes, quanto aos teores de proteína e óleo e genotipados com marcadores moleculares de regiões de QTLs. A partir de cada grupo, estabeleceram-se subgrupos por critérios pré-definidos e avaliou-se a relação entre os parâmetros, tendo-se comparado a divergência média e a variância genética entre os subgrupos. Os subgrupos foram definidos com base nos critérios de diferença em divergência média, homogeneidade e heterogeneidade nos subgrupos e proximidade em uma projeção tridimensional da matriz de distância. As percentagens de concordância entre maiores valores de divergência média e de variância genética para o total de subgrupos de cada grupo inicial foram de 72,5 e de 73,4%, respectivamente. Portanto, nestes genótipos, há relação positiva entre as estimativas de divergência em regiões de QTL e variância genética para os teores de proteína e de óleo dos grãos. As distâncias genéticas com base nos marcadores moleculares de regiões de QTLs são eficientes para a predição da variabilidade genética em genótipos de soja para os teores de proteína e de óleo dos grãos.

Mapeamento de QTLs associados à produção de frutos e sementes em híbridos de Citrus Sunki vs. Poncirus trifoliata

O grupo dos citros apresenta acentuada juvenilidade manifestada pela incapacidade de florescimento, vigor e formação de espinhos. No caso da laranja 'valência', os híbridos de trifoliata, utilizados como porta-enxertos, induzem maior precocidade e produtividade. O número de sementes do fruto (NSF) constitui importante característica da variedade candidata a porta-enxerto de citros. O presente estudo teve como objetivo detectar QTLs associados às características número de frutos por planta (NF) e número de sementes por fruto em uma progênie F1 derivada de um cruzamento entre Citrus sunki e Poncirus trifoliata 'Rubidoux'. Os grupos de ligação (GL) dos parentais foram obtidos usando a estratégia 'pseudotestcross' e marcadores do tipo RAPD. A contagem do número de frutos por planta e o número médio de sementes por fruto foi realizada usando 80 indivíduos da progênie. A detecção dos QTLs foi realizada pelo método de mapeamento por intervalo composto utilizando o programa QTLCartographer. Foram detectados um QTL associado à frutificação e um QTL ligado ao número de sementes situados no GL 4 e GL 5 no mapa de P. trifoliata, respectivamente.

Genetic linkage map of Phaseolus vulgaris and identification of QTLs responsible for resistance to Xanthomonas axonopodis pv. phaseoli

Common bean (Phaseolus vulgaris) cultivars with a high degree of resistance to Xanthomonas axonopodis pv. phaseoli (Xap) are not available in Brazil. Despite many studies, a low degree of resistance to Xap continues to exist due to its complex genetic inheritance, which is not well known. The objectives of this research were to complement a common bean genetic map based on the cross between a susceptible genotype 'HAB-52' and a resistant genotype 'BAC-6', and to map and analyze genomic regions (quantitative trait loci – QTLs) related to Xap resistance. Eleven linkage groups were determined using 143 RAPD markers, covering 1,234.5 cM of the genome. This map was used to detect QTLs associated with Xap resistance on leaves and pods. The averages of disease severity on leaves (represented by the transformed disease index – TDI) and pods (represented by the diameter of lesion on pods – DLP) were added to the data of the linkage map. Five TDI QTLs and only one LDP QTL were detected. The TDI QTLs were placed in the A, B, G and J linkage groups, with phenotypic variations ranging from 12.7 to 71.6%. The DLP QTL explained 12.9% of the phenotypic variation and was mapped in a distinct linkage group. These results indicate that there are different genes involved in the control of resistance on leaves and pods.

Mapping of QTLs related with wood quality and developmental characteristics in hybrids (Eucalyptus grandis x Eucalyptus urophylla)

The present work aimed to characterize and identify QTLs for wood quality and growth traits in E. grandis x E. urophylla hybrids. For this purpose a RAPD linkage map was developed for the hybrids (LOD=3 and r=0.40) containing 52 markers and 12 linkage groups. Traits related to wood quality and growth were evaluated in the QTL analyses. QTL analyses were performed using chi-square tests, single-marker, interval mapping and composite interval mapping analyses. All approaches led to the identification of similar QTLs associated with wood density, cellulose pulp yield and percentage of extractives, which were detected and confirmed by both the interval mapping and composite interval mapping methodologies. Some QTLs regions were confirmed only by the composite interval mapping methodology: percentage of soluble lignin, percentage of insoluble lignin, CBH and total height. Overlapping QTLs regions were detected, and these, can be the result of major genes involved in the regulation and control of the growth traits by epistatic interactions. In order to evaluate the effect of early selection using RAPD molecular data, molecular markers adjacent to QTLs were used genotype selection. The analysis of selection differential values suggests that for all the traits the phenotypic selection at seven years should generate larger genetic gains than early selection assisted by molecular markers and the combination of the strategies should elevate the selection efficiency.

QTLs for shelf life in lettuce co-locate with those for leaf biophysical properties but not with those for leaf developmental traits

Developmental and biophysical leaf characteristics that influence post-harvest shelf life in lettuce, an important leafy crop, have been examined. The traits were studied using 60 informative F-9 recombinant inbed lines (RILs) derived from a cross between cultivated lettuce (Lactuca sativa cv. Salinas) and wild lettuce (L. serriola acc. UC96US23). Quantitative trait loci (QTLs) for shelf life co-located most closely with those for leaf biophysical properties such as plasticity, elasticity, and breakstrength, suggesting that these are appropriate targets for molecular breeding for improved shelf life. Significant correlations were found between shelf life and leaf size, leaf weight, leaf chlorophyll content, leaf stomatal index, and epidermal cell number per leaf, indicating that these pre-harvest leaf development traits confer post-harvest properties. By studying the population in two contrasting environments in northern and southern Europe, the genotype by environment interaction effects of the QTLs relevant to leaf development and shelf life were assessed. In total, 107 QTLs, distributed on all nine linkage groups, were detected from the 29 traits. Only five QTLs were common in both environments. Several areas where many QTLs co-located (hotspots) on the genome were identified, with relatively little overlap between developmental hotspots and those relating to shelf life. However, QTLs for leaf biophysical properties (breakstrength, plasticity, and elasticity) and cell area correlated well with shelf life, confirming that the ideal ideotype lettuce should have small cells with strong cell walls. The identification of QTLs for leaf development, strength, and longevity will lead to a better understanding of processability at a genetic and cellular level, and allow the improvement of salad leaf quality through marker-assisted breeding.

Mapeamento de QTLs para tolerância ao frio e características de importância agronômica em arroz

A temperatura baixa é um estresse comum na cultura do arroz em regiões temperadas, portanto a tolerância ao frio é uma característica altamente desejável em genótipos brasileiros de arroz cultivados no sul do país, onde as temperaturas baixas prejudicam o estabelecimento da lavoura e diminuem o redimento de grãos. O mapeamento molecular é uma estratégia promissora para o estudo e compreensão de de características com controle complexo, tais como a tolerância ao frio em arroz. Com base nisso, os objetivos deste trabalho foram estudar a herança e herdabilidade das características de tolerância ao frio e das características de importância agronômica a serem maepadas, desenvolver um mapa molecular a partir da população segregante F2 proveniente do cruzamento IRGA 417 (Índica) x Quilla 66304 (Japônica) e identificar locos de características quantitativas (QTLs) para a tolerância ao frio no período de germinação e vegetativo e características agronômicas que diferenciam estas duas subespécies. Por fim, investigar a possibilidade de obter indivíduos recombinantes com caracteristicas agronômicas desejáveis e toleância ao frio em uma população F2 do cruzamento entre IRGA 417 x Quilla 66304. As análises das distrbuições de freqüências da geração F2 evidenciaram a dificuldade de estimar o número de genes que controlam as características analisadas, sendo que todas elas apresentam distribuição contínua e segregação transgressiva em relação aos genitores. O mapa foi construído com base em oito marcadores SSR e 42 marcadores do tipo AFLP com uma densidade média de marcador a cada 38,8 cM, sendo o comprimento total do mapa de 581,6 cM. Foram identificados cinco QTLs, sendo que um deles confere tolerância ao frio no período de germinação, e explica 15,9% da variação fenotípica deste caráteer. O outros quatro QTLs identificados foram para largura do grgão (21,3%), esterilidade de espiguetas (61,6%), estatura das plantas (34, 5%) e comprimento do grão (21,9%). A detecção de um QTL associado à tolerância ao frio no período de germinação, e outros QTLs associados à demais características a viabilidade deste tipo de análise. Entretanto, um mapa de ligação enriquecido é nceessário para permitir a detecção de outros QTLs associados às características estudadas. Plantas recombinantes com alto recrescimento de coleóptilo e outrras caracteristicas agronômicas desejáveis foram encontradas, o que evidencia o potencial da população proveniente de IRGA 417 x Quilla 66304 para o melhoramento da tolerância ao frio de genótipos de arroz adaptados ao Sul do Brasil.

Identification of QTLs associated with citrus resistance to Phytophthora gummosis

Citrus gummosis, caused by Phylophthora spp., is an important citrus disease in Brazil. Almost all citrus rootstock varieties are susceptible to it to some degree, whereas resistance is present in Poncirus trifoliata, a closely related species. The objective of this study was to detect QTLs linked to citrus Phylophthora gummosis resistance. Eighty individuals of the F, progeny, obtained by controlled crosses between Sunki mandarin Citrus sunki (susceptible) and Poncirus trifoliata cv. Rubidoux (resistant), were evaluated. Resistance to Phytophthora parasitica was evaluated by inoculating stems of young plants with a disc of fungal mycelia and measuring lesion lengths a month later. Two QTLs linked to gummosis resistance were detected in linkage groups I and 5 of the P. trifoliala map, and one QTL in linkage group 2 of the C sunki map. The phenotypic variation explained by individual QTLs was 14% for C sunki and ranged from 16 to 24% for P. trifoliala. The low character heritability (h(2) = 18.7%) and the detection of more than one QTL associated with citrus Phytophthora gummosis resistance showed that inheritance of the resistance is quantitative.

Mapping QTLs for aluminum tolerance in maize

Aluminum toxicity is one of the major constraints for plant development in acid soils, limiting food production in many countries. Cultivars genetically adapted to acid soils may offer an environmental compatible solution, providing a sustainable agriculture system. The aim of this work was to identify genomic regions associated with Al tolerance in maize, and to quantify the genetic effects on the phenotypic variation. A population of 168 F-3:4 families derived from a cross between two contrasting maize inbred lines for Al tolerance was evaluated using the NSRL and RSRL parameters in nutrient solution containing toxic level of aluminum. Variance analyses indicated that the NSRL was the most reliable phenotypic index to measure Al tolerance in the population, being used for further QTL mapping analysis. RFLP and SSR markers were selected for bulked segregant analysis, and additional SSR markers, flanking the polymorphisms of interest, were chosen in order to saturate the putative target regions. Seven linkage groups were constructed using 17 RFLP and 34 SSR markers. Five QTLs were mapped on chromosomes 2, 6 and 8, explaining 60% of the phenotypic variation. QTL(4) and marker umc043 were located on chromosomes 8 and 5, close to genes encoding for enzymes involved in the organic acids synthesis pathways, a widely proposed mechanism for Al tolerance in plants. QTL(2) was mapped in the same region as Alm2, also associated with Al tolerance in maize. In addition, dominant and additive effects were important in the control of this trait in maize.