It has been reported that an individual’s initial level of response to a drug might be predictive of his or her future risk of becoming dependent, thus basal gene expression profiles underlying those drug responses may be informative for both predicting addiction susceptibility and determining targets for intervention. This dissertation research aims to elucidate genetic risk factors underlying acute alcohol and nicotine dependence phenotypes using mouse genetic models of addiction. Phenotyping, brain region-specific mRNA expression profiling, and genetic mapping of a recombinant inbred panel of over 25 mouse strains were performed in order to identify quantitative trait loci (QTL) harboring candidate genes that may modulate these phenotypes. Previous BXD (B6 x D2) behavioral studies performed in our laboratory identified an ethanol-induced anxiolysis-like QTL (Etanq1) in the light dark box (LDB). We hypothesized that genetic variation within Nin (a gene within the Etanq1 support interval involved in microtubule-anchoring) may modulate anxiolytic-like responses to acute ethanol in the LDB as well as other preclinical models of anxiety, the elevated plus maze (EPM), and marble burying (MB) task. Molecular studies have allowed us to confirm cis regulation of Nin transcript levels in the NAc. To elucidate potential mechanisms mediating Etanq1, the pharmacological tools, diazepam and HZ166 (a benzodiazepine derivative) were utilized to interrogate whether GABAA receptor activation modulates ethanol’s anxiety-like behaviors in the LDB. We show that the LDB phenotype, percent time spent (PTS) in the light following a brief restraint stress, is not being modulated through direct activation of GABAA α2/α3 receptor subunits. To genetically dissect Etanq1 as well as parse the ethanol anxiolytic-like phenotype, we have assayed 8 inbred strains, selected based on genotypes at Nin, in various preclinical models of anxiety. Principal components analysis of these behavioral data suggests that the gene(s) modulating the ethanol anxiolytic-like component in the LDB do not overlap with similar phenotypes in the elevated plus maze (EPM), nor the MB phenotype. Furthermore, site-specific delivery of an sh-Nin lentivirus into the NAc of D2 mice revealed that Nin may modulate one LDB endophenotype, latency to enter the light side of the LDB, which loaded as a part of the “anxiolysis” principal component. These data strongly imply that basal neuronal Nin expression in the NAc is important for acute ethanol anxiolytic-like behavior, perhaps through a novel mechanism involving synaptic remodeling. In separate behavioral QTL mapping studies, we hypothesized that genetic variation regulating expression of Chrna7 modulates the reward-like phenotype, conditioned place preference (CPP), for nicotine. We provide evidence for genetic regulation of Chrna7 across the BXD panel of mice and through pharmacological and genetic behavioral studies, confirm Chrna7 as a quantitative trait gene modulating CPP for nicotine in mice. Microarrays, followed by network analyses, allowed us to identify a genetically co-regulated network within the nucleus accumbens (NAc), differentially expressed in mice null for Chrna7, which was similarly correlated in the BXD panel of mice. Our network and molecular analyses suggest a putative role for Chrna7 in regulating insulin signaling in the NAc, which together, may contribute to the enhanced sensitivity to nicotine observed in strains of mice that lack or have low mRNA levels of Chrna7 in the NAc. Overall, this research has elucidated and confirmed new genetic risk factors underlying alcohol and nicotine dependence phenotypes and has enabled a better understanding of the neurogenomic bases of alcohol and nicotine addiction. Future studies that further investigate the signaling pathways and/or gene interactions involving Nin and Chrna7 may lead the field to new candidates for pharmacotherapies that may be tailored for use in individuals with susceptible genotypes. Supported by NIAAA grants P20AA017828 and R01AA020634 to MFM, NIDA T32DA007027 to WLD, and NIDA R01DA032246 to MFM and MID.

Master of Science / Department of Agronomy / Allan K. Fritz / Fusarium head blight (FHB), caused by Fusarium graminearum, is an important cereal disease in humid and semi-humid wheat growing regions. In recent FHB epidemics in the USA, FHB dramatically reduced wheat yields and grain quality due to mycotoxin contamination. Five types of FHB resistance have been reported, but resistance to disease spread within a spike (Type II) and low deoxynivalenol (DON) accumulation in infected kernels (Type III) have drawn the most attention. A Chinese Spring-Sumai3 chromosome 7A substitution line (CS-SM3-7ADSL) was reported to have a high level of Type II resistance, but quantitative trait locus (QTL) on chromosome 7A has never been mapped. To characterize QTL on chromosome 7A, we developed 191 Chinese Spring-Sumai3-7A chromosome recombinant inbred lines (CRIL) from a cross between Chinese Spring and CS-SM3-7ADSL and evaluated the CRIL in a greenhouse for both types of resistance in three experiments. Two major QTL with Sumai 3 (SM3) origin, conditioning Type II and Type III resistance were mapped in chromosomes 3BS and 7AC. QTL on chromosome 3BS corresponds to Fhb1, previously reported from SM3, whereas 7AC QTL, designated as Fhb5, is a novel QTL identified from SM3 in this study. Fhb5 explains 22% phenotypic variation for Type II resistance and 24% for Type III resistance. Marker Xwmc17 is the closest marker to Fhb5 for both types of resistance. Fhb1 and Fhb5 were additive and together explained 56% variation for Type II and 41% for Type III resistance and resulted in 66% reduction in FHB severity and 84% in DON content. Both QTL showed significant pleiotropy effects on Type II and Type III resistance, suggesting both types of resistance may be controlled by the same gene(s). Haplotype analysis of SM3’s parents revealed that Fhb5 originated from Funo, an Italian cultivar. A survey of worldwide germplasm collection of 400 accessions showed that Fhb5 is present mainly in Chinese cultivars, especially in Funo-related accessions. Further, Fhb5 is the second major QTL from SM3 and have potential to be used in improving wheat cultivars for both types of resistance.

Master of Science / Division of Biology / Michael A. Herman / Caenorhabditis elegans are free-living bacterivorous nematodes that naturally consume bacteria as food source. As an excellent genetic model, C. elegans has proven to be a successful system to study innate immune responses to human pathogens, which resulted in identification of many evolutionarily conserved defense pathways. Most of these studies examined innate immune pathway mutants in a single genetic background in response to monoculture of human pathogens that worms might not necessarily encounter in the wild. While this has led to the successful genetic dissection of these defense pathways, in order to fully understand their biological functions, the relevant ecological and evolutionary context needs to be taken into account. The bacterial environment C. elegans naturally encounter is likely to be highly heterogeneous. While many bacteria are mainly considered as dietary resource for worms, some could be potential pathogens. Worms thus constantly face the challenge to defend against the pathogens mixed in the food. Stenotrophomonas maltophilia is one such bacterium. S. maltophilia is a ubiquitous bacterium that has been found associated with native nematodes. But it can also cause nosocomial infections in human, especially in immune-compromised individuals. Due to its natural resistance to multiple antibiotics, it has been emerging as an opportunistic human pathogen. Our lab isolated a S. maltophilia strain, JCMS, which was found being pathogenic to C. elegans. Both C. elegans strains, N2 (Bristol, England) and CB4856 (Hawaii), showed decreased survivorship when fed on S. maltophilia JCMS compared to E. coli OP50. However, more interestingly, the specific responses towards bacteria are different between strains. This indicates that survivorship of C. elegans is determined by not only genetic and environmental factors, but also genotype by environment (G×E) interactions (GEI). In order to identify the underlying genetic basis, we mapped quantitative trait loci (QTL) in a N2×CB4856 recombinant inbred panel for the survivorship in response to E. coli OP50 and S. maltophilia JCMS.

Breeding programmes that use elite sires with the best estimated breeding values for muscling traits have achieved significant improvement in lamb production in the UK. Further acceleration of the rate of genetic gain for the desirable production traits could be achieved using DNA marker-assisted selection (MAS) breeding strategies. The underlying causal genetic variants associated with improved muscling may be unknown and lying between a cluster of genes known as quantitative trait loci (QTL) or could be single nucleotide polymorphisms (SNP). LoinMAXTM, Texel muscling QTL (TM-QTL) and c.*1232G > A myostatin mutation were genetic variants that reported to be associated with improved muscling characteristics and hence subjected to further analysis in this project. It is essential before incorporating segregating genetic variants in any breeding scheme to comprehensively evaluate their effects on carcass traits. In-vivo scanning (ultrasound scanning (US) and computed tomography scanning (CT)), and carcass video image analyses (VIA) were used in the current studies. Objective VIAprediction weights of the carcass primal cuts could be the backbone of a value-based marketing system that is suggested to replace the current Meat and Livestock Commission (MLC) carcass grades for conformation scores (MLC-C) and fat class (MLC-F). The effect of a single copy of LoinMAXTM QTL (LM-QTL) compared to noncarriers was evaluated in UK crossbred lambs out of Scottish Mule ewes. M. longissimus lumborum (MLL) width, depth and area, as measured by CT scanning, were significantly greater in lambs heterozygous for LM-QTL compared to noncarriers. VIA detected a significant effect of the LM-QTL on the predicted weight of saleable meat yield in the loin primal cut (+2.2%; P < 0.05). The effects of the ovine c.*1232G > A myostatin mutation (MM), found on sheep chromosome 2, on carcass traits in heterozygous crossbred lambs sired by Texel and Poll Dorset rams were studied. Texel crossbred lambs carrying MM had increased loin depth and area. In both crossbred lambs, MM-carriers had significantly higher CT-estimated lean weight and proportion (2 to 4%) and muscle to bone ratios (by ~3%). Poll Dorset heterozygous crossbred animals had higher muscle to fat ratio (28%) and significantly lower fat-related measurements. The c.*1232G > A (MM) mutation as well as TM-QTL effects were evaluated in a different genetic background of Texel x Welsh Mountain crossbreed lambs. Carrying two copies of MM was associated with a significant positive effect on 8 week weight, a negative effect on ultrasound fat depth, a substantial decrease in MLC-fat score, positive impact on VIA-estimated weight of the hind leg, chump and loin primal cuts, as well as the muscularity of the hind leg and loin regions with greater loin muscle width, depth and area. Two copies of MM altered lambs‟ morphological traits with significantly wider carcasses across the shoulders, breast and hind legs and greater areas of the back view of the carcass when measured by VIA. TM-QTL significantly increased US-muscle depth and TM-QTL carriers had significantly greater loin muscle width and area measurements. Comparing TM-QTL genetic groups (homozygote allele carriers (TM/TM), heterozygote carriers of paternal and maternal origin of allele (TM/+ and +/TM, respectively) and homozygote non-carriers (+/+)) and TM-QTL mode of action were then studied. TM/TM carcasses were significantly heavier than non-carriers by 1.6 kg and scored higher conformation values when compared to heterozygote groups only. TM/+ lambs had significantly higher VIA-predicted weight and muscularity in the hind leg and loin, and higher loin dimensions relative to some other genotypic groups. The effect of TM-QTL on some carcass shape measurements was significant. TM-QTL mode of action results on the loin muscling traits supports the earlier reports of polar over dominance. In the light of growing calls to replace the current subjective carcass payment system with the objective VIA system that values the carcass according to the superiority of its cuts, I investigated the ability of US and CT measurements to predict the VIAestimated weights of the carcass primal cuts. Several prediction equations were examined but the best could be achieved when ultrasound measurement, CT linear measurements and live weight were fitted in the model. Since CT scanning information of elite sires is now being used for genetic selection for carcass merit, genetic parameters and genetic relationships between CT scanning measurements and post mortem traits (VIA and MLC-FC) were estimated. However, results were not sufficiently accurate to be of practical use due to lack of data.

Orientador: Lucia Galvão de Albuquerque / Resumo: Registros de características quantitativas e informações genotípicas cole- tadas para cada animal são utilizados para identificar regiões do genoma associadas à variação fenotípica. No entanto, essas investigações são, geralmente, realizadas com base em testes estatísticos de correlação ou associação, que não implicam em causalidade. A fim de explorar amplamente essas informações, métodos poderosos de inferência causal foram desenvolvidos para estimar os efeitos causais entre as variáveis estudadas. Apesar do progresso significativo neste campo, inferir os efeitos causais entre variáveis aleatórias contínuas ainda é um desafio e poucos estudos têm explorado as relações causais em genética quantitativa e no melhoramento animal. Neste contexto, dois estudos foram realizados com os seguintes objetivos: 1) Buscar as relações causais entre as características de carcaça e qualidade de carne usando um modelo de equação estrutural (MEE), sob modelo linear misto em bovinos da raça Nelore, e 2) Reconstruir redes de genes-fenótipos e realizar análise de rede causal por meio da integração de dados fenotípicos, genotípicos e transcriptômicos em bovinos da raça Nelore. Para o primeiro estudo, um total de 4.479 animais com informação fenotípica para o peso da carcaça quente (PCQ), área de olho lombo (AOL), espessura de gordura subcutânea (EGS), força de cisalhamento (FC) e marmoreio (MAR) foram usados. Os animais foram genotipados usando os painéis BovineHD Bead- Chip e GeneSeek Genomic Pro... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Quantitative traits and genotypes information have been collected for each animal and used to identify genome regions related to phenotypes variation. However, these investigations are, usually, performed based on correlation or association statistical tests, which do not imply in causation. In order to fully explore these information, powerful causal inference methods have been developed to estimate causal effects among the variables under study. Despite significant progress in this field infer causal effect among random variables remains a challenge and some few studies have explored causal relationships in quantitative genetics and animal breeding. In this context, two studies were performed with the following objectives: 1) Search for the causal relationship among carcass yield and meat quality traits using a structural equation model (SEM), under linear mixed model context in Nelore cattle, and 2) Reconstruct gene-phenotype networks and perform causal network analysis through the integrating of phenotypic, genotypic, and transcriptomic data in Nelore cattle. For the first study, a total of 4,479 animals with phenotypic information for hot carcass weight (HCW), longissimus muscle area (LMA), backfat thickness (BF), Warner-Bratzler shear force (WBSF), and marbling score (MB) traits were used. Animals were genotyped using BovineHD BeadChip and GeneSeek Genomic Profiler Indicus HD - GGP75Ki. For causal inference using SEM a multistep procedure methodology was used as follow:... (Complete abstract click electronic access below) / Doutor

Von Willebrand Disease (VWD) is the most common bleeding disorder. In addition to known major genes, genetic modifiers, such as ABO blood group, affect quantitative outcome measures for VWD. The data consist of an 854-member Amish pedigree with established linkage of VWD to a locus within the Von Willebrand Factor (VWF) gene on chromosome 12. The DNA sequence of the causative mutation is known. Phenotypic information and genotypic data consisting of VWF mutation status and a genome screen of markers are available for 385 pedigree members. Genetic modifiers of the VWF mutation are investigated using known and new conditional linkage methods that search for modifier genes of a major gene with known mutation. The MCMC-based program LOKI was used to conduct multipoint linkage analysis of VWD outcome measures while controlling for the VWF mutation. Adjustment for the mutation did not eliminate the linkage signal on chromosome 12 in the same location as the VWF mutation. Evidence for QTLs was also found on six other chromosomes. Smod, a score statistic that detects evidence of a genetic modifier conditional on linkage to a major gene, was developed for sib pair data. To limit the modifier gene main effect, Smod was developed so that variance due to the modifier locus is bounded above by the variance of the interaction between major gene and modifier gene. The performance of Smod was compared to other published score statistics. Power to detect linkage to the modifier locus depended on major gene and modifier gene risk allele frequencies, relative contribution of the major gene main effect to the interaction effect, and the upper bound on the modifier gene main effect. The Amish pedigree was broken up into sib pair data and analyzed using Smod and other score statistics. Using these statistics, the strongest evidence for QTLs for VWD was also found on chromosome 12 in the region of the VWF mutation. Combined with the LOKI results, further analysis will help determine if intragenic modification is occurring or if linkage disequilibrium between the mutation and analyzed markers is driving results.

Needle-to-stem unit rate (NESTUR) is a stem growth index of conifer seedling trees that measures the efficiency of stemwood production per unit of needle growth. Five experiments were carried out in this thesis using progenies of two unrelated full-sib radiata pine crosses. The initial experiment (experiment 1) applied the bulked segregant analysis technique to determine whether RAPD analysis could be successfully extended to the development of molecular markers for NESTUR in radiata pine. The NESTUR values of 174 progenies of the full-sib family 12038 x 10946 were determined. Based on the genotypic analysis of the individuals, two quantitative trait loci (QTL) controlling NESTUR were identified at ANOVA P-levels of 0.01-0.001. An absence of RAPD fragment markers generated by primers OPE-06 and OPA-10 was associated with low NESTUR values, while primer UBC-333 generated a 550 bp band that was associated with high NESTUR values. Linkage to components of NESTUR (increments in stem diameter and stem volume) was demonstrated for one of the QTL, while the other was unique to NESTUR, and not shared with the components. There was a significant interaction between the two QTLs. Presence of OPA-101200 locus appeared to inhibit expression of the QTL linked to UBC-333 [subscript 550]. ¶ To further analyse the quantitative trait loci (QTLs) controlling NESTUR, a linkage map was constructed from RAPD markers segregating in 93 haploid progeny of another full sib cross (30040 x 80121) (experiment 2). Two hundred and sixty-two (262) markers were mapped to 14 linkage groups of at least 7 markers, ranging in size from 39 to 183 cM. The 14 linkage groups covered approximately 1511 cM of genetic map distance. ¶ In experiment 3, the linkage map was used to map QTLs controlling NESTUR, as well as increments in seedling stem diameter, volume, and height and needle volume. Altogether, five putative QTLs were detected for NESTUR, with explained variation ranging from 9 to 22%. Of the five QTLs detected, 3 were coincidental with those for stem growth in height, diameter and volume. The two QTL positions that were unique to NESTUR were flanked by QTLs for the component traits. Together, effects of the five QTLs explained 48% of the total phenotypic variation for NESTUR. ¶ Ability of identified markers to predict the phenotype and seedlings with growth potential was assessed in the cross 30040 x 80121, using six RAPD markers associated with NESTUR at ANOVA P-levels of 0.01-0.001 (experiment 4). The correlation between observed NESTUR and predicted values was 0.70. Differences in observed vs. predicted values were not large and did not indicate serious misclassifications, such as classification of an upper ranking individual into the lower group, or vice versa. ¶ Over a two-year growth period, the ability of NESTUR to predict stem growth was strongly affected by seedling age. In contrast, markers linked to NESTUR showed a consistent ability to predict stem growth, irrespective of seedling age. Compared with the top 1% of the original population, seedlings selected for their genotypic values showed a higher stem volume growth of 103% in the first year, and 76% in the second year. ¶ The expression of QTLs for stem volume, stem diameter, height, number of branches, number of whorls, and branches/whorl were compared at 5, 12, and 24 months of age. Two QTLs detected for height showed contrasting expression over two years, one was gradually reduced from LOD of 2.70 to 0.43 and the other increased from 1.12 to 2.45. Compared with the pattern observed for height, LOD scan profiles for diameter and volume showed less temporal change of peaks, suggesting that the genetic control for height growth is probably more unstable than that of diameter. QTLs controlling the phenotype at the time of measurement (ie the final phenotype) showed similar magnitude of effects on that trait's respective increments (or growth rate).

The seed size and shape of lentil are important traits because they determine the market class, cooking time, and can influence quality and yield of milled lentils. Understanding the genetic control of seed size and shape can help breeders develop varieties with improved seed size and shape characteristics such as seed diameter, seed thickness and seed plumpness. The objectives were to determine the heritability of seed size and shape and identify the genomic regions controlling these traits. This involved i) developing a linkage map for the LR-18 population (CDC Robin x 964a-46) using a recently developed single nucleotide polymorphism (SNP) assay; ii) analyzing the LR-18 population for seed size and shape QTLs; iii) analyzing an association mapping panel for seed size and shape QTLs. Phenotyping trials were grown at two different locations in Saskatchewan, Canada. The mapping population was grown in two different years while the association panel was only grown in one. Seed diameter and thickness were measured using sieves and this data were used to calculate seed plumpness. Days to flowering was also recorded to determine if it had any effect on seed size or shape. A linkage map consisting of 537 SNPs, 10 SSRs and 4 morphological markers on seven linkage groups was constructed and used for the QTL analysis. The heritability estimates were high for seed diameter and seed plumpness (0.92 and 0.94, respectively) while for seed thickness and days to flowering they were more moderate (0.60 and 0.45, respectively). QTL analysis revealed QTLs on five of the seven linkage groups. The association mapping study revealed similar heritability estimates of 0.97, 0.62, 0.94, and 0.62 for seed diameter, seed thickness, seed plumpness and flowering time, respectively. There were 31 different significant marker trait associations, however only 5 of those were significant for both locations. Four of those five markers did not map in the LR-18 linkage map so their genomic locations are still to be determined. Results showed that there are key regions in the genome that control seed size and shape and flowering time in lentil. These markers could be used for marker-assisted selection or for further candidate gene analysis.

Domestication is the process by which animals become adapted to the environment provided by humans. The process of domestication has let to a number of correlated behavioural, morphological and physiological changes among many domesticated animal species. An example is the changes of plumage colour in the chicken. Plumage colour is one of the most readily observable traits that make distinction between breeds as well as between strains within a breed. Understanding the genetic architecture of pigmentation traits or indeed any trait is always a great challenge in evolutionary biology. The main aim of this study was to map quantitative trait loci (QTLs) affecting the red and metallic green coloration in the chicken plumage. In this study, a total of 572 F8 intercross chickens between Red Junglefowl and White Leghorn were used. Phenotypic measurements were done using a combination of digital photography and photography manipulating software. Moreover, all birds were genotyped with 657 molecular markers, covering 30 autosomes. The total map distance covered was 11228 cM and the average interval distance was 17 cM. In this analysis, a total of six QTLs (4 for red and 2 for metallic green colour) were detected on four different chromosomes: 2, 3 11 and 14. For red colour, the most significant QTL was detected on chromosome 2 at 165 cM. An additional QTL was also detected on the same chromosome at 540 cM. Two more QTLs were detected on chromosomes 11 and 14 at 24 and 203 cM respectively. Additionally, two epistatic pairs of QTLs were also detected. The identified four QTLs together can explain approximately 36% of the phenotypic variance in this trait. In addition, for metallic green colour, one significant and one suggestive QTLs were detected on chromosomes 2 and 3 at 399 and 247 cM respectively. Moreover, significant epistatic interactions between these two QTLs were detected. Furthermore, these two QTLs together can explain approximately 24% of the phenotypic variance in this trait. These findings suggest that the expression of pigmentation in the chicken plumage is highly influenced by both the epistatic actions and pleiotropic effects of different QTLs located on different chromosomes.

The protein quality of maize can be improved by replacing normal Opaque2 alleles with non-functional recessive alleles opaque2 (o2). The allele o2 produces a severe phenotype with soft endosperm enhancing its protein quality but decreasing its agronomical value. Plant breeders have restored a desirable ratio of hard to soft endosperm in o2 germplasm known as Quality Protein Maize (QPM). Neither the mechanism nor the genetic components by which the modification of the endosperm in QPM lines occurs are well understood. To increase the understanding of the genetics of endosperm modification, a population of 146 recombinant inbred lines derived from a cross between the o2 inbred line B73o2 and the QPM inbred line CML161 was evaluated in two Texas locations from 2004 to 2006. Four traits related to endosperm texture were measured and showed significant effect of the inbred lines, high heritability estimates and high genetic correlations. Relative content of the essential amino acids lysine, tryptophan and methionine were measured and showed significant effects of the lines and considerable high genetic correlations and heritabilities. Negative correlation was observed between endosperm texture traits and amino acid content. Quantitative trait loci (QTL) were mapped for traits related to the modification of endosperm texture and the content of lysine, tryptophan and methionine. QTLs clusters for endosperm texture traits were detected on chromosomes 3, 5, 6 and 8 explaining 62-68% of the variation. QTLs clusters for amino acid contents were located on chromosomes 7 and 8 that explained up to 39% of the observed variation. The product of the O2 gene is a transcription factor that affects the expression of a number of endosperm genes. A group of 29 endosperm genes associated with the O2 activity were evaluated in developing endosperm of the recombinant inbred lines. Genomic regions controlling gene transcript abundance in developing endosperm were identified by expression QTL mapping. Evidence is presented of QTL hot spots that segregate in association with endosperm texture modification or amino acid contents and are associated with the regulation of the expression of a group of endosperm genes.