This work focuses on the evaluation of a bagging EB method in terms of its ability to select a subset of QTL-related markers for accurate EBV prediction. Experiments were performed on several simulated and real datasets consisting of SNP genotypes and phenotypes. The simulated datasets modeled different dominance levels and different levels of background noises. Our results show that the bagging EB method is able to detect most of the simulated QTL, even with large background noises. The average recall of QTL detection was $0.71$. When using the markers detected by the bagging EB method to predict EBVs, the prediction accuracy improved dramatically on the simulation datasets compared to using the entire set of markers. However, the prediction accuracy did not improve much when doing the same experiments on the two real datasets. The best accuracy of EBV prediction we achieved for the dairy dataset is 0.57 and the best accuracy for the beef dataset is 0.73.

No description available.

Master of Science / Agronomy / Guihua Bai / Guorong Zhang / Fusarium head blight (FHB) is one of the most damaging diseases in wheat, which impacts both grain yield and quality drastically. Recently, the disease has become more prevalent in the hard winter wheat (HWW) grown areas of the United States including Oklahoma where FHB has not been reported before. Growing resistant cultivars is the most economical and effective strategy for disease management. To dissect quantitative trait loci (QTL) for FHB resistance in a moderately resistant hard winter wheat (HWW) cultivar, Overland, a population of 186 recombinant inbred lines (RILs) was developed from the cross between Overland and Overley, a susceptible HWW cultivar from Kansas. The RILs were evaluated for FHB type II resistance in one field and three greenhouse experiments and genotyped using genotyping-by-sequencing (GBS) markers. Three FHB resistance QTLs were mapped on Chromosomes 4DL, 4AL, and 5BL. The QTL on 4DL was the most consistent one and explained up to 13% of the phenotypic variation for type II resistance and 14 % for low Fusarium damaged kernels (FDK). Two GBS markers closely linked to the 4DL QTL were successfully converted to Kbioscience competitive allelic specific PCR (KASP) assays and can be used in marker-assisted breeding. In breeding, a single QTL may provide only partial resistance and pyramiding of several resistance QTLs in a cultivar can provide more protection in FHB epidemics. Fhb1 is a major QTL for FHB resistance from a Chinese source and Fhb3 is an alien gene from wild rye grass (Leymus racemosus). To study the effects of these QTLs individually and cumulatively in hard winter wheat backgrounds, they were transferred into two HWW cultivars Overland and Jagger. The results show that Fhb1 significantly increased FHB resistance, but Fhb3 did not. Thus, Fhb3 is not an effective gene for improvement of FHB resistance in HWW.

Linkage between markers and genes that affect a phenotype of interest may be determined by examining differences in marker allele frequency in the extreme progeny of a cross between two inbred lines. This strategy is usually employed when pooling is used to reduce genotyping costs. When the cross progeny are asexual the extreme progeny may be selected by multiple generations of asexual reproduction and selection. In this thesis I will analyse this method of measuring phenotype in asexual cross progeny. The aim is to examine the behaviour of marker allele frequency due to selection over many generations, and also to identify statistically significant changes in frequency in the selected population. I will show that stochasticity in marker frequency in the selected population arises due the finite initial population size. For Mendelian traits, the initial population size should be at least in the low to mid hundreds to avoid spurious changes in marker frequency in the selected population. For quantitative traits the length of time selection is applied for, as well as the initial population size, will affect the stochasticity in marker frequency. The longer selection is applied for, the more chance of spurious changes in marker frequency. Also for quantitative traits, I will show that the presence of epistasis can hinder changes in marker frequency at selected loci, and consequently make identification of selected loci more difficult. I also show that it is possible to detect epistasis from the marker frequency by identifying reversals in the direction of marker frequency change. Finally, I develop a maximum likelihood based statistical model that aims to identify significant changes in marker frequency in the selected population. I will show that the power of this statistical model is high for detecting large changes in marker frequency, but very low for detecting small changes in frequency.

Sorghum downy mildew (SDM) of maize is caused by the oomycete Peronosclerospora sorghi (Weston and Uppal) C. G. Shaw. The disease can cause devastating yield losses in maize (Zea mays L.). Quantitative trait loci (QTLs) mediating resistance to SDM were mapped using both restriction fragment length polymorphisms (RFLPs), and simple sequence repeats (SSRs) in 220 F2 individual maize progeny derived from a cross between two extremes; highly susceptible inbred parent SC-TEP5-19-1-3-1-4-1-1 (white) and highly resistant inbred P345C4S2B46-2-2-1-2-B-B-B (yellow). The phenotypic expression was assessed on F2:3 families in a wide range of environments under natural field infection and in a controlled greenhouse screening method. Heritability estimates of disease reaction ranged from 93.3% in Thailand sit 1 to 48% in Thailand sit 2. One hundred and thirty three polymorphic markers were assigned to the ten chromosomes of maize with LOD scores exceeding 4.9 covering about 1265 cM with an average interval length between markers of 9.5 cM. About 90% of the genome was located within a 10 cM distance to the nearest marker. Three putative QTLs were detected in association with resistance to SDM in different environments using composite interval mapping. Despite environmental and symptom differences, one QTL on chromosome 2 bin 9 had a major effect in all trials and explained up to 70% of the phenotypic variation in Thailand where the highest disease pressure was experienced. Two other QTLs on chromosome 3 bin 5 and chromosome 9 bin 2 had a minor effect, each explaining no more than 4% of the phenotypic variation. These results revealed one major gene and two minor genes that control sorghum downy mildew resistance. These markers should be very useful in breeding programs in facilitating the introgression of the resistance genes into commercial varieties. Marker-assisted selection for these loci should be useful in incorporating SDM resistance genes in maize across environments, even in the absence of the pathogen.

Sorghum (Sorghum bicolor [L.] Moench) is the world's fifth most economically important cereal crop, grown worldwide as a source of food for both humans and livestock. Sorghum is a C4 grass that is well adapted to hot and arid climes and is popular for cultivation on lands of marginal quality. Recent interest in development of biofuels from lignocellulosic biomass has drawn attention to sorghum, which can be cultivated in areas not suitable for more traditional crops, and is capable of generating plant biomass in excess of 40 tons per acre. While the quantity of biomass and low water consumption make sorghum a viable candidate for biofuels growth, the biomass composition is enriched in lignin, which is problematic for enzymatic and chemical conversion techniques. The genetic basis for stem composition was analyzed in sorghum populations using a combination of genetic, genomic, and bioinformatics techniques. Utilizing acetyl bromide extraction, the variation in stem lignin content was quantified across several sorghum cultivars, confirming that lignin content varied considerably among sorghum cultivars. Previous work identifying sorghum reduced-lignin lines has involved the monolignol biosynthetic pathway; all steps in the pathway were putatively identified in the sorghum genome using sequence analysis. A bioinformatics toolkit was constructed to allow for the development of genetic markers in sorghum populations, and a database and web portal were generated to allow users to access previously developed genetic markers. Recombinant inbred lines were analyzed for stem composition using near infrared reflectance spectroscopy (NIR) and genetic maps constructed using restriction site-linked polymorphisms, revealing 34 quantitative trail loci (QTL) for stem composition variation in a BTx642 x RTx7000 population, and six QTL for stem composition variation in an SC56 x RTx7000 population. Sequencing the genome of BTx642 and RTx7000 to a depth of ~11x using Illumina sequencing revealed approximately 1.4 million single nucleotide polymorphisms (SNPs) and 1 million SNPs, respectively. These polymorphisms can be used to identify putative amino acid changes in genes within these genotypes, and can also be used for fine mapping. Plotting the density of these SNPs revealed patterns of genetic inheritance from shared ancestral lines both between the newly sequenced genotypes and relative to the reference genotype BTx623.

In wheat, cooler canopies have been associated with yield under high temperature stress. The objectives of this study were, i) to understand the role of leaf cuticular waxes as physiological adaptive mechanisms during reproductive stage high temperature stress ii) define quantitative trait loci (QTL) regulating leaf cuticular waxes and determine its link with the QTL for reproductive stage heat tolerance iii) define stable QTL associated with leaf cuticular waxes and yield stability across environments. For the first objective, thirteen wheat cultivars were subjected to a 2-day heat treatment at 38 degrees C at 10 days after pollination (DAP). Leaf cuticular waxes, canopy temperature depression and stomatal conductance were estimated during high temperature stress. At maturity the percent reduction in yield components in each cultivar was calculated. The wheat cultivars 'Kauz' and 'Halberd' had significantly high leaf cuticular wax content of 2.91mg/dm^-2 and 2.36mg/dm^-2 respectively and cooler canopies. Leaf cuticular waxes were significantly correlated with leaf temperature depression and reduction in yield components. A set of 121 recombinant inbred lines (RIL) population derived from the cross of heat tolerant wheat cultivar 'Halberd' and heat susceptible wheat cultivar 'Karl 92' was utilized for QTL mapping. The RIL population received a 2-day short-term high temperature stress at 38°C at 10DAP in 2008 and a long-term high temperature stress at 38 degrees C from 10DAP until maturity in 2009 in the greenhouse. The RIL population was also planted in College Station, Texas in 2009 and 2010 and in Uvalde, Texas in 2010. Leaf cuticular wax was estimated at 10DAP and leaf/spike temperatures were recorded during grain filling. Yield components were estimated after harvest. Heat susceptibility indexes for main spike yield components were estimated in the greenhouse. Overall ten significant QTL were identified for leaf cuticular waxes each explaining 8-19 percent of the variation respectively. Stable QTL for leaf cuticular waxes were located on chromosome 5A and 1B and co-localized with QTL for leaf/spike temperature depression and HSI for kernel weight and single kernel weight of main spike. Another QTL on chromosome 1B contributed by Karl92 was found in the greenhouse and field environments and co-localized with a previously identified QTL on 1B for spike non-glaucousness. The results suggest that leaf cuticular waxes may reduce leaf temperatures and improve adaptation during high temperature stress.

Sorghum downy mildew (SDM) of maize is caused by the oomycete Peronosclerospora sorghi (Weston and Uppal) C. G. Shaw. The disease can cause devastating yield losses in maize (Zea mays L.). Quantitative trait loci (QTLs) mediating resistance to SDM were mapped using both restriction fragment length polymorphisms (RFLPs), and simple sequence repeats (SSRs) in 220 F2 individual maize progeny derived from a cross between two extremes; highly susceptible inbred parent SC-TEP5-19-1-3-1-4-1-1 (white) and highly resistant inbred P345C4S2B46-2-2-1-2-B-B-B (yellow). The phenotypic expression was assessed on F2:3 families in a wide range of environments under natural field infection and in a controlled greenhouse screening method. Heritability estimates of disease reaction ranged from 93.3% in Thailand sit 1 to 48% in Thailand sit 2. One hundred and thirty three polymorphic markers were assigned to the ten chromosomes of maize with LOD scores exceeding 4.9 covering about 1265 cM with an average interval length between markers of 9.5 cM. About 90% of the genome was located within a 10 cM distance to the nearest marker. Three putative QTLs were detected in association with resistance to SDM in different environments using composite interval mapping. Despite environmental and symptom differences, one QTL on chromosome 2 bin 9 had a major effect in all trials and explained up to 70% of the phenotypic variation in Thailand where the highest disease pressure was experienced. Two other QTLs on chromosome 3 bin 5 and chromosome 9 bin 2 had a minor effect, each explaining no more than 4% of the phenotypic variation. These results revealed one major gene and two minor genes that control sorghum downy mildew resistance. These markers should be very useful in breeding programs in facilitating the introgression of the resistance genes into commercial varieties. Marker-assisted selection for these loci should be useful in incorporating SDM resistance genes in maize across environments, even in the absence of the pathogen.

Dry beans (Phaseolus vulgaris L.) contain high levels of folates. These compounds are essential vitamins and folate deficiencies may lead to a number of health problems. The objectives of this study were to examine the mode of inheritance of folate content and identify quantitative trait loci (QTL) associated with folate content in dry beans. Inheritance of folate content was studied in the F1 hybrids of one-way diallel crosses among Othello, AC Elk, Redhawk and Taylor, and an F2 population of the cross between Redhawk and Othello. Total folate content and 5 methyltetrahydrofolate (5MTHF) were measured twice within a one hour interval. Significant variation in folate content was observed among the parental genotypes, their F1 hybrids, and the F2 individuals of a cross between Redhawk and Othello, ranging from 147 to 345 µg/100g. Reductions in the 5MTHF content and total folate content values in the second measurement from samples were highly variable for all four parental lines ranging from 5 to 30% and 7 to 33%, respectively. A single marker QTL analysis identified at least three QTL for folate content in the F2 population. For the majority of identified QTL, dominance effects appeared to be the major genetic effect.

Glucosinolates are amino acid derived secondary metabolites found in the order Capparales. It is an important class of phytochemicals involved in plant-microbe, plant-insect, plant-animal and plant-human interactions. It is, therefore, important to understand genetic mechanism of glucosinolate biosynthesis in Brassica for efficient manipulation. In this study, QTL mapping of leaf and seed glucosinolates was performed in B. rapa using two RIL populations, SR-RILs and BU-RILs. QTL mapping was performed using SR-RILs developed from a cross of Chinese cabbage and turnip rapeseed and a genetic map in B.rapa. Genetic map was developed using a total 1,579 molecular markers including 9 markers specific to glucosinolate genes, GSL-ELONG, GSL-PRO, GSL-FMOOX1, and GSL-AOP/ALK. Several QTL for progoitrin, gluconapin, glucoalyssin, glucobrassicanapin, 2-methylpropyl and 4-hydoxyglucobrassicin glucosinolates were identified with phenotype variance between 6 and 54%. Interestingly, a major QTL for 5C aliphatic glucosinolates was co-localized with a candidate Br-GSL-ELONG locus on linkage group A3, displayed co-segregation with co-dominant SCAR marker BrMAM1-1. The Br-GSL-ELONG locus was identified to regulate 20 µmole/g seed 5C glucosinolate biosynthesis. BU-RILs derived from a cross of yellow sarson and USU9 was segregated for glucoerucin, gluconapin and progoitrin 4C aliphatic glucosinolates with 4-hydoxyglucobrassicin. Phenotyping was performed in controlled and field environments for seed glucosinolates and controlled environments for leaf glucosinolates. Genetic map was developed using SRAP markers and glucosinolate gene, GSL-ELONG and GSL-PRO specific 4 loci were integrated on map. Four and three QTL were identified for seed glucoerucin and gluconapin, respectively in both environments with phenotypic variance up to 49%. Additionally, genetic manipulation of glucosinolates was performed by backcross with MAS in B. rapa. Resynthesized B. napus line was backcrossed with B. rapa genotypes, RI16, BAR6 and USU9 for replacement or introgression of glucosinolate genes, GSL-ELONG- and GSL-PRO+. In RI16 genotype, 15 to 25 µmole/g seed 5C glucosinolates reduced in 15 BC3F2 lines those were positive with GSL-ELONG- marker and negative with the A-genome and gene specific marker BrMAM1-1. This suggests that the functional allele has replaced by non-functional from B. oleracea. GSL-PRO+ positive backcross lines in RI16 genotype displayed sinigrin 3C aliphatic glucosinolate in B. rapa. This suggests introgression of GSL-PRO+ in B. rapa.