Doctor of Philosophy / Department of Agronomy / Jianming Yu / Knowledge of the genetic bases of grain quality traits will complement plant breeding efforts to improve the end use value of sorghum (Sorghum bicolor (L.) Moench). The objective of the first experiment was to assess marker-trait associations for 10 grain quality traits through candidate gene association mapping on a diverse panel of 300 sorghum accessions. The 10 grain quality traits were measured using the single kernel characterization system (SKCS) and near-infrared reflectance spectroscopy (NIRS). The analysis of the accessions through 1,290 genome-wide single nucleotide polymorphisms (SNPs) separated the panel into five subpopulations that corresponded to three major sorghum races (durra, kafir, and caudatum), one intermediate race (guinea-caudatum), and one working group (zerazera/caudatum). Association analysis between 333 SNPs in candidate genes/loci and grain quality traits resulted in eight significant marker-trait associations. A SNP in starch synthase IIa (SSIIa) gene was associated with kernel hardness (KH) with a likelihood ratio–based R[superscript]2 (R[subscript]L[subscript]R[superscript]2) value of 0.08. SNPs in starch synthase (SSIIb) gene (R[subscript]L[subscript]R[superscript]2 = 0.10) and loci pSB1120 (R[subscript]L[subscript]R[superscript]2 = 0.09) was associated with starch content. Sorghum is a crop well adapted to the semi arid regions of the world and my harbor genes for drought tolerance. The objective of second experiment was to identify quantitative trait loci (QTLs) for yield potential and drought tolerance. From a cross between Tx436 (food grain type) and 00MN7645 (drought tolerant) 248 recombinant inbred lines (RILs) was developed. Multi-location trials were conducted in 8 environments to evaluate agronomic performance of the RILs under favorable and drought stress conditions. The 248 RILs and their parents were genotyped by genotyping-by-sequencing (GBS). A subset of 800 SNPs was used for linkage map construction and QTL detection. Composite interval mapping identified a major QTLs for grain yield in chromosome 8 and QTL for flowering time in chromosome 9 under favorable conditions. Three major QTLs were detected for grain yield in chromosomes 1, 6, and 8 and two flowering time QTLs on chromosome 1 under drought conditions. Six QTLs were identified for stay green: two on chromosome 4; one each on chromosome 5, 6, 7, and 10 under drought conditions.

Association mapping (AM), based on linkage disequilibrium, is a complementary strategy to traditional quantitative trait loci (QTL) mapping for describing associations between genotypes and phenotypes in crop plants. Yellow endosperm colour, an important quality trait in durum wheat (<i>Triticum turgidum L. var. durum</i>), was studied to determine the potential of AM to (1) identify previously reported QTL using a genome wide scan and (2) to determine allelic association of the phytoene synthase 1 (Psy1) gene using a candidate gene analysis. At present, a number of QTL for endosperm colour have been identified, and phytoene synthase, the initial enzyme of the carotenoid biosynthetic pathway, has been associated with QTL on the group 7 chromosomes which are considered to play a significant role in expression of yellow pigment concentration. CIE 1976 b*, a light reflectance measurement, and water-saturated butanol extracted pigments were assessed on a collection of 93 elite accessions from a variety of geographic origins, and genotyped with 245 markers. Population structure was assessed using genetic distance and Bayesian model based approaches, identifying five sub-populations consistent with breeding origin and pedigree. Association analysis identified significant associations with yellow endosperm colour on all chromosomes, including several previously identified QTL as well as new regions for genomic dissection. Pairwise LD mapping of Psy1-B1 and Psy1-A1 located the genes to chromosomes 7B and 7A respectively, to regions which have previously been identified for yellow pigment concentration QTL. The results of this study indicate that AM can be used to complement traditional QTL mapping techniques, and identify novel QTL for further study.

Early maturity is an important objective for breeding flax adapted to the Western Canada. Crop canopy traits influence seed yield; however, studying its effects is challenging due to the complexity and limited knowledge of the genetics of this trait. The objectives of this research are : i) to characterize flax accessions from the Canadian gene bank collection for early flowering, maturity and canopy traits; ii) to identify SSR markers associated with plant branching and leaf area index (LAI); iii) to use Structural Equation Modeling (SEM) to identify canopy variables with significant effects on yield. The flax core collection, consisting of approximately 381 accessions, was grown at the Kernen Crop Research Farm in 2010 and 2011. Additionally, 17 early and 17 late flowering accessions from the flax core collection were screened and their phenotypic responses in both growth chamber and field environments were measured. A large amount of phenotypic diversity was observed in long day and short day environments in these experiments. Some accessions appeared to be more photosensitive, while others were photoperiod insensitive. The genetic control of canopy traits such as LAI and plant branching were studied using association mapping. Genotyping of the core collection was conducted using 375 SSR markers. Population structure analysis assigned the 381 flax accessions in the core collection into four distinct groups. Model comparison revealed that the mixed linear model reduced spurious marker trait associations. A total of 26 markers were identified to be significantly associated with plant branching and LAI. The simultaneous examination of crop phenology and canopy traits to seed yield was performed using SEM analysis. The results indicated greater plant stand resulted in higher irradiance absorption and which resulted in greater seed yield. Days to flowering had a significant negative effect on seed yield and growing degree days to maturity had a significant effect on seed yield. Plant branching and plant height had a positive non-linear effect on seed yield. This study has provided several insights into molecular approaches and statistical methods to improve flax breeding.

Association mapping (AM), based on linkage disequilibrium, is a complementary strategy to traditional quantitative trait loci (QTL) mapping for describing associations between genotypes and phenotypes in crop plants. Yellow endosperm colour, an important quality trait in durum wheat (<i>Triticum turgidum L. var. durum</i>), was studied to determine the potential of AM to (1) identify previously reported QTL using a genome wide scan and (2) to determine allelic association of the phytoene synthase 1 (Psy1) gene using a candidate gene analysis. At present, a number of QTL for endosperm colour have been identified, and phytoene synthase, the initial enzyme of the carotenoid biosynthetic pathway, has been associated with QTL on the group 7 chromosomes which are considered to play a significant role in expression of yellow pigment concentration. CIE 1976 b*, a light reflectance measurement, and water-saturated butanol extracted pigments were assessed on a collection of 93 elite accessions from a variety of geographic origins, and genotyped with 245 markers. Population structure was assessed using genetic distance and Bayesian model based approaches, identifying five sub-populations consistent with breeding origin and pedigree. Association analysis identified significant associations with yellow endosperm colour on all chromosomes, including several previously identified QTL as well as new regions for genomic dissection. Pairwise LD mapping of Psy1-B1 and Psy1-A1 located the genes to chromosomes 7B and 7A respectively, to regions which have previously been identified for yellow pigment concentration QTL. The results of this study indicate that AM can be used to complement traditional QTL mapping techniques, and identify novel QTL for further study.

Population based association studies in crops that were established by domestication and early breeding can be a valuable basis for the identification of QTLs. A case control design in a population is an ideal way to identify maximum candidate sites contributing to a complex polygenic trait such as drought. In the current study, marker loci associated with drought related QTLs were identified in sugarcane (Saccharum spp), one of the most complex crop genomes, with its polyploid nature (>8), chromosome number (&gt;100) and interspecific origin. The objectives of this investigation were: 1) development of genic markers, which can be used for marker-assisted selection of drought tolerant genotypes of sugarcane. 2) genotypic characterization of sugarcane population at drought related loci using EST-SSR markers. Using 55 microsatellite markers, 56 polymorphisms were scored among 80 modern sugarcane genotypes. Homogeneity of the population was confirmed by determining the distribution of allele frequencies obtained by random genomic microsatellite markers. This analysis was conducted in the STRUCTURE program and the population was divided in 3 subgroups based on the allelic distribution. Phenotypic data to evaluate drought tolerance among the genotypes was collected by measuring chlorophyll content, chlorophyll fluorescence, leaf temperature and leaf relative water content. A generalized linear model in SPSS was used to find association between marker loci and phenotypic data. Markers with significant association (P 0.001 level) with the trait were subjected to linear regression to screen the spurious associations. Based on the results, 21 EST-SSR markers and 11 TRAP markers related to drought-defining physiological parameters were considered as genuine associations in this study. Fifty-six polymorphisms produced by 13 EST-SSR primers were used to produce genetic similarity matrix for 80 genotypes. Dendrogram prepared from this genetic similarity matrix will be useful in selecting parents carrying diversity at drought specific loci.

Maize production is limited agronomically by the availability of water and nutrients during the growing season. Of these two limiting factors, water availability is predicted to increase in importance as climate change and the growing urban landscape continue to stress limited supplies of freshwater. Historically, efforts to breed maize for water-limited environments have been extensive; especially in the areas of root architecture and flowering physiology. As progress has been made and new traits have been discovered and selected for, the different responses to drought stress at specific developmental stages of the maize plant have been selected as a whole when drought tolerance is evaluated. Herein we attempt to define the characteristics of the maize drought response during different developmental stages of the maize plant that can be altered through plant breeding. Towards breeding for drought tolerance, 400 inbred lines from a diversity panel were amplified and sequenced at the ZmLOX4 and ZmLOX5 loci in an effort to characterize their linkage disequilibrium and genetic diversity. Understanding these characteristics is essential for an association mapping study that accompanies this project, searching for novel and natural allelic diversity to improve drought tolerance and aflatoxin resistance in maize. This study is among the first to investigate genetic diversity at important gene paralogs ZmLOX4 and ZmLOX5 believed to be highly conserved among all Eukaryotes. We show very little genetic diversity and very low linkage disequilibrium in these genes, but also identified one natural variant line with knocked out ZmLOX5, a variant line missing ZmLOX5, and five line variants with a duplication of ZmLOX5. Tajima's D test suggests that both ZmLOX4 and ZmLOX5 have both been under neutral selection. Further investigation of haplotype data revealed that ZmLOX12, a member of the ZmLOX family, showed strong LD that extends much further than expected in maize. Linkage disequilibrium patterns at these loci of interest are crucial to quantify for future candidate gene association mapping studies. Knockout and copy number variants of ZmLOX5, while not a surprising find, are under further investigation for crop improvement.

Crown rust, caused by the fungus Puccinia coronata, is a severe disease negatively impacting seed quality and yield in oat (Avena sativa). Host genetic resistance is the primary means for controlling this disease. The most extensive oat map contains nearly 2,500 genetic markers, many of which are restriction and amplified fragment length polymorphic (RFLP or AFLP) markers. However, the use of more abundant single nucleotide polymorphic (SNP) markers combined with diversity arrays technology (DArT) would be more advantageous for marker assisted breeding (MAB) and genome wide selection (GWS) applications due to the availability of high density genotyping technologies. The purpose of using this technology is to improve the competitiveness of oat by producing varieties with durable resistance to crown rust and desirable traits that will benefit oat growers in the U.S. Panels of winter and spring oat were evaluated for resistance to crown rust in four field environments in Texas, Louisiana, Minnesota, and North Dakota during a two-year study in 2010 and 2011. Plants representing 702 elite lines of oat were phenotyped for crown rust resistance and found to have highly diverse responses. The winter oat lines demonstrated the best crown rust resistance and are expected to yield the most QTL to be used in developing durable crown rust resistance. Heritability of crown rust resistance in this study ranged from 0.88 to 0.90 in spring and winter oats, respectively. Crown rust measurements were also found to be repeatable. Repeatability ranged from 0.56 to 0.88 at Castroville, TX in 2011 and 2010, respectively in spring oats and from 0.79 at St. Paul, MN in 2011 to 0.96 at Castroville, TX in 2010 in winter oats. Oat lines contributed by states along the Puccinia pathway in Texas, Louisiana, Minnesota, North Dakota, and Wisconsin on average exhibited the best crown rust resistance as compared to other areas in the country where spring and winter oat are grown. GGE biplot analysis indicated that Castroville, TX was the most representative and most ideal testing location. The above results are expected to increase knowledge of the genetic diversity of the oat germplasm, yield comprehensive genotyping and phenotyping information for North American oat breeding programs, and to promote further use of GWS and MAB for key traits regarding disease resistance in oat. Future work is to conclude the association mapping process by completing genotypic analysis.

Linseed oil (Linum usitatissimum L.) is valued for its food and non-food applications. Although Canada is the world’s largest linseed producer and exporter, linseed remains a minor crop in part because its yield has been stagnating over the last decade compared to other oilseeds. Narrow genetic base, absence of an efficient hybrid production system and limited genomic tools for linseed breeding are the main factors hindering yield and quality improvements. Here, we characterized the Canadian flax core collection of 407 accessions with 448 genome-wide simple sequence repeat markers and, using association mapping (AM), we demonstrated its potential for the improvement of seed quality and agronomic traits. Genetic structure analyses assigned all accessions to two major groups that were weakly differentiated (FST = 0.094). Genetic diversity was abundant in the total panel (5.32 alleles per locus) with weak familial relatedness (mean = 0.287) for most individual pairs. Linkage disequilibrium (LD) decayed relatively quickly with an average genome-wide LD of ~1 cM. AM for seven seed quality traits including oil content (OIL), palmitic acid (PAL), stearic acid (STE), oleic acid (OLE), linoleic acid (LIO), linolenic acid (LIN) and iodine value (IOD) identified nine stable candidate QTL. LIO and LIN QTL co-localized with previously identified QTL and some mapped in the vicinity of genes known to be involved in the fatty acid biosynthesis pathway. AM conducted for nine agronomic traits including yield, bolls per area (BPA), seeds per boll (SPB), thousand seed weight (TSW), start of flowering (FL5%), end of flowering (FL95%), plant height (PH), plant branching (PB) and lodging (LDG) identified twelve significant marker-trait associations for six of the traits. The associated markers explained between 0.5 to 18.5% of the phenotypic variation, with Lu526 and Lu2532 associated with TSW and Lu943 associated with flowering being the most promising for marker-assisted selection. Statistical simulation for five markers associated with TSW indicated that the favorable alleles have additive effects. None of the accessions carried the five favorable alleles but a few breeding lines had four, indicating that further improvement of TSW and yield could be achieved through marker assisted breeding

Loblolly pine (Pinus taeda L.) is an ecologically and economically important southern pine, distributed across the southeastern United States. Its genetic improvement for breeding and deployment is a major goal of the Western Gulf Forest Tree Improvement Program (WGFTIP) hosted by the Texas A&M Forest Service. Rapid advances in genomics and molecular marker technology have created potential for application of Marker Assisted Selection (MAS) and Genomic Selection (GS) for accelerated breeding in forest trees. First-generation selection (FGS) and second- generation selection (SGS) breeding populations of loblolly pine from east Texas were studied to estimate the genetic diversity, population structure, linkage disequilibrium (LD), signatures of selection and association of breeding traits with genetic markers using a genome-wide panel of 4264 single nucleotide polymorphisms (SNPs). Under- standing the genetic basis of local adaptation is crucial to disentangle the dynamics of gene flow, drift and selection and to address climate change. Bayesian mixed linear models and logistic regression were used to associate SNP variation with geography, climate, aridity and growth season length and markers with strong correlations were investigated for biological functions. Relatively high levels of observed (Ho = 0.178–0.198) and expected (He = 0.180-0.198) heterozygosities were found in all populations. The amount of inbreeding was very low, and many populations exhibited a slight excess of heterozygotes. The population substructure was weak, but FST indicated more pronounced differentiation in the SGS populations. As expected for outcrossing natural populations, the genome-wide LD was low, but marker density was insufficient to deduce the decay rate. Numerous associations were found between various phenotypes and SNPs, but few remained significant after false positive correction. Signatures of diversifying and balancing selection were found in markers representing important biological functions. Strong correlations supported by Bayes factors were found between various environmental variables and several SNPs. Logistic regression found hundreds of significant marker-environment associations, but none remained significant after false-positive correction, which was likely too stringent and will require further investigation. Annotations of significant markers implicated them in crucial biological functions. These results present the first step in the application of MAS to the WGFTIP for loblolly pine genetic improvement and will contribute to the knowledgebase necessary for genomic selection technology. Results from environmental association study provide important information for designing breeding strategies to address climate change and for genetic conservation purposes.

Doctor of Philosophy / Department of Agronomy / Geoffrey Morris / Climate change has been anticipated to affect agriculture, with most the profound effect in regions where low input agriculture is being practiced. Understanding of how plants evolved in adaptation to diverse climatic conditions in the presence of local stressors (biotic and abiotic) can be beneficial for improved crop adaptation and yield to ensure food security. Great genetic diversity exists for agroclimatic adaptation in sorghum (Sorghum bicolor L. Moench) but much of it has not been characterized. Thus, limiting its utilization in crop improvement. The application of next-generation sequencing has opened the plant genome for analysis to identify patterns of genome-wide nucleotide variations underlying agroclimatic adaptation. To understand the genetic basis of adaptive traits in sorghum, the genetic architecture of sorghum inflorescence traits was characterized in the first study. Phenotypic data were obtained from multi-environment experiments and used to perform joint linkage and genome-wide association mapping. Mapping results identified previously mapped and novel genetic loci underlying inflorescence morphology in sorghum. Inflorescence traits were found to be under the control of a few large and many moderate and minor effect loci. To demonstrate how our understanding of the genetic basis of adaptive traits can facilitate genomic enabled breeding, genomic prediction analysis was performed with results showing high prediction accuracies for inflorescence traits. In the second study, the sorghum-nested association mapping (NAM) population was used to characterize the genetic architecture of leaf erectness, leaf width, and stem diameter. About 2200 recombinant inbred lines were phenotyped in multiple environments. The obtained phenotypic data was used to perform joint linkage mapping using ~93,000 markers. The proportion of phenotypic variation explained by QTL and their allele frequencies were estimated. Common and moderate effects QTL were found to underlie marker-trait associations. Furthermore, identified QTL co-localized with genes involved in both vegetative and inflorescence development. Our results provide insights into the genetic basis of leaf erectness and stem diameter in sorghum. The identified QTL will also facilitate the development of genomic-enable breeding tools for crop improvement and molecular characterization of the underlying genes Finally, in a third study, 607 Nigerian accessions were genotyped and the resulting genomic data [about 190,000 single nucleotide polymorphisms (SNPs)] was used for downstream analysis. Genome-wide scans of selection and genome-wide association studies (GWAS) were performed and alongside estimates of levels of genetic differentiation and genetic diversity. Results showed that phenotypic variation in the diverse germplasm had been shaped by local adaptation across climatic gradient and can provide plant genetic resources for crop improvement.