VTechWorks Repository :: Browsing by Author "Holliday, Jason A."

Browsing by Author "Holliday, Jason A."

Now showing 1 - 20 of 42

Architecture and Evolution of Xylem-related Gene Coexpression Networks in Poplars
Suren, Haktan (Virginia Tech, 2013-02-14)
With the advent of sequencing technologies, a variety of methods have been available day by day. Each of these methods have helped scientists to for a deeper understanding of the biological function and evolutionary constraints on the relevant genes, which can be gained through the use of modern computational approaches. Numerous approaches have being developed to advance these goals, and interaction network mapping is one of them. This method has been employed to study a variety of organisms to illustrate shared (conserved) or individual (unique) properties, and is mainly based on identifying and visualizing modules of co-expressed genes. As being a very strong candidate for such tools, co-expression gene network was used in this study to indentify the genes in wood formation of Populus trichocarpa with the help of the other novel bioinformatics tools such as Gene Ontology and Cytoscape. In order to booster the accuracy of the findings, we have combined it with an evolutionary approach, synonymous and non-synonymous ratio (dN/dS) of the proteins to show the selective patterns of the genes in a comparative fashion between woody and non-woody plants. This thesis is proposed to help plant scientists to gain insights into the genes that are involved in wood formation. By taking advantage of the computational studies have been done on this paper, one can validate the experiments along with reducing the cumbersomeness of the lab trials on the topic of wood formation in plants
Discovery of New Protein-DNA and Protein-Protein Interactions Associated With Wood Development in Populus trichocarpa
Petzold, Herman E. III (Virginia Tech, 2017-11-09)
The negative effects from rising carbon levels have created the need to find alternative energy sources that are more carbon neutral. One such alternative energy source is to use the biomass derived from forest trees to fulfill the need for a renewable alternative fuel. Through increased understanding and optimization of regulatory mechanisms that control wood development the potential exists to increase biomass yield. Transcription factors (TFs) are DNA-binding regulatory proteins capable of either activation or repression by binding to a specific region of DNA, normally located in the 5-prime upstream promoter region of the gene. In the first section of this work, six DNA promoters from wood formation-related genes were screened by the Yeast One-Hybrid (Y1H) assay in efforts to identify novel interacting TFs involved in wood formation. The promoters tested belong to genes involved in lignin biosynthesis, programmed cell death, and cambial zone associated TFs. The promoters were screened against a mini-library composed of TFs expressed 4-fold or higher in differentiating xylem vs phloem-cambium. The Y1H results identified PtrRAD1 with interactions involving several of the promoters screened. Further testing of PtrRAD1 by Yeast Two-Hybrid (Y2H) assay identified a protein-protein interaction (PPI) with poplar DIVARACATA RADIALIS INTERACTING FACTOR (DRIF1). PtrDRIF1 was then used in the Y2H assay and formed PPIs with MYB/SANT domain proteins, homeodomain family (HD) TFs, and cytoskeletal-related proteins. In the second section of this work, PPIs involving PtrDRIF1s' interaction partners were further characterized. PtrDRIF1 is composed of two separate domains, an N-terminal MYB/SANT domain that interacted with the MYB/SANT domain containing PtrRAD1 and PtrDIVARICATA-like proteins, and a C-terminal region containing a Domain of Unknown Function 3755 (DUF3755). The DUF3755 domain interacted with HD family members belonging to the ancient WOX clade and Class II KNOX domain TFs. In addition, PtrDRIF1 was able to form a complex between PtrRAD1 and PtrWOX13c in a Y2H bridge assay. PtrDRIF1 may function as a regulatory module linking cambial cell proliferation, lignification, and cell expansion during growth. Combined, these findings support a role for PtrDRIF1 in regulating aspects of wood formation that may contribute to altering biomass yield.
Effects of Glyphosate Herbicide on Phytophthora cinnamomi and Mine Soil Microbial Communities
Klopf, Sara K.; Holliday, Jason A.; Badgley, Brian D. (Virginia Tech. Powell River Project, 2019)
The American Chestnut (Castanea denata) was once the dominant hardwood species within the forests of the Appalachians and an important resource for people and wildlife. In the early 1900s, a fungal blight (Cryphonectria parasitica) was introduced from imported ‘Japanese Giant’ nursery trees that caused topkill of American chestnuts (Tallamy 2007). Trees infected with C. parasitica die back, then continually resprout from the roots. Since 1983, the American Chestnut Foundation has been conducting a backcross breeding program to produce hybrid trees resistant to the blight, and more recently, various research institutions have been working to develop transgenic varieties of American chestnut which express genes that may give the trees resistance to the blight. Additional pathogens, such as the fungus-like oomycete which causes root rot (Phytophthora cinnamomi), have furthered threatened C. dentata, and research of transgenic American chestnuts has included the identification of genes that may provide resistance to P. cinnamomi.
Evaluating the Adaptive Genomic Landscape of Remnant and Backcross American Chestnut Populations to Inform Germplasm Conservation
Sandercock, Alexander M. (Virginia Tech, 2023-07-27)
The American chestnut tree (Castanea dentata) is a deciduous tree that largely exists in the eastern United States along the Appalachian Mountain range. Approximately 100 years ago, a fungal pathogen (Cryphonectria parasitica) decimated chestnut populations, resulting in the loss of billions of trees. Disease-resistant American chestnut populations have been developed, but the introgression of wild adaptive diversity into these breeding populations will be necessary to develop locally adapted and disease resistant chestnut trees for reintroduction. In this dissertation, I presented our findings which addressed previous gaps in knowledge regarding the population genomics of wild and backcross American chestnut populations. I 1) estimated the genomic diversity, population structure, and demographic history of remnant wild American chestnut populations; 2) revealed the genomic basis of local climate adaptation in American chestnut, developed a novel method to make tree sampling estimates for germplasm conservation, and defined unique seed zones for American chestnut based on climate and genotype, and 3) determined the amount of wild adaptive diversity captured by the backcross breeding program and made recommendations for their replanting region. These results will inform the development of a breeding plan for the introgression of adaptive diversity into backcross and transgenic chestnut populations.
Evaluation of Genomic Prediction and the Agronomic Performance of Facultative Barley
Reith, Francis Arthur (Virginia Tech, 2024-10-28)
Cultivated barley typically exhibits either a winter growth habit or a spring growth habit. Some cultivars display a facultative growth habit, allowing them to be cultivated as either winter or spring crops. This study evaluated 1,128 elite barley cultivars and breeding lines under fall and spring sowing to determine which lines had a facultative growth habit and the underlying genetics thereof. In the fall of 2021 and subsequent spring of 2022, The first trial focused on identifying genetic factors associated with facultative habit. Results indicated that facultative lines were rare, with the majority exhibiting a winter growth habit. GENOME WIDE SCANS revealed no novel QTL associated with facultative habit, but significant QTLs on chromosome 4H were identified, correlating with the vernalization gene "VRN-H2." Several haplotypes found on chromosome 4H within appear significant and may contribute to differences in facultative habit. Only 28% of facultative lines could be accurately predicted based on genetic data, suggesting that facultative habit is a more complex trait than previously understood. Significant epistatic interactions between chromosome 4H and 4 other chromosomes were discovered. These findings indicate facultative habit is a much more quantitative trait than previously reported. The second trial involved growing the best-performing lines from the first trial under both fall and spring sowing conditions. Winter-sown barley consistently outperformed spring-sown barley in grain yield across all facultative lines. Despite strong performance under spring conditions, yield rankings were inconsistent across both sowing seasons, implying that agronomic performance cannot be reliably predicted across seasons. Notably, the Virginia Tech malt barley line 'Avalon' demonstrated facultative growth but exhibited poor agronomic quality under spring sowing. In contrast, lines such as 'VA22M-20DH1349' and 'VA22M-20DH1182' showed superior performance in both sowing regimes, indicating their potential for future breeding programs and agronomic trials for facultative barley in the American East.
Evidence for extensive heterotrophic metabolism, antioxidant action, and associated regulatory events during winter hardening in Sitka spruce
Collakova, Eva; Klumas, Curtis; Suren, Haktan; Myers, Elijah; Heath, Lenwood S.; Holliday, Jason A.; Grene, Ruth (2013-04-30)
Background Cold acclimation in woody perennials is a metabolically intensive process, but coincides with environmental conditions that are not conducive to the generation of energy through photosynthesis. While the negative effects of low temperatures on the photosynthetic apparatus during winter have been well studied, less is known about how this is reflected at the level of gene and metabolite expression, nor how the plant generates primary metabolites needed for adaptive processes during autumn. Results The MapMan tool revealed enrichment of the expression of genes related to mitochondrial function, antioxidant and associated regulatory activity, while changes in metabolite levels over the time course were consistent with the gene expression patterns observed. Genes related to thylakoid function were down-regulated as expected, with the exception of plastid targeted specific antioxidant gene products such as thylakoid-bound ascorbate peroxidase, components of the reactive oxygen species scavenging cycle, and the plastid terminal oxidase. In contrast, the conventional and alternative mitochondrial electron transport chains, the tricarboxylic acid cycle, and redox-associated proteins providing reactive oxygen species scavenging generated by electron transport chains functioning at low temperatures were all active. Conclusions A regulatory mechanism linking thylakoid-bound ascorbate peroxidase action with “chloroplast dormancy” is proposed. Most importantly, the energy and substrates required for the substantial metabolic remodeling that is a hallmark of freezing acclimation could be provided by heterotrophic metabolism.
Evolutionary Genomics of Populus trichocarpa (Western Poplar)
Bawa, Rajesh Kumar (Virginia Tech, 2017-08-15)
Forest trees are an important pool of biodiversity at the gene, individual and an ecosystem level. This variation is a result of complex environmental interactions, as well as neutral and selective forces acting on populations. Patterns of standing genetic variation are the result of adaption to past and contemporary climate change, but also historical demographic events, and disentangling the role of these forces is a central problem in population genomics. The overall goal of this study is to characterize the relative effects of demography and selection in the genome of Populus trichocarpa, a riparian deciduous tree species of North America. Specifically, I used a variety of methods to summarize patterns of genetic diversity and population structure in P. trichocarpa, and to reconstruct its demographic history. I subsequently incorporated these demographic insights to guide the application of several methods to identify genome-wide targets of natural selection within and among rangewide populations adapted to heterogeneous selection regimes. Results of this study provide insights into the history of divergence and differentiation in P. trichocarpa populations and help us identify the functional genetic variants contributing to phenotypic divergence and fitness of the individuals in it.
Exome resequencing and GWAS for growth, ecophysiology, and chemical and metabolomic composition of wood of Populus trichocarpa
Guerra, Fernando P.; Suren, Haktan; Holliday, Jason A.; Richards, James H.; Fiehn, Oliver; Famula, Randi; Stanton, Brian J.; Shuren, Richard; Sykes, Robert; Davis, Mark F.; Neale, David B. (2019-11-20)
Background Populus trichocarpa is an important forest tree species for the generation of lignocellulosic ethanol. Understanding the genomic basis of biomass production and chemical composition of wood is fundamental in supporting genetic improvement programs. Considerable variation has been observed in this species for complex traits related to growth, phenology, ecophysiology and wood chemistry. Those traits are influenced by both polygenic control and environmental effects, and their genome architecture and regulation are only partially understood. Genome wide association studies (GWAS) represent an approach to advance that aim using thousands of single nucleotide polymorphisms (SNPs). Genotyping using exome capture methodologies represent an efficient approach to identify specific functional regions of genomes underlying phenotypic variation. Results We identified 813 K SNPs, which were utilized for genotyping 461 P. trichocarpa clones, representing 101 provenances collected from Oregon and Washington, and established in California. A GWAS performed on 20 traits, considering single SNP-marker tests identified a variable number of significant SNPs (p-value < 6.1479E-8) in association with diameter, height, leaf carbon and nitrogen contents, and δ15N. The number of significant SNPs ranged from 2 to 220 per trait. Additionally, multiple-marker analyses by sliding-windows tests detected between 6 and 192 significant windows for the analyzed traits. The significant SNPs resided within genes that encode proteins belonging to different functional classes as such protein synthesis, energy/metabolism and DNA/RNA metabolism, among others. Conclusions SNP-markers within genes associated with traits of importance for biomass production were detected. They contribute to characterize the genomic architecture of P. trichocarpa biomass required to support the development and application of marker breeding technologies.
Exome Resequencing Reveals Evolutionary History, Genomic Diversity, and Targets of Selection in the Conifers Pinus taeda and Pinus elliottii
Acosta, Juan J.; Fahrenkrog, Annette M.; Neves, Leandro G.; Resende, Marcio FR R.; Dervinis, Christopher; Davis, John M.; Holliday, Jason A.; Kirst, Matias (Oxford University Press, 2019-02-01)
Loblolly pine (Pinus taeda) and slash pine (Pinus elliottii) are ecologically and economically important pine species that dominate many forest ecosystems in the southern United States, but like all conifers, the study of their genetic diversity and demographic history has been hampered by their large genome size. A small number of studies mainly based on candidate-gene sequencing have been reported for P. taeda to date, whereas none are available for P. elliottii. Targeted exome resequencing has recently enabled population genomics studies for conifers, approach used here to assess genomic diversity, signatures of selection, population structure, and demographic history of P. elliottii and P. taeda. Extensive similarities were revealed between these species: both species feature rapid linkage disequilibrium decay and high levels of genetic diversity. Moreover, genome-wide positive correlations for measures of genetic diversity between the species were also observed, likely due to shared structural genomic constraints. Also, positive selection appears to be targeting a common set of genes in both pines. Demographic history differs between both species, with only P. taeda being affected by a dramatic bottleneck during the last glacial period. The ability of P. taeda to recover from a dramatic reduction in population size while still retaining high levels of genetic diversity shows promise for other pines facing environmental stressors associated with climate change, indicating that these too may be able to adapt successfully to new future conditions even after a drastic population size contraction.
Flowering Gene Homologs Regulate Seasonal Growth Changes in Poplar
Sheng, Xiaoyan (Virginia Tech, 2018-10-19)
The adaptation of trees to temperate and boreal climates depends on their ability to respond to environmental signals that are markers of seasonal changes in order to survive winter and maximize growth. The genus Populus (poplars) is a model system for identifying the genes and molecular mechanisms that regulate growth and dormancy transitions. Photoperiod and temperature regulate both vegetative and floral phenology. FLOWERING LOCUS T (FT) and FLOWERING LOCUS D (FD) are key regulators of flowering time in Arabidopsis and other plants. The distinct functions of three poplar FD-LIKE (FDL) genes and two FTs were explored through gain-of-function, dominant repression, and CRISPR/CAS9-mediated gene editing. We studied trees in controlled environments, including manipulation of daylength and temperature to mimic an annual seasonal growth and dormancy cycle. Our studies showed that the FDL proteins share less than complete functional equivalency. Among the three paralogs, only FDL2.2 promoted precocious flowering, whereas FDL1 and FDL3 appear to have distinct roles in vegetative growth and phenology. Whereas overexpression of any FDL gene delays short day-induced growth cessation and bud set, only FDL3 coordinately altered leaf development and the transition to secondary growth in a photoperiod-dependent manner. For the first time, we demonstrate distinct functions of the two FT paralogs in vegetative phenology. Study of ft1ft2 double mutants and ft1-specific mutants showed that FT1 promotes dormancy release, whereas FT2 is necessary to sustain growth. Collectively, our results reveal that poplar FTs and FDLs have distinct roles in controlling different aspects of vegetative phenology and woody shoot development.
Focused ultrasound extraction (FUSE) for the rapid extraction of DNA from tissue matrices
Holmes, Hal R.; Haywood, Morgan; Hutchison, Ruby; Zhang, Qian; Edsall, Connor; Hall, Timothy L.; Baisch, David; Holliday, Jason A.; Vlaisavljevich, Eli (2020-10-09)
Rapid DNA extraction is a critical barrier for routine and fieldable genetics tests for applications in conservation, such as illegal trafficking and fraudulent mislabelling. Here, we develop a non-thermal focused ultrasound extraction (FUSE) technique that creates a dense cloud of high-pressure acoustic cavitation bubbles to disintegrate targeted tissues into an acellular debris, resulting in the rapid release of entrapped DNA. In this work, we demonstrate the proof-of-concept of the FUSE technique by obtaining species identifiable sequences and shotgun sequencing reads from DNA extracted from Atlantic salmon Salmo salar tissues. Having mitigated the key risks for this technique, we hypothesize future developments with this technology can be applied to accelerate and simplify DNA extraction from exceedingly difficult samples with complex tissue matrices (i.e. fibrous tissue and timber samples) in both laboratory and field settings.
Forest genomics: Advancing climate adaptation, forest health, productivity, and conservation
Isabel, Nathalie; Holliday, Jason A.; Aitken, Sally N. (2020-01)
Forest ecosystems provide important ecological services and resources, from habitat for biodiversity to the production of environmentally friendly products, and play a key role in the global carbon cycle. Humanity is counting on forests to sequester and store a substantial portion of the anthropogenic carbon dioxide produced globally. However, the unprecedented rate of climate change, deforestation, and accidental importation of invasive insects and diseases are threatening the health and productivity of forests, and their capacity to provide these services. Knowledge of genetic diversity, local adaptation, and genetic control of key traits is required to predict the adaptive capacity of tree populations, inform forest management and conservation decisions, and improve breeding for productive trees that will withstand the challenges of the 21st century. Genomic approaches have well accelerated the generation of knowledge of the genetic and evolutionary underpinnings of nonmodel tree species, and advanced their applications to address these challenges. This special issue of Evolutionary Applications features 14 papers that demonstrate the value of a wide range of genomic approaches that can be used to better understand the biology of forest trees, including species that are widespread and managed for timber production, and others that are threatened or endangered, or serve important ecological roles. We highlight some of the major advances, ranging from understanding the evolution of genomes since the period when gymnosperms separated from angiosperms 300 million years ago to using genomic selection to accelerate breeding for tree health and productivity. We also discuss some of the challenges and future directions for applying genomic tools to address long-standing questions about forest trees.
Functional analysis of novel protein-protein interactions involving ROP GTPases in Arabidopsis thaliana and Populus trichocarpa
Jia, Xiaoyan (Virginia Tech, 2013-09-02)
We are using the yeast two-hybrid (Y2H) system to identify novel protein-protein interactions (PPI) relevant to wood formation. Bait proteins for Y2H binary assays and screening against a xylem cDNA prey library were selected from approximately 400 Populus trichocarpa genes that are at least 8-fold more highly expressed in differentiating secondary xylem versus phloem-cambium, and designated here as poplar biomass (PB) genes. Here we report some of the interactions involving selected PB proteins and efforts to characterize their functions in Populus and Arabidopsis. Members of the ROP GTPase family, PB15 in poplar and ROP11 in Arabidopsis, interact with the domain of unknown function (DUF) 620 (DUF620) proteins (e.g., PB129 in poplar). Ectopic co-expression of PB15 and PB129 in Arabidopsis caused outgrowths at the base of flower pedicels and altered leaf morphology. Interestingly, the co-expression phenotype could not be observed in transgenic plants that are only expressing either one of the interacting partners separately. Transgenics altered in expression of PB15 and/or PB129 were prepared in Populus and characterization of transgenic trees will be performed in greenhouse and field. In addition to DUF620 family proteins, ROP11 also interacts with the COP9 subunit CSN5A in Arabidopsis. We confirmed the interaction of ROP11 and CSN5A in Y2H and employed available mutants for ROP11 and CSN5A in Arabidopsis to genetically characterize this interaction. Surprisingly, loss of ROP11 was found to rescue the csn5a-2 pleiotropic phenotype. Ectopic expression of a ROP11 dominant negative mutant in the csn5a-2 background also complemented the stunted growth phenotype. Transcript analysis and gel blot assays showed that CSN5A transcript levels remained unchanged in all rescue lines, whereas CSN5A protein levels increased relative to WT. Taken together, we concluded that ROP11 negatively regulate CSN5A protein level in plant by some as yet unknown mechanism.
Functional analysis of Poplar genes regulating flowering and vegetative growth
Mahendra, Rienzy Ayeshan Rangajeewa (Virginia Tech, 2019-06-24)
Poplar (Populus spp. and hybrids) are used for pulp, paper and solid wood products. Furthermore, poplar is being developed as a dedicated biomass crop for biofuels and biomaterials. Thus, methods to accelerate genetic improvement to improve woody biomass yield, quality and optimal growth on marginal lands are of considerable interest. One approach is to identify genes that could be manipulated through breeding or biotechnology to achieve these goals. I studied two sets of candidate genes for improving biomass, growth and manipulating flowering time. First, I studied the functions of PopNAC154 and PopNAC156, co-orthologs of Arabidopsis SECONDARY CELL WALL NAC DOMAIN2 (SND2), which are putative regulators of wood cell wall synthesis, the source of lignocellulosic biomass. Second, I studied PopCEN1, PopCEN2, and PopBFT, members of the TERMINAL FLOWER1 (TFL1)/CENTRORADIALIS (CEN) gene family that act as flowering repressors in Arabidopsis and many plants. I studied INRA 717-1B (P. tremula x P. alba) transgenics with an artificial microRNA (AmiRNA) downregulating PopNAC154 and PopNAC156 (AmiSND2 trees). In a field trial, AmiSND2 trees showed higher mean height and diameter than wild-type (WT). We also observed that AmiSND2 transgenics showed delayed leaf senescence and leaf drop. After conducting controlled environment studies with AmiSND2 trees, I was able to confirm that downregulation of PopNAC154 and PopNAC156 genes does not alter the short daylength-induced bud set and growth cessation but it delays the low temperature induced leaf senescence and leaf drop. Further I was able to show that down regulation of the PopNAC154 and PopNAC156 genes resulted in significantly higher mean plant heights and delayed bud set compared to the WT plants under low soil nutrient conditions. Wood chemistry data analysis of field grown AmiSND2 trees showed that they have a significantly higher cellulose content a lower lignin content compared to that of the WT. Thus, these results show that downregulating the PopNAC154 and PopNAC156 genes has the potential to increase biomass yield and quality. In a previous study, simultaneous downregulation of PopCEN1 and PopCEN2 genes using RNA interference (RNAi) method caused poplar trees to flower only after two years of growth in the field. I used CRISPR/CAS9 method to knock-out each paralog individually as well as the related gene, PopBFT. The popcen1 mutant trees developed flowers even under in vitro conditions, but popcen2 mutants did not show an obvious phenotype. popbft mutant trees also did not show an obvious phenotype under standard growing conditions. However, when soil nutrient availability was allowed to deplete, the popbft mutants showed lower mean plant height compared to the WT and also showed lower root length and root volume under low Nitrogen conditions in an in vitro assay compared to the WT. These results prove that PopCEN1 gene is directly involved in repressing flowering in poplar and allele-specific mutation should be tested as an approach to accelerate breeding. PopCEN2 and PopBFT might not have a role in regulation of flowering time, and though additional studies are needed, PopBFT appears to have a role in regulating growth in response to nutrient availability.
Genetic Heterogeneity of Residual Variance for Production and Functional Traits in American Angus Cattle
Amorim, Sabrina Thaise (Virginia Tech, 2024-08-14)
Beef cattle are continuously selected for different traits and the success in improving these traits has been remarkable. However, for certain traits, it is essential not only to improve the average performance, but also to control the variation around the mean. There is evidence that residual variance may be under genetic control, which opens the possibility of selecting for uniformity. In this sense, the objectives of the present dissertation were: 1) to investigate the extent of genetic heterogeneity of residual variance at the pedigree level in birth weight (BW), weaning weight (WW), yearling weight (YW), foot angle (FA), and claw set (CS) in American Angus cattle; 2) to compare the results of different genetic heterogeneity models; 3) to evaluate the effectiveness of Box-Cox transformation in continuous traits; and 4) to address limitations and explore alternative solutions for implementing genetic parameters for residual variance in genetic evaluations. The first study investigated the genetic heterogeneity of residual variances for BW, WW, and YW. Three models were compared: a homoscedastic residual variance model (M1), a double hierarchical generalized linear model (DHGLM, M2), and a genetically structured environmental variance model (MCMC, M3). The results showed significant genetic heterogeneity of residual variances in growth traits, suggesting the possibility of selection for uniformity. The genetic coefficient of variation for residual variance ranged from 0.90 to 0.92 in M2 and 0.31 to 0.38 in M3 for BW, 0.64 in M2 and 0.01 to 0.29 in M3 for WW, and 0.67 to 0.63 in M2 and 0.25 to 0.31 in M3 for YW. Low heritability estimates for residual variance were found, particularly in M2 (0.08 for BW, 0.06 for WW, and 0.09 for YW). The study identified both negative and positive genetic correlations between mean and residual variance, depending on the trait and data transformation. Negative correlations suggest the potential to increase trait means while decreasing residual variance. However, positive correlations indicate that the genetic response to selection for uniformity may be limited unless a selection index is used. Data transformation reduced skewness but did not eliminate genetic heterogeneity of residual variances. The Bayesian approach provided higher estimates of additive genetic variance for residual variance compared to DHGLM. Overall, the findings indicate the potential to reduce variability through selection and lay the groundwork for incorporating uniformity of growth traits into breeding goals. The second study focused on the genetic heterogeneity of residual variance for two foot conformation traits, FA and CS. Using 45,667 phenotypic records collected between 2009 and 2021, three models were compared: a traditional homoscedastic residual variance model (M1), a DHGLM (M2), and a genetically structured environmental variance model (M3). Results showed that heritability estimates for FA and CS means were within expected ranges, although lower in M2. Despite low heritability estimates for residual variance (0.07 for FA and 0.05 for CS in M2), significant genetic coefficients of variation were found, suggesting that selection on trait mean would also influence residual variance. Positive genetic correlations between mean and residual variance in M2 and M3 indicate that selection for uniformity is feasible, but may require additional strategies such as selection indices. The study highlights the potential of FA and CS as indicators for breeding programs aimed at improving production uniformity in beef cattle. Our findings suggest that selection for uniformity in growth and foot score traits in beef cattle may be limited by low heritability of residual variance and moderate to high positive genetic correlations between mean and residual variance. This was observed for most of the traits studied. To overcome these challenges, further research is needed, particularly to explore genomic information to improve the prediction accuracy of estimated breeding values (EBV) for residual variance. Although studies of uniformity using genomic data are limited, they have shown improved EBV accuracy for residual variance. Additionally, alternative methods for measuring uniformity, such as different uniformity or resilience indicators, should be considered, especially with advances in digital phenotyping. Precision livestock farming technologies that allow for extensive data collection on various production traits should be integrated into the development of new uniformity indicators. This dissertation provides valuable insights into the genetic heterogeneity of residual variance in American Angus cattle and highlights the complexity of selecting for uniformity while improving mean traits. Continued research with larger data sets, genomic information, and further methodological refinement will be critical to advance these findings to improve uniformity and productivity in beef cattle breeding.
A genome-guided strategy for climate resilience in American chestnut restoration populations
Sandercock, Alexander M.; Westbrook, Jared W.; Zhang, Qian; Holliday, Jason A. (National Academy of Sciences, 2024-07-16)
American chestnut (Castanea dentata) is a deciduous tree species of eastern North America that was decimated by the introduction of the chestnut blight fungus (Cryphonectria parasitica) in the early 20th century. Although millions of American chestnuts survive as root collar sprouts, these trees rarely reproduce. Thus, the species is considered functionally extinct. American chestnuts with improved blight resistance have been developed through interspecific hybridization followed by conspecific backcrossing, and by genetic engineering. Incorporating adaptive genomic diversity into these backcross families and transgenic lines is important for restoring the species across broad climatic gradients. To develop sampling recommendations for ex situ conservation of wild adaptive genetic variation, we coupled whole-genome resequencing of 384 stump sprouts with genotype–environment association analyses and found that the species range can be subdivided into three seed zones characterized by relatively homogeneous adaptive allele frequencies. We estimated that 21 to 29 trees per seed zone will need to be conserved to capture most extant adaptive diversity. We also resequenced the genomes of 269 backcross trees to understand the extent to which the breeding program has already captured wild adaptive diversity, and to estimate optimal reintroduction sites for specific families on the basis of their adaptive portfolio and future climate projections. Taken together, these results inform the development of an ex situ germplasm conservation and breeding plan to target blight-resistant breeding populations to specific environments and provides a blueprint for developing restoration plans for other imperiled tree species.
Genomic Prediction and Genetic Dissection of Yield-Related Traits in Soft Red Winter Wheat
Ward, Brian Phillip (Virginia Tech, 2017-05-02)
In multiple species, genome-wide association (GWA) studies have become an increasingly prevalent method of identifying the quantitative trait loci (QTLs) that underlie complex traits. Despite this, relatively few GWA analyses using high-density genomic markers have been carried out on highly quantitative traits in wheat. We utilized single-nucleotide polymorphism (SNP) data generated via a genotyping-by-sequencing (GBS) protocol to perform GWA on multiple yield-related traits using a panel of 329 soft red winter wheat genotypes grown in four environments. In addition, the SNP data was used to examine linkage disequilibrium and population structure within the testing panel. The results indicated that an alien translocation from the species Triticum timopheevii was responsible for the majority of observed population structure. In addition, a total of 50 significant marker-trait associations were identified. However, a subsequent study cast some doubt upon the reproducibility and reliability of plant QTLs identified via GWA analyses. We used two highly-related panels of different genotypes grown in different sets of environments to attempt to identify highly stable QTLs. No QTLs were shared across panels for any trait, suggesting that QTL-by-environment and QTL-by-genetic background interaction effects are significant, even when testing across many environments. In light of the challenges involved in QTL mapping, prediction of phenotypes using whole-genome marker data is an attractive alternative. However, many evaluations of genomic prediction in crop species have utilized univariate models adapted from animal breeding. These models cannot directly account for genotype-by-environment interaction, and hence are often not suitable for use with lower-heritability traits assessed in multiple environments. We sought to test genomic prediction models capable of more ad-hoc analyses, utilizing highly unbalanced experimental designs consisting of individuals with varying degrees of relatedness. The results suggest that these designs can successfully be used to generate reasonably accurate phenotypic predictions. In addition, multivariate models can dramatically increase predictive accuracy for some traits, though this depends upon the quantity and characteristics of genotype-by-environment interaction.
Genomic Selection and Genome-Wide Association Study in Populus trichocarpa and Pinus taeda
Kaplan, Abdullah (Virginia Tech, 2016-09-20)
Forest tree breeding methods rank among the most efficient ways to increase productivity and quality of forests. With the advent of high-throughput genotyping technology, genome-enabled breeding has started to gain importance and may overcome some weaknesses of traditional tree breeding. Genomic Selection (GS), which involves using genome-wide markers to predict breeding values of individuals in a population, has been proposed for animal and plant breeding programs. GS enables very accurate selection decisions through estimation of genomic estimated breeding values (GEBVs). While the goal of GS is to predict phenotype from genotype, it does not identify the underlying genes that have important roles in a trait. Genome-Wide Association Studies (GWAS) approaches are therefore complementary to GS, enabling identification of these genes, which may be useful for marker-assisted selection in some traits. In this study, we first estimated heritability for several adaptive traits (cold hardiness, dbh, bud flush, height, and bud set) in a population of Populus trichocarpa and for height, diameter, and stem straightness in Pinus taeda. GEBVs accuracies were estimated using a ridge regression–best linear unbiased prediction (rrBLUP) model, and these accuracies were compared with estimated heritabilities. GWAS was also performed for the both imputed and non–imputed data of P. taeda population using TASSEL (Trait Analysis by aSSociation Evolution and Linkage) software, as well as rrBLUP and FFBSKAT (Fast Family-Based Sequence Kernel Association Test) packages in R. Heritabilities ranged from 0.34 to 0.56 for P. trichocarpa and 0.14 to 0.37 for P.taeda. GWAS identified 3244 associations for dbh, 4077 associations for stem straightness, and 5280 SNPs for height (p≤0.05) in TASSEL using the reduced model (marker data only), whereas 2729, 3272 and 3531 associations were found with the full model where we also included population structure as a covariate. FFBSKAT showed a similar number of SNP associations (2989, 3046 and 3058). There was an inflation of SNP associations (~20k) found in rrBLUP, which suggests population structure was not effectively controlled. The GEBVs accuracies ranged from 0.09 and 0.22 for P.trichocarpa and 0.09 to 0.23 for P.taeda using rrBLUP method. Testing the effect of repetation on the accuracy of GEBV for poplar showed that there was no significant difference between the number of cycles. Also, there was no significant difference the accuracy of GEBVs in pine between two different imputation methods, the marker mean value and Beagle software.
Genomics of Climatic Adaptation in Populus Trichocarapa
Zhang, Man (Virginia Tech, 2016-08-10)
Temperate tree species exhibit seasonal growth cycling, and the timing of such transition varies with local climate. Under anthropogenic climate change, the local pattern of growth and dormancy in tree populations is expected to become uncoupled with shifting seasonal environmental signals, particularly temperature. Thus, an understanding of the genetic underpinnings of local adaptation is key to predicting the fate of tree populations in the future. In this thesis, we coupled sampling of range-wide natural accessions of P. trichocarpa with adaptive trait phenotyping and genome-wide genotyping to uncover relationships between genotype, phenotype, and environment. We detected strong correlations between adaptive phenotypes, climate, and geography, which suggested climatic selection driving adaptation of these populations to local environments. We subsequently combined genotype-phenotype association tests with sliding window analysis and identified regions strongly associated with these adaptive traits. We also compared adaptive markers identified in two independent GWAS on samples across latitude and altitude transects and found a set of associated variants shared across both transects. We further scanned the genome with three selection tests to identify regions showing evidence of recent positive and divergent selection. By comparing candidate selection regions across altitude and latitude, we detected a set of overlapping regions showing differentiation across gradients of the same climate variables. We validated the functional imortance of these selection regions by combining GWAS and showed that selection regions contain a strong signature of phenotypic associations. We also studied the distribution of deleterious allels across genome and natural populations, and found that deleterious alleles preferentially accumulate in regions of low recombination and hithihking regions. Finally, marginal populations contained more deleterious alleles compared with central populations, which is likely due to ineffective selection in small populations and recent bottlenecks associated with postglacial recolonization. These findings provide new insights into the genomic architecture underlying climatic adaptation and how selection drives adaptive evolution of tree species.
GWAS on the Attack by Aspen Borer Saperda calcarata on Black Cottonwood Trees Reveals a Response Mechanism Involving Secondary Metabolism and Independence of Tree Architecture
Sepúlveda, Sebastián L.; Neale, David B.; Holliday, Jason A.; Famula, Randi; Fiehn, Oliver; Stanton, Brian J.; Guerra, Fernando P. (MDPI, 2023-05-30)
Black cottonwood (Populus trichocarpa) is a species of economic interest and an outstanding study model. The aspen borer (Saperda calcarata) causes irreversible damage to poplars and other riparian species in North America. The insect can produce multiple effects ranging from the presence of some galleries in the stem to tree death. Despite the ecological and commercial importance of this tree–insect interaction, the genetic mechanisms underlying the response of P. trichocarpa to S. calcarata are scarcely understood. In this study, a common garden trial of P. trichocarpa provenances, established in Davis, California, was assessed at the second year of growth, regarding the infestation of S. calcarata from a natural outbreak. A genome-wide association study (GWAS) was conducted using 629k of exonic SNPs to assess the relationship between genomic variation and insect attack. Tree architecture, in terms of stem number per plant, and the wood metabolome were also included. Insect attack was independent of the number of stems per tree. The performed GWAS identified three significantly associated SNP markers (q-value < 0.2) belonging to the same number of gene models, encoding proteins involved in signal transduction mechanisms and secondary metabolite production, including that of R-mandelonitrile lyase, Chromodomain-helicase-DNA-binding family protein, and Leucine-rich repeat protein. These results are aligned with the current knowledge of defensive pathways in plants and trees, helping to expand the understanding of the defensive response mechanisms of black cottonwood against wood borer insects.

Browsing by Author "Holliday, Jason A."

Results Per Page

Sort Options