Life Sciences Commons^™

The Discovery Of A Novel Bacteria From A Large Co-Assembly Of Metagenomes, Matthew Finkelberg

Masters Theses

In the summer of 2022, a co-assembly of metagenome was created using the microbes found at Barres Woods in Harvard Forest. 14 samples were taken, and sample was split into the organic and mineral layer, which totals 28 Bulk MAGs. Within this Co-assembly, 4 different genomes were found which were designated with the phylum of FCPU426. Three of which were considered medium quality and one being assigned high quality. The novel phyla first appeared in NCBI and GTDB databases in June 2018. The name FCPU426 dates to 2010 and was named based on the 16s amplicon sequencing.

The novel phylum …

Changes In Gene Expression From Long-Term Warming Revealed Using Metatranscriptome Mapping To Fac-Sorted Bacteria, Christopher A. Colvin

Masters Theses

Soil microbiomes play pivotal roles to the health of the environment by maintaining metabolic cycles. One question is how will climate change affect soil bacteria over time and what could the repercussions be. To answer these questions, the Harvard Forest Long-Term Warming Experiment was established to mimic predicted climate change by warming plots of land 5℃ above ambient conditions. In 2017, 14 soil core samples were collected from Barre Woods warming experiment to mark 15 years since the establishment of the soil warming in that location. These samples underwent traditional metatranscriptomics to generate an mRNA library as well as a …

Chemosensory Receptors In Berghia Stephanieae: Bioinformatics And Localization, Kelsi L. Watkins

Masters Theses

Chemosensation is achieved through the binding of chemical signals to chemoreceptor proteins embedded in the membranes of sensory neurons. The molecular identity of these receptors, as well as the downstream processing of chemosensory signals, has been well studied in arthropods and vertebrates. However, very little is known about molluscan chemosensation. The identity of chemoreceptor proteins in the nudibranch mollusc Berghia stephanieae are unknown. Data from other protostome and molluscan studies suggest Berghia may use ionotropic receptors for some forms of chemoreception. This study used a bioinformatics approach to identify potential chemosensory ionotropic receptors in the transcriptome of Berghia. A …

Computational Analysis Of Transcriptional And Post-Transcriptional Feedback Loops In Development And Cell Differentiation, Benjamin Rigel Nordick

Masters Theses

Regulation of gene expression is critical to organism development and health. Positive and negative feedback in networks of gene regulatory interactions are associated with the maintenance of distinct cell states/types or oscillations, respectively. Recent computational studies identified two structures of positive feedback relevant to establishment of cell type: interconnected transcriptional “high-feedback” loops and a purely post-transcriptional feedback loop arising from multiple microRNA molecules cooperatively regulating one mRNA. The goals of this work were to develop methods for analyzing often-unintuitive high-feedback loops in large networks, generalize the two new regulatory structures to consider oscillation, and further explore the ability of post-transcriptional …

Screening For Binding Partners And Protein-Protein Interactions Of A Fungal Transcription Factor- Xdr1, Nishadi Punsara Gallala Gamage

Masters Theses

Clarireedia spp. (formerly Sclerotinia homoeocarpaF.T. Bennett) is the causal agent dollar spot, the most economically important turfgrass disease impacting golf courses in North America. The most effective strategy for dollar spot control is repeated application of multiple classes of fungicides. However, reliance on chemical application has led to resistance to four classes of fungicides as well as multidrug resistance (MDR). Fungi are known to detoxify xenobiotics, like fungicides, through transcriptional regulation of three detoxification phases: modification, conjugation and secretion. Little is known, however, of the protein-protein interactions that facilitate these pathways. Following next-generation RNA sequencing of Clarireedia spp., a …

Molecular Markers Within And Beyond Viburnum: A Single Set Of Ssrs For An Immense Genus, Trinity Paige Hamm

Masters Theses

Comprised of more than 160 species, Viburnum is the largest genus in the Viburnaceae (formerly Adoxaceae) alongside Adoxa and Sambucus. The native range of Viburnum species spans much of the Northern Hemisphere and extends into the mountains of South America and Southeastern Asia. Their wide geographic range has made Viburnum particularly interesting to phylogeographers and phylogeneticists. This diverse genus is also horticulturally valuable with varying traits such as fragrance, fruit color, and bud and inflorescence form. There is sufficient morphological diversity in the genus for there to be more than 70 species and intraspecific hybrids in cultivation, and in …

Metabolic Modeling Of Cystic Fibrosis Airway Microbiota From Patient Samples, Arsh Vyas

Masters Theses

Cystic Fibrosis (CF) is a genetic disorder, found with higher prevalence in the Caucasian population, affecting > 30,000 individuals in the United States and > 70,000 worldwide. Due to the astoundingly high rate of mortality among CF patients being attributed to respiratory failure brought on by chronic bacterial infections and subsequent airway inflammation, there has been a lot of focus on systematically analyzing CF lung airway communities. While it is observed traditionally that Pseudomonas aeruginosa is the most threatening and persistent CF colonizer due to high antibiotic resistance, recent studies have elicited the roles of other pathogens and it has been widely …

Structure-Function Studies Of The Trypanosome Mitochondrial Replication Protein Polib, Raveen Armstrong

Masters Theses

Trypanosoma brucei and related protists are distinguished from all other eukaryotes by an unusual mitochondrial genome known as kinetoplast DNA (kDNA) that is a catenated network composed of minicircles and maxicircles. Replication of this single nucleoid involves a release, replicate, and reattach mechanism for the thousands of catenated minicircles and requires at least three DNA polymerase (POLIB, POLIC and POLID) with similarity to E. coli DNA polymerase I. Like other proofreading replicative DNA polymerases, POLIB has both an annotated polymerase domain and an exonuclease domain. Predictive modelling of POLIB indicates that it has the canonical right hand …

Factors Influencing Primate Hair Microbiome Diversity, Catherine Kitrinos

Masters Theses

Primate hair is both a substrate upon which essential social interactions occur and an important host-pathogen interface. As commensal microbes provide important immune functions for their hosts, understanding the microbial diversity in primate hair could provide insight into primate immunity and disease transmission. While studies of human hair and skin microbiomes show differences in microbial communities across body regions, little is known about the nonhuman primate hair microbiome. In this study, we collected hair samples (n=159) from 8 body regions across 12 nonhuman primate species housed at 3 US institutions to examine 1) the diversity and composition of the primate …

Pedigree And Molecular Assessment Of Relatedness In A Captive Chimpanzee Population, Francesca Golus

Masters Theses

As a crucial component of biodiversity, genetic diversity contributes to variability among individuals, allowing populations of endangered species to be resilient in the face of changing environmental conditions. Zoological institutions have become a cornerstone of conservation efforts and a refuge for endangered species given threats imposed on wild populations by climate change, habitat fragmentation and degradation, and overexploitation. Chimpanzees (Pan troglodytes) are an endangered species experiencing drastic population decline in the wild, yet are common residents in zoos and wildlife sanctuaries. To sustain long-term, genetically viable populations of species of concern, zoos use studbooks and paternity testing to …

Evaluating The Winter Diet Of A Reintroduced Herd Of Elk In The Cumberland Mountains, Tennessee, Using Next-Generation Sequencing Techniques, Dailee L. Metts

Masters Theses

A distinct subspecies of elk (Cervus canadensis), the North American elk (C. canadensis canadensis), once inhabited portions of the southeastern United States, including Tennessee, until their extirpation in the mid 1800s. From 2000 to 2008, 201 Manitoban elk were reintroduced on the North Cumberland Wildlife Management Area (NCWMA). A year-long food habits study using histological analysis of plant material from feces was completed for the NCWMA elk from 2003 to 2004 and has since aided managers in their landscape planning. Since then, more elk have been released onto the area, food plots have been established throughout …

Genome-Wide Characterization Of Lateral Organ Boundaries Domain Genes In Sunflower (Helianthus Annuus L.), Saleh M. Aljurbua

Masters Theses

Lateral Organ Boundaries (LBD) gene family encodes transcription factors found only in plants with essential roles in growth, development, and response to biotic and abiotic factors. The sunflower (Helianthus annuus L., Asteraceae family) LBD genes were studied using different in silico and phylogenetic approaches including four major steps: gene identification, phylogenic analysis, structural analysis, and expression analysis. Sixty-four sunflower LBD genes (HaLBDs) were identified using query search of all Arabidopsis LBD genes against sunflower genome (version r1.2) followed by LBD domain sequence verification. A phylogenetic tree was reconstructed using MEGA-X program. The identified genes were phylogenetically classified into two main …

Carbon Metabolism In Cave Subaerial Biofilms, Victoria E. Frazier

Masters Theses

Subaerial biofilms (SABs) grow at the interface between the atmosphere and rock surfaces in terrestrial and subterranean environments around the world. Multi-colored SABs colonizing relatively dry and nutrient-limited cave surfaces are known to contain microbes putatively involved in chemolithoautotrophic processes using inorganic carbon like carbon dioxide (CO₂) or methane (CH₄). However, the importance of CO₂ and CH₄ to SAB biomass production has not been quantified, the environmental conditions influencing biomass production and diversity have not been thoroughly evaluated, and stable carbon and nitrogen isotope compositions have yet to be determined from epigenic cave SABs. …

Quantitative Trait Loci Analysis Of A Ril Soybean Population To Determine Chromosomal Regions Governing Seed Protein, Oil, And Linolenic Acid Content, Ronald E. Moore

Masters Theses

180 RILs (recombinant inbred lines) segregating for protein, oil, and fatty acids were produced from a cross between TN12-4098 and TN13-4303. These lines were grown across three locations spread horizontally across Tennessee: Research Education Center at Milan (RECM), Highland Rim Research and Education Center (HRREC), and East Tennessee Research and Education Center (ETREC) in 2018 and 2019. 21 quantitative trait loci (QTLs) spanning 7 chromosomes were found using WinQTLCart2.5.

Investigation Of Glutamate Carboxypeptidase Ii Roles In The Folate Cycle Dependent Reproduction And Development Of Caenorhabditis Elegans, Jessica M. Derham

Masters Theses

Glutamate Carboxypeptidase II (GCPII) is a transmembrane, zinc metallopeptidase that is expressed in a wide range of organisms, including roundworms, mice, and humans. In humans, GCPII is primarily expressed in the prostate, kidneys, small intestine, and central nervous system. Within the small intestine, the expected function of GCPII is to aid in the absorption of dietary folate from the intestinal lumen. GCPII cleaves excess glutamates from folate to yield monoglutamated folate which is then readily transported into the enterocyte. Folate can then be used through the one carbon metabolic cycle for the synthesis of nucleotides, conversion of homocysteine to methionine, …

Identification And Analysis Of Genes With Differential And Tissue-Specific Expression In Poplars, Bailey Mitchell

Masters Theses

Poplars have become one of the most frequently studied plant species for their applications in biotechnology, commercial benefits, and ecological importance. Microarray analysis is commonly used to analyze differential gene expression in many organisms. Investigations have examined differential gene expression and performed functional analyses on a single tissue or organ within poplar trees or a closely related set of tissues and organs, but usually do not broadly analyze gene expression across a large number of tissues and organs in poplar. This thesis work aims to generate data on the most differentially expressed genes for the major groups of tissues and …

The Transcriptome Response Of The White-Rot Fungus Trametes Versicolor To Wild-Type And Lignin-Modified Hybrid Poplar, Anbarah Alzabaidi

Masters Theses

Plant biomass is a renewable and sustainable feedstock for biofuel production that can reduce societal dependence on fossil fuels. However, the production of liquid biofuels from the non-starch (i.e. lignocellulosic) material through fermentation technology is limited due to the complexity of the cell wall structure. This necessitates the use of chemical, thermal, and/or mechanical pretreatment technologies, which adds significant capital, operational, and environmental costs. Biological pretreatment strategies have the potential to mitigate these expenses by harnessing the innate ability of specialized bacteria and fungi to deconstruct lignocellulose. White-rot fungi (e.g. Trametes versicolor) have been shown to be …

Patterns Of Morphological Plasticity In Metriaclima Zebra And Danio Rerio Suggest Differently Canalized Phenotypes Due To Form-Function Relationships, Dylan Jockel

Masters Theses

In order to ascertain the degree of compatibility in developmental restructuring and behavioral plasticity between two fish species frequently made subject of laboratory research (Metriaclima zebra & Danio rerio), alternative trophic niche exposure experiments utilizing novel three-prong feeding treatments were conducted to obtain morphometric data, which demonstrated both species do bear some degree of plasticity. The results are somewhat complicated by differences in locality of detectable restructuring, which may be due to disparity in the form-function relationship for each species’ lineage. Each is notable in the manner of respective species’ jaw protrusion, as it is driven by anterior …

A Pipeline For Exogenous Gene Expression And Biomass Analysis In Plants, Bijay Bisunke

Masters Theses

Biological pretreatment of biomass is typically required for the production of liquid biofuels, which are viable alternatives to traditional fossil fuels. While traditional thermomechanical pretreatments are effective at helping to deconstruct lignocellulose, they are often expensive and tend to generate environmentally dangerous waste materials. An alternative to traditional pretreatment strategies is biological pretreatment, which has focused on lignocellulose-degrading organisms, such as white-rot fungi. While direct pretreatment of biomass with white-rot fungi has shown great promise, directly expressing functional white-rot fungi genes in plants (i.e. in planta expression) has the potential to be even more efficient because the biomass itself …

Investigating The Prevalence Of An Insecticide Resistance Gene In Culex Mosquito Populations Of Central Illinois, Kylee R. Noel

Masters Theses

The globalization of society has led to the spread of the vectors, mosquitoes, and diseases. Each year millions of people are killed by diseases vectored by mosquitoes. For example, West Nile Virus alone killed over 2,000 people in North America in 2017; 8 of those deaths being in Illinois. Two prominent vectors of West Nile Virus are Culex pipiens and Culex restuans; which are difficult to differentiate based upon morphological features. In this study, mosquitoes collected from two Illinois counties, Coles County and Champaign County, were first identified based upon morphology and this identification was then compared to probe-based …

Detection Of Beta-Lactamase Variants In Municipal Wastewater And Fresh Water, Sunil Pandey

Masters Theses

The occurrence and spread of antibiotic-resistant genes (ARGs) are pressing public health problems worldwide. A key factor contributing to the spread of ARGs is lateral gene transfer. Wastewater treatment plants (WWTPs) are measured hot spots of microbial diversity and resistance because they receive polluted wastewater from diverse sources and contain a variety of different environments with dense bacterial loads. Due to the overuse of antibiotics the genetic capacities of microbes have profited. This helps every source of resistance gene and every means of horizontal gene transmission to develop the multiple mechanism of resistance to each antibiotic used clinically, agriculturally, or …

Haplotype Diversity And Population Genetic Structure Of Antistrophus Gall Wasps Associated With Two Silphium Species And Implication For Host Mediated Speciation, Jiaxin Deng

Masters Theses

Gall making insects form a special feeding guild of phytophagous animals, and by manipulating host plant tissue differentiation, are able to avoid plant chemical defenses and thus have no need for counter defense mechanisms. Host plant selection is crucial to the evolution of these insects because successful gall formation is largely dependent on host plant ability to respond to stimuli. In Illinois and neighboring states, Antistrophus gall wasps associated with the rosin weed (S. integrifolium) and the cup plant (S. perfoliatum) are morphologically indistinguishable and thus have been treated as belonging to single species. However, the wasps from the host …

Creg1: Its Role As A Master Regulator Of Liver Function, Iffat Jahan

Masters Theses

Mammalian development involves a complex system of regulatory signals and reactions resulting in highly differentiated cell types with specific structure and function. The liver is a major organ that has been studied extensively to understand underlying genetic processes responsible for specification, establishment and maintenance of tissue identity. Hepatoma and hepatoma variant cell lines have been used as models to understand genetic networks responsible for liver function. Whole genome microarray analysis of hepatocyte cell lines has revealed candidate genes that may serve as regulators or master regulators of liver specificity. In two previous studies in our lab, the role of candidate …

A Genome Wide Survey Of The Insertion Sequences In Halanaerobium Hydrogeniformans, A Haloalkaliphilic Anaerobic Bacterium, Kody Austin Bassett

Masters Theses

"Insertion sequences (IS) are the smallest prokaryotic transposable elements. These mobile genes are highly diverse in size and structure, making them difficult to study. The transposition activity of insertion sequences has a significant role in evolution by enabling genomic plasticity via genome rearrangements. Insertion sequences are largely uncharacterized and thus methods to improve the ability to accurately identify, annotate, and infer genomic impact of insertion sequences is limited. A sequential set of methods using readily available genomic and bioinformatic tools was developed to accurately identify insertion sequences. This method was used to perform an entire genome survey of Halanaerobium hydrogeniformans …

Characterization Of Chromosomal Translocations In A Group Of Killifish Species By Using Genome-Wide High-Density Snp Mapping Approach, Naznin Sultana Remex

Masters Theses

"The role of chromosomal rearrangements in reproductive isolation and introgression between species is poorly understood. In heterozygous form, rearrangements may directly interrupt meiotic progression leading to partial sterility/subfertility (underdominance) or may suppress local meiotic segregation (recombination suppression). Such unbalanced meiotic segregation may also result in reproductive isolation and play roles as a driving force of speciation. The objective of this study was to gain insight into the pattern of chromosomal rearrangements in two closely related killifish species in the genus Fundulus (F. notatus, and F. olivaceus) by constructing genetic linkage maps using high-resolution single nucleotide polymorphism (SNP) …

Clpxp-Regulated Proteins Suppress Requirement For Reca In Dam Mutants Of Escherichia Coli K-12, Amie Savakis

Masters Theses

Double strand breaks (DSB) are a common source of DNA damage in both prokaryotes and eukaryotes. If they are not repaired or are repaired incorrectly, they can lead to cell death (bacteria) or cancer (humans). In Escherichia coli, repair of DSB are typically accomplished via homologous recombination and mediated by RecA. This repair pathway, among others, is associated with activation of the SOS response. DNA adenine methyltransferase (dam) mutants have an increased number of DSB and, therefore, are notorious for being RecA-dependent for viability. Here, we show that the synthetic lethality of Δdam/ΔrecA is suppressed when clpP is removed, suggesting …

Transposable Elements In Fusarium Oxysporum & Growth Inhibition Of Fusarium Oxysporum Using Pepper Extracts, Taylor Aguiar

Masters Theses

The following contains two projects focused on the fungal pathogen, Fusarium oxysporum. The first project was purely computational in the examination of transposable elements (TEs), which are mobile sequences with the ability to multiply and move in their host genome. In F. oxysporum, TEs such as miniature impala elements are associated with the secreted in xylem gene that are related to its virulence over its host. The F. oxysporum species complex can be utilized as a model system for the examination of TE content and TE expression during the infection cycle. To find whether TEs play a role …

The Role Of Prrx1 And Snai2 As Master Regulators Of Fibroblast Identity, Huda A. Alzahrani

Masters Theses

Mammalian development involves a complex system of regulatory signals and reactions, leading to the creation of distinct tissues with specific structures and functions controlled by various gene expression profiles. Tissue-specific gene expression is by controlled specific combinations of transcription factors and cofactors that dictate cell differentiation via activation and repression of genes in mammalian cells.

Whole genome microarray analyses of fibroblasts have revealed candidate genes that may serve as master regulators of fibroblast identity, such as Prrx1 and Snai2, that activate downstream genes such as Bmp3, Twist, Shox2, Cfos, Slug, Sema3A, Spp1, and Col1a1. A previous study showed that Prrx1 …

The Transcriptional Response Of Trametes Versicolor To Growth On Maple Chips And Miscanthus Straw, Maha Alanazi

Masters Theses

This thesis research explored the transcriptional response of Trametes versicolor, a white-rot fungus, in response to fundamentally different types of lignocellulosic biomass (miscanthus and maple) and rich medium (malt extract agar). After five weeks of growth by the fungus on the biomass, the fungal RNA was extracted from three biological replicates per biomass type and mRNA was sequenced (approximately 30 million reads per sample). The reads were processed using ArrayStar to covert to RPKM and annotated using JGI's T. versicolor GO annotation file along with NCBI's BLAST. Comparisons were made between average gene expression of the fungus grown on …

Stressing Out: Dynamics Of Chromatin Insulator Body Formation In Eukaryotes Under Osmotic Stress, Shannon Marie Garland

Masters Theses

Higher order genome organization and the role it plays in governing cell dynamics and protein expression has become a widely studied field. Chromatin insulators are important to this organization in their ability to form long range contacts between distant regions on the genome. During times of osmotic stress, insulator proteins leave their binding sites on the DNA to form insulator bodies in the nucleus. This phenomenon relieves the DNA of its structure and is rapidly reversible. Using a variety of immunofluorescent staining methods, this work looked to further characterize this process while attempting to identify molecular mechanisms that mediate chromatin …