Genetics and Genomics Commons^™

Neural Rrp6 Depletion Drives Circrna Accumulation And Motor Impairment In Drosophila Melanogaster Neurons, Joseph Serio

Honors Theses

Circular RNAs (circRNAs) are a class of ubiquitous eukaryotic RNAs created by back splicing. These noncoding RNAs can modulate gene expression and are implicated in aging and neurodegenerative diseases. To limit the contribution of circRNAs to aging and pathobiology, it is essential to understand how turnover of these molecules is regulated, as it is not clear which enzymes are chiefly responsible for degrading circRNAs in neurons. This study evaluates the roles of ribonucleases potentially involved in neural circRNA turnover using the model fly Drosophila melanogaster. A mifepristone-activated GAL4/UAS GeneSwitch expressed RNAi transgenes and inactivated the ribonucleases. Flies were evaluated on …

Sibship Analysis Of Fall And Spring Spawn Gulf Sturgeon Eggs In The Choctawhatchee River, Florida, Lyric Mccullouch

Honors Theses

The Gulf sturgeon (Acipenser oxyrinchus desotoi) is an anadromous species that inhabits rivers along the northern Gulf of Mexico. Facing challenges from overfishing and habitat alteration, the Gulf sturgeon is listed as threatened under the Endangered Species Act. This study investigated the genetic distinctiveness of fall and spring spawning groups of Gulf sturgeon in the Choctawhatchee River, a critical habitat for the species. In this study, I used microsatellite loci to analyze Gulf sturgeon eggs previously collected in the fall and spring of 2019 and 2020. The goals of this study were to verify the genetic identity of …

Uncovering Novel Small Regulatory Rna In Protostome, Sweta Khanal

Dissertations

Small RNAs play pivotal roles in post-transcriptional gene regulation across diverse phylum of protostomes. In this study, we investigate the functional significance of atypical miRNAs, mirtron miR-1017 in Drosophila. Through ectopic expression in neuronal cells, we demonstrate that miR-1017 extends lifespan by targeting its host transcript, acetylcholine receptor Dα2, and influencing its splicing. This novel trans-regulatory function suggests a mechanism for mirtron evolution, highlighting the interplay between splicing and post-transcriptional regulation. Additionally, we profile small RNA populations in the polychaete developmental model Capitella teleta, shedding light on the small RNA landscape in annelid worms. Our analysis reveals a rich …

Sensing The Rhizosphere: Microbial Transcriptome Responses To Root Exudates And Redox-Active Metabolites, Ankita Bhattacharyya

Dissertations

The term “rhizosphere” describes the dynamic interface between plant roots and soil influenced by root exudates. It is a hotspot of microbial activity, plant-microbe, and microbe-microbe interactions. Distinct variations in bacterial diversity are thought to be driven by plant root exudates. However, the molecular details of these processes still remain unclear.

I addressed this gap by focusing on two strains, Pseudomonas synxantha 2-79 and Burkholderia lata 383, representing diverse groups of Gammaproteobacteria comprised of species with environmental, agricultural, and medical significance. In the first part of this project, we used a combination of metabolomics and RNA-seq to characterize transcriptome responses …

Genetic Parameters For Heat And Salinity Stress Tolerance In Northern Gulf Of Mexico Eastern Oyster, Crassostrea Virginica, Paul Mcdonald

Master's Theses

Summer mortalities in Gulf of Mexico eastern oysters have been linked to excessively high temperatures, and chronic exposure to low salinity. In this study, the feasibility of improving tolerance to these two stresses by selection was evaluated in common garden experiments employing a pool of 160 families. Heat stress tolerance was evaluated in seed (3.5 months) and adults (14 months) by applying a sustained temperature of 38-39 °C until all oysters expired. Low salinity challenges on juveniles (9 months) and adults (15 months) applied a salinity of 2.5 psu. Survival duration of individual oysters during challenges was recorded and tissue …

Missing Value Imputation For Single Omics And Multi-Omics Data, Meng Song

Dissertations

The integration analyses of multi-omics data have the advantages of extending our understanding of biological system across multiple omics layers, unraveling the functional mechanism of complex disease development, and refining the discovery of novel drug targets. However, multi-omics studies often face challenges such as data heterogeneity, missing values problem, interpretability, and imbalance classes. Among these challenges, the missing values problem is a critical issue for large cohort studies as not all samples will get a complete measurement for all the omics layers. To address the problem of missing values in multi-omics data, I focused on the imputation of completely missing …

Annotation Of Non-Model Species’ Genomes, Taiya Jarva

Master's Theses

The innovations in high throughput sequencing technologies in recent decades has allowed unprecedented examination and characterization of the genetic make-up of both model and non-model species, which has led to a surge in the use of genomics in fields which were previously considered unfeasible. These advances have greatly expanded the realm of possibilities in the fields of ecology and conservation. It is now possible to the identification of large cohorts of genetic markers, including single nucleotide polymorphisms (SNPs) and larger structural variants, as well as signatures of selection and local adaptation. Markers can be used to identify species, define population …

Comparative Analysis Of Rhizosphere And Endosphere Microbiome Of Different Blueberry Species (Vaccinium Sp.), Niladri Bhowmik

Master's Theses

Blueberries are an important agricultural commodity in all over the United States. Due to its health benefits, there is a huge demand globally, thus expanding the industry. Breeding programs are essential to maintain such industries. Challenges that play a role in contemporary breeding programs are various biotic and abiotic stress factors. Studies have shown that microorganisms are recruited by plants to alleviate them during stressful conditions. Though blueberries have been cultivated for about 100 years, how the microbiome has been affected due to this is poorly understood. We hypothesized that interspecific crosses and artificial selection have significantly changed the microbiome …

Pirna Expression In The Tube Feet Of Lytechinus Variegatus, Reagan Milliet

Honors Theses

Tissue regeneration is an area of research with implications for medicine and animal health. While nearly all living multi-cellular organisms are capable of regeneration, there are major differences in the animal kingdom. Some organisms are capable of regenerating virtually every cell in their bodies. In hopes of engineering tissue regeneration for medical applications, the mechanisms by which organisms regenerate are being widely investigated. To better understand regeneration, the role of P-element Induced WImpy testis (PIWI) proteins are being evaluated. In collaboration with PIWIinteracting RNAs (piRNAs), PIWI proteins have been proven instrumental to transposon silencing and maintenance of the genome in …

Development Of A Novel Environmental Dna (Edna) Tool For Monitoring Vulnerable Freckled Guitarfish, Pseudobatos Lentiginosus, In The Western Central Atlantic, Sarah Toepfer

Honors Theses

Rhino-rays are the most threatened group of elasmobranchs, having experienced widespread declines due to mortalities in fisheries and habitat degradation. Within the Western Central Atlantic, there are two extant species of Rhino-rays, the Critically Endangered Smalltooth Sawfish, Pristis pectinata, and the Vulnerable Freckled Guitarfish, Pseudobatos lentiginosus. Although there is research committed to P. pectinata in this region, less is known about the distribution status of P. lentiginosus. Over the past 50 years, P. lentiginosus have undergone a presumed range contraction in U.S. waters; once found from North Carolina to Texas, and historically common in the north central …

Pirna Expression In Regenerative Tissue Of Octopus Bimaculoides, Bailey Ervin

Honors Theses

Tissue regeneration is present in varying capacities across the animal kingdom. Animals such as Hydra and planarians have the capacity to regenerate entire bodies from extremely small sections of amputated tissue. Others, such as humans, have restricted capacities of regeneration, especially in terms of full appendages and specialized tissues such as cardiac and nervous tissue. One of the primary goals of studying regeneration in other organisms is to achieve the development of regenerative medicine. Interaction of P-element induced WImpy testis (PIWI) proteins and PIWI-interacting RNAs (piRNAs) have been implicated in germline genome maintenance, as well as transposable element silencing. Research …

A Genomic Investigation Of Divergence Between Tuna Species, Pavel V. Dimens

Dissertations

Effective management and conservation of marine pelagic fishes is heavily dependent on a robust understanding of their population structure, their evolutionary history, and the delineation of appropriate management units. The Yellowfin tuna (Thunnus albacares) and the Blackfin tuna (Thunnus atlanticus) are two exploited epipelagic marine species with overlapping ranges in the tropical and sub-tropical Atlantic Ocean. This work analyzed genome-wide genetic variation of both species in the Atlantic basin to investigate the occurrence of population subdivision and adaptive variation. A de novo assembly of the Blackfin tuna genome was generated using Illumina paired-end sequencing data and …

Systematic Overexpression Of Genes Encoded By Mycobacteriophage Waterfoul Reveals Novel Inhibitors Of Mycobacterial Growth, Danielle Heller, Isabel Amaya, Aleem Mohamed, Ilzat Ali, Dmitri Mavrodi, Padraig Deighan, Viknesh Sivanathan

Faculty Publications

Bacteriophages represent an enormous reservoir of novel genes, many of which are unrelated to existing entries in public databases and cannot be assigned a predicted function. Characterization of these genes can provide important insights into the intricacies of phage–host interactions and may offer new strategies to manipulate bacterial growth and behavior. Overexpression is a useful tool in the study of gene-mediated effects, and we describe here the construction of a plasmid-based overexpression library of a complete set of genes for Waterfoul, a mycobacteriophage closely related to those infecting clinically important strains of Mycobacterium tuberculosis and/or Mycobacterium abscessus. The arrayed …

Population Genetics Of Cownose Rays, Rhinoptera Spp. In The Western Atlantic, Helen Weber

Master's Theses

Using molecular tools (e.g., the NADH subunit II mitochondrial gene), this study was the first to document the Brazilian cownose ray, Rhinoptera brasiliensis, within the northwestern Atlantic (NWA), and was the first study to examine population structure in R. brasiliensis within the NWA or northern Gulf of Mexico (GMX), revealing novel insights into the population biology of the animal and extending its range by nearly 1,500 km. This study also examined the sympatrically occurring American cownose ray, R. bonasus, and found population structure between the NWA and the GMX and the NWA and the southwestern Atlantic (SWA). High …

Characterizing Endogenous Dicer Products To Unravel Novel Rnai Biogenesis Pathways, Jacob Oche Peter

Dissertations

ABSTRACT

RNA interference (RNAi) is a pervasive gene regulatory mechanism in eukaryotes based on the action of multiple classes of small RNA (sRNA). Exploiting RNAi pathways in non-model systems have great potential for creating potent RNAi technologies. Here, we accessed RNAi-mediated control of gene expression in the two-spotted spider mite, Tetranychus urticae (T. urticae) using engineered dsRNA designed to modulate the host RNAi pathway and increase RNAi efficacy. Analysis of Dicer (Dcr) generated fragments revealed how exogenous RNAs access the host RNAi pathway in this animal, opening avenues for designing RNAi technology for their control. Further, some organisms …

Investigation Of Dna Variability And Phylogenetic Relationships Of Perlesta (Plecoptera: Perlidae) In Mississippi, James C. Valentine, Mac H. Alford

Master's Theses

The genus Perlesta Banks, 1906 (Plecoptera: Perlidae) consists of 35 species, 33 native to the United States and Canada and two native to China. For over a century these small, brown stonefly adults and freckled yellow nymphs have gone by the name of the type species of the genus, Perlesta placida, but taxonomic work in the genus since 1989 has resulted in the recognition of additional species. These species were mostly recognized and described using morphological characteristics, but two areas that are lacking include (1) linking nymphs to adults and (2) phylogenetic analysis of all species occurring in Mississippi …

Reassessment Of Species Boundaries And Phylogenetic Relationships In The Desmodium Ciliare Complex (Fabaceae) Using Morphological And Dna Data, Joshua Wilkinson

Master's Theses

Desmodium (Beggar's ticks) is a generally weedy genus of approximately 280 species in the angiosperm family Fabaceae (Subfamily Papilionoideae, Tribe Desmodieae) characterized by indehiscent loments constricted into segments. Within the Southeastern U.S., the Desmodium ciliare group is one of two Desmodium species complexes that have been historically difficult, with much argument about the limits of species and the role of hybridization. There are three commonly recognized species within the D. ciliare group, D. ciliare (Muhl. ex Willd.) DC., Desmodium marilandicum (L.) DC., and D. obtusum (Muhl. ex Willd.) DC., with two supposed hybrids in addition to the …

Phylogenetic Origins And Age-Based Proportions Of Malacho (Elops Smithi) Relative To Ladyfish (Elops Saurus): Species On The Move In The Western Gulf Of Mexico, Damon Williford, Nicolette S. Beeken, Joel Anderson, Polly Hajovsky, Roberta Weixelman

Gulf and Caribbean Research

Two species of ladyfish occur in the Gulf of Mexico (GOM), Elops saurus and Elops smithi, that are morphologically indistinguishable except for vertebral counts but can also be identified by mitochondrial DNA haplotypes. Here we expand on previous work, most of which has occurred in Florida, and examine the demography, phylogenetics, geographic distribution, and age—structure of ladyfishes in Texas estuaries. Fishery—independent gill net data demonstrated that ladyfishes increase in abundance from north to south along the Texas coast. The abundance of ladyfishes also increased in Texas waters from 1982–2021, which coincides with recent trends of warmer winters. Genetic data …

Distribution, Abundance, And Population Genetics Of The Alligator Snapping Turtle (Macrochelys Temminckii) In Mississippi, Luke Pearson

Dissertations

The southeastern United States is a freshwater turtle biodiversity hotspot, second only to southeastern Asia, and this region is inhabited by one of the largest freshwater turtles in the world: the alligator snapping turtle (Macrochelys temminckii). This species was heavily harvested from the 1960’s to the 1980’s for the turtle soup industry, apparently contributing to substantial, range-wide population declines. These declines, coupled with anthropogenic impacts on riverine and floodplain habitats, have led to M. temminckii being petitioned for federal listing under the U.S. Endangered Species Act; it is currently under review by the U.S. Fish and Wildlife Service …

Environmental Rnai Pathways In The Two-Spotted Spider Mite, Mosharrof Mondal, Jacob Peter, Obrie Scarbrough, Alex Flynt

Faculty Publications

© 2020, The Author(s).

Background:RNA interference (RNAi) regulates gene expression in most multicellular organisms through binding of small RNA effectors to target transcripts. Exploiting this process is a popular strategy for genetic manipulation and has applications that includes arthropod pest control. RNAi technologies are dependent on delivery method with the most convenient likely being feeding, which is effective in some animals while others are insensitive. The two-spotted spider mite, Tetranychus urticae, is prime candidate for developing RNAi approaches due to frequent occurrence of conventional pesticide resistance. Using a sequencing-based approach, the fate of ingested RNAs was explored to …

Comparative Genome Analysis Of Fungal Antagonists Marinomonas Ostreistagni 398 And M. Spartinae 468., Jessie L. Fields

Honors Theses

Under certain conditions, the Spartina alterniflora and Juncus roemerianus grasses in marshes undergo progressive decline in an event known as Saltwater Marsh Dieback, which may be attributed to the presence of the plant pathogenic fungi Fusarium. The microbiomes of S. alterniflora and J. roemerianus from Deer Island, MS were characterized and Fusarium suppressing bacteria were identified. Among isolates capable of antagonizing Fusarium were Marinomonas ostreistagni 398 and M. spartinae 468. Despite the progress in understanding the diversity of Marinomonas, our ability to explain ecological, metabolic, and biochemical traits of marinomonads at the genomic sequence level remains limited. Analysis …

Human 5’-Tailed Mirtrons Are Processed By Rnasep, Mohammad Farid Zia

Dissertations

Approximately a thousand microRNAs (miRNAs) are documented from human cells. A third appear to transit non-canonical pathways that typically bypass processing by Drosha, the dedicated nuclear miRNA producing enzyme. The largest class of non-canonical miRNAs are mirtrons which eschew Drosha to mature through spliceosome activity. While mirtrons are found in several configurations, the vast majority of human mirtron species are 5’-tailed. For these mirtrons, a 3’ splice site defines the 3’ end of their hairpin precursor while a “tail” of variable length separates the 5’ base of the hairpin from the nearest splice site. How this tail is removed is …

Insecticidal Rna Interference, Thinking Beyond Long Dsrna, Alex S. Flynt

Faculty Publications

Over 20 years ago double-stranded RNA (dsRNA) was described as the trigger of RNAi interference (RNAi)-based gene silencing. This paradigm has held since, especially for insect biopesticide technologies where dsRNAs, similar to those described in 1998, are used to inhibit gene expression. In the intervening years, investigation of RNAi pathways has revealed the small RNA effectors of RNAi are diverse and rapidly evolving. The rich biology of insect small RNAs suggests potential to use multiple RNAi modes for manipulating gene expression. By exploiting different RNAi pathways, the menu of options for pest control can be expanded and could lead to …

Investigating Pirna Expression And Function In Regenerating Tissue Of Segmented Annelid Capitella Teleta, Beatriz Schueng Zancanela

Honors Theses

Regeneration can be observed virtually in all animals and previous studies have identified numerous genes involved in this process. In some invertebrates, the P-element induced wimpy testis (PIWI) genes appear to have an essential role. One such organism is Capitella teleta, a widely accepted model for annelid development. PIWI proteins are associated with small non-coding RNA called PIWI-interacting RNAs or piRNAs, which are involved in transposon silencing in the germline cells of many animals. It was previously believed that these proteins were only expressed in germline cells, however, recent studies have shown expression in somatic tissues as well. The function …

The Diversity And Evolution Of Phenazine Biosynthesis Pathways In Enterobacterales, Christian Leise

Honors Theses

Enterobacterales is an order of Gram-negative bacteria that encompasses plant and animal pathogens and organisms of industrial importance. Some of these bacteria produce secondary metabolites classified as phenazines (Phz). Studies in other groups of microorganisms revealed that phenazines are redox-active and exhibit broad antibacterial, antifungal, and antiparasitic activity. Enterobacterales are known to produce phenazines, but details about the diversity, biochemistry, and function of phenazine metabolites in these organisms are missing. In this work, we screened the National Center for Biotechnology Information (NCBI) GenBank for genome sequences of phenazine-producing (Phz+) Enterobacterales. Additionally, genomes of Phz+ strains Pectobacterium carotovorum cc303 and …

An Examination Of The Influence Of Multi-Scale Processes And Connectivity On The Population And Assemblage Dynamics Of Headwater Fishes, Josh P. Hubbell

Dissertations

Typified by their branching pattern, headwaters are numerically abundant as the density of these habitats increases with increasing distance from the base of a dendritic river system. Connectivity among headwaters is complex, resulting in the spatial isolation of populations. Headwater specialists have evolved a suite of traits that permit these species to permanently reside within these habitats. The spatial configuration and connectivity of headwaters has repercussions for metapopulations and meta-assemblages. I investigated how multi-scale processes and connectivity influenced the patch occupancy, coexistence, movement ecology, population structure, and gene flow of headwater specialists. In chapter two, I used occupancy modeling to …

Assessment Of Genetic Diversity In Largetooth Sawfish, Pristis Pristis, Populations Over The 20th Century, Ann Fearing

Master's Theses

The Critically Endangered Largetooth Sawfish, Pristis pristis, experienced global declines in range and abundance over the past century and Australia is now their last stronghold. This research aimed to understand whether these declines have been accompanied by a reduction in levels of genetic diversity. Using P. pristis tissue samples sourced from natural history specimens, three fragments (616-bp, 386-bp, 141-bp) in the mitochondrial DNA (mtDNA) control region were PCR-amplified. At each fragment, significant genetic structure was found in P. pristis overall (Φ_ST=0.946, N=9; 0.813, N=54; 0.771, N=99). The Indo-West Pacific and Atlantic/Eastern Pacific each harbor genetically distinct lineages of …

Development And Use Of An Environmental Dna Tool To Monitor Recovery Of The Critically Endangered Smalltooth Sawfish, Pristis Pectinata, In The Northwest Atlantic, Ryan N. Lehman

Master's Theses

The Critically Endangered Smalltooth Sawfish, Pristis pectinata, was once common in the tropical and subtropical waters of the Atlantic Ocean; however, following global declines in range and abundance over the past century, individuals were restricted to the waters of south and southwest Florida (SWFL) by about the 1980’s. Recently, public encounter reports have emerged in historically occupied habitats in United States waters, suggesting individuals are present in, or re-occupying, these areas, although the status of P. pectinata outside of SWFL is not currently well understood. Targeted environmental DNA (eDNA) surveys were chosen to assess the occurrence of P …

A Review Of Integrative Imputation For Multi-Omics Datasets, Meng Song, Jonathan Greenbaum, Joseph Luttrell, Weihua Zhou, Chong Wu, Hui Shen, Ping Gong, Chaoyang Zhang, Hong Wen Deng

Faculty Publications

Multi-omics studies, which explore the interactions between multiple types of biological factors, have significant advantages over single-omics analysis for their ability to provide a more holistic view of biological processes, uncover the causal and functional mechanisms for complex diseases, and facilitate new discoveries in precision medicine. However, omics datasets often contain missing values, and in multi-omics study designs it is common for individuals to be represented for some omics layers but not all. Since most statistical analyses cannot be applied directly to the incomplete datasets, imputation is typically performed to infer the missing values. Integrative imputation techniques which make use …

Exploiting Somatic Pirnas In Bemisia Tabaci Enables Novel Gene Silencing Through Rna Feeding, Mosharrof Mondal, Judith K. Brown, Alex Flynt

Faculty Publications

© This article is available under a Creative Commons License (Attribution 4.0 International, as described at https://creativecommons.org/licenses/by/4.0/). RNAi promises to reshape pest control by being nontoxic, biodegradable, and species specific. However, due to the plastic nature of RNAi, there is a significant variability in responses. In this study, we investigate small RNA pathways and processing of ingested RNAi trigger molecules in a hemipteran plant pest, the whitefly Bemisia tabaci. Unlike Drosophila, where the paradigm for insect RNAi technology was established, whitefly has abundant somatic piwi-associated RNAs (piRNAs). Long regarded as germline restricted, piRNAs are common in the …