Matching Items (12)
Description
Acceptance of the plant group Martyniaceae as a distinct family has long been questioned. Previously placed in the family Pedaliaceae, the Martyniaceae have been allied to numerous other families within the order Lamiales. The objectives of this study include the investigation of the placement of the Martyniaceae within the order Lamiales using molecular data (chloroplast DNA sequences), the further examination of the internal relationships of the Martyniaceae using an expanded nuclear and chloroplast sequences data set, and the construction of a taxonomic treatment of the family that includes all published names and taxa in the Martyniaceae. An analysis of the Lamiales using two chloroplast gene regions (ndhF and rps16) reveals that the Martyniaceae should be segregated from the family Pedaliaceae, but is not able to support the placement of any of its putatively-related families as sister to the Martyniaceae. Sequences from 151 taxa of the Lamiales are included in the analysis, including six representatives from the Martyniaceae. An analysis of the Martyniaceae using three chloroplast gene regions (psbA-trnH spacer, trnQ-5'rps16 intergenic spacer, and trnS-trnG-trnG spacer and intron) and the Internal Transcribed Spacer resolves two major clades within the Martyniaceae corresponding to the North American taxa (Martynia and Proboscidea) and the South American taxa (Craniolaria, Holoregmia, and Ibicella). Sequences from all five genera and 15 taxa were included in the analysis. Results from the molecular phylogenetic analyses are incorporated into a revised taxonomic treatment of the family. Five genera and thirteen species are recognized for the family Martyniaceae.
ContributorsGutiérrez, Raúl (Author) / Wojciechowski, Martin F (Thesis advisor) / Pigg, Kathleen B (Committee member) / Landrum, Leslie R (Committee member) / Butterworth, Charlie (Committee member) / Arizona State University (Publisher)
Created2011
Description
Coronaviruses are a medically significant group of viruses that cause respiratory and enteric infections in humans and a broad range of animals. Coronaviruses assemble at the internal membranes of the endoplasmic reticulum- Golgi intermediate compartment (ERGIC). While there is a basic understanding of how viruses assemble at these membranes, the full mechanistic details are not understood. The coronavirus envelope (E) protein is a small multifunctional viroporin protein that plays a role in virus assembly but its function is unknown. The two goals of this study were : 1. To identify and analyze the localization of MHV E and 2. To identify the functions of conserved residues in the tail of the E protein. This study closely examined the localization, dynamics and mobility of the mouse hepatitis virus (MHV) E protein to gain insight into its functions. The results from the first aim of this study showed that the MHV E protein localizes at the site of assembly in the ERGIC-Golgi region based on analysis by immunofluorescence and correlative electron microscopy. A novel tetra-cysteine tagged MHV E protein was used to study the dynamics of the protein in cells. A recombinant MHV E Lumio virus was used to study the trafficking and mobility of the E protein. Live cell imaging and surface biotinylation confirmed that the E protein does not traffic to the cell surface. Fluorescence recovery after photo-bleaching (FRAP) analyses revealed that the E protein is mobile at the site of localization. As a part of the second aim, conserved prolines and tyrosine in the tail of the protein were targeted by site directed mutagenesis and analyzed for functionality. While none of the residues were absolutely essential for localization or virus production, the mutations had varying degrees of effect on envelope formation, protein stability and virus release. Differential scanning calorimetry data suggests that the proline and tyrosine residues enhance interaction with lipids. A wild type (WT) peptide contained the conserved residues was also able to significantly reduce the hexagonal phase transition temperature of lipids, whereas a mutant peptide with alanine substitutions for the residues did not cause a temperature shift. This suggests that the peptide can induce a negative curvature in lipids. The E protein may be playing a role as a scaffold to allow membrane bending to initiate budding or possibly scission. This data, along with the localization data, suggests that the E protein plays a mechanistic role at the site of virus assembly possibly by remodeling the membrane thereby allowing virus budding and/or scission.
ContributorsVenkatagopalan, Pavithra (Author) / Hogue, Brenda G (Thesis advisor) / Jacobs, Bertram L (Committee member) / Roberson, Robert W. (Committee member) / Fromme, Petra (Committee member) / Arizona State University (Publisher)
Created2012
Description
The distinguishing feature of the filamentous fungi is the hyphae - tube-like microscopic cells that exhibit polarized growth via apical extension and allow the fungus to interact with its environment. Fungi elongate at the hyphal apex, through the localized construction of new plasma membrane and cell wall through the exocytosis of secretory vesicles. One population of these vesicles have been identified as chitosomes, containing chitin synthase isoenzymes, which are responsible for the polymerization of N-acetylglucosamine from UDP N-acetylglucosamine into chitin, the primary fibrillar component of the fungal cell wall. The chitosomes, in addition to other vesicles, can be observed aggregating in the hyphal tip in most filamentous fungi. In the Ascomycota and Basidiomycota, this collection of vesicles exhibits discrete organization and has been termed a Spitzenkörper. Although accumulations of vesicles can be observed in the hyphal tip of many growing filamentous fungi, some debate continues as to what precisely defines a Spitzenkörper. This study reports the details of three separate projects: first, to document the effects of deleting a single chitin synthase, CHS-1 and CHS-6 in Neurospora crassa with regards to hyphal ultrastructure, cytoplasmic organization, and growth in comparison to the wild-type. Given the importance of chitin synthesis in fungal cell growth, deletion of a critical chitin synthase presumably impacts cell wall structure, fungal growth and cytoplasmic organization. Second, an examination of the ultrastructure of four zygomycetous fungi - Coemansia reversa, Mortierella verticillata, Mucor indicus, and Gilbertella persicaria has been conducted. Utilization of cryofixation and freeze-substitution techniques for electron microscopy has produced improved preservation of cytoplasmic ultrastructure, particularly at the hyphal apex, allowing detailed analysis of vesicle size, contents, and organization. Lastly, hyphal tip organization was reviewed in a broad range of fungi. Previous studies had either focused on a few select fungi or representative groups. Vesicle organization, composition and size do appear to vary among the classes of fungi, but some trends, like the vesicle crescent in the zygomycetous fungi have been documented.
ContributorsFisher, Karen Elizabeth (Author) / Roberson, Robert W. (Thesis advisor) / Chandler, Douglas (Committee member) / Riquelme, Meritxell (Committee member) / Stutz, Jeam (Committee member) / Wojciechowski, Martin (Committee member) / Arizona State University (Publisher)
Created2015
Description
The remarkable conservation of molecular and intra-/inter-cellular pathways underpinning the fundamental aspects of sugar partitioning in two evolutionarily divergent organisms – a non-vascular moss Physcomitrella patens and a vascular cereal crop Oryza sativa (rice) – forms the basis of this manuscript. Much of our current knowledge pertaining to sugar partitioning in plants mainly comes from studies in thale cress, Arabidopsis thaliana, but how photosynthetic sugar is loaded into the phloem in a crop as important as rice is still debated. Even less is known about the mechanistic aspects of sugar movement in mosses. In plants, sugar either moves passively via intercellular channels called plasmodesmata, or through the cell wall spaces in an energy-consuming process. As such, I first investigated the structure of plasmodesmata in rice leaf minor vein using electron tomography to create as of yet unreported 3D models of these channels in both simple and branched conformations. Contrary to generally held belief, I report two different 3D morphotypes of simple plasmodesmata in rice. Furthermore, the complementary body of evidence in arabidopsis implicates plasma membrane localized Proton Pyrophosphatase (H+-PPase) in the energy-dependent movement of sugar. Within this wider purview, I studied the in situ ultrastructural localization patterns of H+-PPase orthologs in high-pressure frozen tissues of rice and physcomitrella. Were H+-PPases neo-functionalized in the vascular tissues of higher plants? Or are there evolutionarily conserved roles of this protein that transcend the phylogenetic diversity of land plants? I show that H+-PPases are distinctly expressed in the actively growing regions of both rice and physcomitrella. As expected, H+-PPases were also localized in the vascular tissues of rice. But surprisingly, H+-PPase orthologs were also prominently expressed at the gametophyte-sporophyte junction of physcomitrella. Upon immunogold labeling, H+-PPases were found to be predominantly localized at the plasma membrane of the phloem complexes of rice source leaves, and both the vacuoles and plasma membrane of the transfer cells in the physcomitrella haustorium, linking H+-PPases in active sucrose loading in both plants. As such, these findings suggest that the localization and presumably the function of H+-PPases are conserved throughout the evolutionary history of land plants.
ContributorsRegmi, Kamesh Chandra (Author) / Gaxiola, Roberto A (Thesis advisor) / Wojciechowski, Martin F (Committee member) / Pigg, Kathleen B (Committee member) / Roberson, Robert W. (Committee member) / Arizona State University (Publisher)
Created2016
Evolutionary analysis of the CAP superfamily of proteins using amino acid sequences and splice sites
Description
Here I document the breadth of the CAP (Cysteine-RIch Secretory Proteins (CRISP), Antigen 5 (Ag5), and the Pathogenesis-Related 1 (PR)) protein superfamily and trace some of the major events in the evolution of this family with particular focus on vertebrate CRISP proteins. Specifically, I sought to study the origin of these CAP subfamilies using both amino acid sequence data and gene structure data, more precisely the positions of exon/intron borders within their genes. Counter to current scientific understanding, I find that the wide variety of CAP subfamilies present in mammals, where they were originally discovered and characterized, have distinct homologues in the invertebrate phyla contrary to the common assumption that these are vertebrate protein subfamilies. In addition, I document the fact that primitive eukaryotic CAP genes contained only one exon, likely inherited from prokaryotic SCP-domain containing genes which were, by nature, free of introns. As evolution progressed, an increasing number of introns were inserted into CAP genes, reaching 2 to 5 in the invertebrate world, and 5 to 15 in the vertebrate world. Lastly, phylogenetic relationships between these proteins appear to be traceable not only by amino acid sequence homology but also by preservation of exon number and exon borders within their genes.
ContributorsAbraham, Anup (Author) / Chandler, Douglas E. (Thesis advisor) / Buetow, Kenneth H. (Committee member) / Roberson, Robert W. (Committee member) / Arizona State University (Publisher)
Created2016
Description
In the United States, Escherichia coli O157:H7 (E. coli O157:H7) is the most frequent cause of hemolytic uremic syndrome (HUS) and it is also the primary cause of acute renal failure in children. The most common route of the infection is ingestion of contaminated meat or dairy product originating from cattle or vegetables contaminated with bovine manure. Since cattle are the main reservoir for human infection with E. coli O157:H7, the reduction of intestinal colonization by these bacteria in cattle is the best approach to prevent human infections. Intimin is an outer membrane protein of E. coli O157:H7 that plays an important role in adhesion of the bacteria to the host cell. Hence, I proposed to express intimin protein in tomato plants to use it as a vaccine candidate to reduce or prevent intestinal colonization of cattle with E. coli O157:H7. I expressed His-tagged intimin protein in tomato plants and tested the purified plant-derived intimin as a vaccine candidate in animal trials. I demonstrated that mice immunized intranasally with purified tomato-derived intimin produced intimin-specific serum IgG1and IgG2a, as well as mucosal IgA. I further demonstrated that mice immunized with intimin significantly reduced time of the E. coli O157:H7 shedding in their feces after the challenge with these bacteria, as compared to unimmunized mice. Shiga toxin is the major virulence factor that contributes to HUS. Since Shiga toxin B subunit has an important role in the attachment of the toxin to its receptor, I fused intimin to Shiga toxin B subunit to create multivalent subunit vaccine and tested the effects upon immunization of mice with the B subunit when combined with intimin. His-tagged intimin, Shiga toxin B subunit, and Shiga toxin-intimin fusion proteins were expressed in E. coli and purified. I demonstrated that this multivalent fusion protein vaccine candidate elicited intimin- and Shiga toxin B-specific IgG1, IgG2a, and IgA antibodies in mice. I also showed a reduction in the duration of the bacterial shedding after the challenge compared to the control sham-immunized groups.
ContributorsTopal, Emel (Author) / Mason, Hugh S. (Thesis advisor) / Bingham, Scott E. (Committee member) / Mor, Tsafrir (Committee member) / Roberson, Robert W. (Committee member) / Arizona State University (Publisher)
Created2010
Description
The Juglandaceae (walnuts, hickories, pecans) has one of the best-documented fossil records in the Northern Hemisphere. The oldest modern genus, Cyclocarya, today restricted to China, first appears in the late Paleocene (57 ma) of North Dakota, USA. Unlike walnuts and pecans that produce edible fruits dispersed by mammals, Cyclocarya fruits are small nutlets surrounded by a prominent circular wing, and are thought to be wind- or water-dispersed. The current study provides the first evidence that fossil fruits were different from modern forms in the number and organization of their attachment to reproductive branches, and in their anatomical structure. Unlike the modern genus that bears separate pistillate and staminate flowers the fossil fruits had attached pollen-bearing structures. Unisexual pollen catkins are also present, suggesting the fossil Cyclocarya may have differed from its modern relative in this feature. Like several other plants from the late Paleocene Almont/Beicegel Creek floras, Cyclocarya shows a mosaic combination of characters not seen in their modern counterparts. Fossils were collected from the field, and examined for specimens exposed on the weathered rock surface. Specimens from Almont were photographed with reflected light, while those from Beicegel Creek cut into slabs and prepared by etching the rock matrix in 49% hydrofluoric and re-embedding the exposed plant material in cellulose acetate and acetone to make "peels". Selected specimens are cut out, mounted on microscope slides, and studied with light microscopy. These fossil fruits were studied because they are the earliest fossil evidence of Cyclocarya. They are exceptionally preserved and thus provide critical structural evidence for changes in that occurred during the evolution of plants within this lineage. Because Cyclocarya fruits are winged, they might be assumed to be wind-dispersed. Their radial symmetry does not have the aerodynamic qualities typical of wind-dispersed fruits, and may have been dispersed by water.
ContributorsTaylor, Malcom DeWitt (Author) / Pigg, Kathleen B (Thesis advisor) / Wojciechowski, Martin F (Committee member) / Devore, Melanie L (Committee member) / Farmer, Jack (Committee member) / Gill, Anthony (Committee member) / Arizona State University (Publisher)
Created2010
Description
In the face of the sixth mass extinction on Earth, with the flowering plant family Cactaceae assessed as the fifth most endangered plant or animal family by the International Union for the Conservation of Nature (IUCN), it is imperative that all available tools be used to understand the biodiversity, habitat suitability, climate change impacts and population viability of cacti. Within the Cactaceae, Mammillaria Haw and the closely related genus Cochemiea (K. Brandegee) Walton of Baja California, Mexico, are species-rich, with 46 regionally endemic taxa, 12 of which have been assessed as threatened or endangered by the IUCN. This study clarifies the evolutionary relationships in the Mammilloid clade, a complex and species-rich clade in tribe Cacteae, and generic circumscription of the genera Mammillaria Haw. and Cochemiea (K. Brandegee) Walton, estimates divergence times, diversification rates and ancestral ranges and explores habitat suitability and the risk of extinction of a representative species within these genera. The r species, Cochemiea halei (K. Brandegee) Walton, a narrowly distributed island endemic, is assessed using species distribution modeling (SDM) and population viability analysis (PVA). SDM in this study includes projections to two climate change scenarios over the next century, using four representative particle concentration pathways, and the PVA uses habitat-specific deterministic and stochastic models. The results of molecular phylogenetic analyses of the Mammilloid cladde restore the genus Mammillaria to monophyly via new combinations in the genus Cochemiea. The taxa in this study are shown to be of recent origin resulting from rapid diversification and radiation. Geological and climatic forces at multiple scales appear to be responsible for the high degree of biodiversity and endemism of these cacti. SDM shows that C. halei is likely to be stranded in its fragmented island habitat, has a facultative adaptation to ultramafic soils, and faces a 21%–53% contraction of its range on the islands under climate change scenarios. PVA suggests that C. halei is at increased risk of extinction in response to slight decreases in fecundity and persistence. In general, the perspectives in this dissertation fill several gaps in our prior knowledge of the evolution, biogeography, and conservation pressures of an important, species-rich group of cacti, occurring in a region of high biodiversity and endemism.
ContributorsBreslin, Peter (Author) / Wojciechowski, Martin F (Thesis advisor) / Albuquerque, Fabio (Committee member) / Fehlberg, Shannon (Committee member) / Majure, Lucas (Committee member) / Rebman, Jon (Committee member) / Arizona State University (Publisher)
Created2020
Description
The Phoenix Zoo, also known as the Arizona Center for Nature Conservation (PZ), is an Association of Zoos and Aquariums (AZA) accredited zoological institution and among largest-nonprofit, privately-owned zoos in the United States (Smith, 2020). Located within Papago Park in Phoenix (Maricopa County), Arizona, adjacent to the Desert Botanical Garden (DES), the two combine to bring environmental awareness to the Phoenix Metropolitan Region. While the DES specializes in botanical presentation, the ACNC focuses on zoological education. Whereas the flora of DES is well known, that of ACNC has yet to be completely documented. Given its role as a center for public engagement and education, documenting and mapping the floristic diversity of the Phoenix Zoo provides updated botanical information and occurrence records, an important component of understanding biodiversity for the Phoenix area. Between the fall of 2017 and the Spring of 2021, the grounds of the ACNC were walked within the 2-mile perimeter and surrounding exterior within Papago Park. Plant specimens and photographs were collected and archived for later identification using various botanical keys. Species names were verified through updated botanical databases such as Tropicos.org and worldfloraonline.org and compiled into a checklist. A total of 706 species have been identified, and of those 548 specimens have been collected as specimen vouchers. Of these, 120 are of taxa known to be native to the Phoenix Salt River Valley. While approximately 79 of those previously listed taxa native to Papago Park were either not found during this study or were extirpated from the grounds of the ACNC during urbanization of the region. There are 586 exotic taxa, some are common cultivars, while others are new to the region. Data for this survey is publicly available on SEINet, a regional network of North America herbaria (https://swbiodiversity.org/seinet/), as georeferenced voucher specimens, human observations, and photographs. Data is also partially duplicated through the iNaturalist platform (iNaturalist.com).
ContributorsBerry, Zachery R (Author) / Makings, Elizabeth (Thesis advisor) / Pigg, Kathleen B (Thesis advisor) / Wojciechowski, Martin F (Committee member) / Arizona State University (Publisher)
Created2021
Description
The intracellular motility seen in the cytoplasm of angiosperm plant pollen tubes is known as reverse fountain cytoplasmic streaming (i.e., cyclosis). This effect occurs when organelles move anterograde along the cortex of the cell and retrograde down the center of the cell. The result is a displacement of cytoplasmic volume causing a cyclic motion of organelles and bulk liquid. Visually, the organelles appear to be traveling in a backwards fountain hence the name. The use of light microscopy bioimaging in this study has documented reverse fountain cytoplasmic streaming for the first time in fungal hyphae of Rhizopus oryzae and other members in the order Mucorales (Mucoromycota). This is a unique characteristic of the mucoralean fungi, with other fungal phyla (e.g., Ascomycota, Basidiomycota) exhibiting unidirectional cytoplasmic behavior that lacks rhythmic streaming (i.e., sleeve-like streaming). The mechanism of reverse fountain cytoplasmic streaming in filamentous fungi is currently unknown. However, in angiosperm plant pollen tubes it’s correlated with the arrangement and activity of the actin cytoskeleton. Thus, the current work assumes that filamentous actin and associated proteins are directly involved with the cytoplasmic behavior in Mucorales hyphae. From an evolutionary perspective, fungi in the Mucorales may have developed reverse fountain cytoplasmic streaming as a method to transport various organelles over long and short distances. In addition, the mechanism is likely to facilitate driving of polarized hyphal growth.
ContributorsShange, Phakade Mdima (Author) / Roberson, Robert W. (Thesis advisor) / Gile, Gillian (Committee member) / Baluch, Debra (Committee member) / Arizona State University (Publisher)
Created2020