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Developing A Comprehensive Genome-Scale Metabolic Model For The Arabidopsis Thaliana Root System, Lohani Esterhuizen

Department of Chemical and Biomolecular Engineering: Dissertations, Theses, and Student Research

Arabidopsis thaliana (Arabidopsis) is the most well-established model plant to date. Being the first plant to have its genome mapped, studies on Arabidopsis have provided insurmountable insights into the physiological and biochemical nature of plants. Methods that allow us to computationally study the metabolism of organisms include the use of genome-scale metabolic models (GEMs). Despite its popularity, no GEM currently maps the metabolic activity in the root system of Arabidopsis, which is the first organ to face and respond to stress conditions in the soil. This work aims to develop and implement a comprehensive GEM of the Arabidopsis root system …

Investigating The Function Of Protein S-Acyltransferase 2 In Arabidopsis Thaliana Using Crispr Mutants, Nicole Benjamin

Honors Theses and Capstones

Arabidopsis thaliana contains 24 Protein Acyl Transferases (PATs). These enzymes are responsible for catalyzing the addition of a palmitate to a target substrate (palmitoylation). However, beyond its broad enzymatic function, it is unknown how PATs interact on a cellular level. This research focuses on identifying the function of PAT2. CRISPR-Cas9 was used to create deletion mutations that would result in a non-functional PAT. Two plants homozygous for a pat2 deletion were identified: pat2-5 and pat2-6. Preliminary results suggest that pollen grains in both pat2 mutants are larger in both length and width. Additionally, it appears that mutant pollen tends to …

Genome-Wide Identification Of Qtls Associated With Iron Deficiency Tolerance In Soybean Reveals Gata12 As The Integrator Of Iron Signaling In Roots, Amir Maqbool, Ahmet Şahi̇n, Allah Bakhsh, Emre Aksoy

Turkish Journal of Botany

Iron (Fe) is an essential micronutrient required for plant growth and development. However, its deficiency causes substantial yield losses, particularly in alkaline soils. Soybean (Glycine max) serves as an ideal model to study Fe deficiency chlorosis (IDC) due to its inefficient Fe uptake from the soil. Although the quantitative trait loci (QTL) associated with IDC tolerance were determined in soybean, the specific genes within these QTL regions remain unidentified. In this study, it was aimed to identify and analyze the expression levels of genes present in IDC-responsive soybean QTL under Fe deficiency. Through this investigation, 6593 genes were identified within …

Identification Of Putative AtTt2 R2r3-Myb Transcription Factor Orthologues In Tanniferous Tissues Of L. Corniculatus Var. Japonicus Cv Gifu, D. N. Bryant, P. Bailey, P. Morris, Mark P. Robbins, C. Martin, T. Wang

IGC Proceedings (1993-2023)

R2R3-MYB plant transcription factors are sequence–specific DNA-binding proteins, which regulate the expression of specific gene(s) following the R2R3 DNA-binding domain interacting with the corresponding promoter sequence(s). The biosynthetic pathway leading to the production of anthocyanins has been demonstrated to be under MYB transcriptional regulatory control (Cone et al., 1986), while the accumulation of proanthocyanidins (PAs) in Arabidopsis seed coats is determined by the R2R3-MYB AtTT2 (Nesi et al., 2001). Using an informatics approach, partial sequences of putative AtTT2 orthologues have been identified and cloned from the forage legume Lotus corniculatus var. japonicus cv Gifu.

Investigation Of The Function Of Protein Acyl Transferases Using Crispr-Cas9 To Create Null Mutants, Claire E. Christopher

Honors Theses and Capstones

24 Protein Acyl Transferases (PATs) have been identified in the model organism Arabidopsis thaliana. Despite knowing the enzymatic function of these genes (palmitoylation), the specific subcellular pathways, protein targets, and cellular activities of these proteins remain unknown. To investigate the pathways that PAT genes are involved in, deletion mutations were created in several genes and the phenotype of the organisms carrying homozygous mutations was observed. To introduce mutations, the CRISPR/Cas9 system was inserted into the plant’s genome using Agrobacterium tumefaciens to cause deletions in PAT genes of interest. To date, no CRISPR-induced pat mutants have a noticeable phenotype. These …

Monoclonal Antibodies Against Methyl Glucuronoxylan Epitopes For Understanding Xylan Structure And Modification, Utku Avci

Turkish Journal of Botany

Lignocellulosic biomass is an abundant and renewable source that could be utilized for production of many bioproducts. It mainly consists of cellulose, hemicellulose, and lignin, all of which come together to form a recalcitrant wall that impedes production of bioproducts. In order to overcome the recalcitrant nature of the biomass, it is important to fully understand how cell wall components are synthesized and assembled. Xylan is one of the hemicellulosic components of the wall that contributes to recalcitrance. Xylan has substitutions on the backbone that are formed by the orchestration of many enzymes. For example, glucuronoxylan methyltransferases are responsible for …

Genetic Characterization Of Centromere-Mediated Uniparental Genome Elimination In Arabidopsis, Diana Spencer

Electronic Theses and Dissertations

The production of haploids through crossing was first discovered from instances of interspecies crosses in Nicotiana spp. in 1924. Since then, haploid induction crosses has been used to improve plant breeding programs and commercially utilized in a number of crop industries. The generation of doubled haploid instantaneously creates a pure homozygous line, therefore eliminating the need for several generations of inbreeding. There are several pathways to induce haploids in plants: of these methods, centromere-mediated genome elimination pathway engenders the highest haploid induction rate (HIR) with up to 45% in Arabidopsis compared to 15% through phospholipase-mediated haploid induction in maize. Centromere-mediated …

Dynamics Of Redox-Driven Molecular Processes In Local And Systemic Plant Immunity, Philip Berg

Theses and Dissertations

The work here presents two main parts. In the first part, chapters 1 – 3 focus on dynamical systems modeling in plant immunity, whereas chapters 4 – 6 describe contributions to computational modeling and analysis of proteomics and genomics data. Chapter 1 investigates dynamical and biochemical patterns of reversibly oxidized cysteines (RevOxCys) during effector-triggered immunity (ETI) in Arabidopsis, examines the regulatory patterns associated with Arabidopsis thimet oligopeptidase 1 and 2’s (TOP1 and TOP2), roles in the RevOxCys events during ETI, and analyzes the redox phenotype of the top1top2 mutant. The second chapter investigates the peptidome dynamics during ETI …

Determination Of The Functional Role Of Rab-Ggt In Physcomitrium Patens., Hyun Jin Jung

Electronic Theses and Dissertations

Protein prenylation, a common lipid post-translational modification, is required for growth and development in eukaryotes. Rab geranylgeranylation involves the addition of one or two 20-carbon geranylgeranyl moieties to Rab-GTPase target proteins, which regulate intracellular vesicle trafficking. The reaction is carried out by heterodimeric Rab geranylgeranyltransferase (Rab-GGT), which is composed of two associated α- and β-subunits, with the assistance of an additional protein called Rab escort protein (REP). Loss of function of the Rab-GGT α subunit RGTA1 has not been reported in any plant. While knockout of either of the two β subunits RGTB1 or RGTB2 results in …

Genetic And Biochemical Investigation Of Seed Fatty Acid Accumulation In Arabidopsis, Chinedu Charles Nwafor, Delin Li, Ping Qin, Long Li, Wei Zhang, Yuanwei Zhou, Jingjing Xu, Yongtai Yin, Jianbo Cao, Limin He, Fu Xiang, Chao Liu, Liang Guo, Yongming Zhou, Edgar B. Cahoon, Chunyu Zhang

Center for Plant Science Innovation: Faculty and Staff Publications

As a vegetable oil, consisting principally of triacylglycerols, is the major storage form of photosynthetically-fixed carbon in oilseeds which are of significant agricultural and industrial value. Photosynthesis in chlorophyll-containing green seeds, along with photosynthesis in leaves and other green organs, generates ATP and reductant (NADPH and NADH) needed for seed fatty acid production. However, contribution of seed photosynthesis to fatty acid accumulation in seeds have not been well-defined. Here, we report the contribution of seed-photosynthesis to fatty acid production by probing segregating green (photosynthetically-competent) and non-green or yellow (photosynthetically-non-competent) seeds in siliques of an Arabidopsis chlorophyll synthase mutant. Using this …

Inactivation Of The Entire Arabidopsis Group Ii Gh3s Confers Tolerance To Salinity And Water Deficit, Rubén Casanova-Sáez, Eduardo Mateo-Bonmatí, Jan Šimura, Aleš Pěnčík, Ondřej Novák, Paul Staswick, Karin Ljung

Department of Agronomy and Horticulture: Faculty Publications

Indole-3-acetic acid (IAA) controls a plethora of developmental processes. Thus, regulation of its concentration is of great relevance for plant performance. Cellular IAA concentration depends on its transport, biosynthesis and the various pathways for IAA inactivation, including oxidation and conjugation.

Group II members of the GRETCHEN HAGEN 3 (GH3) gene family code for acyl acid amido synthetases catalysing the conjugation of IAA to amino acids. However, the high degree of functional redundancy among them has hampered thorough analysis of their roles in plant development.

In this work, we generated an Arabidopsis gh3.1,2,3,4,5,6,9,17 (gh3oct) mutant to knock out the group II …

Calmodulin Like 38 Is Required For Autophagy Of Hypoxia-Induced Cytoplasmic Rna Granules In Arabidopsis Thaliana, Sterling Field

Doctoral Dissertations

In response to the energy crisis resulting from submergence stress and hypoxia, the model plant Arabidopsis thaliana limits non-essential mRNA translation, and accumulates cytosolic stress granules. Stress granules are phase-separated mRNA-protein particles that partition transcripts for various fates: storage, degradation, or return to translation after stress alleviation. Another response by the plant cell to low oxygen stress is the induction of the turnover pathway autophagy. Stress granule regulation by autophagy occurs by a ‘granulophagy’ pathway in yeast and mammalian systems through which parts or whole stress granules are degraded. Whether this occurs in plants has not been investigated.

A connection …

The Effects Of Ppal-1 In Arabidopsis Gamete Development, Amanda J White, Susana Perez-Martinez, Mark P. Running

The Cardinal Edge

Prenylation is a type of post-translational modification in which a 15- or 20-carbon lipid is added to the carboxyl (C) terminus of the protein. Arabidopsis thaliana contains the PROTEIN PRENYLTRANSFERASE ALPHA SUBUNIT-LIKE (PPAL) gene, which encodes a protein with homology to the α-subunits of the three known prenylation enzymes, PFT, PGGT, and Rab-GGT. We previously identified two mutations in PPAL, one of which is ppal-1, which contains a T-DNA insertion in the fourth intron. We have previously observed that self-fertilizing heterozygous ppal-1 plants produce progeny in which homozygous ppal-1 is underrepresented. This project attempts to ascertain …

Effect Of Volatile Compounds Produced By Selected Bacterial Endophytes In Promoting Plant Growth, Asha Maheshwari, Margaret T. Mmbaga, Bandana Bhusal, Christine A. Ondzighi-Assoume

Agricultural and Environmental Sciences Faculty Research

Bacterial endophytes selected for their capability to suppress diverse fungal pathogens in vitro and in greenhouse studies have been shown to promote plant growth. The effect of volatile compounds emitted by selected bacteria on plant growth in Arabidopsis thaliana, tomato (Solanum lycopersicum), sweet pepper (Capsicum annuum), and cucumbers (Cucumis sativus) was evaluated on container-grown plants nested above bacterial cultures, with roots exposed to the volatiles without direct contact between bacterial cells and the plant roots. Significant increases in plant growth were observed in plant height, root length, leaf size, fresh weight, and chlorophyll content in all plants tested. Although diverse …

Mutations In Several Auxin Biosynthesis Genes And Their Effects On Plant Phenotypes In Arabidopsis, Gabriela Hernandez, Lauren Huebner, Bethany Karlin Zolman

Undergraduate Research Symposium

Auxins are important hormones in plants that regulate growth and development. Disruptions in the auxin biosynthesis pathway result in morphological changes in phenotypes in the model plant Arabidopsis thaliana, including differences in root and leaf formation. Mutations in the Tryptophan Aminotransferase of Arabidopsis (TAA1) and YUCCA (YUC4) genes interfere with the plant's ability to synthesize Indole-3-acetic acid (IAA), the primary auxin involved in plant development. IBR1 and IBR3 act in the multistep conversion of indole-3-butyric acid (IBA) to IAA. ILL2, IAR3, and ILR1 hydrolyze IAA-amino acid conjugates into free IAA. The goal of …

Does Light Quality Influence Arabidopsis Thaliana Growth In Controlled Environments?, Nathan A. Deppe, Daniel A. Little, Kathleen K. Zapf

Purdue Methods for Arabidopsis Growth

This study examines the use of diverse lamp types, with inherently different spectral attributes, to determine light quality influence on Arabidopsis thaliana growth in controlled environments.

The Role Of The Golgi Elmo Proteins In Cell Adhesion In Arabidopsis Thaliana, Wesley James Hudson

Honors Projects

Proper growth and development of plant cells is dependent upon successful cell adhesion between cells, and this is mostly mediated by pectin in the plant cell wall. Previously, the Kohorn Laboratory identified a non-enzymatic Golgi protein named ELMO1 as it is required for cell adhesion, likely acting as a scaffold for cell wall polymer synthesis. Plants with mutant ELMO1 demonstrate a weak defective cellular adhesion phenotype as well as reduced mannose content in the cell wall. ELMO1 has homologous proteins in at least 29 different vascular plants. These homologues have 2 possible deletions in their amino acid sequence, but protein …

Arp2/3-Independent Wave/Scar Pathway And Class Xi Myosin Control Sperm Nuclear Migration In Flowering Plants, Mohammad F. Ali, Umma Fatema, Xiongbo Peng, Samuel W. Hacker, Daisuke Maruyama, Meng-Xiang Sun, Tomokazu Kawashima

Plant and Soil Sciences Faculty Publications

After eukaryotic fertilization, gamete nuclei migrate to fuse parental genomes in order to initiate development of the next generation. In most animals, microtubules control female and male pronuclear migration in the zygote. Flowering plants, on the other hand, have evolved actin filament (F-actin)-based sperm nuclear migration systems for karyogamy. Flowering plants have also evolved a unique double-fertilization process: two female gametophytic cells, the egg and central cells, are each fertilized by a sperm cell. The molecular and cellular mechanisms of how flowering plants utilize and control F-actin for double-fertilization events are largely unknown. Using confocal microscopy live-cell imaging with a …

Dissecting The Regulatory Network Of Sphingolipid Biosynthesis In Plants, Ariadna Gonzalez-Solis

Department of Biochemistry: Dissertations, Theses, and Student Research

Sphingolipids are a diverse group of lipids recognized as important components of cellular membranes and regulators of processes during development and in response to environmental stresses. Serine palmitoyltransferase (SPT) catalyzes the first step in sphingolipid biosynthesis and is a primary regulatory point for homeostasis. ORM proteins have been identified as negative regulators of SPT activity, however the mechanistic details of the regulation are only beginning to be understood. In this work, we show that ORM1 and ORM2 are essential for life cycle completion in Arabidopsis thaliana. Furthermore, the study of a structural ORM1 variant provided information about a transmembrane …

Characterizing The Role Of Β-Amylase3 In Cold Stress Response And Recovery In Arabidopsis Thaliana, Isabelle G. Houston

Senior Honors Projects, 2020-current

Starch is a polymer of glucose that is used as an energy store in plants. Mobilization of starch has implications in abiotic stress survival and recovery. While the importance of carbon and energy allocation in plant survival has been explored, the specific roles of starch degrading enzymes in plant responses to stress are still unclear. β-Amylase3, or BAM3, is the principle starch degrading enzyme at night and is transcriptionally upregulated in response to cold stress in the plant Arabidopsis thaliana. Using single and quadruple knockout mutant plants, I aimed to clarify the role of BAM3 in the response to …

Expression Profiling And Mutant Analysis Reveal Complex Regulatory Networks Involved In Arabidopsis Response To Biotic, Abiotic And Hormonal Stresses, Arjun Sham Valiyaveettil Shamgopal

Biology Dissertations

Biotic and abiotic stress factors consistently threat during different stages of the plant’s life cycle. A complex network of signaling molecules is required to overcome stresses, which is linked to general physiological and molecular processes. Botrytis cinerea is a necrotrophic fungal pathogen that causes grey mold disease on many crops. Apart from B. cinerea, several biotic, abiotic and hormonal stresses have detrimental effects on plant growth and productivity. The aim of this study is to identify the genetic regulations of individual stress types using comparative microarray analyses, and to find out the role of differentially expressed genes in Arabidopsis- …

Investigating The Role Of Protein Phosphatase 2a In The Salt Stress Response In Arabidopsis Thaliana, Daniel J. Dirocco

Honors Theses and Capstones

Protein Phosphatase 2A (PP2A) is a ubiquitous enzyme in eukaryotes that regulates a large array of cellular signaling processes. PP2A is composed of three subunits: catalytic C subunit, regulatory B subunit, and scaffolding/regulatory A subunit. In Arabidopsis thaliana, the A subunit has three isoforms - A1, A2, and A3 - that are highly conserved at the protein level, indicating that these proteins may be functionally interchangeable. In comparison to wildtype plants, seedlings with a mutation in the A1 gene have roots with obvious root cell file rotation or twisting under conditions of moderate salt stress. Twisted root cells result …

Defensive Manoeuvres Of Nhx1 And Sos1 Co/Overexpression In Plant Salt Tolerance, Necla Pehli̇van Gedi̇k, Li Sun, Neelam Mishra

Turkish Journal of Botany

Excessive accumulation of salts is one of the most important factors affecting the production quality of plants. The difficulty to develop salt stress tolerant crops has prevented the security of the global food system. The classical crop breeding approach that uses random mutagenesis/recombination is time-consuming in the face of an ever-increasing human population and extreme weather pattern shifts. Today, technologies introducing foreign genetic material into plants have the best potential to assist in crop breeding improvement due to its high efficiency, accuracy, low risk of off-target effects, and minimal labour compared to classical methods. The notion of Na+/H+ antiporters in …

Lorelei Localization And Ovule Ultrastructure In Arabidopsis Thaliana, Juleen May Dickson

Theses and Dissertations

Communication between the male and female gametophyte is vital to successful fertilization during sexual reproduction in plants. One of the proteins known to be important for communication between the male and female gametophyte is LORELEI (LRE). Several studies have shown that there are defects in pollen tube guidance and synergid degeneration, however this is the first study that shows that cell wall thickness in the female gametophyte may also be affected. Previous confocal studies have documented that LRE is present both in the filiform apparatus and found in puncta throughout the cytoplasm. This study confirmed this, but our studies suggest …

A Clade-Specific Arabidopsis Gene Connects Primary Metabolism And Senescence, Dallas C. Jones, Wenguang Zheng, Sheng Huang, Chuanlong Du, Xuefeng Zhao, Ragothaman M. Yennamalli, Taner Z. Sen, Dan Nettleton, Eve S. Wurtele, Ling Li

Dan Nettleton

Nearly immobile, plants have evolved new components to be able to respond to changing environments. One example is Qua Quine Starch (QQS, AT3G30720), an Arabidopsis thaliana-specific orphan gene that integrates primary metabolism with adaptation to environment changes. SAQR (Senescence-Associated and QQS-Related, AT1G64360), is unique to a clade within the family Brassicaceae; as such, the gene may have arisen about 20 million years ago. SAQR is up-regulated in QQS RNAi mutant and in the apx1 mutant under light-induced oxidative stress. SAQR plays a role in carbon allocation: overexpression lines of SAQR have significantly decreased starch content; …

Root Hair Single Cell Type Specific Profiles Of Gene Expression And Alternative Polyadenylation Under Cadmium Stress, Jingyi Cao, Congting Ye, Guijie Hao, Carole Dabney-Smith, Arthur G. Hunt, Qingshun Q. Li

Plant and Soil Sciences Faculty Publications

Transcriptional networks are tightly controlled in plant development and stress responses. Alternative polyadenylation (APA) has been found to regulate gene expression under abiotic stress by increasing the heterogeneity at mRNA 3′-ends. Heavy metals like cadmium pollute water and soil due to mining and industry applications. Understanding how plants cope with heavy metal stress remains an interesting question. The Arabidopsis root hair was chosen as a single cell model to investigate the functional role of APA in cadmium stress response. Primary root growth inhibition and defective root hair morphotypes were observed. Poly(A) tag (PAT) libraries from single cell types, i.e., root …

Pharmacologically Induced Meiosis Apomeiosis Interconversions In Boechera, Arabidopsis And Vigna, Lei Gao

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Apomixis is a clonal propagation method that produces offspring identical to the mother plant. With this feature, superior traits could be maintained over generations. However, our knowledge about apomixis is limited. In this study, we analyzed several apomictic Boechera embryologically to learn the details of apomixis. Meanwhile, we designed chemical treatments to successfully induce sex in apomictic plants and apomixis in sexual plants. Our experiments suggest that sex and apomixis coexist in plants and that sexual and apomictic reproduction are switchable by treating with specific chemicals.

Identification And Characterization Of Salinity Tolerance Genes By Activation Tagging In Arabidopsis, Jawaher Alkahtani

Graduate Theses and Dissertations

Salinity often affects irrigated areas in arid and semi-arid regions of the world. The existence and accumulation of soluble salts in the soil layers limit the growth of crops essential for our food. Salt stress dramatically affects plant growth, plant development, as well as crop yield. Arabidopsis thaliana is the plant model that provides a comprehensive knowledge of plant development, genetics and physiology, and response to abiotic stresses such as salinity. The redundancy of genes due to duplication, even in the simple model genome of Arabidopsis, limits the value of knockout (KO) mutagenesis to provide complete information on gene function. …

Interplay Of Dgat1, Pdat1 And Dgat2 Enzymes In Plant Triacylglycerol Assembly, Anushobha Regmi

Master's Theses

Acyl-CoA:diacylglycerol acyltransferase (DGAT) catalyzes the transesterification of fatty acid from acyl-CoA to diacylglycerol (DAG) forming triacylglycerol (TAG, a.k.a oils and fats). Most plants have at least two unrelated DGAT genes, DGAT1 and DGAT2. Plants predominantly express only one during oil synthesis; the reason, however is not clear. A few studies have indicated that each enzyme prefers DAG and acyl-CoA substrates with different fatty acid compositions. Industrially desirable seed oil composition can be obtained through genetic engineering by replacing the endogenous enzyme with one that has different substrate selectivity. In Arabidopsis thaliana, DGAT1 and another unrelated enzyme PDAT1 are essential …

Enhancing Resolution Of Natural Methylome Reprogramming Behavior In Plants, Robersy Sanchez, Xiaodong Yang, Hardik Kundariya, Jose R. Barreras, Yashitola Wamboldt, Sally A. Mackenzie

Department of Agronomy and Horticulture: Faculty Publications

We have developed a novel methylome analysis procedure, Methyl-IT, based on information thermodynamics and signal detection. Methylation analysis involves a signal detection problem, and the method was designed to discriminate methylation regulatory signal from background noise induced by thermal fluctuations. Comparison with three commonly used programs and various available datasets to furnish a comparative measure of resolution by each method is included. To confirm results, methylation analysis was integrated with RNAseq and network enrichment analyses. Methyl-IT enhances resolution of genome methylation behavior to reveal network-associated responses, offering resolution of gene pathway influences not attainable with previous methods.