Life Sciences Commons^™

Isolation Of Arabidopsis Thaliana Plants Homozygous For An Insertional Inactivation Mutation Within Atprp4., Sydney Raitz, Timothy D. Trott

Research in Biology

The AtPRP4 gene in Arabidopsis thaliana has been shown to function in several specific parts of the plant’s cell wall. It is shown to be expressed in the seeds, radicles, roots, leaves, inflorescences, and embryos of Arabidopsis thaliana. These patterns have suggested unique functions for ATPRP4 in determining cell-type-specific wall structure during the development of a plant as well as contributing to defense reactions against physical damage to the plant and pathogen infection within the plant. In this study, a simple DNA prep was performed on the true leaves of Arabidopsis thaliana. Subsequent PCR reactions were performed using …

Cell-Type-Specific Profiling Of The Arabidopsis Thaliana Membrane Protein-Encoding Genes, Sergio Alan Cervantes-Pérez, Marc Libault

Department of Agronomy and Horticulture: Faculty Publications

Membrane proteins work in large complexes to perceive and transduce external signals and to trigger a cellular response leading to the adaptation of the cells to their environment. Biochemical assays have been extensively used to reveal the interaction between membrane proteins. However, such analyses do not reveal the unique and complex composition of the membrane proteins of the different plant cell types. Here, we conducted a comprehensive analysis of the expression of Arabidopsis membrane proteins in the different cell types composing the root. Specifically, we analyzed the expression of genes encoding membrane proteins interacting in large complexes. We found that …

Cell-Type-Specific Profiling Of The Arabidopsis Thaliana Membrane Protein-Encoding Genes, Sergio Alan Cervantes-Pérez, Marc Libault

Department of Agronomy and Horticulture: Faculty Publications

Membrane proteins work in large complexes to perceive and transduce external signals and to trigger a cellular response leading to the adaptation of the cells to their environment. Biochemical assays have been extensively used to reveal the interaction between membrane proteins. However, such analyses do not reveal the unique and complex composition of the membrane proteins of the different plant cell types. Here, we conducted a comprehensive analysis of the expression of Arabidopsis membrane proteins in the different cell types composing the root. Specifically, we analyzed the expression of genes encoding membrane proteins interacting in large complexes. We found that …

Isolation Of Arabidopsis Thaliana Plants Homozygous For An Insertional Inactivation Mutation Within Atprp4., Sydney Raitz, Timothy D. Trott

Faculty Works

The AtPRP4 gene in Arabidopsis thaliana has been shown to function in several specific parts of the plant’s cell wall. It is shown to be expressed in the seeds, radicles, roots, leaves, inflorescences, and embryos of Arabidopsis thaliana. These patterns have suggested unique functions for ATPRP4 in determining cell-type-specific wall structure during the development of a plant as well as contributing to defense reactions against physical damage to the plant and pathogen infection within the plant. In this study, a simple DNA prep was performed on the true leaves of Arabidopsis thaliana. Subsequent PCR reactions were performed using …

The Influence Of Singlet Oxygen And Loss Of Function Of Fatty Acid Desaturase 7 In The Chloroplast On Aphid Resistance In Arabidopsis Thaliana, Hillary Donna Fischer

Graduate Theses and Dissertations

Fatty Acid Desaturase 7 (FAD7) is a chloroplast-localized enzyme that alters the fatty acid content of photosynthetic membranes, and that negatively regulates plant defenses against aphids. Previous studies in the model organism Arabidopsis (Arabidopsis thaliana) have shown that loss-of-function mutations in FAD7 decrease population growth of the green peach aphid (GPA; Myzus persicae Sulzer). This study further characterized the effects of a fad7 null mutant on aphids, and investigated the role of reactive oxygen species (ROS), including singlet oxygen (1O2), in plant responses to aphid resistance in fad7 as well as in wild type plants and a mutant with heightened …

Genome-Wide Occupancy Of Polycomb Group Proteins And Chromatin Remodeler Splayed And Their Interplay In Arabidopsis, Jie Shu

Electronic Thesis and Dissertation Repository

The Polycomb group (PcG) proteins form two protein complexes, Polycomb repressive complex 1 (PRC1) and PRC2, which are key epigenetic regulators in eukaryotes. PRC2 represses gene expression by catalyzing trimethylation of histone H3 lysine 27 (H3K27me3). In Arabidopsis thaliana, CURLY LEAF (CLF) and SWINGER (SWN) are two major H3K27 methyltransferases, playing essential roles in plant growth and development. Despite their importance, genome-wide occupancy profiles of CLF and SWN have not yet been determined and compared. In this thesis, I generated transgenic lines expressing GFP-tagged CLF/SWN and determined the genome-wide distributions of CLF and SWN in Arabidopsis seedlings. I also …

Defective Aba-Mediated Sugar Signalling Pathway In An Established Arabidopsis Thaliana Cell Suspension Culture Explains Its Stay-Green Phenotype At High Sugar Concentrations, Avery Mccarthy

Electronic Thesis and Dissertation Repository

An unusual sugar insensitive phenotype was identified in an established cell suspension culture of Arabidopsis thaliana. We characterized the physiology, biochemistry and genetics of the sugar insensitive cell culture, in order to identify factors contributing to the phenotype. Chlorophyll levels of the cell suspension culture were insensitive to high sucrose (6-15% w/v) and maintained a green phenotype. Immunoblotting indicated that levels of key photosynthetic proteins (PsaA, Lhcb2 and Rubisco) increased as a function of external sucrose concentration. The green cell culture was photosynthetically competent based on light-dependent, CO₂-saturated rates of O₂ evolution as well as Fv/Fm …

Nonpolar Residues In The Presumptive Pore‐Lining Helix Of Mechanosensitive Channel Msl10 Influence Channel Behavior And Establish A Nonconducting Function, Grigory Maksaev, Jennette K. Shoots, Simran Ohri, Elizabeth S. Haswell

Biology Faculty Publications & Presentations

Mechanosensitive (MS) ion channels provide a universal mechanism for sensing and responding to increased membrane tension. MscS‐like (MSL) 10 is a relatively well‐studied MS ion channel from Arabidopsis thaliana that is implicated in cell death signaling. The relationship between the amino acid sequence of MSL10 and its conductance, gating tension, and opening and closing kinetics remains unstudied. Here, we identify several nonpolar residues in the presumptive pore‐lining transmembrane helix of MSL10 (TM6) that contribute to these basic channel properties. F553 and I554 are essential for wild type channel conductance and the stability of the open state. G556, a glycine residue …

Development Of Ifox-Hunting As A Functional Genomic Tool And Demonstration Of Its Use To Identify Early Senescence-Related Genes In The Polyploid Brassica Napus, Juan Ling, Renjie Li, Chinedu Charles Nwafor, Junluo Cheng, Maoteng Li, Qing Xi, Jian Wu, Lu Gan, Qingyong Yang, Chao Liu, Ming Chen, Yongming Zhou, Edgar B. Cahoon, Chunyu Zhang

Center for Plant Science Innovation: Faculty and Staff Publications

Functional genomic studies of many polyploid crops, including rapeseed (Brassica napus), are constrained by limited tool sets. Here we report development of a gain-of-function platform, termed ‘iFOX (inducible Full-length cDNA OvereXpressor gene)-Hunting’, for inducible expression of B. napus seed cDNAs in Arabidopsis. A Gateway-compatible plant gene expression vector containing a methoxyfenozide-inducible constitutive promoter for transgene expression was developed. This vector was used for cloning of random cDNAs from developing B. napus seeds and subsequent Agrobacterium-mediated transformation of Arabidopsis. The inducible promoter of this vector enabled identification of genes upon induction that are otherwise lethal when constitutively overexpressed and …

Non-Canonical Signaling From The Etr1 And Etr2 Ethylene Receptors In Arabidopsis Thaliana, Arkadipta Bakshi

Doctoral Dissertations

The gaseous phytohormone ethylene regulates several physiological and developmental processes in higher plants. There are five ethylene receptor isoforms that mediate the responses to ethylene in the model plant Arabidopsis thaliana. Prior research has shown that these five ethylene receptor isoforms in Arabidopsis have both overlapping and non-overlapping roles in regulating diverse responses such as growth in air, growth recovery after removal of ethylene, and ethylene stimulated nutational bending. Functional divergence of ETR1 has been determined in controlling some of these traits and in some of these cases, ETR1 subfunctionalization requires the receiver domain. Using homology modeling and sequence …

Selection Of Efficient Taq Dna Polymerase To Optimize T-Dna Genotyping Method For Rapid Detection Of Mutant Arabidopsis Thaliana Plants, Inna B. Chastukhina, Liliia R. Nigmatullina, Lia R. Valeeva, Eugene V. Shakirov

Biological Sciences Faculty Research

Plants harbor homologues of various animal genes involved in phosphorus metabolism, telomere biology and other cellular processes. Compared to experiments with many other multicellular organisms, research in the model plant Arabidopsis thaliana takes advantage of short generation time and an ever increasing arsenal of genetic and transgenic tools, including large collections of T-DNA knockout and activation lines. The availability of thousands of publicly available transgenic Arabidopsis lines provides a unique opportunity to address a number of important biological questions. However, identification of individual T-DNA mutant plants from a pool of seeds provided by a biological stock distribution center remains a …

Plastid Osmotic Stress Influences Cell Differentiation At The Plant Shoot Apex, Margaret E. Wilson, Matthew Mixdorf, R Howard Berg, Elizabeth S. Haswell

Biology Faculty Publications & Presentations

The balance between proliferation and differentiation in the plant shoot apical meristem is controlled by regulatory loops involving the phytohormone cytokinin and stem cell identity genes. Concurrently, cellular differentiation in the developing shoot is coordinated with the environmental and developmental status of plastids within those cells. Here, we employ an Arabidopsis thaliana mutant exhibiting constitutive plastid osmotic stress to investigate the molecular and genetic pathways connecting plastid osmotic stress with cell differentiation at the shoot apex. msl2 msl3 mutants exhibit dramatically enlarged and deformed plastids in the shoot apical meristem, and develop a mass of callus tissue at the shoot …

Glucan Synthase-Like 8: A Key Player In Early Seedling Development In Arabidopsis, Behnaz Saatian

Electronic Thesis and Dissertation Repository

Plants’ cell walls have unique chemical composition and features which enable them to play essential roles during plant development as shaping the cells and providing intercellular communication between adjacent cells. Polysaccharides, including callose, and glycoproteins are known as the main constituents of the cell wall. Callose, a linear β-1,3-glucan polymer, is accumulated at the cell plate during cytokinesis, in plasmodesmata, where it regulates cell-to-cell communication, in dormant phloem, where it seals sieve plates after mechanical injury and pathogen attack, and in male and female gametophytes. GLUCAN SYNTHASE-LIKE (GSL) genes in Arabidopsis comprise a family of 12 members. A …

Dioxygenase-Encoding Atdao1 Gene Controls Iaa Oxidation And Homeostasis In Arabidopsis, Silvana Porco, Aleš Pěnčík, Afaf Rashed, Ute Voβ, Rubén Casanova-Sáez, Anthony Bishop, Agata Golebiowska, Rahul Bhosale, Ranjan Swarup, Kamal Swarup, Pavlína Penáková, Ondrej Novák, Paul E. Staswick, Peter Hedden, Andrew L. Phillips, Kris Vissenberg, Malcolm J. Bennett, Karin Ljung

Department of Agronomy and Horticulture: Faculty Publications

Auxin represents a key signal in plants, regulating almost every aspect of their growth and development. Major breakthroughs have been made dissecting the molecular basis of auxin transport, perception, and response. In contrast, how plants control the metabolism and homeostasis of the major form of auxin in plants, indole-3- acetic acid (IAA), remains unclear. In this paper, we initially describe the function of the Arabidopsis thaliana gene DIOXYGENASE FOR AUXIN OXIDATION 1 (AtDAO1). Transcriptional and translational reporter lines revealed that AtDAO1 encodes a highly root-expressed, cytoplasmically localized IAA oxidase. Stable isotope-labeled IAA feeding studies of loss and …

Transcriptional Regulation Of Specialized Metabolites In Arabidopsis Thaliana And Catharanthus Roseus, Craig M. Schluttenhofer

Theses and Dissertations--Plant and Soil Sciences

For millennia humans have utilized plant specialized metabolites for health benefits, fragrances, poisons, spices, and medicine. Valued metabolites are often produced in small quantities and may command high prices. Understanding when and how the plant synthesizes these compounds is important for improving their production. Phytohormone signaling cascades, such as jasmonate (JA) activate or repress transcription factors (TF) controlling expression of metabolite biosynthetic genes. TFs regulating specialized metabolite biosynthetic genes can be manipulated to engineer plants with increased metabolite production.

WRKY transcription factor are known components of both JA signaling cascades and regulation of specialized metabolism. The presence of WRKY binding …

Mechanosensitive Channel Msl8 Regulates Osmotic Forces During Pollen Hydration And Germination, Eric S. Hamilton, Gregory S. Jensen, Grigory Maksaev, Andrew Katims, Ashley M. Sherp, Elizabeth S. Haswell

Biology Faculty Publications & Presentations

Pollen grains undergo dramatic changes in cellular water potential as they deliver the male germ line to female gametes, and it has been proposed that mechanosensitive ion channels may sense the resulting mechanical stress. Here, we identify and characterize MscS-like 8 (MSL8), a pollen-specific, membrane tension–gated ion channel required for pollen to survive the hypoosmotic shock of rehydration and for full male fertility. MSL8 negatively regulates pollen germination but is required for cellular integrity during germination and tube growth. MSL8 thus senses and responds to changes in membrane tension associated with pollen hydration and germination. These data further suggest that …

Detecting Genomic Regions Responsible For Resistance In Arabidopsis, Valeria Cancino, Anjali Iyer-Pascuzzi, Rucha Karve

The Summer Undergraduate Research Fellowship (SURF) Symposium

Ralstonia solanacearum is a soil-borne plant root colonizing pathogen and the casual agent of bacterial wilt (BW) disease. BW leads to severe yield loss in a wide variety of agricultural commodity crops, such as tomato, banana, and pepper. In this study, we look at the plant-pathogen interaction between Ralstonia solanacearum and various ecotypes of Arabidopsis thaliana with the goal of finding resistant ecotypes. To identify resistant ecotypes, seeds are first sterilized and left to soak in the dark. Then the seeds are plated on agar media, transferred to a growth chamber, and allowed to grow for 5 days. On day …

Calmodulin-Like Protein 38: A Component Of Ribonucleoprotein Particles During Hypoxic Stress Responses In Arabidopsis, Ansul Lokdarshi

Doctoral Dissertations

Waterlogging stress leads to a crisis in energy metabolism and the accumulation of toxic metabolites due to the hypoxic and/or anoxic environment associated with this condition. To respond and adapt to this situation, higher plants employ an integrated genetic program that leads to the induction of anaerobic response polypeptide genes that encode metabolic and signaling proteins involved in altering metabolic flow and other adaptive responses. The study presented here shows that the Arabidopsis thaliana calmodulin-like protein CML38 is calcium sensor protein that serves as a member of the core anaerobic response gene family and is involved in modulating the survival …

Identification Of Putative Plant Defense Genes Using A Novel Hydroponic Co-Cultivation Technique For Studying Plant-Pathogen Interaction, Naeem Nathoo

Electronic Thesis and Dissertation Repository

Previous work on identifying the molecular mechanisms mediating plant-pathogen interactions and reciprocal host responses have little emphasis on developing models that closely resemble host-microbe interaction in planta. This work establishes an amalgamated model of interaction wherein successful pathogens elicit and overcome host defenses activated by microbial signatures and virulence factors. Using a hydroponic co-cultivation model, we assessed the responses of Arabidopsis thaliana Col-0 to Agrobacterium tumefaciens C58 to ameliorate limitations of previous approaches. Comparisons of differential gene expression between directly and indirectly affected host sites by microarray analysis revealed both reactive and pro-active defense responses, respectively. Selected homozygous single-gene …

Arabidopsis Chromatin Remodeler Brahma: Its Functional Interplay With Polycomb Proteins And The Ref6 Histone Demethylase, Chenlong Li

Electronic Thesis and Dissertation Repository

BRAHMA (BRM) is a SWI/SNF-type chromatin remodeling ATPase that plays an important role in regulation of gene expression. Tri-methylation of lysine 27 on histone H3 (H3K27me3) is a histone modification that is associated with transcriptionally repressed genes and catalyzed by Polycomb Group (PcG) proteins. BRM has been proposed to antagonize the function of PcG proteins but the underlying molecular mechanism is unclear. To understand how BRM regulates the function of PcG proteins during plant development, a genome-wide analysis of H3K27me3 in brm mutant was performed using chromatin immunoprecipitation followed by next generation sequencing (ChIP-seq). Loss of BRM leads to increased …

Overexpression Of Patatin-Related Phospholipase Aiiiβ Altered The Content And Composition Of Sphingolipids In Arabidopsis, Maoyin Li, Jennifer E. Markham, Xuemin Wang

Department of Biochemistry: Faculty Publications

In plants, fatty acids are primarily synthesized in plastids and then transported to the endoplasmic reticulum (ER) for synthesis of most of the complex membrane lipids, including glycerolipids and sphingolipids. The first step of sphingolipid synthesis, which uses a fatty acid and a serine as substrates, is critical for sphingolipid homeostasis; its disruption leads to an altered plant growth. Phospholipase As have been implicated in the trafficking of fatty acids from plastids to the ER. Previously, we found that overexpression of a patatin-related phospholipase, pPLAIIIβ, resulted in a smaller plant size and altered anisotropic cell expansion. Here, we determined the …

Regulation Of Mor By Different Abiotic Stresses In Arabidopsis Thaliana, Luke Stepan, Rucha Karve, Anjali Iyer-Pascuzzi

The Summer Undergraduate Research Fellowship (SURF) Symposium

The climate is changing and as a consequence the environment is becoming hotter and drier. How different plants will react to these changes is unknown. Identification of genes involved in stress tolerance can help predict plant-environment interactions and lead to stress tolerant plants. The MOR gene (Modulator Of Root ROS, ROS = Reactive Oxygen Species) in the model plant Arabidopsis thaliana encodes a transcription factor that may regulate stress responses, as mor mutants are drought tolerant. We hypothesized that MOR expression changes in response to different abiotic stress stimuli. We tested MOR expression in response to salt (NaCl), abscisic acid …

Subset Of Heat-Shock Transcription Factors Required For The Early Response Of Arabidopsis To Excess Light, Hou-Sung Jung, Peter A. Crisp, Gonzalo M. Estavillo, Benjamin Cole

Dartmouth Scholarship

Sunlight provides energy for photosynthesis and is essential for nearly all life on earth. However, too much or too little light or rapidly fluctuating light conditions cause stress to plants. Rapid changes in the amount of light are perceived as a change in the reduced/oxidized (redox) state of photosynthetic electron transport components in chloroplasts. However, how this generates a signal that is relayed to changes in nuclear gene expression is not well understood. We modified redox state in the reference plant, Arabidopsis thaliana, using either excess light or low light plus the herbicide DBMIB (2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone), a well-known inhibitor of photosynthetic …

Identification Of Cyclophilin Gene Family In Soybean And Characterization Of Gmcyp1, Hemanta Raj Mainali

Electronic Thesis and Dissertation Repository

I identified members of the Cyclophilin (CYP) gene family in soybean (Glycine max) and characterized the GmCYP1, one of the members of soybean CYP. CYPs belong to the immunophilin superfamily with peptidyl-prolyl cis-trans isomerase (PPIase) activity. PPIase catalyzes the interconversion of the cis- and trans-rotamers of the peptidyl-prolyl amide bond of peptides. After extensive data mining, I identified 62 different CYP genes in soybean (GmCYP1 to GmCYP62), of which 8 are multi-domain proteins and 54 are single domain proteins. At least 25% of the GmCYP genes are expressed in soybean. GmCYP1 …

Functional Annotation, Transcriptional Characterization And Enzymatic Contributions Of Essential Amino Acid Biosynthesis-Related Genes, Teresa J. Clark

Masters Theses

In plants, essential amino acid biosynthesis predominantly or exclusively occurs in the plastid. The plastid in the heterokont alga Nannochloropsis oceanica is surrounded by four membranes, which add great complexity to intracellular trafficking and communication. N. oceanica genes in essential amino acid biosynthesis were functionally annotated. The biosynthesis pathways resemble the pathways in Arabidopsis thaliana, but the gene content seems to be simpler in N. oceanica.

In addition, two A. thaliana mutants with loss-of-function mutations in the aspartate kinase-homoserine dehydrogenase 2 (AK-HSDH2) gene were characterized. These ak-hsdh2 mutants demonstrate unexpected accumulation of aspartate-derived amino acids (ADAAs), particularly threonine, in …

Effects Of Ethylene On Secondary Xylem Formation In Arabidopsis Thaliana, Haley M. Rupp

Dissertations, Master's Theses and Master's Reports - Open

Ethylene has myriad roles as a plant hormone, ranging from senescence and defending against pathogen attacks to fruit ripening and interactions with other hormones. It has been shown to increase cambial activity in poplar, but the effect on wood formation in Arabidopsis hypocotyl has not previously been studied. The Auxin-Regulated Gene involved in Organ Size (ARGOS), which increases organ size by lengthening the time for cell division, was found to be upregulated by ethylene. We tested the effect of ethylene treatment at 10 and 100 µM ACC on three genotypes of Arabidopsis, Col0 (wild-type), an ARGOS deficient mutant (argos), and …

The Clubroot Pathogen (Plasmodiophora Brassicae) Influences Auxin Signaling To Regulate Auxin Homeostasis In Arabidopsis, Linda Jahn, Stefanie Mucha, Sabine Bergmann, Cornelia Horn, Paul E. Staswick, Bianka Steffens, Johannes Siemens, Jutta Ludwig-Müller

Department of Agronomy and Horticulture: Faculty Publications

The clubroot disease, caused by the obligate biotrophic protist Plasmodiophora brassicae, affects cruciferous crops worldwide. It is characterized by root swellings as symptoms, which are dependent on the alteration of auxin and cytokinin metabolism. Here, we describe that two different classes of auxin receptors, the TIR family and the auxin binding protein 1 (ABP1) in Arabidopsis thaliana are transcriptionally upregulated upon gall formation. Mutations in the TIR family resulted in more susceptible reactions to the root pathogen. As target genes for the different pathways we have investigated the transcriptional regulation of selected transcriptional repressors (Aux/IAA) and transcription …

The Effects Of Cytokinin On The Transcriptional Regulation Of Pin Expression In Arabidopsis Thaliana, Elizabeth Burgess

Honors Theses and Capstones

The processes of cell division and differentiation are critical to the development of any multicellular organism. During the formation of plant roots these processes take place at a region of the root tip called the meristem. Cytokinin and auxin are two plant growth hormones that influence this process. Although these two growth hormones are both necessary they also appear in many ways to have an antagonistic relationship. As meristematic root cells undergo differentiation they cease dividing. It has been proposed that the size of the root meristem and thus the overall rate of root growth are determined by the balance …

Host Pathogen Interactions: Is Arabidopsis Thaliana Remembered By Its Nemesis Pseudomonas Syringae?, Daniel Z. Kreiser

Lawrence University Honors Projects

Plants contain innate immune systems that deter pathogen infection. Pattern recognition receptors bind microbe-associated molecular patterns (MAMPs), triggering immunity. MAMPs are proteins exclusive to pathogens that are typically indispensable for their survival. For this reason, MAMPs cannot be mutated or removed without causing pathogen death. However, this does not necessitate constitutive expression of MAMPs. In this study, the MAMP response of Arabidopsis thaliana was utilized to determine differential detection of MAMPs expressed by Pseudomonas syringe pv. tomato DC3000 when pretreated with A. thaliana. Results demonstrated that more MAMPs are detected when P. syringae had previously encountered A. thaliana, …

No Accession-Specific Effect Of Rhizosphere Soil Communities On The Growth And Competition Of Arabidopsis Thaliana Accessions, Anna G. Aguilera, Adán Colón-Carmona, Rick Kesseli, Jeffrey S. Dukes

Biology Faculty Publication Series

Soil communities associated with specific plant species affect individual plants' growth and competitive ability. Limited evidence suggests that unique soil communities can also differentially influence growth and competition at the ecotype level. Previous work with Arabidopsis thaliana has shown that accessions produce distinct and reproducible rhizosphere bacterial communities, with significant differences in both species composition and relative abundance. We tested the hypothesis that soil communities uniquely affect the growth and reproduction of the plant accessions with which they are associated. Specifically, we examined the growth of four accessions when exposed to their own soil communities and the communities generated by …