Biotechnology Commons^™

Agrobacterium-Mediated Transformation Of Hemp (Cannabis Sativa L.)., Carthel Tyler Newton

Masters Theses

Agrobacterium­-mediated transformation has long been utilized as a method of producing stable, transgenic plants. This project describes an attempt to develop a system of producing stably transformed hemp (C. sativa (L.)) plants. Because of its recalcitrance to in vitro tissue culture and regeneration, transformation studies of hemp have been severely limited. In this study, I combined media components that have been shown to improve tissue culture health and regeneration. Additionally, I used hemp cultivars and explant sources that have been reported to function well in in vitro systems. The results showed that the improved media components did not …

A Robust Agroinfiltration Method, Bryce N. Trull

Masters Theses

Stable transformation of soybean (Glycine max) is a markedly slow and laborious process. Thus, a tool that enables rapid evaluation of genetic elements in planta is critical to advance complex research and genetic engineering in soybean. To that end, a substantially robust agroinfiltration method was innovated in this work. Agroinfiltration is a technique that leverages Agrobacterium-mediated transient transformation to deliver genetic elements to the cells of whole plant tissues, usually leaves. Several factors were found to be relevant to successful soybean leaf agroinfiltration, including genotype, surfactant, developmental stage, and Agrobacterium culture medium. The research represents not only …

Sequencing Antisense Transcriptome Of Populus Through Antisense/Sense Transcript Pair Enrichment, Emma V. Burke

Dissertations, Master's Theses and Master's Reports

Past transcriptome research on plants focused primarily on protein coding genes, and only recently researchers began looking into the non-protein coding regions that may play significant roles in gene regulation. Antisense RNA transcripts that are found naturally in the cell share complementary sequence with sense transcripts and have been shown to regulate expression of their sense counterparts. Since antisense RNA has been largely under-studied and difficult to sequence because of their low relative abundance, new methods are needed to target antisense RNA for efficient genome-wide profiling. To address this gap in methods to efficiently and cost effectively enrich antisense RNA …

Characterization Of Algerian Apricots (Prunus Armeniaca) Using Morphological And Pomological Markers, Kaouther Boutiti, Ines Bellil, Douadi Khelifi

Karbala International Journal of Modern Science

The aim of this study was to evaluate the diversity of an Algerian apricot germplasm. This Algerian apricot was characterized by a green-yellow skin, a red ground color, and a light orange flesh color in general. Besides, highly positive and negative significant correlations were revealed between the studied characters. Whereby, the principal component analysis explained 81% of the variability. Fruit, stone and leaves dimensions were the main features that explained evidentially the majority of variability. Moreover, the cluster analysis divided the accessions into two major groups. Thus, Algerian accessions selected in this study may have the potential to be used …

Characterizing Drought Adaptations, Phenotypic Plasticity, And Fixed Gene Expression Patterns Within Quercus, James K. Rauschendorfer

Dissertations, Master's Theses and Master's Reports

This dissertation was written on topics related to the genus Quercus with a primary focus on Quercus ellipsoidalis (northern pin oak) and Quercus rubra (northern red oak). Within this dissertation are chapters related to the setup of experimental common gardens within the Ford and Kellogg experimental forest, a literature review describing drought adaptations of Quercus sect. Lobatae (red oak group), identification of transcription factors within the Q. robur (English oak) and Q. rubra genomes, a study comparing leaf trait phenotypic plasticity of Q. ellipsoidalis and Q. rubra, and an RNA-seq experiment studying ecological speciation between Q. ellipsoidalis and Q. …

First Detection Of Naturally Introgressed Bt Cry1ab In Asian Corn Borer [Ostrinia Furnacalis (Guenée)] - Resistant Traditional Maize (Silangan) In The Philippines, Merdelyn T. Caasi-Lit, Eureka Teresa C. Ocampo, Artemio Salazar, Jefferson Paril, Bryan Novio, Bernard Panabang, Angelyn Marta Marmeto

The Philippine Agricultural Scientist

Resistance against insect pests is one of the most promising and highly favored characteristics of an economically important crop. In the Philippines, the Asian corn borer (ACB), Ostrinia furnacalis (Guenée), is the most destructive insect pest of maize. The discovery of Bacillus thuringiensis and the Bt toxin, which led to the development of Bt corn, effectively controlled this pest. In this study, four traditional varieties of maize that showed resistance against ACB were tested for the presence of Bt Cry1Ab protein. Out of these four, a traditional variety from San Carlos City, Pangasinan, which was initially thought to have natural …

Epinasty Accompanies Potassium Nitrate-Induced Flower Bud Break In 'Carabao' Mango [Research Note], Ulysses Besas, Calixto Protacio

The Philippine Agricultural Scientist

The hypothesis that potassium nitrate application promotes epinasty in ‘Carabao’ mango through increased ethylene production was investigated. The degree of drooping, apical primordium growth as affected by shoot ethylene and levels after potassium nitrate spraying were monitored. Evidence presented established for the first time in ‘Carabao’ mango that epinasty occurred just before flower emergence, coinciding with visible bud break, and extended up to 18 days, marking floral anthesis. The possible reasons could be the higher levels of ethylene at 0.31% and earlier increase of ethylene in the shoot apex at four hours and apical primordium growth. Increased ethylene levels at …

Spectral Lights Based Treatment Enhanced Biomass Accumulation And Secondary Metabolites Production In Callus Culture Of Citrus Reticulata, Faiza Anum, Naveed Iqbal Raja, Tahira Sultana, Abeer Kazmi, Amir Ali, Bushra Qayyum, Anza Afzal, Aneela Nijibat, Zia-Ur-Rehman Mashwani, Abeer Kazmi

The Philippine Agricultural Scientist

Citrus reticulata is a common fruit, belongs to the family Rutaceae. It contains terpenes, phenolics, flavonoids, and vitamin C in conjugation with sugars, amino acids, minerals, organic acids and other biologically important compounds. Application of light elicitation strategies in in vitro plant cultures is acquiring more interest for their potential in feasible biomass accumulation and therapeutically important secondary metabolites production. The present study was designed to investigate the effects of different monochromatic lights (white, red, blue, green, and yellow) on the proliferation and secondary metabolites accumulation in callus cultures of Citrus reticulata raised in response to treatment with AgNPs and …

Marker-Assisted Selection To Determine The Introgression Of Rpv-3 Mediated Downy Mildew Resistance In 'Chambourcin' X 'Caberenet Sauvignon' Grapevine Population, Kavya Sri Tummala

MSU Graduate Theses

Downy mildew caused by the fungus, Plasmopara viticola in the grapes is one of the major diseases affecting crop yield throughout the world. Breeding grapes via conventional methods for the downy mildew resistance will take approximately 25-30 years to fully evaluate and release new cultivars. Because of the long breeding cycle in woody plant species like grape, developing DNA markers linked to genes or quantitative trait loci (QTLs) for horticultural traits can greatly accelerate the breeding process and allow a much more accurate selection of progeny. This study aims to determine the presence of resistance in the 267 F1 genotypes …

Drought-Stress-Inducible Synthetic Promoters In Rice (Oryza Sativa), Paolo Montano Tagaloguin

Masters Theses

Drought can potentially affect the global rice supply. Recent climate modeling studies projected a more frequent and intense drought scenario, especially in top producing and exporting countries worldwide. Several drought-tolerant genes had been identified in recent years, and transgenic approaches had been employed to develop drought-tolerant rice. However, the constitutive expression of some drought-tolerant genes resulted in undesirable phenotypes and metabolic burden. The use of an inducible synthetic promoter is advantageous in preventing the possible pleiotropic effects from constitutively expressing a drought-tolerant gene. In this study, rationally designed synthetic promoters based on poplar (Populus trichocarpa) cis-motifs were …

Resistance To Aflatoxin Accumulation In Maize Mediated By Host-Induced Gene Silencing Of Aspergillus Flavus Alkaline Protease And O-Methyltransferase-A Genes, Olanike Omotola Omolehin

LSU Doctoral Dissertations

Aspergillus flavus is a soil-borne fungal pathogen that infects maize and produces aflatoxins. In the current study, portions of the alkaline protease (alk) and the O-methyl transferase (omtA) genes, which are key in A. flavus virulence and aflatoxin biosynthesis, respectively, were targeted for suppression through an RNAi (RNA interference) approach known as Host-Induced Gene Silencing (HIGS). Separate RNAi vectors were designed to carry regions of the alk and omtA gene fragments (Alk-RNAi and OmtA-RNAi) and introduced into B104 maize zygotic embryos. Eight and six transformation events were positive for the alk and omtA transgene, respectively. …

An Analysis Of Crispr-Cas Gene Editing In Agriculture, Ashley Laliberte

Honors Scholar Theses

The CRISPR-Cas system is a promising form of gene editing, especially for the agriculture industry. The ability to make single-nucleotide edits within a gene of interest, without the need to introduce foreign DNA, is a powerful tool for designing healthier and more efficient crops and food animals. This system provides opportunity for increased nutritional value, decreased food waste, and more economically and environmentally sustainable food production. Though this biotechnology is facing mechanistic limitations due to off-target effects and inefficient homology-directed repair, vast improvements have already been made to improve its efficacy. The CRISPR-Cas system is already the most advanced form …

Functional Characterization Of Petiolule-Like Pulvinus (Plp) Gene In Abscission Zone Development In Medicago Truncatula And Its Application To Genetic Improvement Of Alfalfa, Juan Du, Shaoyun Lu, Maofeng Chai, Chuanen Zhou, Liang Sun, Yuhong Tang, Jin Nakashima, Jaydeep Kolape, Zhaozhu Wen, Marjan Behzadirad, Tianxiu Zhong, Juan Sun, Yunwei Zhang, Zeng-Yu Wang

Nebraska Center for Biotechnology: Faculty and Staff Publications

Alfalfa (Medicago sativa L.) is one of the most important forage crops throughout the world. Maximizing leaf retention during the haymaking process is critical for achieving superior hay quality and maintaining biomass yield. Leaf abscission process affects leaf retention. Previous studies have largely focused on the molecular mechanisms of floral organ, pedicel and seed abscission but scarcely touched on leaf and petiole abscission. This study focuses on leaf and petiole abscission in the model legume Medicago truncatula and its closely related commercial species alfalfa. By analysing the petiolule-like pulvinus (plp) mutant in M. truncatula at phenotypic level (breakstrength and …

The Balancing Act Of Cytokinin In Environmental Stress Tolerance, Sumudu Sandeepani Karunadasa

Theses and Dissertations--Plant and Soil Sciences

Cytokinin, long known as a phytohormone that regulates plant growth and development, has been recently recognized as an important regulator of stress responses. However, our current knowledge about the mechanisms by which cytokinin regulates stress responses is fragmentary, as many of the studies in this field yielded conflicting results. Most of the work described here has focused on analyses of the molecular mechanisms of cytokinin-dependent regulation of growth and development under stress conditions, with an emphasis on the role of cytokinin-dependent regulation of protein synthesis in development and stress tolerance.

One of the important contributions of this study is the …

Field Evaluation Of Tobacco Engineered For High Leaf-Oil Accumulation, James Perry

Theses and Dissertations--Plant and Soil Sciences

The biofuel market is dominated by ethanol and biodiesel derived from cellulosic and lipid-based biomass crops. This is largely due to the relatively low costs and reliability of production. At present, production of non-food plant-derived oils for biofuel production in the U.S. is minimal. A research team from the Commonwealth Scientific and Industrial Research Organization (CSIRO), an independent Australian federal government research institution, has developed an efficient transgenic system to engineer oil production in tobacco leaves. This novel system is comprised of multiple transgenes that direct the endogenous metabolic flux of oil precursors towards triacylglycerol (TAG) production. Additional genes were …

Application Of Short Tandem Target Mimic (Sttm) Technique For Functional Analysis Of Micro-Rna396 In Transgenic Poplar Trees, Surattana Boonsai

Dissertations, Master's Theses and Master's Reports

Short Tandem Target Mimic (STTM) is a commonly technique used for functional studies of a number of genes in several plant model systems. However, very little is known about application of STTM technique in tree species. In this study, STTM was applied to knock down microRNA396 (miR396) in transgenic poplar trees for the first time. STTM396 expression resulted in dramatic decrease in miR396 expression levels leading to taller plants with larger leaves and larger leaf cell size. Additionally, an expression analysis of growth regulating factor genes (GRFs) that are members of miR396 target gene family showed up-regulation of GRF07 gene …

The Use Of Potato (Solanum Tuberosum) Suspension Cells For Rapid Screening Of Chloroplast Transformation Vectors, Lindsey Shover

EURēCA: Exhibition of Undergraduate Research and Creative Achievement

Chloroplast genetic engineering is unique in that it allows for high levels of gene expression while providing a means of natural bio-confinement. Although the chloroplast genome sequence of over 800 plant species is known, the elements that naturally regulate chloroplast gene expression are poorly understood. Recently, our lab has developed a 264 part modular cloning kit that contains known chloroplast regulatory elements. This kit can be used for the construction of novel chloroplast transformation cassettes; however, functional testing of these cassettes is currently hindered by standard chloroplast transformation technologies. Therefore, the aim of this experiment was to develop a rapid …

Towards The Development Of A Sustainable Soya Bean-Based Feedstock For Aquaculture, Hyunwoo Park, Steven Weier, Fareha Razvi, Pamela A. Peña, Neil A. Sims, Jennica Lowell, Cory Hungate, Karma Kissinger, Gavin Key, Paul Fraser, Jonathan Napier, Edgar B. Cahoon, Thomas Clemente

Center for Plant Science Innovation: Faculty and Staff Publications

Soya bean (Glycine max (L.) Merr.) is sought after for both its oil and protein components. Genetic approaches to add value to either component are ongoing efforts in soya bean breeding and molecular biology programmes. The former is the primary vegetable oil consumed in the world. Hence, its primary usage is in direct human consumption. As a means to increase its utility in feed applications, thereby expanding the market of soya bean coproducts, we investigated the simultaneous displacement of marine ingredients in aquafeeds with soya bean-based protein and a high Omega-3 fatty acid soya bean oil, enriched with alpha-linolenic …

Neutral And Adaptive Genetic Variation In North American Hardwood Tree Species, Sudhir Khodwekar

Dissertations, Master's Theses and Master's Reports

'Northern Hardwoods,’ are a characteristic composition of multiple tree species and a part of the rich and diverse northeastern forests. Hardwoods e.g. oaks (Quercus: Fagaceae and Sugar maple), serve as foundation species and offer ecological and economical gains to wildlife and humans. Most of the forest trees go through many biotic and abiotic stresses, for example climate change in their long life span. To overcome these threats and to adapt to changing conditions, tree species need to maintain variation especially in adaptive genes. Therefore, it is necessary to analyze genetic diversity within forest tree populations and take appropriate …

Nanocomposite Adhesive Of English Ivy (Hedera Helix): Bioproduction, Nanoparticle Isolation, And Molecular Analysis, Jason Neil Burris

Doctoral Dissertations

English ivy naturally produces organic nanoparticles from its adventitious root hairs, and possesses characteristics that may allow them to replace metal-based nanoparticles in common applications, such as sunscreen. At the onset of the research, it was hypothesized that a physical mechanism of attachment, similar to the gecko footpad, was used to generate the adhesive force for attachment; however, through the results obtained from recent work, it is clear that a biochemical mechanism is involved in the generation of the strength of adhesion. Therefore, the goal of this research was to provide a better understanding of the genetic basis of nanoparticle …

Bio-Piracy: Creating Proprietary Rights In Plant Genetic Resources, James O. Odek

Journal of Intellectual Property Law

No abstract provided.

Efficiency Of The Nuclease I-Scei In Excising Selectable Marker Genes From The Plant Genome, Elliot E. Pruett, Soumen Nandy, Vibha Srivastava

Discovery, The Student Journal of Dale Bumpers College of Agricultural, Food and Life Sciences

Gene stacking is a method used in biotechnology by which multiple genes can be placed at a single genomic site, thereby simplifying plant breeding. In this approach, DNA nucleases are used for excising selectable marker genes (SMG), which are the unneeded components of transgenic plants. The goal of this project is to evaluate the effectiveness of the nuclease I-SceI in excising DNA in plants. Specifically, this study tests heat-inducible I-SceI through the use of a heat-shock promoter (HS) in order to control SMG excision by heat application. The DNA plasmid containing a visual marker gene flanked by I-SceI target sites …

Anaerobic Detoxification Of Acetic Acid In A Thermophilic Ethanologen, A Joe Shaw, Bethany B. Miller, Stephen R. Rogers, William Robert Kenealy, Alex Meola, Ashwini Bhandiwad, W Ryan Sillers, Indraneel Shikhare, David Hogsett, Christopher Herring

Dartmouth Scholarship

The liberation of acetate from hemicellulose negatively impacts fermentations of cellulosic biomass, limiting the concentrations of substrate that can be effectively processed. Solvent-producing bacteria have the capacity to convert acetate to the less toxic product acetone, but to the best of our knowledge, this trait has not been transferred to an organism that produces ethanol at high yield. We have engineered a five-step metabolic pathway to convert acetic acid to acetone in the thermophilic anaerobe Thermoanaerobacterium saccharolyticum.

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The Exometabolome Of Clostridium Thermocellum Reveals Overflow Metabolism At High Cellulose Loading, Evert K. Holwerda, Philip G. Thorne, Daniel G. Olson, Daniel Amador-Noguez, Nancy L. Engle, Timothy J. Tschaplinski, Johannes P. Van Dijken, Lee R. Lynd

Dartmouth Scholarship

BackgroundClostridium thermocellum is a model thermophilic organism for the production of biofuels from lignocellulosic substrates. The majority of publications studying the physiology of this organism use substrate concentrations of ≤10 g/L. However, industrially relevant concentrations of substrate start at 100 g/L carbohydrate, which corresponds to approximately 150 g/L solids. To gain insight into the physiology of fermentation of high substrate concentrations, we studied the growth on, and utilization of high concentrations of crystalline cellulose varying from 50 to 100 g/L by C. thermocellum. .

Tracking The Cellulolytic Activity Of Clostridium Thermocellum Biofilms, Alexandru Dumitrache, Gideon M. Wolfaardt, David Allen, Steven N. Liss, Lee R. Lynd

Dartmouth Scholarship

Microbial cellulose conversion by Clostridium thermocellum 27405 occurs predominantly through the activity of substrate-adherent bacteria organized in thin, primarily single cell-layered biofilms. The importance of cellulosic surface exposure to microbial hydrolysis has received little attention despite its implied impact on conversion kinetics.

Progress And Future Prospects Of Developing Transgenics In Mango, Rajesh Pati

Rajesh Pati

Mango (Mangifera indica L.) is the most important fruit crop of the world. India ranks first among world’s mango producing countries accounting for 57.18 % of the total world mango production of 19.22 million tones. With the development of rDNA technology, plant breeders have access to a large number of genes that can be integrated into the plant genome.

Bioconfinement Of A Putatively Sterile Nicotiana Hybrid And Development Of Tools For Assessing Gene Flow, John Hollis Rice

Masters Theses

Production of transgenic crops in open field environments is an ongoing concern of due to the potential for gene flow. New transgenic crops, such as plant-made-pharmaceuticals may generate additional concerns about effects of adventitious transgenes. Use of a bioconfinement strategy may alleviate any consequences by preventing gene flow. The following chapters discuss previous and current research on gene flow, testing of a Nicotiana hybrid system for bioconfinement efficiency, and development of methods for transgene detection. The candidate ‘platform plant’ that was tested is a Nicotiana hybrid (Nicotiana tabacum ‘TN 90’ × Nicotiana glauca) previously identified to be sexually …

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 …

A Morphological Analysis Of A Hybrid Swarm Of Native Ulmus Rubra And Introduced U. Pumila (Ulmaceae) In Southeastern Nebraska, Christian Elowsky, Ingrid E. Jordon-Thaden, Robert B. Kaul

Nebraska Center for Biotechnology: Faculty and Staff Publications

The parental species and hybrid swarm of native Ulmus rubra Muhl. and the introduced, naturalized and weedy U. pumila L. were investigated in a 65-km transect in Lancaster, Saunders, and Butler counties in Nebraska. Thirty-two trees of U. rubra, 32 of U. pumila, and 50 of the hybrid swarm were sampled for leaves and buds and subsampled for flowers and fruits. Leaves were measured for petiole length, blade length, width, primary and secondary teeth per cm, number of secondary teeth per primary tooth, and texture. Buds were scored for color and distribution of trichomes. Flowers were sampled for stamen counts …

Epigenetics Of Stress Adaptation In Arabidopsis: The Case Of Histone Modifications, Behnaz Najafi Majdabadi Farahani

Electronic Thesis and Dissertation Repository

Changes in the epigenetic status of plants, which contribute to changes in gene expression, in response to abiotic stress are well documented. However, their stability and transmission to subsequent generations, and their incorporation into plant stress adaptation are still a matter for debate. Using chromatin immuno-precipitation and Next Generation Sequencing (ChIP-seq), we compared genome wide enrichment of two histone marks, H3K9ac and H3K4me2, in the progeny of salt stressed and control Arabidopsis plants. Data showed less enrichment of the H3K9ac in the chromatin of the progeny of salt stressed plants, but no changes were detected in the enrichment of the …