Digital Commons Network^™

Metabolomics-Based Analysis Of Miniature Flask Contents Identifies Tobacco Mixture Use Among The Ancient Maya, Mario Zimmermann, Korey J. Brownstein, Luis Pantoja Díaz, Iliana Ancona Aragón, Scott R. Hutson, Barry Kidder, Shannon Tushingham, David R. Gang

Anthropology Faculty Publications

A particular type of miniature ceramic vessel locally known as “veneneras” is occasionally found during archaeological excavations in the Maya Area. To date, only one study of a collection of such containers successfully identified organic residues through coupled chromatography–mass spectrometry methods. That study identified traces of nicotine likely associated with tobacco. Here we present a more complete picture by analyzing a suite of possible complementary ingredients in tobacco mixtures across a collection of 14 miniature vessels. The collection includes four different vessel forms and allows for the comparison of specimens which had previously formed part of museum exhibitions with recently …

Characteristics Of Cercospora Nicotianae With Reduced Sensitvity To Azoxystrobin, William Barrett Barlow

Theses and Dissertations--Plant Pathology

Burley and dark tobacco production are important to agriculture in Kentucky, worth $145.1 and $94.4 million in 2020, respectively. The price received for a tobacco crop is influenced by leaf quality, determined by multiple characteristics including leaf damage. Frogeye leaf spot (FLS), caused by Cercospora nicotianae, has historically been a minor disease of tobacco. However, when FLS infections reach the upper canopy of tobacco, lesions are seen as damage that negatively influence leaf quality. Fungicides are regularly used to manage foliar diseases of tobacco, and the only systemic fungicide active ingredient labeled for tobacco is azoxystrobin. Azoxystrobin belongs to the …

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 …

Alteration Of Tobacco Alkaloid Content Through Modification Of Specific Cytochrome P450 Genes, Ralph E. Dewey, Balazs Siminszky, Steven W. Bowen, Lily Gavilano

Plant and Soil Sciences Faculty Patents

Compositions and methods for reducing the level of nornicotine and N'-nitrosonomicotine (NNN) in Nicotiana plants and plant parts thereof are provided. The compositions comprise isolated polynucleotides and polypeptides for cytochrome P450s that are involved in the metabolic con- version of nicotine to nornicotine in these plants. Expression cassettes, vectors, plants, and plant parts thereof comprising inhibitory sequences that target expression or function of the disclosed cytochrome P450 polypeptides are also provided. Methods for the use of these novel sequences to inhibit expression or function of cytochrome P450 polypeptides involved in this metabolic conversion are also provided. The methods find use …

Inhibition Of Fusarium Oxysporum F. Sp. Nicotianae Growth By Phenylpropanoid Pathway Intermediates, Timothy E. Shull, Jasmina Kurepa, Robert D. Miller, Natalia Martinez-Ochoa, Jan A. Smalle

Plant and Soil Sciences Faculty Publications

Fusarium wilt in tobacco caused by the fungus Fusarium oxysporum f. sp. nicotianae is a disease‑management challenge worldwide, as there are few effective and environmentally benign chemical agents for its control. This challenge results in substantial losses in both the quality and yield of tobacco products. Based on an in vitro analysis of the effects of different phenylpropanoid intermediates, we found that the early intermediates trans‑cinnamic acid and para‑coumaric acid effectively inhibit the mycelial growth of F. oxysporum f. sp. nicotianae strain FW316F, whereas the downstream intermediates quercetin and caffeic acid exhibit no fungicidal properties. Therefore, our in …

Alteration Of Tobacco Alkaloid Content Through Modification Of Specific Cytochrome P450 Genes, Ralph E. Dewey, Balazs Siminszky, Steven W. Bowen, Lily Gavilano

Plant and Soil Sciences Faculty Patents

Compositions and methods for reducing the level of nornicotine and N'-nitrosonomicotine (NNN) in Nicotiana plants and plant parts thereof are provided. The compositions comprise isolated polynucleotides and polypeptides for cytochrome P450s that are involved in the metabolic conversion of nicotine to nornicotine in these plants. Expression cassettes, vectors, plants, and plant parts thereof comprising inhibitory sequences that target expression or function of the disclosed cytochrome P450 polypeptides are also provided. Methods for the use of these novel sequences to inhibit expression or function of cytochrome P450 polypeptides involved in this metabolic conversion are also provided. The methods find use in …

Oxidative Stress-Induced Formation Of Covalently Linked Ribulose-1,5-Bisphosphate Carboxylase/Oxygenase Large Subunit Dimer In Tobacco Plants, Jasmina Kurepa, Jan A. Smalle

Plant and Soil Sciences Faculty Publications

Objective: Many abiotic stresses cause the excessive accumulation of reactive oxygen species known as oxidative stress. While analyzing the effects of oxidative stress on tobacco, we noticed the increased accumulation of a specific protein in extracts from plants treated with the oxidative-stress inducing herbicide paraquat which promotes the generation of reactive oxygen species primarily in chloroplasts. The primary objectives of this study were to identify this protein and to determine if its accumulation is indeed a result of oxidative stress.

Results: Here we show that the paraquat-induced protein is a covalently linked dimer of the large subunit of ribulose-1,5-bisphosphate carboxylase …

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 …

Agronomic And Chemical Performance Of Field-Grown Tobacco Engineered For Triterpene And Methylated Triterpene Metabolism, Zuodong Jiang, Chase Kempinski, Santosh Kumar, Scott Kinison, Kristin Linscott, Eric Nybo, Sarah Janze, Constance Wood, Joseph Chappell

Pharmaceutical Sciences Faculty Publications

Squalene is a linear intermediate to nearly all classes of triterpenes and sterols and is itself highly valued for its use in wide range of industrial applications. Another unique linear triterpene is botryococcene and its methylated derivatives generated by the alga Botryococcus braunii race B, which are progenitors to fossil fuel deposits. Production of these linear triterpenes was previously engineered into transgenic tobacco by introducing the key steps of triterpene metabolism into the particular subcellular compartments. In this study, the agronomic characteristics (height, biomass accumulation, leaf area), the photosynthetic capacity (photosynthesis rate, conductance, internal CO₂ levels) and triterpene content …

Alteration Of Tobacco Alkaloid Content Through Modification Of Specific Cytochrome P450 Genes, Ralph E. Dewey, Balazs Siminszky, Steven W. Bowen, Lily Gavilano

Plant and Soil Sciences Faculty Patents

Compositions and methods for reducing the level of nornicotine and N′-nitrosonornicotine (NNN) in Nicotiana plants and plant parts thereof are provided. The compositions comprise isolated polynucleotides and polypeptides for cytochrome P450s that are involved in the metabolic conversion of nicotine to nornicotine in these plants. Expression cassettes, vectors, plants, and plant parts thereof comprising inhibitory sequences that target expression or function of the disclosed cytochrome P450 polypeptides are also provided. Methods for the use of these novel sequences to inhibit expression or function of cytochrome P450 polypeptides involved in this metabolic conversion are also provided. The methods find use in …

Chemical Topping Burley Tobacco, Mitchell D. Richmond

Theses and Dissertations--Plant and Soil Sciences

The act of topping tobacco (Nicotiana tabacum L.) involves the removal of the terminal bud or inflorescence of the tobacco plant. This practice ordinarily is accomplished by manually removing the top of each tobacco plant in an entire field which is labor intensive and costly. Chemical topping utilizes sucker control products to inhibit the terminal bud and axillary bud growth without manually removing the top of the tobacco plant. There were several research objectives in order to determine the utility of a chemical topping system: 1) determine if burley tobacco could be chemically topped with currently registered suckercide products …

Reducing Biomass Recalcitrance By Heterologous Expression Of A Bacterial Peroxidase In Tobacco (Nicotiana Benthamiana), Ayalew Ligaba-Osena, Bertrand Hankoua, Kay Dimarco, Robert Pace, Mark Crocker, Jesse Mcatee, Nivedita Nagachar, Ming Tien, Tom L. Richard

Center for Applied Energy Research Faculty and Staff Publications

Commercial scale production of biofuels from lignocellulosic feed stocks has been hampered by the resistance of plant cell walls to enzymatic conversion, primarily owing to lignin. This study investigated whether DypB, the lignin-degrading peroxidase from Rodococcus jostii, depolymerizes lignin and reduces recalcitrance in transgenic tobacco (Nicotiana benthamiana). The protein was targeted to the cytosol or the ER using ER-targeting and retention signal peptides. For each construct, five independent transgenic lines were characterized phenotypically and genotypically. Our findings reveal that expression of DypB in the cytosol and ER does not affect plant development. ER-targeting increased protein accumulation, and …

Co-Opting Atp-Generating Glycolytic Enzyme Pgk1 Phosphoglycerate Kinase Facilitates The Assembly Of Viral Replicase Complexes, K. Reddisiva Prasanth, Chingkai Chuang, Peter D. Nagy

Plant Pathology Faculty Publications

The intricate interactions between viruses and hosts include exploitation of host cells for viral replication by using many cellular resources, metabolites and energy. Tomato bushy stunt virus (TBSV), similar to other (+)RNA viruses, induces major changes in infected cells that lead to the formation of large replication compartments consisting of aggregated peroxisomal and ER membranes. Yet, it is not known how TBSV obtains the energy to fuel these energy-consuming processes. In the current work, the authors discovered that TBSV co-opts the glycolytic ATP-generating Pgk1 phosphoglycerate kinase to facilitate the assembly of new viral replicase complexes. The recruitment of Pgk1 into …

Genome Sequence Variation In The Constricta Strain Dramatically Alters The Protein Interaction And Localization Map Of Potato Yellow Dwarf Virus, Chanyong Jang, Renyuan Wang, Joseph Wells, Fabian Leon, Mark Farman, John Hammond, Michael M. Goodin

Plant Pathology Faculty Publications

The genome sequence of the constricta strain of Potato yellow dwarf virus (CYDV) was determined to be 12 792 nt long and organized into seven ORFs with the gene order 3′-N-X-P-Y-M-G-L-5′, which encodes the nucleocapsid, phospho, movement, matrix, glyco, and RNA-dependent RNA polymerase proteins, respectively, except for X, which is of unknown function. Cloned ORFs for each gene, except L, were used to construct a protein interaction and localization map (PILM) for this virus, which shares greater than 80 % amino acid similarity in all ORFs except X and P with the sanguinolenta strain of this species (SYDV). Protein localization …

The Effect Of Sazetidine-A And Other Nicotinic Ligands On Nicotine Controlled Goal-Tracking In Female And Male Rats, S. Charntikov, A. M. Falco, K. Fink, Linda P. Dwoskin, R. A. Bevins

Pharmaceutical Sciences Faculty Publications

Nicotine is the primary addictive component of tobacco products and its complex stimulus effects are readily discriminated by humans and non-human animals. Previous preclinical research investigating directly the nature of the nicotine stimulus has been limited to male rodents. The current study began to address this significant gap in the literature by training female and male rats to discriminate 0.4 mg/kg nicotine from saline in the discriminated goal-tracking task. In this task, access to sucrose was intermittently available on nicotine session. On saline session, intermixed with nicotine sessions on separate days, sucrose was not available. Both sexes acquired the discrimination …

Using Exogenous Hormone Application To Suppress Axillary Shoot Development In Tobacco, W. Jesse Carmack

Theses and Dissertations--Plant and Soil Sciences

The variability in the number of basal axillary shoots (ground suckers) among all tobacco (Nicotiana tabacum L.) varieties, has increased since the hydroponic transplant production system became the standard. The larger root ball of hydroponically produced transplants compared to traditionally produced transplants potentially generates a difference in the ratio of auxin (inhibits axillary shoot formation) to cytokinin (promotes lateral branching), that induces basal axillary shoot development. Starting in 2014, studies were conducted to investigate whether the addition of synthetic auxins or cytokinins to hydroponic transplant production could prevent ground sucker formation. Different tobacco cultivars, with high or low ground sucker …

Developmental Toxicity Of Nicotine: A Transdisciplinary Synthesis And Implications For Emerging Tobacco Products, Lucinda J. Enland, Kjersti Aagaard, Michele Bloch, Kevin Conway, Kelly Cosgrove, Rachel Grana, Thomas J. Gould, Dorothy Hatsukami, Frances Jensen, Denise Kandel, Bruce Lanphear, Frances Leslie, James R. Pauly, Jenae Neiderhiser, Mark Rubinstein, Theodore A. Slotkin, Eliot Spindel, Laura Stroud, Lauren Wakschlag

Pharmaceutical Sciences Faculty Publications

While the health risks associated with adult cigarette smoking have been well described, effects of nicotine exposure during periods of developmental vulnerability are often overlooked. Using MEDLINE and PubMed literature searches, books, reports and expert opinion, a transdisciplinary group of scientists reviewed human and animal research on the health effects of exposure to nicotine during pregnancy and adolescence. A synthesis of this research supports that nicotine contributes critically to adverse effects of gestational tobacco exposure, including reduced pulmonary function, auditory processing defects, impaired infant cardiorespiratory function, and may contribute to cognitive and behavioral deficits in later life. Nicotine exposure during …

Use Of Several Natural Products From Selected Nicotiana Species To Prevent Black Shank Disease In Tobacco, Antoaneta B. Kroumova, Ivan Artiouchine, George J. Wagner

Kentucky Tobacco Research and Development Center Faculty Publications

Black shank is a major annual disease threat to all types of tobacco worldwide. It is caused by the fungus Phytophthora parasitica var. nicotianae (PPN). The major tobacco growing areas in US - Kentucky, Tennessee and North Carolina can experience devastating losses, reaching in some fields up to 100%. Thus far, the main approaches to control this disease have been creation of resistant varieties, fungicide treatments, and crop rotation. Some fungicides are reported to have negative effects on the environment. The goal of this work was to test the antifungal activity of several natural products that are synthesized by certain …

Expression Of A Constitutively Active Nitrate Reductase Variant In Tobacco Reduces Tobacco-Specific Nitrosamine Accumulation In Cured Leaves And Cigarette Smoke, Jianli Lu, Leichen Zhang, Ramsey S. Lewis, Lucien Bovet, Simon Goepfert, Anne M. Jack, James D. Crutchfield, Huihua Ji, Ralph E. Dewey

Kentucky Tobacco Research and Development Center Faculty Publications

Burley tobaccos (Nicotiana tabacum) display a nitrogen-use-deficiency phenotype that is associated with the accumulation of high levels of nitrate within the leaf, a trait correlated with production of a class of compounds referred to as tobacco-specific nitrosamines (TSNAs). Two TSNA species, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) and N-nitrosonornicotine (NNN), have been shown to be strong carcinogens in numerous animal studies. We investigated the potential of molecular genetic strategies to lower nitrate levels in burley tobaccos by overexpressing genes encoding key enzymes of the nitrogen-assimilation pathway. Of the various constructs tested, only the expression of a constitutively active nitrate reductase (NR) …

Aphid Performance Changes With Plant Defense Mediated By Cucumber Mosaic Virus Titer, Xiaobin Shi, Yang Gao, Shuo Yan, Xin Tang, Xuguo Zhou, Deyong Zhang, Yong Liu

Entomology Faculty Publications

Background: Cucumber mosaic virus (CMV) causes appreciable losses in vegetables, ornamentals and agricultural crops. The green peach aphid, Myzus persicae Sulzer (Aphididae) is one of the most efficient vectors for CMV. The transmission ecology of aphid-vectored CMV has been well investigated. However, the detailed description of the dynamic change in the plant-CMV-aphid interaction associated with plant defense and virus epidemics is not well known.

Results: In this report, we investigated the relationship of virus titer with plant defense of salicylic acid (SA) and jasmonic acid (JA) during the different infection time and their interaction with aphids in CMV-infected tobacco plants. …

Field Evaluation Of Burley Lines Containing Alleles Minimizing Nicotine To Nornicotine Conversion, Cameron G. Shelton

Theses and Dissertations--Plant and Soil Sciences

CYP82e4, CYP82e5, and CYP82e10 mutant alleles that minimize the conversion of nicotine to nornicotine have been introgressed into numerous existing low converting (LC) burley varieties and parental lines developed by the Kentucky-Tennessee tobacco breeding program. A backcross breeding protocol was utilized, with the objective being the creation of "e3" varieties that differed from their LC counterparts only for nornicotine and nitroso-nornicotine content. Field studies were conducted in Kentucky and Tennessee during the 2013 growing season, with 17 prospective parental lines and 20 prospective commercial varieties grown and compared to their original counterparts. Most of the e3 lines were not morphologically …

Genetic Dissection Of Disease Resistance To The Blue Mold Pathogen, Peronospora Tabacina, In Tobacco, Xia Wu, Dandan Li, Yinguang Bao, David Zaitlin, Robert D. Miller, Shengming Yang

Plant and Soil Sciences Faculty Publications

Tobacco blue mold, caused by the obligately biotrophic oomycete pathogen Peronospora tabacina D.B. Adam, is a major foliar disease that results in significant losses in tobacco-growing areas. Natural resistance to P. tabacina has not been identified in any variety of common tobacco. Complete resistance, conferred by RBM1, was found in N. debneyi and was transferred into cultivated tobacco by crossing. In the present study, we characterized the RBM1-mediated resistance to blue mold in tobacco and show that the hypersensitive response (HR) plays an important role in the host defense reactions. Genetic mapping indicated that the disease resistance …

Alteration Of Tobacco Alkaloid Content Through Modification Of Specific Cytochrome P450 Genes, Ralph E. Dewey, Balazs Siminszky, Steven W. Bowen, Lily Gavilano

Plant and Soil Sciences Faculty Patents

Compositions and methods for reducing the level of nomicotine and N'-nitrosonornicotine (NNN) in Nicotiana plants and plant parts thereof are provided. The compositions comprise isolated polynucleotides and polypeptides for cytochrome P450s that are involved in the metabolic conversion of nicotine to nornicotine in these plants. Expression cassettes, vectors, plants, and plant parts thereof comprising inhibitory sequences that target expression or function of the disclosed cytochrome P450 polypeptides are also provided. Methods for the use of these novel sequences to inhibit expression or function of cytochrome P450 polypeptides involved in this metabolic conversion are also provided. The methods find use in …

In Vivo Localization Of Iris Yellow Spot Tospovirus (Bunyaviridae)-Encoded Proteins And Identification Of Interacting Regions Of Nucleocapsid And Movement Proteins, Diwaker Tripathi, Gaurav Raikhy, Michael M. Goodin, Ralf G. Dietzgen, Hanu R. Pappu

Plant Pathology Faculty Publications

BACKGROUND: Localization and interaction studies of viral proteins provide important information about their replication in their host plants. Tospoviruses (Family Bunyaviridae) are economically important viruses affecting numerous field and horticultural crops. Iris yellow spot virus (IYSV), one of the tospoviruses, has recently emerged as an important viral pathogen of Allium spp. in many parts of the world. We studied the in vivo localization and interaction patterns of the IYSV proteins in uninfected and infected Nicotiana benthamiana and identified the interacting partners.

PRINCIPAL FINDINGS: Bimolecular fluorescence complementation (BiFC) analysis demonstrated homotypic and heterotypic interactions between IYSV nucleocapsid …

Engineering Triterpene Metabolism In Tobacco, Zuodong Jiang

Theses and Dissertations--Plant and Soil Sciences

Terpenes comprise a large diverse class of natural products and many of them attract interest because of their physiological function, therapeutic and industrial values. Triterpene oils including squalene (C30), botrycococcene (C30) and their methylated derivatives (C31-C37) generated by the green algae Botryococcus braunii race B, which have recently received significant attention because of their utility for advanced biofuels. However, the slow growth habit of B. braunii makes it impractical as a robust biofuel production system. In this thesis, we firstly evaluated the potential of generating high levels of triterpene (C30) production in tobacco plants by diverting carbon flux from cytosolic …

Soil Management And Nitrogen Dynamics In Burley Tobacco Rotations, Congming Zou

Theses and Dissertations--Plant and Soil Sciences

Agronomic practices, including tillage, crop rotation and N fertilization, have been developed to efficiently manage soil N dynamics and crop N nutrition. These practices can affect soil organic carbon (SOC) and soil total nitrogen (STN) sequestration, and consequently influence soil nitrogen mineralization (SNM) and crop N nutrition. However, little research has been systematically and simultaneously conducted to examine the effect of agronomic management on (1) SOC and STN stocks; (2) SNM; and (3) crop N nutrition. Burley tobacco (Nicotiana tobacum L.) is a N demanding crop and subject to inefficiency in N fertilization. Moreover, conservation tillage and rotation have been …

Limitations And Benefits To Cultivating Tobacco, Edwin L. Ritchey, Gary K. Palmer

Plant and Soil Sciences Research Report

Historically, between-row cultivation for weed control has been an integral part of tobacco production. In one season, a grower might perform five or more cultivations and several hand weeding operations for adequate weed control. Early herbicide chemistries would control some weeds, but not all, forcing the producer to cultivate and hand weed. The development of better herbicides improved the range of weeds controlled, sometimes to the point that cultivation for weed control was not warranted. Many producers are so accustomed to cultivating they have forgotten why cultivation was necessary. The decision to cultivate should largely be based on the presence …

Construction Of A Sonchus Yellow Net Virus Minireplicon: A Step Toward Reverse Genetic Analysis Of Plant Negative-Strand Rna Viruses, Uma Ganesan, Jennifer N. Bragg, Min Deng, Sharon Marr, Mi Yeon Lee, Shasha Qian, Manling Shi, Justin Kappel, Cole Peters, Yeon Lee, Michael M. Goodin, Ralf G. Dietzgen, Zhenghe Li, Andrew O. Jackson

Plant Pathology Faculty Publications

Reverse genetic analyses of negative-strand RNA (NSR) viruses have provided enormous advances in our understanding of animal viruses over the past 20 years, but technical difficulties have hampered application to plant NSR viruses. To develop a reverse genetic approach for analysis of plant NSR viruses, we have engineered Sonchus yellow net nucleorhabdovirus (SYNV) minireplicon (MR) reporter cassettes for Agrobacterium tumefaciens expression in Nicotiana benthamiana leaves. Fluorescent reporter genes substituted for the SYNV N and P protein open reading frames (ORFs) exhibited intense single-cell foci throughout regions of infiltrated leaves expressing the SYNV MR derivatives and the SYNV nucleocapsid (N), phosphoprotein …

Alternative Methods Of Control For Phytophthora Nicotianae Of Tobacco, Anna M. Holdcroft

Theses and Dissertations--Plant Pathology

Kentucky is the nation’s leading producer of burley tobacco and the crop’s most economically important disease is black shank, caused by Phytophthora nicotianae (Pn). Current management is effective, however, problems with expense and pathogen persistence are issues. Two alternative methods for control of Pn were examined: biofumigation and soil application of an organic, yeast fermentation‐derived product (Soil‐Set). Field studies in 2009 and 2010 found no effect on populations of fungi, disease severity of Pn, and yield between mustard‐ and wheat‐amended plots. Experiments in the greenhouse suggested that survival of Pn was impacted by biomass rather than biofumigation. Biofumigation is not …

Development And Functional Analysis Of Novel Genetic Promoters Using Dna Shuffling, Hybridization And A Combination Thereof, Rajiv Ranjan, Sunita Patro, Bhubaneswar Pradhan, Alok Kumar, Indu B. Maiti, Nrisingha Dey

Kentucky Tobacco Research and Development Center Faculty Publications

BACKGROUND: Development of novel synthetic promoters with enhanced regulatory activity is of great value for a diverse range of plant biotechnology applications.

METHODOLOGY: Using the Figwort mosaic virus full-length transcript promoter (F) and the sub-genomic transcript promoter (FS) sequences, we generated two single shuffled promoter libraries (LssF and LssFS), two multiple shuffled promoter libraries (LmsFS-F and LmsF-FS), two hybrid promoters (FuasFScp and FSuasFcp) and two hybrid-shuffled promoter libraries (LhsFuasFScp and LhsFSuasFcp). Transient expression activities of approximately 50 shuffled promoter clones from each of these libraries were assayed in tobacco (Nicotiana tabacum cv. Xanthi) protoplasts. It was observed that most …