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Sample records for modified tobacco plants

  1. Transgenic Tobacco Plants With Efficient Insect Resistance

    Institute of Scientific and Technical Information of China (English)

    李太元; 田颖川; 秦晓峰; 莽克强; 李文谷; 何永刚; 沈蕾

    1994-01-01

    Insecticidal protein gene CryIA(c)from Bacillus thuringiensis HD-1(B.t.toxin gene)with 5’-end modified and 3’-end deleted to 4 different lengths were inserted downstream of 35S promoterwith double enhancer and"Ω’"fragment of TMV-RNA cDNA in the binary vector pBin438 to constructthe chimeric expression vector of B.t.toxin gene.Leave stripes of tobacco plant var.NC89 widelygrown in China were transformed with A.tumefaciens LBA4404 harbouring the above expression vectorsrespectively,and kanamycin resistant tobacco plants were regenerated.Insect test with tobacco budwormH.assulta showed that insect-resistant transform.ants could be obtained from the regenerated plantstransformed with B.t.genes of different lengths though highest percentage(~50%)of plants with ahigh morality(90%-100%)to the testing insects is among those transformed with 1.8-kb toxin gene.Genetic,molecular and biological analyses of T1 and T2 progenies of plants with high efficient insect re-sistance showed that B.t.toxin gene and the character of insect resistance have been inherited in the pro-genies.Insect-resistant homozygotes D8-14 and D19-8 have been selected for small-scale field tests.

  2. PHYTOREMEDIATION OF PERCHLORATE BY TOBACCO PLANTS

    Science.gov (United States)

    Previous studies have shown that tobacco plants are tolerant of perchlorate and will accumulate perchlorate in the plant tissues. The objective of this research was to determine the effectiveness of tobacco plants in phytoremediation, a technology that employs plants to degrade,...

  3. PHYTOREMEDIATION OF PERCHLORATE BY TOBACCO PLANTS

    Science.gov (United States)

    Previous studies have shown that tobacco plants are tolerant of perchlorate and will accumulate perchlorate in the plant tissues. The objective of this research was to determine the effectiveness of tobacco plants in phytoremediation, a technology that employs plants to degrade,...

  4. Improved phytoaccumulation of cadmium by genetically modified tobacco plants (Nicotiana tabacum L.). Physiological and biochemical response of the transformants to cadmium toxicity

    Energy Technology Data Exchange (ETDEWEB)

    Gorinova, N. [AgroBioInstitute, 8 Dragan Tzankov Blvd., 1164 Sofia (Bulgaria)]. E-mail: noraig60@yahoo.co.uk; Nedkovska, M. [AgroBioInstitute, 8 Dragan Tzankov Blvd., 1164 Sofia (Bulgaria); Todorovska, E. [AgroBioInstitute, 8 Dragan Tzankov Blvd., 1164 Sofia (Bulgaria); Simova-Stoilova, L. [Institute of Plant Physiology, Bulgarian Academy of Sciences, 1113 Sofia (Bulgaria); Stoyanova, Z. [Institute of Plant Physiology, Bulgarian Academy of Sciences, 1113 Sofia (Bulgaria); Georgieva, K. [Institute of Plant Physiology, Bulgarian Academy of Sciences, 1113 Sofia (Bulgaria); Demirevska-Kepova, K. [Institute of Plant Physiology, Bulgarian Academy of Sciences, 1113 Sofia (Bulgaria); Atanassov, A. [AgroBioInstitute, 8 Dragan Tzankov Blvd., 1164 Sofia (Bulgaria); Herzig, R. [Phytotech-Foundation PT-F, Quartiergasse 12, CH 3013 Bern (Switzerland)

    2007-01-15

    The response of tobacco plants (Nicotiana tabacum L.)-non-transformed and transformed with a metallothionein gene MThis from Silene vulgaris L. - to increase cadmium supply in the nutrient solution was compared. The transgenic plants accumulated significantly more Cd both in the roots and the leaves. Visual toxicity symptoms and disturbance in water balance were correlated with Cd tissue content. Treatment with 300 {mu}M CdCl{sub 2} resulted in inhibition of photosynthesis and mobilization of the ascorbate-glutathione cycle. Treatment with 500 {mu}M CdCl{sub 2} led to irreversible damage of photosynthesis and oxidative stress. An appearance of a new peroxidase isoform and changes in the leaf polypeptide pattern were observed at the highest Cd concentration. The level of non-protein thiols gradually increased following the Cd treatment both in transgenic and non-transformed plants. - Genetic transformation of Nicotiana tabacum L. by metallothionein gene improved phytoaccumulation of cadmium.

  5. Infection of Plants by Tobacco Mosaic Virus.

    Science.gov (United States)

    McDaniel, Larry; Maratos, Marina; Farabaugh, Joan

    1998-01-01

    Provides three exercises that introduce high school and college students to a common strain of the tobacco mosaic virus and the study of some basic biological processes. Activities involve inoculation of plants and observing and recording symptom development in infected plants. (DDR)

  6. ACCUMULATION OF PERCHLORATE IN TOBACCO PLANTS: DEVELOPMENT OF A PLANT KINETIC MODEL

    Science.gov (United States)

    Previous studies have shown that tobacco plants are tolerant of perchlorate and will accumulate perchlorate in plant tissues. This research determined the uptake, translocation, and accumulation of perchlorate in tobacco plants. Three hydroponics growth studies were completed u...

  7. ACCUMULATION OF PERCHLORATE IN TOBACCO PLANTS: DEVELOPMENT OF A PLANT KINETIC MODEL

    Science.gov (United States)

    Previous studies have shown that tobacco plants are tolerant of perchlorate and will accumulate perchlorate in plant tissues. This research determined the uptake, translocation, and accumulation of perchlorate in tobacco plants. Three hydroponics growth studies were completed u...

  8. Allergenicity assessment of genetically-modified tobacco expressing salt tolerance cbl gene.

    Science.gov (United States)

    Verma, Alok Kumar; Kumar, Sandeep; Chaudhari, Bhushan P; Tuteja, Narendra; Das, Mukul; Dwivedi, Premendra D

    2014-09-01

    It is mandatory to assess the allergenic potential of genetically modified (GM) crops before their commercialization. Recently, a transgene [Calcineurin B-like (CBL) protein] has been introduced into tobacco plant to make the crop salt resistance. Therefore, it was felt necessary to assess the allergenic potential of the cbl gene product, which was introduced and expressed in Nicotiana tabacum (tobacco) plant and compared the allergenic effects with the wild-type (WT) counterpart. Bioinformatic analysis revealed that there was no significant sequence homology with known allergens. Also, no difference between the protein digestibility profiles of GM and WT tobacco was found. Rapid digestion of CBL protein (Mol Wt 35 kDa) by simulated gastric fluid (SGF) indicated reduced chances of this protein to induce allergenicity. In addition, BALB/c mice sensitized by intraperitoneal administration of WT and GM tobacco protein showed comparable levels of clinical score, specific IgE, IgG1, histamine level, similar effect on different organs as well as IgE binding proteins. These findings indicate that insertion of cbl gene in tobacco did not cause any additional allergic risk to consumer and the GM and native tobacco proteins behave similarly in both in vitro and in vivo situations even after genetic modification.

  9. Amyloid-like protein inclusions in tobacco transgenic plants.

    Directory of Open Access Journals (Sweden)

    Anna Villar-Piqué

    Full Text Available The formation of insoluble protein deposits in human tissues is linked to the onset of more than 40 different disorders, ranging from dementia to diabetes. In these diseases, the proteins usually self-assemble into ordered β-sheet enriched aggregates known as amyloid fibrils. Here we study the structure of the inclusions formed by maize transglutaminase (TGZ in the chloroplasts of tobacco transplastomic plants and demonstrate that they have an amyloid-like nature. Together with the evidence of amyloid structures in bacteria and fungi our data argue that amyloid formation is likely a ubiquitous process occurring across the different kingdoms of life. The discovery of amyloid conformations inside inclusions of genetically modified plants might have implications regarding their use for human applications.

  10. Expression of a Modified Crylle Gene in E.Coli and in Transgenic Tobacco Confers Resistance to Corn Borer

    Institute of Scientific and Technical Information of China (English)

    Yun-Jun LIU; Fu-Ping SONG; Kang-Lai HE; Yuan YUAN; Xiao-Xia ZHANG; Peng GAO; Jian-Hua WANG; Guo-Ying WANG

    2004-01-01

    The wild-type Crylle gene from Bacillus thuringiensis was modified for its efficient expression in transgenic plants. Modified Cry1 Ie gene (designated as Cry1 Iem) was cloned into prokaryotic expression vector pET28b and its expression in E. coli was confirmed by SDS-PAGE analysis. Bioassays using crude expression products in E. coli revealed that Cry1 Iem protein had a similar toxicity to corn borer as wild-type Cry1 Ie. Cry1 Iem gene was then inserted downstream of the maize ubiquitin-1 promoter in plant expression vector p3301. Transgenic tobacco plants carrying Cry1 Iem showed insecticidal activity against com borer.

  11. Loss of alpha-tocopherol in tobacco plants with decreased geranylgeranyl reductase activity does not modify photosynthesis in optimal growth conditions but increases sensitivity to high-light stress.

    Science.gov (United States)

    Grasses, T; Grimm, B; Koroleva, O; Jahns, P

    2001-08-01

    The enzyme geranylgeranyl reductase (CHL P) catalyses the reduction of geranylgeranyl diphosphate to phytyl diphosphate in higher-plant chloroplasts and provides phytol for both chlorophyll (Chl) and tocopherol synthesis. The reduction in CHL P activity in transgenic tobacco (Nicotiana tabacum L.) plants is accompanied by the reduction in total Chl and tocopherol content and the accumulation of geranylgeranylated Chl (ChlGG). The photosynthetic performance and the susceptibility to photo-oxidative stress have been investigated in these transgenic plants. The reduced total Chl content in Chl P antisense plants resulted in the reduction of electron transport chains per leaf area without a concomitant effect on the stoichiometry, composition and activity of both photosystems. However, Chl P antisense plants were much more sensitive to light stress. Analyses of Chl fluorescence quenching indicated an increased photoinhibitory quenching at the expense of the pH-dependent fluorescence quenching after short illumination (15 min) at moderate light intensities. Prolonged illumination (up to 1 h) at saturating light intensities induced an increased photoinactivation from which the Chl P antisense plants could not recover or could only partially recover during a subsequent low light phase. Our data imply that the presence of ChlGG has no influence on harvesting and transfer of light energy in either photosystem. However, the reduced tocopherol content of the thylakoid membrane is a limiting factor for defensive reactions to photo-oxidative stress.

  12. Ethyl gallate displays elicitor activities in tobacco plants.

    Science.gov (United States)

    Goupil, Pascale; Benouaret, Razik; Richard, Claire

    2017-09-29

    Alkyl gallates showed elicitor activities on tobacco in both whole plants and cell suspensions. Methyl gallate (MG), ethyl gallate (EG) and propyl gallate (PG) infiltration into tobacco leaves induced hypersensitive reaction-like lesions and topical production of autofluorescent compounds revealed under UV light. When sprayed on tobacco plants at 5 mM, EG promoted upregulation of defence-related genes such as the antimicrobial PR1, -1,3-glucanase PR2, chitinase PR3 and osmotin PR5 target genes. Tobacco BY-2 cells challenged with EG underwent cell death in 48 h, significantly reduced in the presence of the protease inhibitor aprotinin. The three alkyl gallates all caused alkalinisation of the BY-2 extracellular medium, whereas gallic acid did not trigger any pH variation. Using EGTA or LaCl3, we showed that Ca2+ mobilisation occurred in BY-2 cells elicited with EG. Overall, our findings are the first evidence of alkyl gallate elicitor properties with early perception events on plasma membrane, potential hypersensitive reactions and PR-related downstream defence responses in tobacco.

  13. Bacterial pathogen phytosensing in transgenic tobacco and Arabidopsis plants.

    Science.gov (United States)

    Liu, Wusheng; Mazarei, Mitra; Rudis, Mary R; Fethe, Michael H; Peng, Yanhui; Millwood, Reginald J; Schoene, Gisele; Burris, Jason N; Stewart, C Neal

    2013-01-01

    Plants are subject to attack by a wide range of phytopathogens. Current pathogen detection methods and technologies are largely constrained to those occurring post-symptomatically. Recent efforts were made to generate plant sentinels (phytosensors) that can be used for sensing and reporting pathogen contamination in crops. Engineered phytosensors indicating the presence of plant pathogens as early-warning sentinels potentially have tremendous utility as wide-area detectors. We previously showed that synthetic promoters containing pathogen and/or defence signalling inducible cis-acting regulatory elements (RE) fused to a fluorescent protein (FP) reporter could detect phytopathogenic bacteria in a transient phytosensing system. Here, we further advanced this phytosensing system by developing stable transgenic tobacco and Arabidopsis plants containing candidate constructs. The inducibility of each synthetic promoter was examined in response to biotic (bacterial pathogens) or chemical (plant signal molecules salicylic acid, ethylene and methyl jasmonate) treatments using stably transgenic plants. The treated plants were visualized using epifluorescence microscopy and quantified using spectrofluorometry for FP synthesis upon induction. Time-course analyses of FP synthesis showed that both transgenic tobacco and Arabidopsis plants were capable to respond in predictable ways to pathogen and chemical treatments. These results provide insights into the potential applications of transgenic plants as phytosensors and the implementation of emerging technologies for monitoring plant disease outbreaks in agricultural fields.

  14. Plants having modified response to ethylene

    Science.gov (United States)

    Meyerowitz, E.M.; Chang, C.; Bleecker, A.B.

    1997-11-18

    The invention includes transformed plants having at least one cell transformed with a modified ETR nucleic acid. Such plants have a phenotype characterized by a decrease in the response of at least one transformed plant cell to ethylene as compared to a plant not containing the transformed plant cell. Tissue and/or temporal specificity for expression of the modified ETR nucleic acid is controlled by selecting appropriate expression regulation sequences to target the location and/or time of expression of the transformed nucleic acid. The plants are made by transforming at least one plant cell with an appropriate modified ETR nucleic acid, regenerating plants from one or more of the transformed plant cells and selecting at least one plant having the desired phenotype. 31 figs.

  15. 76 FR 36544 - Scientific Evaluation of Modified Risk Tobacco Product Applications; Public Workshop; Request for...

    Science.gov (United States)

    2011-06-22

    ... produced by the consumption of that tobacco product, that may affect a disease or health-related condition... HUMAN SERVICES Food and Drug Administration Scientific Evaluation of Modified Risk Tobacco Product... of public workshop; request for comments. The Food and Drug Administration (FDA), Center for...

  16. Differential mercury volatilization by tobacco organs expressing a modified bacterial merA gene

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    Mercury pollution is a major environmental problem accompanying industrial activities. Most of the mercury released ends up and retained in the soil as complexes of the toxic ionic mercury (Hg2+), which then can be converted by microbes into the even more toxic methylmercury which tends to bioaccumulate. Mercury detoxification of the soil can also occur by microbes converting the ionic mercury into the least toxic metallic mercury (Hg0) form, which then evaporates. The remediation potential of transgenic plants carrying the MerA gene from E. Coli encoding mercuric ion reductase could be evaluated. A modified version of the gene, optimized for plant codon preferences (merApe9, Rugh et al. 1996), was introduced into tobacco by Agrobacterium-mediated leaf disk transformation. Transgenic seeds were resistant to HgCl2 at 50 μM, and some of them (10-20%) could germinate on media containing as much as 350 μM HgCl2, while the control plants were fully inhibited or died on 50μM HgCl2. The rate of elemental mercury evolution from Hg2+ (added as HgCl2) was 5-8 times higher for transgenic plants than the control. Mercury volatilization by isolated organs standardized for fresh weight was higher (up to 5 times) in the roots than in shoots or the leaves. The data suggest that it is the root system of the transgenic plants that volatilizes most of the reduced mercury (Hg0). It also suggests that much of the mercury need not enter the vascular system to be transported to the leaves for volatilization. Transgenic plants with the merApe9 gene may be used to mercury detoxification for environmental improvement in mercury-contaminated regions more efficiently than it had been predicted based on data on volatilization of whole plants via the upper parts only (Rugh et al. 1996).

  17. Genetically modified soybean plants and their ecosystem

    Directory of Open Access Journals (Sweden)

    Milošević Mirjana B.

    2004-01-01

    Full Text Available Transgenic plants are developed by introgressing new genes using methods of molecular genetics and genetic engineering. The presence of these genes in plant genome is identified on the basis of specific oligonucleotides primers, and the use of PCR (Polymerase Chain Reaction and DNA fragments multiplication. Genetically modified plants such as soybean constitute a newly created bioenergetic potential whose gene expression can cause disturbance of the biological balance ecosystem, soil structure and soil microbiological activity. Genetically modified plants may acquire monogenic or polygenic traits causing genetic and physiological changes in these plants, which may elicit a certain reaction of the environment including changes of microbiological composition of soil rhizosphere. The aim of introgressing genes for certain traits into a cultivated plant is to enhance its yield and intensify food production. There are more and more genetically modified plant species such as soybean, corn, potato, rice and others and there is a pressure to use them as human food and animal feed. Genetically modified soybean plants with introgressed gene for resistance to total herbicides, such as Round-up, are more productive than non-modified herbicide-sensitive soybeans.

  18. Transgenic Tobacco Expressing a Modified VP6 Gene Protects Mice Against Rotavirus Infection

    Institute of Scientific and Technical Information of China (English)

    Jiang-Li DONG; Bo ZHOU; Gang SHENG; Tao WANG

    2005-01-01

    Elevated expression of the rotavirus VP6 antigen in transgenic plants is a critical factor in the development of a safe and effective rotavirus vaccine. Using codon optimization, a gene that encodes the inner capsid protein VP6 of the human group A rotavirus was synthesized (sVP6). The VP6 and sVp6genes were transformed into tobacco (Nicotiana tabacum L.) plants using Agrobacterium tumefaciens. The expression level of the sVP6 gene in transgenic plants was 3.8-34-fold higher than that of controls containing the non-modified VP6 gene, accounting for up to 0.34% of the total soluble protein (TSP). Then, BALB/c female mice that had been gavaged weekly with 10 mg TSP containing 34 μg VP6 protein, in which VP6-specific serum IgG and mucosal IgA antibodies were investigated. The severity and duration of diarrhea caused by simian rotavirus SA-11 challenge were reduced significantly in passively immunized pups, which indicates that anti-VP6 antibodies generated in orally immunized female mice can be passed onto pups and provide heterotypic protection. An edible vaccine based on the VP6 of human rotavirus group A could provide a means to protect children and young animals from severe acute diarrhea.

  19. Composition of hydroponic medium affects thorium uptake by tobacco plants.

    Science.gov (United States)

    Soudek, Petr; Kufner, Daniel; Petrová, Sárka; Mihaljevič, Martin; Vaněk, Tomáš

    2013-08-01

    The ability of thorium uptake as well as responses to heavy metal stress were tested in tobacco cultivar La Burley 21. Thorium was accumulated preferentially in the root system. The presence of citric, tartaric and oxalic acids in hydroponic medium increased thorium accumulation in all plant organs. On the other hand, the addition of diamines and polyamines, the important antioxidants in plants, resulted in decrease of thorium accumulation, especially in the root system. Negative correlation was found between putrescine concentration and thorium accumulation. Nevertheless, the most important factor influencing the accumulation of thorium was the absence of phosphate ions in a hydroponic medium that caused more than 10-fold increase of thorium uptake in all plant parts. Accumulation and distribution of thorium was followed in six cultivars and 14 selected transformants. Cultivar La Barley 21 represented an average between the tested genotypes, having a very good distribution ratio between roots, stems and leaves. Copyright © 2013 Elsevier Ltd. All rights reserved.

  20. Production of Bioactive Recombinant Bovine Chymosin in Tobacco Plants

    Directory of Open Access Journals (Sweden)

    Zheng-Yi Wei

    2016-04-01

    Full Text Available Chymosin (also known as rennin plays an essential role in the coagulation of milk in the cheese industry. Chymosin is traditionally extracted from the rumen of calves and is of high cost. Here, we present an alternative method to producing bovine chymosin from transgenic tobacco plants. The CYM gene, which encodes a preprochymosin from bovine, was introduced into the tobacco nuclear genome under control of the viral 35S cauliflower mosaic promoter. The integration and transcription of the foreign gene were confirmed with Southern blotting and reverse transcription PCR (RT-PCR analyses, respectively. Immunoblotting analyses were performed to demonstrate expression of chymosin, and the expression level was quantified by enzyme-linked immunosorbent assay (ELISA. The results indicated recombinant bovine chymosin was successfully expressed at an average level of 83.5 ng/g fresh weight, which is 0.52% of the total soluble protein. The tobacco-derived chymosin exhibited similar native milk coagulation bioactivity as the commercial product extracted from bovine rumen.

  1. Tobacco

    Science.gov (United States)

    ... these countries are low- or middle-income countries. Mass media campaigns can also reduce tobacco consumption by influencing ... have aired at least 1 strong anti-tobacco mass media campaign within the last 2 years. Ad bans ...

  2. Altered phenotypes in plants transformed with chimeric tobacco peroxidase genes

    Energy Technology Data Exchange (ETDEWEB)

    Lagrimini, L.M.

    1990-12-31

    Peroxidases have been implicated in a variety of secondary metabolic reactions including lignification, cross-linking of cell wall polysaccharides, oxidation of indole-3-acetic acid, regulation of cell elongation, wound-healing, phenol oxidation, and pathogen defense. However, due to the many different isoenzymes and even more potential substrates, it has proven difficult to verify actual physiological roles for peroxidase. We are studying the molecular biology of the tobacco peroxidase genes, and have utilized genetic engineering techniques to produce transgenic plants which differ only in their expression of an individual peroxidase isoenzyme. Many of the in planta functions for any individual isoenzyme may be predicted through the morphological and physiological analysis of transformed plants.

  3. Altered phenotypes in plants transformed with chimeric tobacco peroxidase genes

    Energy Technology Data Exchange (ETDEWEB)

    Lagrimini, L.M.

    1990-01-01

    Peroxidases have been implicated in a variety of secondary metabolic reactions including lignification, cross-linking of cell wall polysaccharides, oxidation of indole-3-acetic acid, regulation of cell elongation, wound-healing, phenol oxidation, and pathogen defense. However, due to the many different isoenzymes and even more potential substrates, it has proven difficult to verify actual physiological roles for peroxidase. We are studying the molecular biology of the tobacco peroxidase genes, and have utilized genetic engineering techniques to produce transgenic plants which differ only in their expression of an individual peroxidase isoenzyme. Many of the in planta functions for any individual isoenzyme may be predicted through the morphological and physiological analysis of transformed plants.

  4. Fungal endophytes: modifiers of plant disease.

    Science.gov (United States)

    Busby, Posy E; Ridout, Mary; Newcombe, George

    2016-04-01

    Many recent studies have demonstrated that non-pathogenic fungi within plant microbiomes, i.e., endophytes ("endo" = within, "phyte" = plant), can significantly modify the expression of host plant disease. The rapid pace of advancement in endophyte ecology warrants a pause to synthesize our understanding of endophyte disease modification and to discuss future research directions. We reviewed recent literature on fungal endophyte disease modification, and here report on several emergent themes: (1) Fungal endophyte effects on plant disease span the full spectrum from pathogen antagonism to pathogen facilitation, with pathogen antagonism most commonly reported. (2) Agricultural plant pathosystems are the focus of research on endophyte disease modification. (3) A taxonomically diverse group of fungal endophytes can influence plant disease severity. And (4) Fungal endophyte effects on plant disease severity are context-dependent. Our review highlights the importance of fungal endophytes for plant disease across a broad range of plant pathosystems, yet simultaneously reveals that complexity within plant microbiomes presents a significant challenge to disentangling the biotic environmental factors affecting plant disease severity. Manipulative studies integrating eco-evolutionary approaches with emerging molecular tools will be poised to elucidate the functional importance of endophytes in natural plant pathosystems that are fundamental to biodiversity and conservation.

  5. Genetically modified plants: public and scientific perceptions.

    Science.gov (United States)

    Rastogi Verma, Smita

    2013-01-01

    The potential of genetically modified plants to meet the requirements of growing population is not being recognized at present. This is a consequence of concerns raised by the public and the critics about their applications and release into the environment. These include effect on human health and environment, biosafety, world trade monopolies, trustworthiness of public institutions, integrity of regulatory agencies, loss of individual choice, and ethics as well as skepticism about the real potential of the genetically modified plants, and so on. Such concerns are enormous and prevalent even today. However, it should be acknowledged that most of them are not specific for genetically modified plants, and the public should not forget that the conventionally bred plants consumed by them are also associated with similar risks where no information about the gene(s) transfer is available. Moreover, most of the concerns are hypothetical and lack scientific background. Though a few concerns are still to be disproved, it is viewed that, with proper management, these genetically modified plants have immense potential for the betterment of mankind. In the present paper, an overview of the raised concerns and wherever possible reasons assigned to explain their intensity or unsuitability are reviewed.

  6. Genetically modified plants in practical applications

    OpenAIRE

    Říhová, Barbora

    2010-01-01

    Genetic engineering (GI) of plants is a very current topic, and more and more controversial, since it is becoming an inseparable part of our lives. GI has, among other things, a great potential to help solve the current problem of hunger and malnutrition in certain parts of the world. The goal of this project is to clarify what genetically modified (GM) plants are, to present the possibilities of their practical use, to explain methods of preparation and to consider their advantages and event...

  7. Expression of E. coli heat-labile enterotoxin B subunit in transgenic tobacco plants

    Institute of Scientific and Technical Information of China (English)

    LIU Hong-li; ZHANG Zheng; LI Wen-sheng; ZHENG Jing; KONG Ling-hong; WANG Yi-li; SI Lü-sheng

    2005-01-01

    Objective: To construct plant transformation vector containing Escherichia coli heat-labile enterotoxin B subunit (LT-B) gene and generate LT-B transgenic tobacco plants. Methods: The LT-B coding sequence was amplified from pMMB68 by PCR, subcloned into middle vector pUCmT and binary vector pBI121 to obtain plant expression vector pBI-LTB, in which LT-B expression was controlled under the Cauliflower mosaic virus (CaMV) 35S promoter. The tobacco plants (Nicotiana tobacum L. Cuttivar Xanthi) were transformed by co-cultivating leaf discs method via Agrobacterium tumefaciens LBA4404 harboring the plant expression vector. The regenerated transgenic tobacco plants were selected by kanamycin and confirmed by PCR, Southern blot, Western blot and ELISA. Results: LT-B gene integrated in the tobacco genomic DNA and were expressed in 9 strains of transgenic tobacco plants. The yield was varied from 3.36-10.56 ng/mg total soluble tobacco leaf protein. Conclusion: The plant binary expression vector pBILTB was constructed successfully, and transgenic LT-B tobacco plants was generated, and confirmed by Southern blot. The protein LT-B expressed by engineered plants was identified by Western blot analysis and had the expected molecular weight of LT-B pentamer protein. This result is an important step close to developing an edible vaccine and supplying a mucasal immunoajuvant, which will contribute to the prevention of mucosa-route evading pathogen.

  8. 75 FR 2879 - Use of Tobacco Marketing Descriptors to Convey Modified Risk; Request for Comments

    Science.gov (United States)

    2010-01-19

    ... information will be used to further FDA's efforts to reduce misleading and deceptive advertising practices..., labeling, or advertising of which uses the descriptors `light', `mild', or `low' or similar descriptors... label, labeling, or advertising of modified risk tobacco products. III. Comments Interested persons may...

  9. Plant Flavonoid Content Modified by Domestication.

    Science.gov (United States)

    Chacón-Fuentes, Manuel; Parra, Leonardo; Lizama, Marcelo; Seguel, Ivette; Urzúa, Alejandro; Quiroz, Andrés

    2017-07-21

    Plant domestication can modify and weaken defensive chemical traits, reducing chemical defenses in plants and consequently their resistance against pests. We characterized and quantified the major defensive flavonols and isoflavonoids present in both wild and cultivated murtilla plants (Ugni molinae Turcz), established in a common garden. We examined their effects on the larvae of Chilesia rudis (Butler) (Lepidoptera: Arctiidae). Insect community and diversity indices were also evaluated. We hypothesized that domestication reduces flavonoid contents and modifies C. rudis preference, the insect community, and diversity. Methanolic extracts were obtained from leaves of U. molinae plants and analyzed by high performance liquid chromatography. Results showed higher insect numbers (86.48%) and damage index (1.72 ± 0.16) in cultivated plants. Four new first records of insects were found associated with U. molinae. Diversity indices, such as Simpson, Shannon, and Margalef, were higher in cultivated plants than in wild plants. Furthermore, eight isoflavonoids were identified in U. molinae leaves for the first time. The five flavonols showed higher concentrations in wild U. molinae leaves (89.8 µg/g) than in cultivated plants (75.2 µg/g); however, no differences were found in isoflavonoids between wild and cultivated plants. The larvae of C. rudis consumed more leaf material of cultivated plants than wild plants in choice (3.8 vs. 0.8 mm2) and no-choice (7.5 vs. 3.0 mm2) assays. Our study demonstrates that domestication in U. molinae reduces the amount of flavonoids in leaves, increasing the preference of C. rudis and the insect community. © The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  10. Plant coexistence can enhance phytoextraction of cadmium by tobacco (Nicotiana tabacum L.) in contaminated soil

    Institute of Scientific and Technical Information of China (English)

    Ling Liu; Yuefang Li; Jianjun Tang; Liangliang Hu; Xin Chen

    2011-01-01

    A mesocosm experiment was conducted to investigate whether plant coexistence affects cadmium (Cd) uptake by plant in contaminated soil. Tobacco (Nicotiana tabacum L. var. K326) and Japanese clover (Kummerowia striata (Thunb.) Schindl.) were used.Cadmium was applied as 3CdSO4.8H2O in solution at three levels (0, 1, and 3 mg/kg soil) to simulate an unpolluted soil and soils that were slightly and moderately polluted with Cd. Tobacco (crop), Japanese clover (non-crop), and their combination were grown under each Cd treatment. Compared to monoculture and under all Cd treatments, co-planting with Japanese clover did not affect tobacco biomass but significantly increased Cd concentration in all tobacco tissues and enhanced Cd accumulation in tobacco shoots and roots.Compared to monoculture, co-planting reduced soil pH and increased Cd bioavailability. For tobacco, co-planting with Japanese clover increased the Cd bioconcentration factor (BCF) in Cd contaminated soil. Japanese clover also accumulated substantial quantities of Cd in shoots and roots. Thus, total Cd uptake by the plants was much greater with co-planting than with monoculture. The results suggested that phytoextraction can be effectively increased through tobacco co-planting with Japanese clover in mildly Cd-contaminated soil.

  11. Lung cancer biomarkers for the assessment of modified risk tobacco products: an oxidative stress perspective.

    Science.gov (United States)

    Lowe, Frazer J; Luettich, Karsta; Gregg, Evan O

    2013-05-01

    Manufacturers have developed prototype cigarettes yielding reduced levels of some tobacco smoke toxicants, when tested using laboratory machine smoking under standardised conditions. For the scientific assessment of modified risk tobacco products, tests that offer objective, reproducible data, which can be obtained in a much shorter time than the requirements of conventional epidemiology are needed. In this review, we consider whether biomarkers of biological effect related to oxidative stress can be used in this role. Based on published data, urinary 8-oxo-7,8-dihydro-2-deoxyguanosine, thymidine glycol, F2-isoprostanes, serum dehydroascorbic acid to ascorbic acid ratio and carotenoid concentrations show promise, while 4-hydroxynonenal requires further qualification.

  12. Sequence modification of merB gene and high organo-mercurial resistance of transgenic tobacco plants

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    Mercury pollution has caused severe damage to environment and great attention has been paid to its control. Phytoremediation may become one of the most efficient measures to recover the polluted soil since it is economical, highly efficient and friendly to environment. In this report, plant genetic engineering methods were employed to modify the DNA sequence of merB genes that catalyze the conversion of organomercurals into ionic mercury. The modified merBhe genes were introduced into tobacco by Agrobacterium, and the resultant transgenic plants were verified by Southern and Northern hybridization. High level of organomercurial resistance was detected on progenies of transgenic plants, some of which were resistant to PMA (phenyl mercury acetate) of 2.5 ?mol/L whereas 0.1 ?mol/L PMA killed the seedlings of wild-type tobacco in soiless culrure. With the increase of PMA concentration, the inhibition of the seedling growth became apparent. This result makes it possible to breed mercury-resistant tobacco for phytoremediation of mercury-polluted soil.

  13. Transgenic tobacco plants harboring tomato proteinase inhibitor II gene and their insect resistance

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    The plant expression vectors pBCT2 and pBT2 were constructed with the cDNA sequence (tin2) and genomic DNA sequence (tin2i) of tomato proteinase inhibitor II gene respectively. Then the two expression vectors were transferred into tobacco via the Agrobacterium tumefaciens strain LBA4404, and transgenic tobacco plants were generated. Molecular analysis and trypsin activity assay showed that both cDNA and genomic DNA were expressed properly in the transgenic plants. Insecticidal activities in these transgenic plants indicated that transgenic tobacco plants carrying tin2i sequence were more resistant to 2-instar larvae of Heliothis armigera Hubner than those carrying tin2 sequence. Therefore the intron of tin2i sequence might be a contributor to insecticidal activity of the transgenic tobacco.

  14. Obtaining High Pest-resistant Tobacco Plants Carrying B.t. insecticidal Gene

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    To increase the expression level of CryIA(c) gene in transgenic plants, a plant expression vector pBinMoBc carrying the CryIA(c) gene under control of chimeric OM promoter and Ω factor was constructed. As a control, pBinoBc carrying the CryIA(c) gene with the CaMV 35S promoter was also constructed. The vectors were transferred into tobacco plants respectively via Agrobacterium-mediated transformation. ELISA assay showed that the expression level of the CryIA(c) gene in pBinMoBc transgenic tobacco plants was 2.44-times that in pBinoBc transgenic tobacco plants, and it could be up to 0.255% of total soluble proteins. Bioassay showed that pBinMoBc transgenic tobacco plants had more notable insecticidal effect than pBinoBc transgenic tobacco plants. The above results showed that the chimeric OM promoter was a stronger promoter than CaMV 35S promoter that was widely used in plant genetic engineering, and this is very useful in pest-resistant plant genetic engineering.

  15. Expression of Human Papillomavirus Type 16 L1 Protein in Transgenic Tobacco Plants

    Institute of Scientific and Technical Information of China (English)

    Hong-Li LIU; Wen-Sheng LI; Ting LEI; Jing ZHENG; Zheng ZHANG; Xiao-Fei YAN; Zhe-Zhi WANG; Yi-Li WANG; Lü-Sheng SI

    2005-01-01

    To develop a plant expression system for the production of the human papillomavirus type 16(HPV16) vaccine, we investigated whether the HPV16 L1 protein can be expressed in tobacco plants and whether it can be used as the cheapest form of edible vaccine. The HPV16 L1 coding sequence was amplified by PCR using specific primers from the plasmid pGEM-T-HPV16 containing the template sequence, and subcloned into the intermediate vector pUCmT and binary vector pBI121 consecutively to obtain the plant expression plasmid pBI-L1. The T-DNA regions of the pBI-L1 binary vector contained the constitutive Cauliflower mosaic virus (CaMV) 35S promoter and the neomycin phosphotransferase npt Ⅱ gene, which allowed the selection of transformed plants using kanamycin. The tobacco plants were transformed by cocultivating them, using the leaf disc method, with Agrobacterium tumefaciens LBA4404, which harbored the plant expression plasmid. The regenerated transgenic tobacco plants were selected using kanamycin, and confirmed by PCR. The results of the Southern blot assay also showed that the HPV16 L1 gene was integrated stably into the genome of the transformed tobacco plants. The Western blot analysis showed that the transformed tobacco leaves could express the HPV16 L1 protein. Furthermore, it was demonstrated by ELISA assay that the expressed protein accounted for 0.034%-0.076% of the total soluble leaf protein, was able to form 55 nm virus-like particles compatible with HPV virus-like particle (VLP), and induced mouse erythrocyte hemagglutination in vitro. The present results indicate that the HPV16 L1 protein can be expressed in transgenic tobacco plants and the expressed protein possesses the natural features of the HPV 16L1 protein, implying that the HPV16 L1 transgenic plants can be potentially used as an edible vaccine.

  16. [Transgenic tobacco plants with ribosome inactivating protein gene cassin from Cassia occidentalis and their resistance to tobacco mosaic virus].

    Science.gov (United States)

    Ruan, Xiao-Lei; Liu, Li-Fang; Li, Hua-Ping

    2007-12-01

    Cassin, the new gene of ribosome-inactivating protein (RIP) isolated from Cassia occidentalis, was inserted into expression vector pBI121 to produce plant expression vector pBI121-cassin (Figs.1, 2). pBI121-cassin was introduced into tobacco cultivar 'K326' by the Agrobacteriurm tumefaciens transformation method and more than 100 independent transformants were obtained. Southern blot hybridization analysis showed that a single gene locus was inserted into the chromosome of the transgenic tobacco lines (Fig.5) and PCR analysis of segregation population of progeny indicated that the inheritance of transgene was dominant in transgenic lines (Fig.4, Table 1). Results of RT-PCR and Northern blot hybridization analysis showed that transgene could be transcribed correctly (Figs.5, 6) . Three self-pollination lines of transgenic T(1) and T(2) were challenged with TMV at different concentration titers by mechanical inoculation. The transgenic lines exhibited different levels of resistance to TMV with the nontransgenic plants. After both titers of TMV concentration were inoculated, transgenic lines were considered as the highly resistant type with a delay of 4-13 d in development of symptoms and 10%-25% of test plants were infected, while nontransgenic control plants were susceptible typical symptoms on the newly emerged leaves (Table 2). One T(2) line, T(2)-8-2-1, was regarded as an immune type because it did not show any symptoms during 70 d and all plants were shown to be virus free by ELISA tests.

  17. Genetically modified food -The dilemma of Africa

    African Journals Online (AJOL)

    STORAGESEVER

    2008-05-02

    May 2, 2008 ... Key words: Benefits, concerns, food security, genetically modified plant. INTRODUCTION .... health effects, environmental safety and conservation, intellectual .... including coffee, tobacco, cocoa, coconut, palm oil, sugar.

  18. Effects of Mechanical Stress and Plant Density on Mechanical Characteristics, Growth, and Lifetime Reproduction of Tobacco Plants

    NARCIS (Netherlands)

    Anten, N.P.R.; Casado-Garcia, R.; Nagashima, H.

    2005-01-01

    Plastic increases in stem elongation in dense vegetation are generally believed to be induced by canopy shading, but because plants protect each other from wind, shielding (reduced mechanical stress) could also play a role. To address this issue, tobacco Nicotiana tabacum plants were subjected to

  19. A genetically modified tobacco mosaic virus that can produce gold nanoparticles from a metal salt precursor.

    Directory of Open Access Journals (Sweden)

    Andrew John Love

    2015-11-01

    Full Text Available We genetically modified tobacco mosaic virus (TMV to surface display a characterized peptide with potent metal ion binding and reducing capacity (MBP TMV, and demonstrate that unlike wild type (WT TMV, this construct can lead to the formation of discrete 10-40 nm gold nanoparticles when mixed with 3 mM potassium tetrachloroaurate. Using a variety of analytical physicochemical approaches it was found that these nanoparticles were crystalline in nature and stable. Given that the MBP TMV can produce metal nanomaterials in the absence of chemical reductants, it may have utility in the green production of metal nanomaterials.

  20. A comparative study of Tam3 and Ac transposition in transgenic tobacco and petunia plants

    NARCIS (Netherlands)

    Haring, Michel A.; Gao, Jie; Volbeda, Tjeerd; Rommens, Caius M.T.; Nijkamp, H. John J.; Hille, Jacques

    1989-01-01

    Transposition of the Anthirrinum majus Tam3 element and the Zea mays Ac element has been monitored in petunia and tobacco plants. Plant vectors were constructed with the transposable elements cloned into the leader sequence of a marker gene. Agrobacterium tumefaciens-mediated leaf disc transformatio

  1. Secretive expression of Aspergillus fumigatus phytase in tobacco improves phosphorus nutrition in plant

    Institute of Scientific and Technical Information of China (English)

    Wang Yan; Gao Xiaorong; Su Qiao; An Lijia

    2007-01-01

    To generate transgenic plants capable of utilizing exogenous phytate,an Aspersgillus fumigatus phytase gene(fphyA) was constitutively expressed in tobacco and recombinant enzyme was secreted from plant roots into the rhizosphere using the signal sequence from tobacco calretieulin.After 40 days of plant growth in hydroponic media,phytase activities in leaves,stems,roots and growth media of transgenic plants were 8.6-fold,7.4-fold,12.6-fold and 14.3-fold higher than those of wild-type plants.Signifi-cant improvements in plant growth and phosphoms(P)utilization were observed in the transgenic plants.When phytate was supplied as the sole P source.45-day-old transgenic tobaccos accumulated 1.0-fold and 0.5-fold more shoot and root biomass than wild-type tobaccos.with a concomitant of 1.7-fold increase in total P concentration.These results indicate that secretive expression of the A.fumigatus phytase improves acquisition and use of P from phytate in plants.

  2. Construction of a new plant expression vector containing two insect resistant genes and its expression in transgenic tobacco plants

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    A new plant expression vector (pBS29K-BA) containing two insect resistant genes, a synthetic chimeric gene BtS29K encoding the activated insecticidal protein Cry1Ac and a gene API-BA encoding the arrowhead (Sagittaria sagittifolia L.) proteinase inhibitor (API) A and B, is constructed. Transgenic tobacco plants expressing these two genes are obtained through Agrobacterium-mediated transformation of tobacco leaf discs. The average expression levels of Cry1Ac and API-BA proteins in transgenic plants are of 3.2 μg and 4.9 μg per gram fresh leaf respectively. The results of insecticidal assay of transgenic plants indicate that the pBS29K-BA transformed plants are more resistant to insect damage than the plants expressing the Cry1Ac gene or API-BA gene alone.

  3. Humans Have Antibodies against a Plant Virus: Evidence from Tobacco Mosaic Virus

    Science.gov (United States)

    Liu, Ruolan; Vaishnav, Radhika A.; Roberts, Andrew M.; Friedland, Robert P.

    2013-01-01

    Tobacco mosaic virus (TMV), a widespread plant pathogen, is found in tobacco (including cigarettes and smokeless tobacco) as well as in many other plants. Plant viruses do not replicate or cause infection in humans or other mammals. This study was done to determine whether exposure to tobacco products induces an immune response to TMV in humans. Using a sandwich ELISA assay, we detected serum anti-TMV antibodies (IgG, IgG1, IgG3, IgG4, IgA, and IgM) in all subjects enrolled in the study (20 healthy smokers, 20 smokeless-tobacco users, and 20 non-smokers). Smokers had a higher level of serum anti-TMV IgG antibodies than non-smokers, while the serum level of anti-TMV IgA from smokeless tobacco users was lower than smokers and non-smokers. Using bioinformatics, we also found that the human protein TOMM40L (an outer mitochondrial membrane 40 homolog – like translocase) contains a strong homology of six contiguous amino acids to the TMV coat protein, and TOMM40L peptide exhibited cross-reactivity with anti-TMV antibodies. People who smoke cigarettes or other tobacco products experience a lower risk of developing Parkinson’s disease, but the mechanism by which this occurs is unclear. Our results showing molecular mimicry between TMV and human TOMM40L raise the question as to whether TMV has a potential role in smokers against Parkinson’s disease development. The potential mechanisms of molecular mimicry between plant viruses and human disease should be further explored. PMID:23573274

  4. Uncoupling Auxin and Ethylene Effects in Transgenic Tobacco and Arabidopsis Plants.

    Science.gov (United States)

    Romano, CP; Cooper, ML; Klee, HJ

    1993-01-01

    Overproduction of auxin in transgenic plants also results in the overproduction of ethylene. Plants overproducing both auxin and ethylene display inhibition of stem elongation and growth, increased apical dominance, and leaf epinasty. To determine the relative roles of auxin and ethylene in these processes, transgenic tobacco and Arabidopsis plants expressing the auxin-overproducing tryptophan monooxygenase transgene were crossed to plants expressing an ethylene synthesis-inhibiting 1-aminocyclopropane-1-carboxylate deaminase transgene. Tobacco and Arabidopsis plants with elevated auxin and normal levels of ethylene were obtained by this strategy. Transgenic auxin-overproducing Arabidopsis plants were also crossed with the ethylene-insensitive ein1 and ein2 mutants. Analysis of these plants indicates that apical dominance and leaf epinasty are primarily controlled by auxin rather than ethylene. However, ethylene is partially responsible for the inhibition of stem elongation observed in auxin-overproducing tobacco. Finally, these data show that auxin overproduction can be effectively uncoupled from ethylene overproduction in transgenic plants to enable direct manipulation of plant morphology for agronomic and horticultural purposes. PMID:12271061

  5. Enhanced tolerance and remediation of anthracene by transgenic tobacco plants expressing a fungal glutathione transferase gene

    Energy Technology Data Exchange (ETDEWEB)

    Dixit, Prachy; Mukherjee, Prasun K.; Sherkhane, Pramod D.; Kale, Sharad P. [Nuclear Agriculture and Biotechnology Division, Bhabha Atomic Research Centre, Mumbai 400085 (India); Eapen, Susan, E-mail: eapenhome@yahoo.com [Nuclear Agriculture and Biotechnology Division, Bhabha Atomic Research Centre, Mumbai 400085 (India)

    2011-08-15

    Highlights: {yields} Transgenic plants expressing a TvGST gene were tested for tolerance, uptake and degradation of anthracene. {yields} Transgenic plants were more tolerant to anthracene and take up more anthracene from soil and solutions compared to control plants. {yields} Using in vitro T{sub 1} seedlings, we showed that anthracene-a three fused benzene ring compound was phytodegraded to naphthalene derivatives, having two benzene rings. {yields} This is the first time that a transgenic plant was shown to have the potential to phytodegrade anthracene. - Abstract: Plants can be used for remediation of polyaromatic hydrocarbons, which are known to be a major concern for human health. Metabolism of xenobiotic compounds in plants occurs in three phases and glutathione transferases (GST) mediate phase II of xenobiotic transformation. Plants, although have GSTs, they are not very efficient for degradation of exogenous recalcitrant xenobiotics including polyaromatic hydrocarbons. Hence, heterologous expression of efficient GSTs in plants may improve their remediation and degradation potential of xenobiotics. In the present study, we investigated the potential of transgenic tobacco plants expressing a Trichoderma virens GST for tolerance, remediation and degradation of anthracene-a recalcitrant polyaromatic hydrocarbon. Transgenic plants with fungal GST showed enhanced tolerance to anthracene compared to control plants. Remediation of {sup 14}C uniformly labeled anthracene from solutions and soil by transgenic tobacco plants was higher compared to wild-type plants. Transgenic plants (T{sub 0} and T{sub 1}) degraded anthracene to naphthalene derivatives, while no such degradation was observed in wild-type plants. The present work has shown that in planta expression of a fungal GST in tobacco imparted enhanced tolerance as well as higher remediation potential of anthracene compared to wild-type plants.

  6. Expression of Cryptogein in tobacco plants exhibits enhanced disease resistance and tolerance to salt stress

    Institute of Scientific and Technical Information of China (English)

    JIANG Donghua; CHEN Xujun; WU Kunlu; GUO Zejian

    2004-01-01

    Cryptogein (Crypt), an elicitin secreted from Phytophthora cryptogea, was used for genetic engineering of biotic and abiotic resistance plants. We generated transgenic tobacco plants harboring a rice phenylalanine ammonia-lyase (PAL) promoter and Crypt fusion gene (PAL::Crypt) or the mutated Crypt (mutation of the lysine at the position 13 to valine) under the control CaMV35S promoter (CaMV35S::CryK13V). T2 progeny of the transgenic plants showed significantly enhanced disease resistance to pathogens of fungal Phytophthora parasitica var nicotiana (Ppn) and Alternaria alternata, and bacterial Pseudomonas syringae pv tabaci. The amount of mRNA accumulation of Crypt and CryK13V was quite low in the transgenic lines analyzed by Northern blot, and was detected by a reverse transcription PCR method. Plants harboring PAL::Crypt construct showed faster and stronger induction of PR-1a gene after Ppn inoculation than that in the wild-type plants. The results suggested that the inducible PAL promoter could rapidly respond to pathogen attack and efficiently suppress the pathogen infection. Furthermore, the enhanced tolerance to salt stress in both of the Crypt and CryK13V expressing tobacco plants was also observed compared with that in the control plants. The constitutive expression of PR and transcription factor genes in the transformants was probably associated with the salt tolerance. The above observations suggested that a cross-talk between biotic and abiotic stresses existed in tobacco plants.

  7. [Construction and analysis of transgenic plants of Nicotiana tabacum L. expressing a bacterial gene for beta-1,3-glucanase. I. Construction of vector plasmids for transfer into plants and expression of a modified gene for beta-1,3-glucanase from Clostridium thermocellum in tobacco protoplasts].

    Science.gov (United States)

    Darbinian, N S; Popov, Iu G; Mochul'skiĭ, A V; Volkova, L V; Piruzian, E S; Vasilevko, V T

    1996-02-01

    We constructed two vectors, pC27-glc and pC29-glc, that allow expression of the beta-1,3-glucanase gene (glc) in plant cells. The glc gene was previously cloned from anaerobic thermophilous bacterium Clostridium thermocellum. To increase the efficiency of expression, the N-terminal fragment of the glc gene encoding bacterial transient peptide was deleted, and hybrid variants of lacZ-glc were obtained. Analysis of expression of the hybrid genes in Escherichia coli showed that deletion of the fragment corresponding to 31 amino acids (a.a.) of beta-glucanase affected neither activity nor thermostability of the enzyme. The modified gene was subcloned into two vectors, pC27 and pC29, in which its expression was controlled by the TR2' promoter of the 2' gene of T-DNA and the rbcS promoter from Arabidopsis, respectively. Each of the resulting plasmids, pC27-glc and pC29-glc, was transfected into protoplasts of Nicotiana plumbaginifolia. Both the plasmids were shown to allow a high level of activity of the thermostable beta-1,3-glucanase. We plan to use the vectors obtained for transformation of agrobacteria and construction of transgenic plants.

  8. Hyperspectral remote sensing applications for monitoring and stress detection in cultural plants: viral infections in tobacco plants

    Science.gov (United States)

    Krezhova, Dora; Petrov, Nikolai; Maneva, Svetla

    2012-09-01

    The objectives of this study were to reveal the presence of viral infections in two varieties of tobacco plants (Nicotiana tabacum L.) as well as to discriminate the levels of the disease using hyperspectral leaf reflectance. Data sets were collected from two tobacco cultivars, Xanthi and Rustica, known as most widespread in Bulgaria. Experimental plants were grown in a greenhouse under controlled conditions. At growth stage 4-6 expanded leaf plants of cultivar Xanthi were inoculated with Potato virus Y (PVY) while the Rustica plants were inoculated with Tomato spotted wilt virus (TSWV). These two viruses are worldwide distributed and cause significant yield losses in many economically important crops. In the course of time after inoculation the concentration of the viruses in plant leaves was assessed by erological analysis via DAS-ELISA and RT-PCR techniques. Hyperspectral reflectance data were collected by a portable fibreoptics spectrometer in the visible and near-infrared spectral ranges (450-850 nm). As control plants healthy untreated tobacco plants were used. The significance of the differences between reflectance spectra of control and infected leaves was analyzed by means of Student's t-criterion at pred (640-680 nm), red edge (690-720 nm) and near infrared (720-780 nm) spectral ranges. Changes in SRC were found for both viral treatments and comparative analysis showed that the influence of PVY was stronger. The discrimination of disease intensity was achieved by derivative analysis of the red edge position.

  9. Metabolic flux analysis in complex isotopolog space. Recycling of glucose in tobacco plants.

    Science.gov (United States)

    Ettenhuber, Christian; Radykewicz, Tanja; Kofer, Waltraud; Koop, Hans-Ulrich; Bacher, Adelbert; Eisenreich, Wolfgang

    2005-02-01

    Tobacco plants grown in vitro were supplied with a mixture of [U-13C6]glucose and unlabelled sucrose via the root system. After 20 days, leaves were harvested and extracted with water. Glucose was isolated from the extract and was analysed by 13C NMR spectroscopy. All 13C signals appeared as complex multiplets due to 13C-13C coupling. The abundance of 21 isotopologous glucose species was determined from the 13C NMR signal integrals by numerical deconvolution using a genetic algorithm. The relative fractions of specific isotopologs in the overall excess of 13C-labelled specimens establish flux contributions via glycolysis/glucogenesis, pentose phosphate pathway, citric acid cycle and Calvin cycle including 13CO2 refixation. The fluxes were modelled and reconstructed in silico by a novel rule-based approach yielding the contributions of circular pathways and the degree of multiple cycling events. The data indicate that the vast majority of the proffered [U-13C6]glucose molecules had been modified by catabolism and subsequent glucogenesis from catabolic fragments, predominantly via passage through the citric acid cycle and the pentose phosphate pathway.

  10. Spectral reflectance, chlorophyll fluorescence and virological investigations of tobacco plants (Nicotiana tabacum L.) infected with Tobacco mosaic virus (TMV)

    Science.gov (United States)

    Krezhova, Dora; Hristova, Dimitrina; Iliev, Ilko; Yanev, Tony

    Application of multispectral remote sensing techniques to plant condition monitoring has been adopted for various purposes. Remote sensing is a reliable tool for detecting signs of vege-tation stress and diseases. Spectral reflectance and chlorophyll fluorescence are functions of tissue optical properties and biological status of the plants, and illumination conditions. The mean reflectance spectrum depends on the relative composition of all the pigments in the leaf including chlorophylls, carotenoids etc. Chlorophyll fluorescence results from the primary re-actions of photosynthesis and during the last decade it finds widening application as a means for revelation of stress and diseases. The changes in chlorophyll function take place before the alteration in chlorophyll content to occur so that changes in the fluorescence signal arise before any visible signs are apparent. The aim of our investigations was to study the development and spreading out of a viral infection on the leaves of two cultivars tobacco plants (Nicotiana tabacum L.) infected with Tobacco mosaic virus (TMV). We applied two remote sensing tech-niques (spectral reflectance and chlorophyll fluorescence measurements) for evaluation of the changes in the optical properties of the plants in accordance to their physiological status. The serological analyses via the Double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) were made with appropriate kits (Leowe, Germany) for quantitative assessment of the concentration of viruses in the plants. The tobacco plants were grown in green house under controlled conditions. The first cultivar Nevrocop 1146 is known as resistive to the TMV, i.e. it shows hypersensitive response. The second cultivar named Krumovgrad is normally sen-sitive to the TMV. At growth stage 4-6 expanded leaf, up to one leaf from 20 plants for each cultivar were inoculated with TMV. The leaves opposite to the infected ones formed the group of control (untreated) leaves. The

  11. Tobacco plants over-expressing the sweet orange tau glutathione transferases (CsGSTUs) acquire tolerance to the diphenyl ether herbicide fluorodifen and to salt and drought stresses.

    Science.gov (United States)

    Lo Cicero, Luca; Madesis, Panagiotis; Tsaftaris, Athanasios; Lo Piero, Angela Roberta

    2015-08-01

    The glutathione transferases (GSTs) are members of a superfamily of enzymes with pivotal role in the detoxification of both xenobiotic and endogenous compounds. In this work, the generation and characterization of transgenic tobacco plants over-expressing tau glutathione transferases from Citrus sinensis (CsGSTU1 and CsGSTU2) and several cross-mutate forms of these genes are reported. Putative transformed plants were verified for the presence of the transgenes and the relative quantification of transgene copy number was evaluated by Taqman real time PCR. The analysis of gene expression revealed that transformed plants exhibit high levels of CsGSTU transcription suggesting that the insertion of the transgenes occurred in transcriptional active regions of the tobacco genome. In planta studies demonstrate that transformed tobacco plants gain tolerance against fluorodifen. Simultaneously, the wild type CsGSTU genes were in vitro expressed and their kinetic properties were determined using fluorodifen as substrate. The results show that CsGSTU2 follows a Michaelis-Menten hyperbolic kinetic, whereas CsGSTU1 generates a sigmoid plot typical of the regulatory enzymes, thus suggesting that when working at sub-lethal fluorodifen concentrations CsGSTU2 can counteract the herbicide injury more efficiently than the CsGSTU1. Moreover, the transgenic tobacco plant over-expressing CsGSTs exhibited both drought and salinity stress tolerance. However, as we show that CsGSTUs do not function as glutathione peroxidase in vitro, the protective effect against salt and drought stress is not due to a direct scavenging activity of the oxidative stress byproducts. The transgenic tobacco plants, which are described in the present study, can be helpful for phytoremediation of residual xenobiotics in the environment and overall the over-expression of CsGSTUs can be helpful to develop genetically modified crops with high resistance to abiotic stresses.

  12. Uptake of Cadmium by Flue-Cured Tobacco Plants: Exploring Bioavailability

    Science.gov (United States)

    Holzer, I.; Robarge, W. P.; Vann, M. C.

    2015-12-01

    Scientific understanding of cadmium (Cd) cycling in North Carolina tobacco plants and soils has lagged, even as production of flue-cured tobacco remains an important part of the NC economy ($903 million in 2014). Cd is considered a tobacco contaminant. When tobacco is burned, Cd can exist as a fine aerosol and subsequent inhalation is linked to cancer. Tobacco root exudates enhance Cd uptake, even though the Cd concentration in NC soils is plants is crucial to understanding Cd bioavailability and implementing soil remediation efforts. The objective of this study was to develop a Cd mass balance for flue-cured tobacco grown under field conditions in NC. Whole plant samples were collected at transplanting and every 2 weeks thereafter until harvest. Individual plants were segregated into root, stalk and individual leaves (n = 15 whole plants/sampling date; composite samples were taken early in the growing season). After recording dry mass, samples were analyzed using ion-coupled plasma optical emission spectrometry or ion-coupled plasma mass spectrometry. Lower leaves contained the highest Cd concentrations ( 7-10 mg/kg). Leaves occupying the upper 50% of the plant had Cd concentrations of 2 mg/kg. Uptake rate was greatest from day 27 to 66 ( 21.5 μg Cd/day). Selective Cd uptake appears evident between day 27 and 43, but overall the relative rate of Cd uptake was similar to other trace metals and micronutrients. Cd distribution within the plants mirrored the distribution of calcium, a macronutrient. Of the 8 mg of soil extractable Cd (0.075 mg/kg) in the rooting zone, 15.0% (1203 μg) is removed by uptake. Of this 15%, 64.2% (772.2 μg) is exported at harvest, and 35.8% (430.8 μg; lower leaves, roots, stalks) is returned to the soil. This study must be replicated to account for seasonal and soil variations. These results do inform selection of tobacco strains that limit uptake of trace metals, particularly Cd.

  13. Acquiring transgenic tobacco plants with insect resistance and glyphosate tolerance by fusion gene transformation.

    Science.gov (United States)

    Sun, He; Lang, Zhihong; Zhu, Li; Huang, Dafang

    2012-10-01

    The advantages of gene 'stacking' or 'pyramiding' are obvious in genetically modified (GM) crops, and several different multi-transgene-stacking methods are available. Using linker peptides for multiple gene transformation is considered to be a good method to meet a variety of needs. In our experiment, the Bt cry1Ah gene, which encodes the insect-resistance protein, and the mG ( 2 ) -epsps gene, which encodes the glyphosate-tolerance protein, were connected by a 2A or LP4/2A linker. Linker 2A is a peptide from the foot-and-mouth disease virus (FMDV) that has self-cleavage activity. LP4 is a peptide from Raphanus sativus seeds that has a recognition site and is cleaved by a protease. LP4/2A is a hybrid peptide that contains the first 9 amino acids of LP4 and 20 amino acids from 2A. We used the linker peptide to construct four coordinated expression vectors: pHAG, pHLAG, pGAH and pGLAH. Two single gene expression vectors, pSAh and pSmG(2), were used as controls. The six expression vectors and the pCAMBIA2301 vector were transferred into tobacco by Agrobacterium tumefaciens-mediated transformation, and 529 transformants were obtained. Molecular detection and bioassay detection data demonstrated that the transgenic tobaccos possessed good pest resistance and glyphosate tolerance. The two genes in the fusion vector were expressed simultaneously. The plants with the genes linked by the LP4/2A peptide showed better pest resistance and glyphosate tolerance than the plants with the genes linked by 2A. The expression level of the two genes linked by LP4/2A was not significantly different from the single gene vector. Key message The expression level of the two genes linked by LP4/2A was higher than those linked by 2A and was not significantly different from the single gene vector.

  14. Expression of peanut Iron Regulated Transporter 1 in tobacco and rice plants confers improved iron nutrition.

    Science.gov (United States)

    Xiong, Hongchun; Guo, Xiaotong; Kobayashi, Takanori; Kakei, Yusuke; Nakanishi, Hiromi; Nozoye, Tomoko; Zhang, Lixia; Shen, Hongyun; Qiu, Wei; Nishizawa, Naoko K; Zuo, Yuanmei

    2014-07-01

    Iron (Fe) limitation is a widespread agricultural problem in calcareous soils and severely limits crop production. Iron Regulated Transporter 1 (IRT1) is a key component for Fe uptake from the soil in dicot plants. In this study, the peanut (Arachis hypogaea L.) AhIRT1 was introduced into tobacco and rice plants using an Fe-deficiency-inducible artificial promoter. Induced expression of AhIRT1 in tobacco plants resulted in accumulation of Fe in young leaves under Fe deficient conditions. Even under Fe-excess conditions, the Fe concentration was also markedly enhanced, suggesting that the Fe status did not affect the uptake and translocation of Fe by AhIRT1 in the transgenic plants. Most importantly, the transgenic tobacco plants showed improved tolerance to Fe limitation in culture in two types of calcareous soils. Additionally, the induced expression of AhIRT1 in rice plants also resulted in high tolerance to low Fe availability in calcareous soils.

  15. Improvement of Drought Tolerance in Transgenic Tobacco Plants by a Dehydrin-Like Gene Transfer

    Institute of Scientific and Technical Information of China (English)

    SHEN Ye; JIA Wei-long; ZHANG Yan-qin; HU Yuan-lei; WU Qi; LIN Zhong-ping

    2004-01-01

    A full-length cDNA of dehydrin BcDh2 from Boea crassifolia and its antisense nucleotide sequence have been transferred into tobacco (Nicotiana tabacum) NC89 under the control of a caulifower mosaic virus 35S promoter. Under a progressive water stress, photosynthetic rate, transpiration rate and stomatal conductance of the sense and antisense plants reduced, and those of the control reduced much more. Photosynthetic rate, transpiration rate and stomatal conductance of all plants tested increased significantly 24 hours later after recoveried water supply, and those of the sense and antisense plants were higher than control. These indicated that overexpression of a dehydrin gene in tobacco may improve tolerance to water stress for plants, however, antisense BcDh2 gene in transgenic plant did not influence physiological conditions. The results of germination experiment of the transgenic seeds showed that on MS medium with different concentration PEG (8000), sense seed could more endure drought than control, while antisense seed was sensitive to drought. The results suggested that the overexpression of a dehydrin gene in tobacco might improve the tolerance to water stress for plants.

  16. Improvement of Drought Tolerance in Transgenic Tobacco Plants by aDehydrin-Like Gene Transfer

    Institute of Scientific and Technical Information of China (English)

    SHENYe; JIAWei-long; ZHANGYan-qin; HUYuan-lei; WUQi; LINZhongping

    2004-01-01

    A full-length cDNA of dehydrin BcDh2 from Boea crassifolia and its antisense nucleotide sequence have been transferred into tobacco (Nicotiana tabacum) NC89 under the control of a caulifower mosaic virus 35S promoter. Under a progressive water stress, photosynthetic rate, transpiration rate and stomatal conductance of the sense and antisense plants reduced, and those of the control reduced much more. Photosynthetic rate, transpiration rate and stomatal conductance of all plants tested increased significantly 24 hours later after recoveried water supply, and those of the sense and antisense plants were higher than control. These indicated that overexpression of a dehydrin gene in tobacco may improve tolerance to water stress for plants, however, antisense BcDh2 gene in transgenic plant did not influence physiological conditions. The results of germination experiment of the transgenic seeds showed that on MS medium with different concentration PEG (8000), sense seed could more endure drought than control, while antisense seed was sensitive to drought. The results suggested that the overexpression of a dehydrin gene in tobacco might improve the tolerance to water stress for plants.

  17. Suppression of infectious TMV genomes expressed in young transgenic tobacco plants.

    Science.gov (United States)

    Siddiqui, S A; Sarmiento, C; Valkonen, S; Truve, E; Lehto, K

    2007-12-01

    Full-length cDNAs of the wild-type (wt) Tobacco mosaic virus (TMV) and of the coat protein gene-deleted (DeltaCP) derivative of wt-TMV, under control of the 35S promoter and downstream ribozyme sequence to produce accurate viral transcripts, were transformed to tobacco plants to analyze plant-virus interactions through different stages of plant development. Surprisingly, young wt-TMV transgenics accumulated only very low levels of viral RNA, remained free of symptoms, and were moderately resistant against exogenous inoculations. This early resistance caused significant stress to the plants, as indicated by reduced growth. Approximately 7 to 8 weeks after germination, the resistance was broken and plants developed typical wt-TMV symptoms, with high accumulation of the viral RNAs and proteins. The DeltaCP-TMV plants likewise were initially resistant to the endogenous inoculum and were stunted, although to a lesser extent than the wt-TMV plants. The resistance was broken at the same time as in the wt-TMV plants, but the mutant replicated to much lower levels and produced much milder symptoms than the wt virus. TMV-specific small interfering RNAs as well as increased transgene methylation were detected in the plants only after the resistance break, indicating that the resistance in the young plants was not due to RNA silencing.

  18. Expression of bacterial chloramphenicol acetyltransferase gene in tobacco plants mediated by TMV-RNA

    OpenAIRE

    Takamatsu, Nobuhiko; Ishikawa, Masayuki; Meshi, Tetsuo; Okada, Yoshimi

    1987-01-01

    We have constructed three tobacco mosaic virus (TMV) cDNA derivatives by modification of the full-length cDNA clone from which infectious TMV-RNA can be transcribed in vitro. A coatless TMV construct lacks most of the coat protein gene and chimeric TMV constructs retain the bacterial chloramphenicol acetyltransferase (CAT) gene in place of the coat protein gene. When in vitro transcripts from these cDNA derivatives were inoculated on the local lesion tobacco plants, TMV-specific lesions were ...

  19. Novel AroA from Pseudomonas putida Confers Tobacco Plant with High Tolerance to Glyphosate

    Science.gov (United States)

    Yan, Hai-Qin; Chang, Su-Hua; Tian, Zhe-Xian; Zhang, Le; Sun, Yi-Cheng; Li, Yan; Wang, Jing; Wang, Yi-Ping

    2011-01-01

    Glyphosate is a non-selective broad-spectrum herbicide that inhibits 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS, also designated as AroA), a key enzyme in the aromatic amino acid biosynthesis pathway in microorganisms and plants. Previously, we reported that a novel AroA (PpAroA1) from Pseudomonas putida had high tolerance to glyphosate, with little homology to class I or class II glyphosate-tolerant AroA. In this study, the coding sequence of PpAroA1 was optimized for tobacco. For maturation of the enzyme in chloroplast, a chloroplast transit peptide coding sequence was fused in frame with the optimized aroA gene (PparoA1optimized) at the 5′ end. The PparoA1optimized gene was introduced into the tobacco (Nicotiana tabacum L. cv. W38) genome via Agrobacterium-mediated transformation. The transformed explants were first screened in shoot induction medium containing kanamycin. Then glyphosate tolerance was assayed in putative transgenic plants and its T1 progeny. Our results show that the PpAroA1 from Pseudomonas putida can efficiently confer tobacco plants with high glyphosate tolerance. Transgenic tobacco overexpressing the PparoA1optimized gene exhibit high tolerance to glyphosate, which suggest that the novel PpAroA1 is a new and good candidate applied in transgenic crops with glyphosate tolerance in future. PMID:21611121

  20. [Soil nutrients spatial variability and soil fertility suitability in Qujing tobacco-planting area].

    Science.gov (United States)

    Li, Qiang; Zhou, Ji-heng; Yang, Rong-sheng; Zhang, Zheng-yan; Xie, Yan; Zhang, Yi-yang; Huang, Kua-ke; Li, Wei

    2011-04-01

    By adopting GPS technique, 2088 sampling sites were installed in the tobacco-planting area of Qujing City, Yunnan Province, with 0-20 cm soil samples collected to determine their main nutrients contents. The overall characteristics and spatial variability of the tobacco soil nutrients were analyzed by classic statistics and geo-statistics, and the soil fertility suitability in planting tobacco was evaluated by the methods of fuzzy mathematics. In the study area, soil pH and soil organic matter, available S, and water-soluble Cl contents were appropriate, soil total N and alkalihydrolyzable N contents were too high, soil available K, Ca, Mg, Cu, Fe, Zn, Mo, and Mn contents were abundant, soil available P content was at medium level, while soil total P and K and available B contents were insufficient. All the nutrient indices presented anisotropic distribution, among which, the spatial variability of soil available P and B was mainly caused by random factors, and that of other nutrients was caused by the co-effects of structural and random factors. The spatial distribution map of soil fertility suitability index (SFI) showed that there was no the excellent grade region for tobacco-planting, good grade region accounted for 8.0%, general grade region accounted for 51.6%, moderate grade region accounted for 39.0%, and low grade region accounted for 1.4%.

  1. Modifying plants for biofuel and biomaterial production.

    Science.gov (United States)

    Furtado, Agnelo; Lupoi, Jason S; Hoang, Nam V; Healey, Adam; Singh, Seema; Simmons, Blake A; Henry, Robert J

    2014-12-01

    The productivity of plants as biofuel or biomaterial crops is established by both the yield of plant biomass per unit area of land and the efficiency of conversion of the biomass to biofuel. Higher yielding biofuel crops with increased conversion efficiencies allow production on a smaller land footprint minimizing competition with agriculture for food production and biodiversity conservation. Plants have traditionally been domesticated for food, fibre and feed applications. However, utilization for biofuels may require the breeding of novel phenotypes, or new species entirely. Genomics approaches support genetic selection strategies to deliver significant genetic improvement of plants as sources of biomass for biofuel manufacture. Genetic modification of plants provides a further range of options for improving the composition of biomass and for plant modifications to assist the fabrication of biofuels. The relative carbohydrate and lignin content influences the deconstruction of plant cell walls to biofuels. Key options for facilitating the deconstruction leading to higher monomeric sugar release from plants include increasing cellulose content, reducing cellulose crystallinity, and/or altering the amount or composition of noncellulosic polysaccharides or lignin. Modification of chemical linkages within and between these biomass components may improve the ease of deconstruction. Expression of enzymes in the plant may provide a cost-effective option for biochemical conversion to biofuel.

  2. Comparative analysis of the plant mRNA-destabilizing element, DST, in mammalian and tobacco cells.

    Science.gov (United States)

    Feldbrügge, M; Arizti, P; Sullivan, M L; Zamore, P D; Belasco, J G; Green, P J

    2002-05-01

    The labile SAUR transcripts from higher plants contain a conserved DST sequence in their 3'-untranslated regions. Two copies of a DST sequence from soybean are sufficient to destabilize reporter transcripts in cultured tobacco cells whereas variants bearing mutations in the conserved ATAGAT or GTA regions are inactive. To investigate the potential for conserved recognition components in mammalian and plant cells, we examined the function of this instability determinant in mouse NIH3T3 fibroblasts and tobacco BY2 cells. In fibroblasts, a tetrameric DST element from soybean accelerated deadenylation and decay of a reporter transcript. However, a version mutated in the ATAGAT region was equally effective in this regard, and a tetrameric DST element from Arabidopsis was inactive. In contrast, the soybean DST element was more active as an mRNA instability element than the mutant version and the Arabidopsis element, when tested as tetramers in tobacco cells. Hence, the plant DST element is not recognized in animal cells with the same sequence requirements as in plant cells. Therefore, its mode of recognition appears to be plant-specific.

  3. 3-D nasal cultures: Systems toxicological assessment of a candidate modified-risk tobacco product.

    Science.gov (United States)

    Iskandar, Anita R; Mathis, Carole; Martin, Florian; Leroy, Patrice; Sewer, Alain; Majeed, Shoaib; Kuehn, Diana; Trivedi, Keyur; Grandolfo, Davide; Cabanski, Maciej; Guedj, Emmanuel; Merg, Celine; Frentzel, Stefan; Ivanov, Nikolai V; Peitsch, Manuel C; Hoeng, Julia

    2017-01-01

    In vitro toxicology approaches have evolved from a focus on molecular changes within a cell to understanding of toxicity-related mechanisms in systems that can mimic the in vivo environment. The recent development of three dimensional (3-D) organotypic nasal epithelial culture models offers a physiologically robust system for studying the effects of exposure through inhalation. Exposure to cigarette smoke (CS) is associated with nasal inflammation; thus, the nasal epithelium is relevant for evaluating the pathophysiological impact of CS exposure. The present study investigated further the application of in vitro human 3-D nasal epithelial culture models for toxicological assessment of inhalation exposure. Aligned with 3Rs strategy, this study aimed to explore the relevance of a human 3-D nasal culture model to assess the toxicological impact of aerosols generated from a candidate modified risk tobacco product (cMRTP), the Tobacco Heating System (THS) 2.2, as compared with smoke generated from reference cigarette 3R4F. A series of experimental repetitions, where multiple concentrations of THS2.2 aerosol and 3R4F smoke were applied, were conducted to obtain reproducible measurements to understand the cellular/molecular changes that occur following exposure. In agreement with "Toxicity Testing in the 21st Century - a Vision and a Strategy", this study implemented a systems toxicology approach and found that for all tested concentrations the impact of 3R4F smoke was substantially greater than that of THS2.2 aerosol in terms of cytotoxicity levels, alterations in tissue morphology, secretion of pro-inflammatory mediators, impaired ciliary function, and increased perturbed transcriptomes and miRNA expression profiles.

  4. Plant Cell Division Analyzed by Transient Agrobacterium-Mediated Transformation of Tobacco BY-2 Cells.

    Science.gov (United States)

    Buschmann, Henrik

    2016-01-01

    The continuing analysis of plant cell division will require additional protein localization studies. This is greatly aided by GFP-technology, but plant transformation and the maintenance of transgenic lines can present a significant technical bottleneck. In this chapter I describe a method for the Agrobacterium-mediated genetic transformation of tobacco BY-2 cells. The method allows for the microscopic analysis of fluorescence-tagged proteins in dividing cells in within 2 days after starting a coculture. This transient transformation procedure requires only standard laboratory equipment. It is hoped that this rapid method would aid researchers conducting live-cell localization studies in plant mitosis and cytokinesis.

  5. The quantitative soil quality assessment tobacco plant in Sindoro mountainous zone

    Directory of Open Access Journals (Sweden)

    Supriyadi

    2014-04-01

    Full Text Available The long-term cultivation of tobacco (Nicotiana tabacum plant in the Sindoro mountainous zone of Central Java has resulted in soil quality degradation that could affect economic development in the region if sustainable production practices are not identified. The objective of the study was to identify appropriate indicators for assessing soil quality on tobacco plant. The quantitative soil quality indicators were total organic-C, pH, available P and available K (chemical, soil depth, bulk density, AWC (available water capacity and soil aggregate stability (physical, and qCO2 (soil respiration, MBC (microbial biomass carbon (biological. The decreases in the soil aggregate stability, available water capacity, cation exchange capacity, soil respiration, microbial biomass carbon and total organic-C; or increases in bulk density (compaction, available P, available K and total nitrogen indicated the decrease in soil quality due to long-term tobacco production. The result of this research showed that the change of soil quality had occurred in Sindoro Mountain. The Soil Quality Index (SQI for three land use systems in Sindoro mountain (forest, mixed farm, and tobacco were 0.60, 0.47, and 0.57, respectively. The comparison of these rates with soil quality classes showed that the soil quality presented moderate to good level of quality; class SQI.

  6. Effect of salicylic acid on tobacco (Nicotiana rustica plant under drought conditions

    Directory of Open Access Journals (Sweden)

    Ghader Habibi

    2015-09-01

    Full Text Available Drought stress impact photosynthesis and stomatal conductance, and may reduce the overall production capacity of plants. Since exogenous application of salicylic acid (SA can partially alleviatebe increased the negative effects tolerance of drought stress by improve the metabolism pathways and increase the net photosynthesison plant photosynthesis and metabolism, the main objective of this study was to clarify the roles of SA in enhancing 28 days tobacco (Nicotiana rustica Basmas tolerance to drought stress (50% FC. The results indicated that foliar application of SA (0.5 mM influenced negatively net CO2 assimilation rate and stomatal conductance and led to reduction of shoot and root dry masses. In contrast, the stress did not reduce significantly the maximal quantum yield of photosystem II (PSII. This that can be explained by enhancement of efficiency for dissipation of excess photon energy in the PSII antenna, determined as non-photochemical quenching, and consequently further protection of PSII from photodamage. Thus, under more drought stress, the reduction of photosynthesis of tobacco plants was due mainly to reduction of stomatal conductance. Under water-deficient conditions, plants showed an increase in chlorophyll a and amino acids concentrations in the leaves when treated with SA while this change for net photosynthesis was negligible. Our results indicated showed that the foliar application of SA had no ameliorative effect on tobacco growth under drought stress, because its effect on elevation of transpiration rate did not increase net photosynthesis under drought condition.

  7. Transgenic tobacco revealing altered bacterial diversity in the rhizosphere during early plant development.

    Science.gov (United States)

    Andreote, Fernando D; Mendes, Rodrigo; Dini-Andreote, Francisco; Rossetto, Priscilla B; Labate, Carlos A; Pizzirani-Kleiner, Aline A; van Elsas, Jan Dirck; Azevedo, João L; Araújo, Welington L

    2008-05-01

    The rhizosphere constitutes a complex niche that may be exploited by a wide variety of bacteria. Bacterium-plant interactions in this niche can be influenced by factors such as the expression of heterologous genes in the plant. The objective of this work was to describe the bacterial communities associated with the rhizosphere and rhizoplane regions of tobacco plants, and to compare communities from transgenic tobacco lines (CAB1, CAB2 and TRP) with those found in wild-type (WT) plants. Samples were collected at two stages of plant development, the vegetative and flowering stages (1 and 3 months after germination). The diversity of the culturable microbial community was assessed by isolation and further characterization of isolates by amplified ribosomal RNA gene restriction analysis (ARDRA) and 16S rRNA sequencing. These analyses revealed the presence of fairly common rhizosphere organisms with the main groups Alphaproteobacteria, Betaproteobacteria, Actinobacteria and Bacilli. Analysis of the total bacterial communities using PCR-DGGE (denaturing gradient gel electrophoresis) revealed that shifts in bacterial communities occurred during early plant development, but the reestablishment of original community structure was observed over time. The effects were smaller in rhizosphere than in rhizoplane samples, where selection of specific bacterial groups by the different plant lines was demonstrated. Clustering patterns and principal components analysis (PCA) were used to distinguish the plant lines according to the fingerprint of their associated bacterial communities. Bands differentially detected in plant lines were found to be affiliated with the genera Pantoea, Bacillus and Burkholderia in WT, CAB and TRP plants, respectively. The data revealed that, although rhizosphere/rhizoplane microbial communities can be affected by the cultivation of transgenic plants, soil resilience may be able to restore the original bacterial diversity after one cycle of plant

  8. Is Chloroplast Movement in Tobacco Plants Influenced Systemically after Local Illumination or Burning Stress?

    Institute of Scientific and Technical Information of China (English)

    Jan Naus; Monika Rolencova; Vladimira Hlavackova

    2008-01-01

    Chloroplast movement has been studied In many plants mainly in relation to the local light, mechanical or stress effects. Here we investigated possible systemic responses of chloroplast movement to local light or burning stress in tobacco plants (Nicotiana tabacum cv. Samsun). Chloroplast movement was measured using two independent methods: one with a SPAD 502 Chlorophyll meter and another by collimated transmittance at a selected wavelength (676 nm). A sensitive pedodic movement of chloroplasts was used in high or low (2 000 or 50 μmol/m2 per s photosynthetically active radiation, respectively) cold white light with periods of 50 or 130 min. Measurements were carried out in the irradiated area, in the non-irradiated area of the same leaf or in the leaf located on the stem below the irradiated or burned one. No significant changes in systemic chloroplast movement in non-irradiated parts of the leaf and in the non-treated leaf were detected. Our data indicate that chloroplast movement in tobacco is dependent dominantly on the intensity and spectral composition of the incident light and on the local stimulation and state of the target tissue. No systemic signal was strong enough tovoke a detectable systemic response in chloroplast movement in distant untreated tissues of tobacco plants.

  9. Is chloroplast movement in tobacco plants influenced systemically after local illumination or burning stress?

    Science.gov (United States)

    Naus, Jan; Rolencová, Monika; Hlavácková, Vladimíra

    2008-10-01

    Chloroplast movement has been studied in many plants mainly in relation to the local light, mechanical or stress effects. Here we investigated possible systemic responses of chloroplast movement to local light or burning stress in tobacco plants (Nicotiana tabacum cv. Samsun). Chloroplast movement was measured using two independent methods: one with a SPAD 502 Chlorophyll meter and another by collimated transmittance at a selected wavelength (676 nm). A sensitive periodic movement of chloroplasts was used in high or low (2 000 or 50 micromol/m(2) per s photosynthetically active radiation, respectively) cold white light with periods of 50 or 130 min. Measurements were carried out in the irradiated area, in the non-irradiated area of the same leaf or in the leaf located on the stem below the irradiated or burned one. No significant changes in systemic chloroplast movement in non-irradiated parts of the leaf and in the non-treated leaf were detected. Our data indicate that chloroplast movement in tobacco is dependent dominantly on the intensity and spectral composition of the incident light and on the local stimulation and state of the target tissue. No systemic signal was strong enough to evoke a detectable systemic response in chloroplast movement in distant untreated tissues of tobacco plants.

  10. Ammonium formation and assimilation in P(SARK)∷IPT tobacco transgenic plants under low N.

    Science.gov (United States)

    Rubio-Wilhelmi, María del Mar; Sanchez-Rodriguez, Eva; Rosales, Miguel Angel; Blasco, Begoña; Rios, Juan Jose; Romero, Luis; Blumwald, Eduardo; Ruiz, Juan Manuel

    2012-01-15

    Wild Type (WT) and transgenic tobacco plants expressing isopentenyltransferase (IPT), a gene encoding the enzyme regulating the rate-limiting step in cytokinins (CKs) synthesis, were grown under limited nitrogen (N) conditions. We analyzed nitrogen forms, nitrogen metabolism related-enzymes, amino acids and photorespiration related-enzymes in WT and P(SARK)∷IPT tobacco plants. Our results indicate that the WT plants subjected to N deficiency displayed reduced nitrate (NO₃⁻) assimilation. However, an increase in the production of ammonium (NH₄⁺), by the degradation of proteins and photorespiration led to an increase in the glutamine synthetase/glutamate synthase (GS/GOGAT) cycle in WT plants. In these plants, the amounts of amino acids decreased with N deficiency, although the relative amounts of glutamate and glutamine increased with N deficiency. Although the transgenic plants expressing P(SARK)∷IPT and growing under suboptimal N conditions displayed a significant decline in the N forms in the leaf, they maintained the GS/GOGAT cycle at control levels. Our results suggest that, under N deficiency, CKs prevented the generation and assimilation of NH₄⁺ by increasing such processes as photorespiration, protein degradation, the GS/GOGAT cycle, and the formation of glutamine.

  11. Tobacco mosaic virus infection results in an increase in recombination frequency and resistance to viral, bacterial, and fungal pathogens in the progeny of infected tobacco plants.

    Science.gov (United States)

    Kathiria, Palak; Sidler, Corinne; Golubov, Andrey; Kalischuk, Melanie; Kawchuk, Lawrence M; Kovalchuk, Igor

    2010-08-01

    Our previous experiments showed that infection of tobacco (Nicotiana tabacum) plants with Tobacco mosaic virus (TMV) leads to an increase in homologous recombination frequency (HRF). The progeny of infected plants also had an increased rate of rearrangements in resistance gene-like loci. Here, we report that tobacco plants infected with TMV exhibited an increase in HRF in two consecutive generations. Analysis of global genome methylation showed the hypermethylated genome in both generations of plants, whereas analysis of methylation via 5-methyl cytosine antibodies demonstrated both hypomethylation and hypermethylation. Analysis of the response of the progeny of infected plants to TMV, Pseudomonas syringae, or Phytophthora nicotianae revealed a significant delay in symptom development. Infection of these plants with TMV or P. syringae showed higher levels of induction of PATHOGENESIS-RELATED GENE1 gene expression and higher levels of callose deposition. Our experiments suggest that viral infection triggers specific changes in progeny that promote higher levels of HRF at the transgene and higher resistance to stress as compared with the progeny of unstressed plants. However, data reported in these studies do not establish evidence of a link between recombination frequency and stress resistance.

  12. Regeneration of fertile plants from isolated tobacco zygotes by in vitro culture

    Institute of Scientific and Technical Information of China (English)

    HE Yuchi; SUN Mengxiang; YANG Hongyuan

    2004-01-01

    Living zygotes of tobacco (Nicotiana Tabacum L.) SR-1 were isolated and cultured in vitro by the microculture technique. Fertile plants were regenerated from the calli derived from cultured zygotes via organogenesis. Ovules were collected 120 h after pollination and used as feeder. MS combined with KM8p was selected as basic medium in the experiment. Zygotes isolated from ovules 108 h after pollination turned out to be suitable material for in vitro culture. Over 80% such zygotes could divide and around 10% of them could grow into calli and regenerate fertile plants.

  13. [Construction of plant expression vectors harboring a peptide antibiotic-apidaecin gene and resistance analysis of the transgenic tobacco].

    Science.gov (United States)

    Wang, H; Sun, C; Peng, X X

    2001-07-01

    Two plant expression vectors(pBinPRHbI and pBinPRSIHbI) were constructed: Firstly, apidaecin gene were fused to the signal peptide coding sequencing of a PR-protein, and cloned into a binary vector pBin438 to form pBinPRHbI. Then, the cassette consisting of 35S promoter, PR signal peptide coding sequencing and apidaecin gene was cut off from pBinPRHbI and inserted into another plant expression vector pBinPRSI to produce a bivalent plant expression vector pBinPRSIHbI. pBinPRSI was constructed previously in our lab and contained PR signal peptide and Shiva-I fusion gene under control of 35S promoter. The three plant expression vectors were introduced into tobacco by Agrobacterium-mediated transformation. The positive rate of PCR was 95% in all putative transgenic plants. Results from Southern blot indicated that foreign genes were integrated into tobacco genome and RT-PCR analysis proved that the foreign gene was transcribed in transgenic tobacco. The transgenic tobacco showed higher resistance to P. syringae pv tabaci, the causal agent of tobacco wild fire disease, than their original cultivars. From the disease index, the transgenic plants carrying apidaecin and Shiva-I genes had highest resistance among three kinds of transgenic plants, and the plants carrying Shiva-I gene alone had lowest resistance.

  14. Principal Component Analysis of Chlorophyll Content in Tobacco, Bean and Petunia Plants Exposed to Different Tropospheric Ozone Concentrations

    Directory of Open Access Journals (Sweden)

    Borowiak Klaudia

    2014-06-01

    Full Text Available Three plant species were assessed in this study - ozone-sensitive and -resistant tobacco, ozone-sensitive petunia and bean. Plants were exposed to ambient air conditions for several weeks in two sites differing in tropospheric ozone concentrations in the growing season of 2009. Every week chlorophyll contents were analysed. Cumulative ozone effects on the chlorophyll content in relation to other meteorological parameters were evaluated using principal component analysis, while the relation between certain days of measurements of the plants were analysed using multivariate analysis of variance. Results revealed variability between plant species response. However, some similarities were noted. Positive relations of all chlorophyll forms to cumulative ozone concentration (AOT 40 were found for all the plant species that were examined. The chlorophyll b/a ratio revealed an opposite position to ozone concentration only in the ozone-resistant tobacco cultivar. In all the plant species the highest average chlorophyll content was noted after the 7th day of the experiment. Afterwards, the plants usually revealed various responses. Ozone-sensitive tobacco revealed decrease of chlorophyll content, and after few weeks of decline again an increase was observed. Probably, due to the accommodation for the stress factor. While during first three weeks relatively high levels of chlorophyll contents were noted in ozone-resistant tobacco. Petunia revealed a slow decrease of chlorophyll content and the lowest values at the end of the experiment. A comparison between the plant species revealed the highest level of chlorophyll contents in ozone-resistant tobacco.

  15. TcCYS4, a cystatin from cocoa, reduces necrosis triggered by MpNEP2 in tobacco plants.

    Science.gov (United States)

    Santana, L S; Costa, M G C; Pirovani, N M; Almeida, A F; Alvim, F C; Pirovani, C P

    2014-09-26

    In Brazil, most cocoa bean production occurs in Southern Bahia. Witches' broom disease arrived in this area in 1989 and has since caused heavy losses in production. The disease is caused by the basidiomycete fungus Moniliophthora perniciosa, a hemibiotrophic fungus that produces the necrosis and ethylene-inducting protein (MpNEP2) during infection; this protein can activate cysteine proteases and induce programmed cell death. Cysteine proteases can be modulated by cystatin. In this study, we overexpressed TcCYS4, a cocoa cystatin, in tobacco plants and evaluated the effect on MpNEP2 in model plants. Tccys4 cDNA was cloned into the pCAMBIA 1390 vector and inserted into the tobacco plants via Agrobacterium tumefaciens. Transgene expression was analyzed by reverse transcription-quantitative PCR and Western blot analysis. Transcript and protein levels in Tcccys4:tobacco lines were 8.9- and 1.5-fold higher than in wild-type plants (wt). Tcccys4:tobacco lines showed no change in growth compared to wt plants. CO2 net assimilation (A) increased in Tcccys4:tobacco lines compared to wt plants. Only one line showed statistically significant stomatal conductance (gs) and transpiration rate (E) changes. MpNEP2 was infiltered into the foliar mesophyll of Tcccys4:tobacco lines and wt plants, and necrotic lesions were attenuated in lines highly expressing Tccys4. Our results suggest that cocoa cystatin TcCYS4 affects MpNEP2 activity related to the progression of programmed cell death in tobacco plants. This may occur through the action of cystatin to inhibit cysteine proteases activated by MpNEP2 in plant tissues. Further studies are necessary to examine cystatin in the Theobroma cacao-M. perniciosa pathosystem.

  16. Characterization of AtSTOP1 orthologous genes in tobacco and other plant species.

    Science.gov (United States)

    Ohyama, Yoshinao; Ito, Hiroki; Kobayashi, Yuriko; Ikka, Takashi; Morita, Akio; Kobayashi, Masatomo; Imaizumi, Ryujiro; Aoki, Toshio; Komatsu, Kenji; Sakata, Yoichi; Iuchi, Satoshi; Koyama, Hiroyuki

    2013-08-01

    Aluminum (Al) and proton (H⁺) tolerances are essential traits for plants to adapt to acid soil environments. In Arabidopsis (Arabidopsis thaliana), these tolerances are mediated by a zinc-finger transcription factor, SENSITIVE TO PROTON RHIZOTOXICITY1 (AtSTOP1), which regulates the transcription of multiple genes critical for tolerance to both stressors. Here, the functions of orthologous proteins (STOP1-like proteins) in other plant species were characterized by reverse genetics analyses and in planta complementation assays. RNA interference of a gene for NtSTOP1 repressed Al and H⁺ tolerances of tobacco (Nicotiana tabacum) roots. Tobacco roots released citrate in response to Al, concomitant with the up-regulated transcription of an ortholog of an Al tolerance gene encoding a citrate-transporting multidrug and toxic compound extrusion protein. The RNA interference repression of NtSTOP1 blocked this process and also repressed the transcription of another orthologous gene for Al tolerance, ALUMINUM SENSITIVE3, which encodes a prokaryote-type transporter. These results demonstrated that NtSTOP1 regulates Al tolerance in tobacco through the transcriptional regulation of these genes. The in planta complementation assays revealed that other plant species, including woody plants, a legume, and a moss (Physcomitrella patens), possess functional STOP1-like proteins that can activate several H⁺ and Al-tolerance genes in Arabidopsis. Knocking out the gene encoding the STOP1-like protein decreased the Al tolerance of P. patens. Together, our results strongly suggest that transcriptional regulation by STOP1-like proteins is evolutionarily conserved among land plants and that it confers the ability to survive in acid soils through the transcriptional regulation of Al- and H⁺-tolerance genes.

  17. [Safety assessment of foods derived from genetically modified plants].

    Science.gov (United States)

    Pöting, A; Schauzu, M

    2010-06-01

    The placing of genetically modified plants and derived food on the market falls under Regulation (EC) No. 1829/2003. According to this regulation, applicants need to perform a safety assessment according to the Guidance Document of the Scientific Panel on Genetically Modified Organisms of the European Food Safety Authority (EFSA), which is based on internationally agreed recommendations. This article gives an overview of the underlying legislation as well as the strategy and scientific criteria for the safety assessment, which should generally be based on the concept of substantial equivalence and carried out in relation to an unmodified conventional counterpart. Besides the intended genetic modification, potential unintended changes also have to be assessed with regard to potential adverse effects for the consumer. All genetically modified plants and derived food products, which have been evaluated by EFSA so far, were considered to be as safe as products derived from the respective conventional plants.

  18. The symptom difference induced by Tobacco mosaic virus and Tomato mosaic virus in tobacco plants containing the N gene is determined by movement protein gene

    Institute of Scientific and Technical Information of China (English)

    YU; Cui; HU; Dongwei; DONG; Jiahong; CUI; Xiaofeng; WU; Jun

    2004-01-01

    Tobacco mosaic virus (TMV) and Tomato mosaic virus (ToMV) are two closely related viruses in the genus Tobamovirus, but they induce obviously different sizes of necrotic lesions in tobacco plants containing the N gene. Comparison of the symptoms produced by TMV, ToMV and a chimaeric virus (T/OMP), in which the TMV movement protein (MP) gene was replaced by the ToMV MP gene, showed T/OMP caused necrotic lesions that were similar in size to those of ToMV in tobacco plants containing the N gene. The coat protein and MP of the three viruses accumulated in planta with similar levels, and the replication level of TMV and T/OMP in protoplasts also had no difference. Comparison of the activities of defense-related enzymes (PAL, POD and PPO) induced by the three viruses also showed that the variability of enzyme activity induced by T/OMP was similar to that induced by TMV, but different from that induced by ToMV. The results indicate that the size difference of necrotic lesions induced by TMV and ToMV in tobacco plants containing the N gene results from the functional difference of their MP genes.

  19. Shoot Apex Demand Determines Assimilate and Nutrients Partitioning and Nutrient-uptake Rate in Tobacco Plants

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Our previous experiment revealed that apex-removed plants have larger root systems but a lower K+-uptake rates than intact tobacco plants.Since the apex is not only e center of growth and metabolism,but also an important place of auxin synthesis and export,the aims of this study were to distinguish whether the apex demand or auxin synthesized in the apex regulates assimilate and nutrients partitioning within plant,and to explain the reason for the lower K+-uptake rate of the apex-ramoved plant.In comparison with the control plant,covering the shoot apex with a black transparent plastic bag reduced net increases In dry matter and nutrients;however,the distribution of the dry matter and nutrients between shoot and roots and nutrient-uptake rates were not changed.Removal of the shoot apex shifted the dry mass and nutrients distributions to roots,and reduced the rate of nutrient uptake.Application of 1-naphthylacetic acid(NAA) could partly replace the role of the removed apex,stimulated assimilate and nutrient deposition into the treated tissue,and enhanced the reduced plasma membrane ATPase activity of roots to the control level.However,treatment of the apex-removed plants with NAA could not rescue the reduced nutrient uptake rate and the shifted assimilates and nutrients partitioning caused by excision of the apex.Higher nutrient uptake rate of the intact plants could not be explained by root growth parameters,such as total root surface area and number of root tips.The results from the present study indicate that strong apex demand determined assimilatas and nutrients partitioning and nutrient-uptake rate in tobacco(Nicotiana tabacum)plants.

  20. Environmental tobacco exposure is associated with vaccine modified measles in junior high school students.

    Science.gov (United States)

    Suzuki, Shuichi; Sato, Kazuki; Watanabe, Hiroko; Nezu, Yoko; Nishimuta, Toshiyuki

    2015-11-01

    Vaccine modified measles (VMM) affects individuals with attenuated vaccine induced immunity. An outbreak of measles occurred in a junior high school, starting from an unvaccinated eighth-grade student who developed natural measles and affected a majority of students who were immunized with a low potent strain of measles vaccine (TD97). To determine whether environmental tobacco smoke (ETS) exposure was associated with the development of VMM in this population, a questionnaire was used asking whether students had VMM symptoms during the outbreak and the smoking status of family members. VMM was defined in the study population as occurrence of fever and/or erythema, along with documented history of measles vaccination. A total of 513 students (85.9%) responded. Overall, the presence of in-house smokers did not differ between VMM students (49.3%) and non-VMM students (50.2%). However, in the ninth grade, presence of an in-house smoker was significantly higher in the family of VMM students (54.0%) than in non-VMM students (36.6%) (P = 0.044). Urinary cotinine levels were also measured in selected students (n = 37). Among families with at least one smoker, urinary cotinine levels were significantly higher in VMM students than in non-VMM students (P = 0.032). Furthermore, a multivariable logistic regression analysis showed that a high urinary cotinine level (>10 ng/mg creatinine; 13.5 percentile) was associated with the development of VMM. Our findings suggest that a high level of ETS exposure may be associated with an increased risk of VMM in a population with attenuated vaccine induced immunity against measles.

  1. Ozone response of tomato plants infected with cucumber mosaic virus and/or tobacco mosaic virus

    Energy Technology Data Exchange (ETDEWEB)

    Ormrod, D.P.; Kemp, W.G.

    1979-10-01

    The sensitivity of three tomato cultivars to several concentrations of ozone was evaluated after prior sequential inoculations with tobacco mosaic virus (TMV) and/or cucumber mosaic virus (CMV). Ozone injury in inoculated and uninoculated tomatoes varied from slight to severe depending on the virus, cultivar, ozone concentration and virus incubation period. The frequency of increased ozone injury was about twice as great as that of suppressed injury on infected plants. Ozone injury occurred more frequently in TMV-inoculated plants than in those inoculated with CMV. There were more increases than decreases in ozone injury after 7 or 14 days of virus infection, but mainly decreases in injury after 21 days infection. Growth was significantly reduced in plants exposed to ozone after a 21-day virus incubation period, particularly when they were inoculated with both viruses.

  2. From pesticides to genetically modified plants : history, economics and politics

    NARCIS (Netherlands)

    Zadoks, J.C.; Waibel, H.

    2000-01-01

    Two technologies of crop protection are compared, crop protection by pesticides and by Genetically Modified Plants (GMPs). The history of pesticides provides lessons relevant to the future of GMPs; (1) high pesticide usage is counter-productive, (2) the technology requires intensive regulation and

  3. From pesticides to genetically modified plants : history, economics and politics

    NARCIS (Netherlands)

    Zadoks, J.C.; Waibel, H.

    2000-01-01

    Two technologies of crop protection are compared, crop protection by pesticides and by Genetically Modified Plants (GMPs). The history of pesticides provides lessons relevant to the future of GMPs; (1) high pesticide usage is counter-productive, (2) the technology requires intensive regulation and (

  4. Overexpression of the Synthetic Chimeric Native-T-phylloplanin-GFP Genes Optimized for Monocot and Dicot Plants Renders Enhanced Resistance to Blue Mold Disease in Tobacco (N. tabacum L.

    Directory of Open Access Journals (Sweden)

    Dipak K. Sahoo

    2014-01-01

    Full Text Available To enhance the natural plant resistance and to evaluate the antimicrobial properties of phylloplanin against blue mold, we have expressed a synthetic chimeric native-phylloplanin-GFP protein fusion in transgenic Nicotiana tabacum cv. KY14, a cultivar that is highly susceptible to infection by Peronospora tabacina. The coding sequence of the tobacco phylloplanin gene along with its native signal peptide was fused with GFP at the carboxy terminus. The synthetic chimeric gene (native-phylloplanin-GFP was placed between the modified Mirabilis mosaic virus full-length transcript promoter with duplicated enhancer domains and the terminator sequence from the rbcSE9 gene. The chimeric gene, expressed in transgenic tobacco, was stably inherited in successive plant generations as shown by molecular characterization, GFP quantification, and confocal fluorescent microscopy. Transgenic plants were morphologically similar to wild-type plants and showed no deleterious effects due to transgene expression. Blue mold-sensitivity assays of tobacco lines were performed by applying P. tabacina sporangia to the upper leaf surface. Transgenic lines expressing the fused synthetic native-phyllopanin-GFP gene in the leaf apoplast showed resistance to infection. Our results demonstrate that in vivo expression of a synthetic fused native-phylloplanin-GFP gene in plants can potentially achieve natural protection against microbial plant pathogens, including P. tabacina in tobacco.

  5. High levan accumulation in transgenic tobacco plants expressing the Gluconacetobacter diazotrophicus levansucrase gene.

    Science.gov (United States)

    Banguela, Alexander; Arrieta, Juan G; Rodríguez, Raisa; Trujillo, Luis E; Menéndez, Carmen; Hernández, Lázaro

    2011-06-10

    Bacterial levansucrase (EC 2.4.1.10) converts sucrose into non-linear levan consisting of long β(2,6)-linked fructosyl chains with β(2,1) branches. Bacterial levan has wide food and non-food applications, but its production in industrial reactors is costly and low yielding. Here, we report the constitutive expression of Gluconacetobacter diazotrophicus levansucrase (LsdA) fused to the vacuolar targeting pre-pro-peptide of onion sucrose:sucrose 1-fructosyltransferase (1-SST) in tobacco, a crop that does not naturally produce fructans. In the transgenic plants, levan with degree of polymerization above 10(4) fructosyl units was detected in leaves, stem, root, and flowers, but not in seeds. High levan accumulation in leaves led to gradual phenotypic alterations that increased with plant age through the flowering stage. In the transgenic lines, the fructan content in mature leaves varied from 10 to 70% of total dry weight. No oligofructans were stored in the plant organs, although the in vitro reaction of transgenic LsdA with sucrose yielded β(2,1)-linked FOS and levan. Transgenic lines with levan representing up to 30mgg(-1) of fresh leaf weight produced viable seeds and the polymer accumulation remained stable in the tested T1 and T2 progenies. The lsdA-expressing tobacco represents an alternative source of highly polymerized levan.

  6. Expression of the major mugwort pollen allergen Art v 1 in tobacco plants and cell cultures: problems and perspectives for allergen production in plants.

    Science.gov (United States)

    Siegert, Marc; Pertl-Obermeyer, Heidi; Gadermaier, Gabriele; Ferreira, Fatima; Obermeyer, Gerhard

    2012-03-01

    An economic and cheap production of large amounts of recombinant allergenic proteins might become a prerequisite for the common use of microarray-based diagnostic allergy assays which allow a component-specific diagnosis. A molecular pharming strategy was applied to express the major allergen of Artemisia vulgaris pollen, Art v 1, in tobacco plants and tobacco cell cultures. The original Art v 1 with its endogenous signal peptide which directs Art v 1 to the secretory pathway, was expressed in transiently transformed tobacco leaves but was lost in stable transformed tobacco plants during the alternation of generations. Using a light-regulated promoter and "hiding" the recombinant Art v 1 in the ER succeeded in expression of Art v 1 over three generations of tobacco plants and in cell cultures generated from stable transformed plants. However, the amounts of the recombinant allergen were sufficient for analysis but not high enough to allow an economic production. Although molecular pharming has been shown to work well for the production of non-plant therapeutic proteins, it might be less efficient for closely related plant proteins.

  7. An Arabidopsis mitochondrial uncoupling protein confers tolerance to drought and salt stress in transgenic tobacco plants.

    Directory of Open Access Journals (Sweden)

    Kevin Begcy

    Full Text Available BACKGROUND: Plants are challenged by a large number of environmental stresses that reduce productivity and even cause death. Both chloroplasts and mitochondria produce reactive oxygen species under normal conditions; however, stress causes an imbalance in these species that leads to deviations from normal cellular conditions and a variety of toxic effects. Mitochondria have uncoupling proteins (UCPs that uncouple electron transport from ATP synthesis. There is evidence that UCPs play a role in alleviating stress caused by reactive oxygen species overproduction. However, direct evidence that UCPs protect plants from abiotic stress is lacking. METHODOLOGY/PRINCIPAL FINDINGS: Tolerances to salt and water deficit were analyzed in transgenic tobacco plants that overexpress a UCP (AtUCP1 from Arabidopsis thaliana. Seeds of AtUCP1 transgenic lines germinated faster, and adult plants showed better responses to drought and salt stress than wild-type (WT plants. These phenotypes correlated with increased water retention and higher gas exchange parameters in transgenic plants that overexpress AtUCP1. WT plants exhibited increased respiration under stress, while transgenic plants were only slightly affected. Furthermore, the transgenic plants showed reduced accumulation of hydrogen peroxide in stressed leaves compared with WT plants. CONCLUSIONS/SIGNIFICANCE: Higher levels of AtUCP1 improved tolerance to multiple abiotic stresses, and this protection was correlated with lower oxidative stress. Our data support previous assumptions that UCPs reduce the imbalance of reactive oxygen species. Our data also suggest that UCPs may play a role in stomatal closure, which agrees with other evidence of a direct relationship between these proteins and photosynthesis. Manipulation of the UCP protein expression in mitochondria is a new avenue for crop improvement and may lead to crops with greater tolerance for challenging environmental conditions.

  8. Disease-tolerance of transgenic tobacco plants expressing Ah-AMP gene of Amaranthus hypochondriacus

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    An antimicrobial peptide gene from Amaranthus hypochondriacus, Ah-AMP, was amplified by PCR and cloned. Sequence analysis results revealed that this gene is 261 bp in length encoding a precursor polypeptide of 87 amino acid residues. Ah-AMP gene was inserted in the binary vector pBin438 to construct a plant expression vector pBinAH916. Leave explants of Nicotiana tabacum var. SR1 were transformed with Agrobacterium tumefaciens LBA4404 harboring the above expression vector. Results from PCR, Southern and Northern blot analyses confirmed that the Ah-AMP gene had been integrated into the tobacco genome and was transcribed at mRNA level. Two bacterial-resistant transgenic plants were selected by inoculating the plants with Pseudomonas solanacearum and statistic analysis of two T1 lines showed that the resistance increased by 2.24 and 1.62 grade and the disease index decreased by 49.6% and 37.3% respectively when compared with the non-transformed control plants SR1. The results from challenging the plants with inoculums of Phytophthora parasitica showed that the symptom development was delayed and disease index was significantly reduced. These results suggest that Ah-AMP gene may be a potentially valuable gene for genetic engineering of plant for disease-resistance.

  9. Exploration of horizontal gene transfer between transplastomic tobacco and plant-associated bacteria.

    Science.gov (United States)

    Demanèche, Sandrine; Monier, Jean-Michel; Dugat-Bony, Eric; Simonet, Pascal

    2011-10-01

    The likelihood of gene transfer from transgenic plants to bacteria is dependent on the transgene copy number and on the presence of homologous sequences for recombination. The large number of chloroplast genomes in a plant cell as well as the prokaryotic origin of the transgene may thus significantly increase the likelihood of gene transfer from transplastomic plants to bacteria. In order to assess the probability of such a transfer, bacterial isolates, screened for their ability to colonize decaying tobacco plant tissue and possessing DNA sequence similarity to the chloroplastic genes accD and rbcL flanking the transgene (aadA), were tested for their ability to take up extracellular DNA (broad host-range pBBR1MCS-3-derived plasmid, transplastomic plant DNA and PCR products containing the genes accD-aadA-rbcL) by natural or electrotransformation. The results showed that among the 16 bacterial isolates tested, six were able to accept foreign DNA and acquire the spectinomycin resistance conferred by the aadA gene on plasmid, but none of them managed to integrate transgenic DNA in their chromosome. Our results provide no indication that the theoretical gene transfer-enhancing properties of transplastomic plants cause horizontal gene transfer at rates above those found in other studies with nuclear transgenes.

  10. Chloride regulates leaf cell size and water relations in tobacco plants.

    Science.gov (United States)

    Franco-Navarro, Juan D; Brumós, Javier; Rosales, Miguel A; Cubero-Font, Paloma; Talón, Manuel; Colmenero-Flores, José M

    2016-02-01

    Chloride (Cl(-)) is a micronutrient that accumulates to macronutrient levels since it is normally available in nature and actively taken up by higher plants. Besides a role as an unspecific cell osmoticum, no clear biological roles have been explicitly associated with Cl(-) when accumulated to macronutrient concentrations. To address this question, the glycophyte tobacco (Nicotiana tabacum L. var. Habana) has been treated with a basal nutrient solution supplemented with one of three salt combinations containing the same cationic balance: Cl(-)-based (CL), nitrate-based (N), and sulphate+phosphate-based (SP) treatments. Under non-saline conditions (up to 5 mM Cl(-)) and no water limitation, Cl(-) specifically stimulated higher leaf cell size and led to a moderate increase of plant fresh and dry biomass mainly due to higher shoot expansion. When applied in the 1-5 mM range, Cl(-) played specific roles in regulating leaf osmotic potential and turgor, allowing plants to improve leaf water balance parameters. In addition, Cl(-) also altered water relations at the whole-plant level through reduction of plant transpiration. This was a consequence of a lower stomatal conductance, which resulted in lower water loss and greater photosynthetic and integrated water-use efficiency. In contrast to Cl(-), these effects were not observed for essential anionic macronutrients such as nitrate, sulphate, and phosphate. We propose that the abundant uptake and accumulation of Cl(-) responds to adaptive functions improving water homeostasis in higher plants.

  11. Composting of tobacco plant waste by manual turning and forced aeration system

    Directory of Open Access Journals (Sweden)

    Nonglak Saithep

    2009-05-01

    Full Text Available The efficiency of tobacco plant waste composting, by the manual turning and the forced aeration system, was compared. Tobacco plant waste, cow manure, urea fertiliser, and a compost inoculum mixture at a 100:10:0.2:0.01 ratio respectively, with 60% (w/v moisture content, were set up in piling forms. The piles of the manual turning system were provided with turning aeration by hand at intervals of 7 days during the composting process. For the forced aeration system, each pile was aerated by a 3-HP air pump with a flow rate of 19 litres min-1 for 15 minutes every morning and evening. The completely randomised design of turned and force-aerated piles was performed in triplicate. The composting activity of both systems during the composting period was measured by several parameters: temperature, pH, moisture content, C/N ratio, growth of microorganisms, cellulase activity, and nicotine degradation in the set-up piles. Both systems had similar temperature, pH, and moisture content conditions in the piles during the composting process. However, the forced aeration system was more advantageous for the growth of mesophilic and thermophilic microorganisms, for cellulase activity from cellulase-producing microorganisms, and for nicotine degradation, when compared to the manual turning system. In conclusion, the forced aeration system was more efficient than the manual turning system in composting and is a viable alternative method for the composting process.

  12. THE ACTIVITY OF ARABIDOSPIS DLL PROMOTER IN TRANSGENIC TOBACCO PLANTS UNDER WATER STRESS CONDITIONS

    Directory of Open Access Journals (Sweden)

    Zuzana Polóniová

    2014-02-01

    Full Text Available In this work we used the Cre/loxP recombination system to study the activity of the Arabidopsis DLL promoter under water stress treatment. For this, the T-DNA containing the Cre/loxP self-excision recombination cassette was introduced into tobacco genome via A. tumefaciens LBA 4404. The expression of the cre gene was regulated by the DLL promoter. On activity of the DLL the Cre recombinase was expected to remove Cre/loxP cassette. Transgenic nature of regenerated transgenic T0 tobacco plantlets was proved by GUS and PCR analyses. The selected 10 transgenic T0 plants were subjected to the water stress analyses under in vitro as well as under in vivo conditions. The osmotic stress experiments were performed with 10 % PEG and 100 mmol.l-1 mannitol (individually. The activity of the DLL was evaluated after 24 hours. For drought stress experiments, the watering was withheld for 10 days. The activity of the DLL was monitored using PCR approach. Under given abiotic stress conditions, no activity of the DLL was observed. The DLL promoter remained stable. It points out the DLL as the promoter with precise control of the gene expression with wide usability in plant biotechnology.

  13. Contribution of AM inoculation and cattle manure to lead and cadmium phytoremediation by tobacco plants.

    Science.gov (United States)

    Wang, Fa Yuan; Shi, Zhao Yong; Xu, Xiao Feng; Wang, Xu Gang; Li, You Jun

    2013-04-01

    Lead and cadmium are both highly toxic pollutants and pose potential risks to the environment and human health. Arbuscular mycorrhizal (AM) inoculation and organic amendments may make a potential contribution to phytoremediation of these toxic metals, but their effects remain unclear. We conducted a pot culture experiment to study the contribution of AM inoculation and/or cattle manure to phytoremediation of two soils artificially polluted with 0, 350, 500 and 1000 mg Pb per kg soil or 0, 1, 10, 100 mg Cd per kg soil using tobacco plants. Results showed that AM colonization was greatly reduced when exposed to more heavy metals especially Cd, whereas organic amendment alleviated metal stress and showed protective effects. In general, AM inoculation and cattle manure, singly or in combination, all significantly increased tobacco growth and Pb and Cd accumulation in shoots and roots, while decreased DTPA-extractable Pb and Cd concentrations in soil, and combination treatments (MN) produced the most pronounced positive effects. Improved plant P nutrition, higher soil pH and lower available metal concentrations contributed by AM inoculation and/or organic amendment may be the main strategies to alleviate metal toxicity and enhance phytoremediation efficiency. Our results indicate that AM fungi and organic manure play a synergistic positive role both in phytoextraction and phytostabilization of Cd and Pb.

  14. 77 FR 20026 - Draft Guidance for Industry: Modified Risk Tobacco Product Applications; Availability; Agency...

    Science.gov (United States)

    2012-04-03

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF HEALTH AND... or to reduce exposure to harmful substances associated with tobacco products, manufacturers be... any proposed advertising and labeling; The conditions for using the product; The formulation of the...

  15. Bioindication with tobacco plants and grass culture at a long-term station in the city of Munich, Germany

    Energy Technology Data Exchange (ETDEWEB)

    Koehler, J. [Schloss Steinenhausen, Kulmbach (Germany). Aussenstelle Nordbayern; Peichl, L. [LfU Augsburg (Germany)

    2002-07-01

    At Bavarian long-term stations, biomonitoring of ozone and metal air pollution is carried out with tobacco plants and standardised grass culture. The stations provide background data of the city of Munich and of typical rural regions. The percentage of damaged leaf area of tobacco plants is definitely lower at the city station than at the rural stations, because there is more reduction potential (NO{sub x}) in the city. In standardised grass culture, the contents of metals specific for traffic emissions are higher at the city station, whereas the accumulation of other metals is in the same range like at all background stations. (orig.)

  16. Suitable Region for Flue-cured Tobacco (Nicotiana glauca L.) Planting Based on Spatial Scene Similarity%Suitable Region for Flue-cured Tobacco (Nicotiana glauca L.) Planting Based on Spatial Scene Similarity

    Institute of Scientific and Technical Information of China (English)

    Junxiang DONG; Danhuai GUO; Xiaodong SHAO

    2012-01-01

    [Objective] The aim was to establish a model based on spatial scene similarity, for which soil, slope, transport, water conservancy, light, social economic factors in suitable planting areas were all considered. A new suitable planting area of flue-cured tobacco was determined by comparison and analysis, with consideration of excellent area. [Method] Totaling thirty natural factors were chosen, which were clas- sified into nine categories, from Longpeng Town (LP) and Shaochong Town (SC) in Shiping County in Honghe Hani and Yi Autonomous Prefecture. [Result] According to weights, the factors from high to low were as follows: soil〉light〉elevation〉slope〉 water conservancy〉transport〉baking facility〉planting plans over the years〉others. The similarity of geographical conditions in the area was 0.894 3, which indicated that the planting conditions in the two regions are similar. If farmer population in unit area, farmland quantity for individual farmer, labors in every household, activity in planting flue-cured tobacco and work of local instructor were considered, the weights of different factors were as follows: farmer population in unit area〉farmland quantity for individual farmer〉farmers' activity in planting flue-cured tobacco〉educational back- ground〉labor force in every household〉instructor〉population of farmers' children at- tending school. The similarity of geographical conditions was 0.703 1, which indicated that it is none-natural factors that influence yield and quality of flue-cured tobacco. [Conclusion] According to analysis on suitable planting area of flue-cured tobacco based on assessment of spatial scene similarity, similarity of growing conditions in two spatial scenes can be analyzed and evaluated, which would promote further exploration on, influencing factors and effects on tobacco production.

  17. Guidance on the environmental risk assessment of genetically modified plants

    DEFF Research Database (Denmark)

    Bartsch, Detlef; Chueca, Cristina; De-Schrijver, Adinda

    . This document describes the six steps for the ERA of GM plants, as indicated in Directive 2001/18/EC, starting with (1) problem formulation including hazard identification; (2) hazard characterisation; (3) exposure characterisation; (4) risk characterisation; (5) risk management strategies; and (6) an overall...... assessment; (5) impact of the specific cultivation, management and harvesting techniques; including consideration of the production systems and the receiving environment(s); (6) effects on biogeochemical processes; and (7) effects on human and animal health. Each specific area of concern is considered......This document provides guidance for the environmental risk assessment (ERA) of genetically modified (GM) plants submitted within the framework of Regulation (EC) No. 1829/2003 on GM food and feed or under Directive 2001/18/EC on the deliberate release into the environment of genetically modified...

  18. Effects of root-zone acidity on utilization of nitrate and ammonium in tobacco plants

    Science.gov (United States)

    Henry, L. T.; Raper, C. D. Jr; Raper CD, J. r. (Principal Investigator)

    1989-01-01

    Tobacco (Nicotiana tabacum L., cv. 'Coker 319') plants were grown for 28 days in flowing nutrient culture containing either 1.0 mM NO3- or 1.0 mM NH4+ as the nitrogen source in a complete nutrient solution. Acidities of the solutions were controlled at pH 6.0 or 4.0 for each nitrogen source. Plants were sampled at intervals of 6 to 8 days for determination of dry matter and nitrogen accumulation. Specific rates of NO3- or NH4+ uptake (rate of uptake per unit root mass) were calculated from these data. Net photosynthetic rates per unit leaf area were measured on attached leaves by infrared gas analysis. When NO3- [correction of NO-] was the sole nitrogen source, root growth and nitrogen uptake rate were unaffected by pH of the solution, and photosynthetic activity of leaves and accumulation of dry matter and nitrogen in the whole plant were similar. When NH4+ was the nitrogen source, photosynthetic rate of leaves and accumulation of dry matter and nitrogen in the whole plant were not statistically different from NO3(-) -fed plants when acidity of the solution was controlled at pH 6.0. When acidity for NH4(+) -fed plants was increased to pH 4.0, however, specific rate of NH4+ uptake decreased by about 50% within the first 6 days of treatment. The effect of acidity on root function was associated with a decreased rate of accumulation of nitrogen in shoots that was accompanied by a rapid cessation of leaf development between days 6 and 13. The decline in leaf growth rate of NH4(+) -fed plants at pH 4.0 was followed by reductions in photosynthetic rate per unit leaf area. These responses of NH4(+) -fed plants to increased root-zone acidity are characteristic of the sequence of responses that occur during onset of nitrogen stress.

  19. Expression of a chitinase gene from Metarhizium anisopliae in tobacco plants confers resistance against Rhizoctonia solani.

    Science.gov (United States)

    Kern, Marcelo Fernando; Maraschin, Simone de Faria; Vom Endt, Débora; Schrank, Augusto; Vainstein, Marilene Henning; Pasquali, Giancarlo

    2010-04-01

    The chit1 gene from the entomopathogenic fungus Metarhizium anisopliae, encoding the endochitinase CHIT42, was placed under the control of the CaMV 35S promoter, and the resulting construct was transferred to tobacco. Seventeen kanamycin-resistant transgenic lines were recovered, and the presence of the transgene was confirmed by polymerase chain reactions and Southern blot hybridization. The number of chit1 copies was determined to be varying from one to four. Copy number had observable effects neither on plant growth nor development. Substantial heterogeneity concerning production of the recombinant chitinase, and both general and specific chitinolytic activities were detected in leaf extracts from primary transformants. The highest chitinase activities were found in plants harboring two copies of chit1 inserts at different loci. Progeny derived from self-pollination of the primary transgenics revealed a stable inheritance pattern, with transgene segregation following a mendelian dihybrid ratio. Two selected plants expressing high levels of CHIT42 were consistently resistant to the soilborne pathogen Rhizoctonia solani, suggesting a direct relationship between enzyme activity and reduction of foliar area affected by fungal lesions. To date, this is the first report of resistance to fungal attack in plants mediated by a recombinant chitinase from an entomopathogenic and acaricide fungus.

  20. Enhanced whitefly resistance in transgenic tobacco plants expressing double stranded RNA of v-ATPase A gene.

    Directory of Open Access Journals (Sweden)

    Nidhi Thakur

    Full Text Available BACKGROUND: Expression of double strand RNA (dsRNA designed against important insect genes in transgenic plants have been shown to give protection against pests through RNA interference (RNAi, thus opening the way for a new generation of insect-resistant crops. We have earlier compared the efficacy of dsRNAs/siRNAs, against a number of target genes, for interference in growth of whitefly (Bemisia tabaci upon oral feeding. The v-ATPase subunit A (v-ATPaseA coding gene was identified as a crucial target. We now report the effectiveness of transgenic tobacco plants expressing siRNA to silence v-ATPaseA gene expression for the control of whitefly infestation. METHODOLOGY/PRINCIPAL FINDINGS: Transgenic tobacco lines were developed for the expression of long dsRNA precursor to make siRNA and knock down the v-ATPaseA mRNA in whitefly. Molecular analysis and insecticidal properties of the transgenic plants established the formation of siRNA targeting the whitefly v-ATPaseA, in the leaves. The transcript level of v-ATPaseA in whiteflies was reduced up to 62% after feeding on the transgenic plants. Heavy infestation of whiteflies on the control plants caused significant loss of sugar content which led to the drooping of leaves. The transgenic plants did not show drooping effect. CONCLUSIONS/SIGNIFICANCE: Host plant derived pest resistance was achieved against whiteflies by genetic transformation of tobacco which generated siRNA against the whitefly v-ATPaseA gene. Transgenic tobacco lines expressing dsRNA of v-ATPaseA, delivered sufficient siRNA to whiteflies feeding on them, mounting a significant silencing response, leading to their mortality. The transcript level of the target gene was reduced in whiteflies feeding on transgenic plants. The strategy can be taken up for genetic engineering of plants to control whiteflies in field crops.

  1. Enhanced whitefly resistance in transgenic tobacco plants expressing double stranded RNA of v-ATPase A gene.

    Science.gov (United States)

    Thakur, Nidhi; Upadhyay, Santosh Kumar; Verma, Praveen C; Chandrashekar, Krishnappa; Tuli, Rakesh; Singh, Pradhyumna K

    2014-01-01

    Expression of double strand RNA (dsRNA) designed against important insect genes in transgenic plants have been shown to give protection against pests through RNA interference (RNAi), thus opening the way for a new generation of insect-resistant crops. We have earlier compared the efficacy of dsRNAs/siRNAs, against a number of target genes, for interference in growth of whitefly (Bemisia tabaci) upon oral feeding. The v-ATPase subunit A (v-ATPaseA) coding gene was identified as a crucial target. We now report the effectiveness of transgenic tobacco plants expressing siRNA to silence v-ATPaseA gene expression for the control of whitefly infestation. Transgenic tobacco lines were developed for the expression of long dsRNA precursor to make siRNA and knock down the v-ATPaseA mRNA in whitefly. Molecular analysis and insecticidal properties of the transgenic plants established the formation of siRNA targeting the whitefly v-ATPaseA, in the leaves. The transcript level of v-ATPaseA in whiteflies was reduced up to 62% after feeding on the transgenic plants. Heavy infestation of whiteflies on the control plants caused significant loss of sugar content which led to the drooping of leaves. The transgenic plants did not show drooping effect. Host plant derived pest resistance was achieved against whiteflies by genetic transformation of tobacco which generated siRNA against the whitefly v-ATPaseA gene. Transgenic tobacco lines expressing dsRNA of v-ATPaseA, delivered sufficient siRNA to whiteflies feeding on them, mounting a significant silencing response, leading to their mortality. The transcript level of the target gene was reduced in whiteflies feeding on transgenic plants. The strategy can be taken up for genetic engineering of plants to control whiteflies in field crops.

  2. The expression of a xylanase targeted to ER-protein bodies provides a simple strategy to produce active insoluble enzyme polymers in tobacco plants.

    Directory of Open Access Journals (Sweden)

    Immaculada Llop-Tous

    Full Text Available BACKGROUND: Xylanases deserve particular attention due to their potential application in the feed, pulp bleaching and paper industries. We have developed here an efficient system for the production of an active xylanase in tobacco plants fused to a proline-rich domain (Zera of the maize storage protein γ-zein. Zera is a self-assembling domain able to form protein aggregates in vivo packed in newly formed endoplasmic reticulum-derived organelles known as protein bodies (PBs. METHODOLOGY/PRINCIPAL FINDINGS: Tobacco leaves were transiently transformed with a binary vector containing the Zera-xylanase coding region, which was optimized for plant expression, under the control of the 35S CaMV promoter. The fusion protein was efficiently expressed and stored in dense PBs, resulting in yields of up to 9% of total protein. Zera-xylanase was post-translationally modified with high-mannose-type glycans. Xylanase fused to Zera was biologically active not only when solubilized from PBs but also in its insoluble form. The resistance of insoluble Zera-xylanase to trypsin digestion demonstrated that the correct folding of xylanase in PBs was not impaired by Zera oligomerization. The activity of insoluble Zera-xylanase was enhanced when substrate accessibility was facilitated by physical treatments such as ultrasound. Moreover, we found that the thermostability of the enzyme was improved when Zera was fused to the C-terminus of xylanase. CONCLUSION/SIGNIFICANCE: In the present work we have successfully produced an active insoluble aggregate of xylanase fused to Zera in plants. Zera-xylanase chimeric protein accumulates within ER-derived protein bodies as active aggregates that can easily be recovered by a simple density-based downstream process. The production of insoluble active Zera-xylanase protein in tobacco outlines the potential of Zera as a fusion partner for producing enzymes of biotechnological relevance. Zera-PBs could thus become efficient and low

  3. Over-expression of Arabidopsis CAP causes decreased cell expansion leading to organ size reduction in transgenic tobacco plants.

    Science.gov (United States)

    Barrero, Roberto A; Umeda, Masaaki; Yamamura, Saburo; Uchimiya, Hirofumi

    2003-04-01

    Cyclase-associated proteins (CAP) are multifunctional proteins involved in Ras-cAMP signalling and regulation of the actin cytoskeleton. It has recently been demonstrated that over-expression of AtCAP1 in transgenic arabidopsis plants causes severe morphological defects owing to loss of actin filaments. To test the generality of the function of AtCAP1 in plants, transgenic tobacco plants over-expressing an arabidopsis CAP (AtCAP1) under the regulation of a glucocorticoid-inducible promoter were produced. Over-expression of AtCAP1 in transgenic tobacco plants led to growth abnormalities, in particular a reduction in the size of leaves. Morphological alterations in leaves were the result of reduced elongation of epidermal and mesophyll cells.

  4. Effects of air pollutants on plant cell tissue cultures. [Tobacco, rose soybean, periwinkle, and morning glory

    Energy Technology Data Exchange (ETDEWEB)

    1967-01-01

    Experiments were conducted to determine morphological and physiological effects of air pollutants on plant tissue cultures. Several cultures will be exposed to polluted atmospheres for various periods and observed for effects. The cultures which have been developed for this purpose are: tobacco pith, rose stem, soybean stem, periwinkle, and morning glory. Exposures will follow two regimens: a relatively high concentration of pollutant for a short duration and a low concentration for a long duration. Effects of pollutants on cell morphology will be observed microscopically. Effects on cell physiology may include altered respiratory quotients which will be determined by Warburg respirometry techniques. The design of an apparatus that is being developed to mix a pollutant with air and deliver it to the cultures is described.

  5. RNA Interference towards the Potato Psyllid, Bactericera cockerelli, Is Induced in Plants Infected with Recombinant Tobacco mosaic virus (TMV.

    Directory of Open Access Journals (Sweden)

    Hada Wuriyanghan

    Full Text Available The potato/tomato psyllid, Bactericera cockerelli (B. cockerelli, is an important plant pest and the vector of the phloem-limited bacterium Candidatus Liberibacter psyllaurous (solanacearum, which is associated with the zebra chip disease of potatoes. Previously, we reported induction of RNA interference effects in B. cockerelli via in vitro-prepared dsRNA/siRNAs after intrathoracic injection, and after feeding of artificial diets containing these effector RNAs. In order to deliver RNAi effectors via plant hosts and to rapidly identify effective target sequences in plant-feeding B. cockerelli, here we developed a plant virus vector-based in planta system for evaluating candidate sequences. We show that recombinant Tobacco mosaic virus (TMV containing B. cockerelli sequences can efficiently infect and generate small interfering RNAs in tomato (Solanum lycopersicum, tomatillo (Physalis philadelphica and tobacco (Nicotiana tabacum plants, and more importantly delivery of interfering sequences via TMV induces RNAi effects, as measured by actin and V-ATPase mRNA reductions, in B. cockerelli feeding on these plants. RNAi effects were primarily detected in the B. cockerelli guts. In contrast to our results with TMV, recombinant Potato virus X (PVX and Tobacco rattle virus (TRV did not give robust infections in all plants and did not induce detectable RNAi effects in B. cockerelli. The greatest RNA interference effects were observed when B. cockerelli nymphs were allowed to feed on leaf discs collected from inoculated or lower expanded leaves from corresponding TMV-infected plants. Tomatillo plants infected with recombinant TMV containing B. cockerelli actin or V-ATPase sequences also showed phenotypic effects resulting in decreased B. cockerelli progeny production as compared to plants infected by recombinant TMV containing GFP. These results showed that RNAi effects can be achieved in plants against the phloem feeder, B. cockerelli, and the TMV-plant

  6. RNA Interference towards the Potato Psyllid, Bactericera cockerelli, Is Induced in Plants Infected with Recombinant Tobacco mosaic virus (TMV).

    Science.gov (United States)

    Wuriyanghan, Hada; Falk, Bryce W

    2013-01-01

    The potato/tomato psyllid, Bactericera cockerelli (B. cockerelli), is an important plant pest and the vector of the phloem-limited bacterium Candidatus Liberibacter psyllaurous (solanacearum), which is associated with the zebra chip disease of potatoes. Previously, we reported induction of RNA interference effects in B. cockerelli via in vitro-prepared dsRNA/siRNAs after intrathoracic injection, and after feeding of artificial diets containing these effector RNAs. In order to deliver RNAi effectors via plant hosts and to rapidly identify effective target sequences in plant-feeding B. cockerelli, here we developed a plant virus vector-based in planta system for evaluating candidate sequences. We show that recombinant Tobacco mosaic virus (TMV) containing B. cockerelli sequences can efficiently infect and generate small interfering RNAs in tomato (Solanum lycopersicum), tomatillo (Physalis philadelphica) and tobacco (Nicotiana tabacum) plants, and more importantly delivery of interfering sequences via TMV induces RNAi effects, as measured by actin and V-ATPase mRNA reductions, in B. cockerelli feeding on these plants. RNAi effects were primarily detected in the B. cockerelli guts. In contrast to our results with TMV, recombinant Potato virus X (PVX) and Tobacco rattle virus (TRV) did not give robust infections in all plants and did not induce detectable RNAi effects in B. cockerelli. The greatest RNA interference effects were observed when B. cockerelli nymphs were allowed to feed on leaf discs collected from inoculated or lower expanded leaves from corresponding TMV-infected plants. Tomatillo plants infected with recombinant TMV containing B. cockerelli actin or V-ATPase sequences also showed phenotypic effects resulting in decreased B. cockerelli progeny production as compared to plants infected by recombinant TMV containing GFP. These results showed that RNAi effects can be achieved in plants against the phloem feeder, B. cockerelli, and the TMV-plant system will

  7. Molecular sensing of bacteria in plants. The highly conserved RNA-binding motif RNP-1 of bacterial cold shock proteins is recognized as an elicitor signal in tobacco.

    Science.gov (United States)

    Felix, Georg; Boller, Thomas

    2003-02-21

    To detect microbial infection multicellular organisms have evolved sensing systems for pathogen-associated molecular patterns (PAMPs). Here, we identify bacterial cold shock protein (CSP) as a new such PAMP that acts as a highly active elicitor of defense responses in tobacco. Tobacco cells perceive a conserved domain of CSP and synthetic peptides representing 15 amino acids of this domain-induced responses at subnanomolar concentrations. Central to the elicitor-active domain is the RNP-1 motif KGFGFITP, a motif conserved also in many RNA- and DNA-binding proteins of eukaryotes. Csp15-Nsyl, a peptide representing the domain with highest homology to csp15 in a protein of Nicotiana sylvestris exhibited only weak activity in tobacco cells. Crystallographic and genetic data from the literature show that the RNP-1 domain of bacterial CSPs resides on a protruding loop and exposes a series of aromatic and basic side chains to the surface that are essential for the nucleotide-binding activity of CSPs. Similarly, these side chains were also essential for elicitor activity and replacement of single residues in csp15 with Ala strongly reduced or abolished activity. Most strikingly, csp15-Ala10, a peptide with the RNP-1 motif modified to KGAGFITP, lacked elicitor activity but acted as a competitive antagonist for CSP-related elicitors. Bacteria commonly have a small family of CSP-like proteins including both cold-inducible and noninducible members, and Csp-related elicitor activity was detected in extracts from all bacteria tested. Thus, the CSP domain containing the RNP-1 motif provides a structure characteristic for bacteria in general, and tobacco plants have evolved a highly sensitive chemoperception system to detect this bacterial PAMP.

  8. Modified artificial diet for rearing of tobacco budworm, Helicoverpa armigera, using the Taguchi method and Derringer's desirability function.

    Science.gov (United States)

    Assemi, H; Rezapanah, M; Vafaei-Shoushtari, R; Mehrvar, A

    2012-01-01

    With the aim to improve the mass rearing feasibility of tobacco budworm, Helicoverpa armigera Hübner (Lepidoptera: Noctuidae), design of experimental methodology using Taguchi orthogonal array was applied. To do so, the effect of 16 ingredients of an artificial diet including bean, wheat germ powder, Nipagin, ascorbic acid, formaldehyde, oil, agar, distilled water, ascorbate, yeast, chloramphenicol, benomyl, penicillin, temperature, humidity, and container size on some biological characteristics of H. armigera was evaluated. The selected 16 factors were considered at two levels (32 experiments) in the experimental design. Among the selected factors, penicillin, container size, formaldehyde, chloramphenicol, wheat germ powder, and agar showed significant effect on the mass rearing performance. Derringer's desirability function was used for simultaneous optimization of mass rearing of tobacco budworm, H. armigera, on a modified artificial diet. Derived optimum operating conditions obtained by Derringer's desirability function and Taguchi methodology decreased larval period from 19 to 15.5 days (18.42 % improvement), decreased the pupal period from 12.29 to 11 days (10.49 % improvement), increased the longevity of adults from 14.51 to 21 days (44.72 % improvement), increased the number of eggs/female from 211.21 to 260, and increased egg hatchability from 54.2% to 72% (32.84 % improvement). The proposed method facilitated a systematic mathematical approach with a few well-defined experimental sets.

  9. Cytoprotective effect of recombinant human erythropoietin produced in transgenic tobacco plants.

    Directory of Open Access Journals (Sweden)

    Farooqahmed S Kittur

    Full Text Available Asialo-erythropoietin, a desialylated form of human erythropoietin (EPO lacking hematopoietic activity, is receiving increased attention because of its broader protective effects in preclinical models of tissue injury. However, attempts to translate its protective effects into clinical practice is hampered by unavailability of suitable expression system and its costly and limit production from expensive mammalian cell-made EPO (rhuEPO(M by enzymatic desialylation. In the current study, we took advantage of a plant-based expression system lacking sialylating capacity but possessing an ability to synthesize complex N-glycans to produce cytoprotective recombinant human asialo-rhuEPO. Transgenic tobacco plants expressing asialo-rhuEPO were generated by stably co-expressing human EPO and β1,4-galactosyltransferase (GalT genes under the control of double CaMV 35S and glyceraldehyde-3-phosphate gene (GapC promoters, respectively. Plant-produced asialo-rhuEPO (asialo-rhuEPO(P was purified by immunoaffinity chromatography. Detailed N-glycan analysis using NSI-FTMS and MS/MS revealed that asialo-rhuEPO(P bears paucimannosidic, high mannose-type and complex N-glycans. In vitro cytoprotection assays showed that the asialo-rhuEPO(P (20 U/ml provides 2-fold better cytoprotection (44% to neuronal-like mouse neuroblastoma cells from staurosporine-induced cell death than rhuEPO(M (21%. The cytoprotective effect of the asialo-rhuEPO(P was found to be mediated by receptor-initiated phosphorylation of Janus kinase 2 (JAK2 and suppression of caspase 3 activation. Altogether, these findings demonstrate that plants are a suitable host for producing cytoprotective rhuEPO derivative. In addition, the general advantages of plant-based expression system can be exploited to address the cost and scalability issues related to its production.

  10. Ecological Risk Assessment of Genetically Modified Higher Plants (GMHP)

    DEFF Research Database (Denmark)

    Kjær, C.; Damgaard, C.; Kjellsson, G.

    Preface This publication is a first version of a manual identifying the data needs for ecological risk assessment of genetically modified higher plants (GMHP). It is the intention of the authors to stimulate further discussion of what data are needed in order to conduct a proper ecological risk...... assessment of GM plants when application for placing on the market is made. It is our hope that both the scientific community, the biotechnological industry and the regulatory bodies will participate in the process of improving the present draft, so that it can develop into a useful tool for both...... the industry as well as the national regulatory bodies. Furthermore, we hope that these efforts will improve the transparency of risk assessment and harmonisation of the requirements for data. The report suggests a structured way to identify the data need for risk assessment of GMHPs. It does not discuss...

  11. Ecological Risk Assessment of Genetically Modified Higher Plants (GMHP)

    DEFF Research Database (Denmark)

    Kjær, C.; Damgaard, C.; Kjellsson, G.

    of the project Biotechnology: elements in environmental risk assessment of genetically modified plants. December 1999 Christian Kjær Introduction In ecological risk assessment of transgenic plants, information on a wide range of subjects is needed for an effective and reliable assessment procedure...... the actual risk assessment procedures and the risk evaluation, which must proceed the data collection. The report use the terminology ecological risk assessment rather than environmental risk assessment because at present this work does not include bio-geochemical effects and environmental impacts from...... for uncertainties in the extrapolation from limited laboratory studies to the species rich field environment. The relationship between the size of the safety factor and the number of species is therefore an issue of the risk assessment. Some of the issues raised in this report overlap with data needs...

  12. Inhibition of brome mosaic virus (BMV) amplification in protoplasts from transgenic tobacco plants expressing replicable BMV RNAs.

    Science.gov (United States)

    Kaido, M; Mori, M; Mise, K; Okuno, T; Furusawa, I

    1995-11-01

    Transgenic tobacco plants (V123 plants) expressing a set of full-length brome mosaic virus (BMV) genomic RNAs from the cauliflower mosaic virus 35S promoter were produced. The accumulation level of BMV RNAs in V123 plant cells was approximately 1% of that in nontransgenic tobacco protoplasts inoculated with BMV RNAs. The level of BMV RNA in V123 protoplasts did not increase after inoculating the protoplasts with BMV RNAs, whereas V123 protoplasts supported the accumulation of cucumber mosaic virus (CMV) RNAs to a level similar to that in non-transgenic tobacco protoplasts after inoculation with CMV RNA. Such BMV-specific resistance was also observed in protoplasts from V12 plants expressing full-length BMV RNA1 and RNA2, both of which are required and sufficient for BMV RNA replication. On the other hand, protoplasts from M12 plants, expressing truncated BMV RNA1 and RNA2 in which the 3' 200 nucleotides required for BMV RNA replication were deleted, exhibited weaker resistance to infection with BMV RNA than V12 protoplasts, although the accumulation level of truncated BMV RNA1 and RNA2 in M12 protoplasts was higher than that of BMV RNA1 and RNA2 in V12 protoplasts. These results suggest that expression of BMV RNA replicons is involved in the induction of resistance, rather than high-level accumulation of BMV RNAs and/or their encoded proteins.

  13. Systems Toxicology Assessment of the Biological Impact of a Candidate Modified Risk Tobacco Product on Human Organotypic Oral Epithelial Cultures.

    Science.gov (United States)

    Zanetti, Filippo; Sewer, Alain; Mathis, Carole; Iskandar, Anita R; Kostadinova, Radina; Schlage, Walter K; Leroy, Patrice; Majeed, Shoaib; Guedj, Emmanuel; Trivedi, Keyur; Martin, Florian; Elamin, Ashraf; Merg, Céline; Ivanov, Nikolai V; Frentzel, Stefan; Peitsch, Manuel C; Hoeng, Julia

    2016-08-15

    Cigarette smoke (CS) has been reported to increase predisposition to oral cancer and is also recognized as a risk factor for many conditions including periodontal diseases, gingivitis, and other benign mucosal disorders. Smoking cessation remains the most effective approach for minimizing the risk of smoking-related diseases. However, reduction of harmful constituents by heating rather than combusting tobacco, without modifying the amount of nicotine, is a promising new paradigm in harm reduction. In this study, we compared effects of exposure to aerosol derived from a candidate modified risk tobacco product, the tobacco heating system (THS) 2.2, with those of CS generated from the 3R4F reference cigarette. Human organotypic oral epithelial tissue cultures (EpiOral, MatTek Corporation) were exposed for 28 min to 3R4F CS or THS2.2 aerosol, both diluted with air to comparable nicotine concentrations (0.32 or 0.51 mg nicotine/L aerosol/CS for 3R4F and 0.31 or 0.46 mg/L for THS2.2). We also tested one higher concentration (1.09 mg/L) of THS2.2. A systems toxicology approach was employed combining cellular assays (i.e., cytotoxicity and cytochrome P450 activity assays), comprehensive molecular investigations of the buccal epithelial transcriptome (mRNA and miRNA) by means of computational network biology, measurements of secreted proinflammatory markers, and histopathological analysis. We observed that the impact of 3R4F CS was greater than THS2.2 aerosol in terms of cytotoxicity, morphological tissue alterations, and secretion of inflammatory mediators. Analysis of the transcriptomic changes in the exposed oral cultures revealed significant perturbations in various network models such as apoptosis, necroptosis, senescence, xenobiotic metabolism, oxidative stress, and nuclear factor (erythroid-derived 2)-like 2 (NFE2L2) signaling. The stress responses following THS2.2 aerosol exposure were markedly decreased, and the exposed cultures recovered more completely compared

  14. [Enhanced resistance to phytopathogenic bacteria in transgenic tobacco plants with synthetic gene of antimicrobial peptide cecropin P1].

    Science.gov (United States)

    Zakharchenko, N S; Rukavtsova, E B; Gudkov, A T; Bur'ianov, Ia I

    2005-11-01

    Plasmids with a synthetic gene of the mammalian antimicrobial peptide cecropin P1 (cecP1) controlled by the constitutive promoter 35S RNA of cauliflower mosaic virus were constructed. Agrobacterial transformation of tobacco plants was conducted using the obtained recombinant binary vector. The presence of gene cecP1 in the plant genome was confirmed by PCR. The expression of gene cecP1 in transgenic plants was shown by Northern blot analysis. The obtained transgenic plants exhibit enhanced resistance to phytopathogenic bacteria Pseudomonas syringae, P. marginata, and Erwinia carotovora. The ability of transgenic plants to express cecropin P1 was transmitted to the progeny. F1 and F2 plants had the normal phenotype (except for a changed coloration of flowers) and retained the ability to produce normal viable seeds upon self-pollination. Lines of F1 plants with Mendelian segregation of transgenic traits were selected.

  15. [Construction and analysis of transgenic plants of Nicotiana tabacum L. expressing a bacterial gene for beta-1,3-glucanase. II. Transgenic tobacco plants expressing the bacterial beta-glucanase gene from Clostridium thermocellum,--a model for studying the differential expression of stress response-related genes].

    Science.gov (United States)

    Darbinian, N S; Popov, Iu G; Mochul'skiĭ, A V; Oming, D; Piruzian, E S; Vasilevko, V T

    1996-02-01

    The modified hybrid beta-1,3-glucanase gene (glc) of Clostridium thermocellum was expressed in tobacco Nicotiana tabacum. The glc gene was cloned into two plasmids, pC27-glc and pC29-glc, in which its expression was controlled by the TR2' promoter of the 2' gene of T-DNA and the rbcS promoter of Arabidopsis, respectively. These constructions were used for transformation of agrobacteria followed by transfer into plants. In transformed plants, each plasmid caused a high level of activity of thermostable bacterial glucanase not observed in reference plants. The plants obtained were used to study activation of some defense-related genes induced by their interaction with either tobacco mosaic virus (TMV) or a pathogenic fungus.

  16. A plant mitochondrial sequence transcribed in transgenic tobacco chloroplasts is not edited

    Energy Technology Data Exchange (ETDEWEB)

    Sutton, C.A.; Hanson, M.R. [Cornell Univ., Ithaca, NY (United States); Zoubenko, O.V.; Maliga, P. [State Univ. of New Jersey, Piscataway, NJ (United States)

    1995-03-01

    RNA editing occurs in two higher-plant organelles, chloroplasts, and mitochondria. Because chloroplasts and mitochondria exhibit some similarity in editing site selection, we investigated whether mitochondrial RNA sequences could be edited in chloroplasts. We produced transgenic tobacco plants that contained chimeric genes in which the second exon of a Petunia hybrida mitochondrial coxII gene was under the control of chloroplast gene regulatory sequences. coxII transcripts accumulated to low or high levels in transgenic chloroplasts containing chimeric genes with the plastid ribosomal protein gene rps16 or the rRNA operon promoter, respectively. Exon 2 of coxII was chosen because it carries seven editing sites and is edited in petunia mitochondria even when located in an abnormal context in an aberrant recombined gene. When editing of the coxII transcripts in transgenic chloroplasts was examined, no RNA editing at any of the usual sites was detected, nor was there any novel editing at any other sites. These results indicate that the RNA editing mechanisms of chloroplasts and mitochondria are not identical but must have at least some organelle-specific components. 33 refs., 5 figs.

  17. Engineering of benzylglucosinolate in tobacco provides proof-of-concept for dead-end trap crops genetically modified to attract Plutella xylostella (diamondback moth).

    Science.gov (United States)

    Møldrup, Morten E; Geu-Flores, Fernando; de Vos, Martin; Olsen, Carl E; Sun, Joel; Jander, Georg; Halkier, Barbara A

    2012-05-01

    Glucosinolates are biologically active natural products characteristic of crucifers, including oilseed rape, cabbage vegetables and the model plant Arabidopsis thaliana. Crucifer-specialist insect herbivores, like the economically important pest Plutella xylostella (diamondback moth), frequently use glucosinolates as oviposition stimuli. This suggests that the transfer of a glucosinolate biosynthetic pathway to a non-crucifer would stimulate oviposition on an otherwise non-attractive plant. Here, we demonstrate that stable genetic transfer of the six-step benzylglucosinolate pathway from A. thaliana to Nicotiana tabacum (tobacco) results in the production of benzylglucosinolate without causing morphological alterations. Benzylglucosinolate-producing tobacco plants were more attractive for oviposition by female P. xylostella moths than wild-type tobacco plants. As newly hatched P. xylostella larvae were unable to survive on tobacco, these results represent a proof-of-concept strategy for rendering non-host plants attractive for oviposition by specialist herbivores with the long-term goal of generating efficient dead-end trap crops for agriculturally important pests.

  18. Improvement of organic phosphate acquisition in transgenic tobacco plants by overexpression of a soybean phytase gene Sphyl

    Institute of Scientific and Technical Information of China (English)

    Li GUO; Yuxin ZHAO; Shuhua ZHANG; Haina ZHANG; Kai XIAO

    2009-01-01

    Due to the huge amount in the soil, phytate is an important potential source for providing the plants with available phosphorus (Pi) by the involved catalytic reaction of phytase. In this study, a construct fusing the open reading frame (ORF) of Sphyl into corresponding positions in the fragment of binary expression vector pBI121 was created and used to transform tobacco. Molecular identification by PCR and RT-PCR indicated the target gene Sphyl in the transgenic tobacco plants was transcribed under the regulation of an upstream promoter. Compared with the control plants, the phytase activities in all the transgenic plants were increased, with the increased range consistent with the expression levels in the transgenic plants. Under the growth conditions with phytate as the sole phosphorus source, the transgenic line 1 plants displayed a high expression level of Sphyl and shows notable improved growth performance, such as higher fresh weight and dry weight, as well as higher total P content and more accumulative P amount per plant than CK. This clearly indicated that overexpression of Sphyl could improve the phosphorus acquisition by the extruded Sphyl phytase in the rhizosphere, where this enzyme could catalyze the degradation of the phytate and release the available Pi for plants. The Sphyl gene seemed to have a potential value in the creation of new crop cultivars with high phosphorus use efficiency.

  19. The importance of lipid modified proteins in plants.

    Science.gov (United States)

    Hemsley, Piers A

    2015-01-01

    Membranes have long been known to act as more than physical barriers within and between plant cells. Trafficking of membrane proteins, signalling from and across membranes, organisation of membranes and transport through membranes are all essential processes for plant cellular function. These processes rely on a myriad array of proteins regulated in a variety of manners and are frequently required to be directly associated with membranes. For integral membrane proteins, the mode of membrane association is readily apparent, but many peripherally associated membrane proteins are outwardly soluble proteins. In these cases the proteins are frequently modified by the addition of lipids allowing direct interaction with the hydrophobic core of membranes. These modifications include N-myristoylation, S-acylation (palmitoylation), prenylation and GPI anchors but until recently little was truly known about their function in plants. New data suggest that these modifications are able to act as more than just membrane anchors, and dynamic S-acylation in particular is emerging as a means of regulating protein function in a similar manner to phosphorylation. This review discusses how these modifications occur, their impact on protein function, how they are regulated, recent advances in the field and technical approaches for studying these modifications.

  20. Transgenic tobacco plants overexpressing a grass PpEXP1 gene exhibit enhanced tolerance to heat stress.

    Directory of Open Access Journals (Sweden)

    Qian Xu

    Full Text Available Heat stress is a detrimental abiotic stress limiting the growth of many plant species and is associated with various cellular and physiological damages. Expansins are a family of proteins which are known to play roles in regulating cell wall elongation and expansion, as well as other growth and developmental processes. The in vitro roles of expansins regulating plant heat tolerance are not well understood. The objectives of this study were to isolate and clone an expansin gene in a perennial grass species (Poa pratensis and to determine whether over-expression of expansin may improve plant heat tolerance. Tobacco (Nicotiana tabacum was used as the model plant for gene transformation and an expansin gene PpEXP1 from Poa pratensis was cloned. Sequence analysis showed PpEXP1 belonged to α-expansins and was closely related to two expansin genes in other perennial grass species (Festuca pratensis and Agrostis stolonifera as well as Triticum aestivum, Oryza sativa, and Brachypodium distachyon. Transgenic tobacco plants over-expressing PpEXP1 were generated through Agrobacterium-mediated transformation. Under heat stress (42°C in growth chambers, transgenic tobacco plants over-expressing the PpEXP1 gene exhibited a less structural damage to cells, lower electrolyte leakage, lower levels of membrane lipid peroxidation, and lower content of hydrogen peroxide, as well as higher chlorophyll content, net photosynthetic rate, relative water content, activity of antioxidant enzyme, and seed germination rates, compared to the wild-type plants. These results demonstrated the positive roles of PpEXP1 in enhancing plant tolerance to heat stress and the possibility of using expansins for genetic modification of cool-season perennial grasses in the development of heat-tolerant germplasm and cultivars.

  1. Efficient gene silencing mediated by tobacco rattle virus in an emerging model plant physalis.

    Directory of Open Access Journals (Sweden)

    Ji-Si Zhang

    Full Text Available The fruit of Physalis has a berry and a novelty called inflated calyx syndrome (ICS, also named the 'Chinese lantern'. Elucidation of the underlying developmental mechanisms of fruit diversity demands an efficient gene functional inference platform. Here, we tested the application of the tobacco rattle virus (TRV-mediated gene-silencing system in Physalis floridana. First, we characterized the putative gene of a phytoene desaturase in P. floridana (PfPDS. Infecting the leaves of the Physalis seedlings with the PfPDS-TRV vector resulted in a bleached plant, including the developing leaves, floral organs, ICS, berry, and seed. These results indicated that a local VIGS treatment can efficiently induce a systemic mutated phenotype. qRT-PCR analyses revealed that the bleaching extent correlated to the mRNA reduction of the endogenous PfPDS. Detailed comparisons of multiple infiltration and growth protocols allowed us to determine the optimal methodologies for VIGS manipulation in Physalis. We subsequently utilized this optimized VIGS methodology to downregulate the expression of two MADS-box genes, MPF2 and MPF3, and compared the resulting effects with gene-downregulation mediated by RNA interference (RNAi methods. The VIGS-mediated gene knockdown plants were found to resemble the mutated phenotypes of floral calyx, fruiting calyx and pollen maturation of the RNAi transgenic plants for both MPF2 and MPF3. Moreover, the two MADS-box genes were appeared to have a novel role in the pedicel development in P. floridana. The major advantage of VIGS-based gene knockdown lies in practical aspects of saving time and easy manipulation as compared to the RNAi. Despite the lack of heritability and mosaic mutation phenotypes observed in some organs, the TRV-mediated gene silencing system provides an alternative efficient way to infer gene function in various developmental processes in Physalis, thus facilitating understanding of the genetic basis of the evolution

  2. Efficient gene silencing mediated by tobacco rattle virus in an emerging model plant physalis.

    Science.gov (United States)

    Zhang, Ji-Si; Zhao, Jing; Zhang, Shaohua; He, Chaoying

    2014-01-01

    The fruit of Physalis has a berry and a novelty called inflated calyx syndrome (ICS, also named the 'Chinese lantern'). Elucidation of the underlying developmental mechanisms of fruit diversity demands an efficient gene functional inference platform. Here, we tested the application of the tobacco rattle virus (TRV)-mediated gene-silencing system in Physalis floridana. First, we characterized the putative gene of a phytoene desaturase in P. floridana (PfPDS). Infecting the leaves of the Physalis seedlings with the PfPDS-TRV vector resulted in a bleached plant, including the developing leaves, floral organs, ICS, berry, and seed. These results indicated that a local VIGS treatment can efficiently induce a systemic mutated phenotype. qRT-PCR analyses revealed that the bleaching extent correlated to the mRNA reduction of the endogenous PfPDS. Detailed comparisons of multiple infiltration and growth protocols allowed us to determine the optimal methodologies for VIGS manipulation in Physalis. We subsequently utilized this optimized VIGS methodology to downregulate the expression of two MADS-box genes, MPF2 and MPF3, and compared the resulting effects with gene-downregulation mediated by RNA interference (RNAi) methods. The VIGS-mediated gene knockdown plants were found to resemble the mutated phenotypes of floral calyx, fruiting calyx and pollen maturation of the RNAi transgenic plants for both MPF2 and MPF3. Moreover, the two MADS-box genes were appeared to have a novel role in the pedicel development in P. floridana. The major advantage of VIGS-based gene knockdown lies in practical aspects of saving time and easy manipulation as compared to the RNAi. Despite the lack of heritability and mosaic mutation phenotypes observed in some organs, the TRV-mediated gene silencing system provides an alternative efficient way to infer gene function in various developmental processes in Physalis, thus facilitating understanding of the genetic basis of the evolution and development

  3. Effects of Transgenic Tobacco Plants Expressing ACA Gene from Amaranthus caudatus on the Population Development of Myzus persicae

    Institute of Scientific and Technical Information of China (English)

    GUOHong-Nian; JIAYan-Tao; ZHOUYong-Gang; ZHANGZhen-Shan; OUYANGQing; JIANGYing; TIANYing-Chuan

    2004-01-01

    To investigate the possible function of the agglutinin from Amaranthus caudatus L. (ACA) in plant defending against insect pests, ACA cDNA was cloned by RT-PCR and the 5' and 3' sequences were confirmed by rapid amplification of cDNA ends (RACE). The phloem-specific expression vector of ACA gene, pBCACAc, was constructed based on the plant binary vector pBC438 and transfered into tobacco plants via Agrobacterium-mediated transformation method. Results from PCR and Southern blotting analysis showed that AOA gene was integrated into the genomes of transformed plants and the transgene integration varied from one to four estimated copies per genome. Western blotting analysis indicated that ACA gene was transcribed and translated in the transgenic plants. The bioassay of Myzus persicae Sulzer on detached leaves demonstrated that the 78% transgenic tobacco plants displayed an average aphid-resistant rate of more than 75%. Some apterous progeny of M. persicae were found dead on the resistant plants. These results indicate that ACA gene should be an effective aphid-resistant gene and could be valuable for application in crop breeding for aphid resistance.

  4. Inhibition of cell proliferation, cell expansion and differentiation by the Arabidopsis SUPERMAN gene in transgenic tobacco plants.

    Science.gov (United States)

    Bereterbide, A; Hernould, M; Castera, S; Mouras, A

    2001-11-01

    Plant development depends upon the control of growth, organization and differentiation of cells derived from shoot and root meristems. Among the genes involved in flower organ determination, the cadastral gene SUPERMAN controls the boundary between whorls 3 and 4 and the growth of the adaxial outer ovule integument by down-regulating cell divisions. To determine the precise function of this gene we overexpressed ectopically the Arabidopsis thaliana (L.) Heynh. SUPERMAN gene in tobacco (Nicotiana tabacum L.). The transgenic plants exhibited a dwarf phenotype. Histologically and cytologically detailed analyses showed that dwarfism is correlated with a reduction in cell number, which is in agreement with the SUPERMAN function in Arabidopsis. Furthermore, a reduction in cell expansion and an impairment of cell differentiation were observed in tobacco organs. These traits were observed in differentiated vegetative and floral organs but not in meristem structures. A potential effect of the SUPERMAN transcription factor in the control of gibberellin biosynthesis is discussed.

  5. Enhanced production of resveratrol derivatives in tobacco plants by improving the metabolic flux of intermediates in the phenylpropanoid pathway.

    Science.gov (United States)

    Jeong, Yu Jeong; An, Chul Han; Woo, Su Gyeong; Park, Ji Hye; Lee, Ki-Won; Lee, Sang-Hoon; Rim, Yeonggil; Jeong, Hyung Jae; Ryu, Young Bae; Kim, Cha Young

    2016-09-01

    The biosynthesis of flavonoids such as anthocyanin and stilbenes has attracted increasing attention because of their potential health benefits. Anthocyanins and stilbenes share common phenylpropanoid precursor pathways. We previously reported that the overexpression of sweetpotato IbMYB1a induced anthocyanin pigmentation in transgenic tobacco (Nicotiana tabacum) plants. In the present study, transgenic tobacco (Nicotiana tabacum SR1) plants (STS-OX and ROST-OX) expressing the RpSTS gene encoding stilbene synthase from rhubarb (Rheum palmatum L. cv. Jangyeop) and the RpSTS and VrROMT genes encoding resveratrol O-methyltransferase from frost grape (Vitis riparia) were generated under the control of 35S promoter. Phenotypic alterations in floral organs, such as a reduction in floral pigments and male sterility, were observed in STS-OX transgenic tobacco plants. However, we failed to obtain STS-OX and ROST-OX plants with high levels of resveratrol compounds. Therefore, to improve the production of resveratrol derivatives in plants, we cross-pollinated flowers of STS-OX or ROST-OX and IbMYB1a-OX transgenic lines (SM and RSM). Phenotypic changes in vegetative and reproductive development of SM and RSM plants were observed. Furthermore, by HPLC and LC-MS analyses, we found enhanced production of resveratrol derivatives such as piceid, piceid methyl ether, resveratrol methyl ether O-hexoside, and 5-methyl resveratrol-3,4'-O-β-D-diglucopyranoside in SM and RSM cross-pollinated lines. Here, total contents of trans- and cis-piceids ranged from approximately 104-240 µg/g fresh weight in SM (F2). Collectively, we suggest that coexpression of RpSTS and IbMYB1a via cross-pollination can induce enhanced production of resveratrol compounds in plants by increasing metabolic flux into stilbenoid biosynthesis.

  6. Revisiting Plant Plasma Membrane Lipids in Tobacco: A Focus on Sphingolipids.

    Science.gov (United States)

    Cacas, Jean-Luc; Buré, Corinne; Grosjean, Kevin; Gerbeau-Pissot, Patricia; Lherminier, Jeannine; Rombouts, Yoann; Maes, Emmanuel; Bossard, Claire; Gronnier, Julien; Furt, Fabienne; Fouillen, Laetitia; Germain, Véronique; Bayer, Emmanuelle; Cluzet, Stéphanie; Robert, Franck; Schmitter, Jean-Marie; Deleu, Magali; Lins, Laurence; Simon-Plas, Françoise; Mongrand, Sébastien

    2016-01-01

    The lipid composition of plasma membrane (PM) and the corresponding detergent-insoluble membrane (DIM) fraction were analyzed with a specific focus on highly polar sphingolipids, so-called glycosyl inositol phosphorylceramides (GIPCs). Using tobacco (Nicotiana tabacum) 'Bright Yellow 2' cell suspension and leaves, evidence is provided that GIPCs represent up to 40 mol % of the PM lipids. Comparative analysis of DIMs with the PM showed an enrichment of 2-hydroxylated very-long-chain fatty acid-containing GIPCs and polyglycosylated GIPCs in the DIMs. Purified antibodies raised against these GIPCs were further used for immunogold-electron microscopy strategy, revealing the distribution of polyglycosylated GIPCs in domains of 35 ± 7 nm in the plane of the PM. Biophysical studies also showed strong interactions between GIPCs and sterols and suggested a role for very-long-chain fatty acids in the interdigitation between the two PM-composing monolayers. The ins and outs of lipid asymmetry, raft formation, and interdigitation in plant membrane biology are finally discussed.

  7. Plant scents modify innate colour preference in foraging swallowtail butterflies.

    Science.gov (United States)

    Yoshida, Mina; Itoh, Yuki; Ômura, Hisashi; Arikawa, Kentaro; Kinoshita, Michiyo

    2015-07-01

    Flower-visiting insects exhibit innate preferences for particular colours. A previous study demonstrated that naive Papilio xuthus females prefer yellow and red, whereas males are more attracted to blue. Here, we demonstrate that the innate colour preference can be modified by olfactory stimuli in a sexually dimorphic manner. Naive P. xuthus were presented with four coloured discs: blue, green, yellow and red. The innate colour preference (i.e. the colour first landed on) of the majority of individuals was blue. When scent from essential oils of either orange flower or lily was introduced to the room, females' tendency to select the red disc increased. Scents of lavender and flowering potted Hibiscus rosa-sinensis, however, were less effective. Interestingly, the odour of the non-flowering larval host plant, Citrus unshiu, shifted the preference to green in females. In males, however, all plant scents were less effective than in females, such that blue was always the most favoured colour. These observations indicate that interactions between visual and olfactory cues play a more prominent role in females. © 2015 The Author(s) Published by the Royal Society. All rights reserved.

  8. Plant scents modify innate colour preference in foraging swallowtail butterflies

    Science.gov (United States)

    Yoshida, Mina; Itoh, Yuki; Ômura, Hisashi; Arikawa, Kentaro; Kinoshita, Michiyo

    2015-01-01

    Flower-visiting insects exhibit innate preferences for particular colours. A previous study demonstrated that naive Papilio xuthus females prefer yellow and red, whereas males are more attracted to blue. Here, we demonstrate that the innate colour preference can be modified by olfactory stimuli in a sexually dimorphic manner. Naive P. xuthus were presented with four coloured discs: blue, green, yellow and red. The innate colour preference (i.e. the colour first landed on) of the majority of individuals was blue. When scent from essential oils of either orange flower or lily was introduced to the room, females’ tendency to select the red disc increased. Scents of lavender and flowering potted Hibiscus rosa-sinensis, however, were less effective. Interestingly, the odour of the non-flowering larval host plant, Citrus unshiu, shifted the preference to green in females. In males, however, all plant scents were less effective than in females, such that blue was always the most favoured colour. These observations indicate that interactions between visual and olfactory cues play a more prominent role in females. PMID:26179802

  9. Expression of chimeric P450 genes encoding flavonoid-3', 5'-hydroxylase in transgenic tobacco and petunia plants(1).

    Science.gov (United States)

    Shimada, Y; Nakano-Shimada, R; Ohbayashi, M; Okinaka, Y; Kiyokawa, S; Kikuchi, Y

    1999-11-19

    Flavonoid-3',5'-hydroxylase (F3'5'H), a member of the cytochrome P450 family, is the key enzyme in the synthesis of 3', 5'-hydroxylated anthocyanins, which are generally required for blue or purple flowers. A full-length cDNA, TG1, was isolated from prairie gentian by heterologous hybridization with a petunia cDNA, AK14, which encodes F3'5'H. To investigate the in vivo function of TG1 and AK14, they were subcloned into a plant expression vector and expressed under the control of the CaMV35S promoter in transgenic tobacco or petunia, both of which originally lack the enzyme. Transgenic petunia plants had a dramatic change in flower color from pink to magenta with a high content of 3',5'-hydroxylated anthocyanins. In contrast, transgenic tobacco plants had minimal color change with at most 35% 3',5'-hydroxylated anthocyanin content. These results indicate that the products of TG1 and AK14 have F3'5'H activity in planta and that interspecific gene transfer alters anthocyanin pigment synthesis. The difference in apparent F3'5'H activity between tobacco and petunia is discussed.

  10. Immunogenicity of nuclear-encoded LTB:ST fusion protein from Escherichia coli expressed in tobacco plants.

    Science.gov (United States)

    Rosales-Mendoza, Sergio; Soria-Guerra, Ruth E; Moreno-Fierros, Leticia; Govea-Alonso, Dania O; Herrera-Díaz, Areli; Korban, Schuyler S; Alpuche-Solís, Ángel G

    2011-06-01

    Enterotoxigenic Escherichia coli (ETEC) is one of the main causative agents of diarrhea in infants and for travelers. Inclusion of a heat-stable (ST) toxin into vaccine formulations is mandatory as most ETEC strains can produce both heat-labile (LT) and ST enterotoxins. In this study, a genetic fusion gene encoding for an LTB:ST protein has been constructed and transferred into tobacco via Agrobacterium tumefaciens-mediated transformation. Transgenic tobacco plants carrying the LTB:ST gene are then subjected to GM1-ELISA revealing that the LTB:ST has assembled into pentamers and displays antigenic determinants from both LTB and ST. Protein accumulation of up to 0.05% total soluble protein is detected. Subsequently, mucosal and systemic humoral responses are elicited in mice orally dosed with transgenic tobacco leaves. This has suggested that the plant-derived LTB:ST is immunogenic via the oral route. These findings are critical for the development of a plant-based vaccine capable of eliciting broader protection against ETEC and targeting both LTB and ST. Features of this platform in comparison to transplastomic approaches are discussed.

  11. Frequency and distance of transposition of a modified Dissociation element in transgenic tobacco

    NARCIS (Netherlands)

    Biezen, E.A. van der; Cardol, E.F.; Chung, H.Y.; Nijkamp, H.J.J.; Hille, J.

    1996-01-01

    Effective transposon tagging with the Ac/Ds system in heterologous plant species relies on the accomplishment of a potentially high transposon-induced mutation frequency. The primary parameters that determine the mutation frequency include the transposition frequency and the transposition distance.

  12. A novel 5-enolpyruvylshikimate-3-phosphate synthase shows high glyphosate tolerance in Escherichia coli and tobacco plants.

    Directory of Open Access Journals (Sweden)

    Gaoyi Cao

    Full Text Available A key enzyme in the shikimate pathway, 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS is the primary target of the broad-spectrum herbicide glyphosate. Identification of new aroA genes coding for EPSPS with a high level of glyphosate tolerance is essential for the development of glyphosate-tolerant crops. In the present study, the glyphosate tolerance of five bacterial aroA genes was evaluated in the E. coli aroA-defective strain ER2799 and in transgenic tobacco plants. All five aroA genes could complement the aroA-defective strain ER2799, and AM79 aroA showed the highest glyphosate tolerance. Although glyphosate treatment inhibited the growth of both WT and transgenic tobacco plants, transgenic plants expressing AM79 aroA tolerated higher concentration of glyphosate and had a higher fresh weight and survival rate than plants expressing other aroA genes. When treated with high concentration of glyphosate, lower shikimate content was detected in the leaves of transgenic plants expressing AM79 aroA than transgenic plants expressing other aroA genes. These results suggest that AM79 aroA could be a good candidate for the development of transgenic glyphosate-tolerant crops.

  13. Dissociation of a population of Pectobacterium atrosepticum SCRI1043 in tobacco plants: formation of bacterial emboli and dormant cells.

    Science.gov (United States)

    Gorshkov, Vladimir; Daminova, Amina; Ageeva, Marina; Petrova, Olga; Gogoleva, Natalya; Tarasova, Nadezhda; Gogolev, Yuri

    2014-05-01

    The population dynamics of Pectobacterium atrosepticum SCRI1043 (Pba) within tobacco plants was monitored from the time of inoculation until after long-term preservation of microorganisms in the remnants of dead plants. We found and characterised peculiar structures that totally occlude xylem vessels, which we have named bacterial emboli. Viable but non-culturable (VBN) Pba cells were identified in the remnants of dead plants, and the conditions for resuscitation of these VBN cells were established. Our investigation shows that dissociation of the integrated bacterial population during plant colonisation forms distinct subpopulations and cell morphotypes, which are likely to perform specific functions that ensure successful completion of the life cycle within the plant.

  14. Influence of soil properties on yield and quality of tobacco plant in Akhisar region of Turkey

    Directory of Open Access Journals (Sweden)

    Sezai Delibacak

    2014-12-01

    Full Text Available The research was carried out in Akhisar environs where tobacco was very popular in the period of 2004-2005. In this study, 9 fields were selected which are known to show differences in terms of the quality and efficiency in the villages called Hacıosmanlar Arabacıbozköy, Dereköy, Mecidiye and Süleymanlı. In order to find out the differences caused by the efficiency, the some properties of soils were examined. The relationships between yield and quality of tobacco and some soil properties were determined by correlation tests. After two years of the study, total alkoloid (nicotine, total reducing sugar, total nitrogen, and raw ash were measured as 0.126-1.410%, 7.81-33.71%, 0.45-3.24 %, 8.49-30.01%, respectively. The yield and total reducing sugar were decreased by increasing bulk density as an important soil property. On the other side raw ash content of tobacco increased. It is recommended that low raw ash and high sugar content are required for tobacco quality. With this content, The yield and quality of tobacco can increase with taken some necessary measurement for decreasing bulk density. The nicotin content of tobacco increased with increasing available Mg, Na and Cu content in soil. On the other side, the raw ash content in tobacco decreased with increasing total salt and available Fe, Zn and Mn in soil. It was determined that there was a positif relationship between salt in soil and reducing sugar in soil which is another quality factor for tobacco. In the research, some results were reached as mentioned above. However, further studies must be carried out in the next years to determine relationships between soil properties and yield and quality of tobacco. It can be possible to improve yield and quality of tobacco with using these relations for producers.

  15. Effect of calcium carbonate on cadmium and nutrients uptake in tobacco (Nicotiana tabacum L.) planted on contaminated soil.

    Science.gov (United States)

    Zeng, Wei-Ai; Li, Fan; Zhou, Hang; Qin, Xiao-Li; Zou, Zi-Jin; Tian, Tao; Zeng, Min; Liao, Bo-Han

    2016-01-01

    In the present study, calcium carbonate (CaCO3) was applied to Cd-contaminated soil at rates of 0, 0.5 and 1.0 g kg(-1). The effect of CaCO3 on soil pH, organic matter, available Cd, exchangeable Cd and level of major nutrients in a tobacco field and on accumulation of various elements in tobacco plants was determined. The results showed that CaCO3 application significantly increased the pH level, available P and exchangeable Ca but decreased organic matter, available Cd, exchangeable Cd, available heavy metals (Fe, Mn, Zn and Cu) and available K in soil. Additionally, CaCO3 application substantially reduced Cd accumulation in tobacco roots, stems, upper leaves, middle leaves and lower leaves, with maximum decrease of 22.3%, 32.1%, 24.5%, 22.0% and 18.2%, respectively. There were large increase in total Ca and slight increases in total N and K but decrease to varying degrees in total Fe, Cu and Zn due to CaCO3 application. CaCO3 had little effect on total P and Mn levels in tobacco leaves.

  16. Host Recovery and Reduced Virus Level in the Upper Leaves after Potato virus Y Infection Occur in Tobacco and Tomato but not in Potato Plants

    Directory of Open Access Journals (Sweden)

    Xianzhou Nie

    2015-02-01

    Full Text Available In this study, the recovery phenomenon following infection with Potato virus Y (PVY was investigated in tobacco (Nicotiana tobaccum, tomato (Solanum lycopersicum and potato (Solanum tuberosum plants. In tobacco plants, infection of severe strains of PVY (PVYN or PVYN:O induced conspicuous vein clearing and leaf deformation in the first three leaves above the inoculated leaves, but much milder symptoms in the upper leaves. The recovery phenotype was not obvious in tobacco plants infected with PVY strain that induce mild symptoms (PVYO. However, regardless of the virus strains, reduction in PVY RNA levels was similarly observed in the upper leaves of these plants. Removal of the first three leaves above the inoculated leaves interfered with the occurrence of recovery, suggesting that the signal(s mediating the recovery is likely generated in these leaves. In PVYN or PVYN:O but not in PVYO-infected tobacco plants, the expression of PR-1a transcripts were correlated with the accumulation level of PVY RNA. Reduced level of PVY RNA in the upper leaves was also observed in infected tomato plants, whereas such phenomenon was not observed in potato plants. PVY-derived small RNAs were detected in both tobacco and potato plants and their accumulation levels were correlated with PVY RNA levels. Our results demonstrate that the recovery phenotype following PVY infection is host-specific and not necessarily associated with the expression of PR-1a and generation of PVY small RNAs.

  17. Oil from Tobacco Leaves: FOLIUM - Installation of Hydrocarbon Accumulating Pathways in Tobacco Leaves

    Energy Technology Data Exchange (ETDEWEB)

    None

    2012-01-01

    PETRO Project: LBNL is modifying tobacco to enable it to directly produce fuel molecules in its leaves for use as a biofuel. Tobacco is a good crop for biofuels production because it is an outstanding biomass crop, has a long history of cultivation, does not compete with the national food supply, and is highly responsive to genetic manipulation. LBNL will incorporate traits for hydrocarbon biosynthesis from cyanobacteria and algae, and enhance light utilization and carbon uptake in tobacco, improving the efficiency of photosynthesis so more fuel can be produced in the leaves. The tobacco-generated biofuels can be processed for gasoline, jet fuel or diesel alternatives. LBNL is also working to optimize methods for planting, cultivating and harvesting tobacco to increase biomass production several-fold over the level of traditional growing techniques.

  18. Transgenic tobacco plants expressing BoRS1 gene from Brassica oleracea var. acephala show enhanced tolerance to water stress

    Indian Academy of Sciences (India)

    Dongqin Tang; Hongmei Qian; Lingxia Zhao; Danfeng Huang; Kexuan Tang

    2005-12-01

    Water stress is by far the leading environmental stress limiting crop yields worldwide. Genetic engineering techniques hold great promise for developing crop cultivars with high tolerance to water stress. In this study, the Brassica oleracea var. acephala BoRS1 gene was transferred into tobacco through Agrobacterium-mediated leaf disc transformation. The transgenic status and transgene expression of the transgenic plants was confirmed by polymerase chain reaction (PCR) analysis, Southern hybridization and semi-quantitative one step RT-PCR analysis respectively. Subsequently, the growth status under water stress, and physiological responses to water stress of transgenic tobacco were studied. The results showed that the transgenic plants exhibited better growth status under water stress condition compared to the untransformed control plants. In physiological assessment of water tolerance, transgenic plants showed more dry matter accumulation and maintained significantly higher levels of leaf chlorophyll content along with increasing levels of water stress than the untransformed control plants. This study shows that BoRS1 is a candidate gene in the engineering of crops for enhanced water stress tolerance.

  19. Populus euphratica XTH overexpression enhances salinity tolerance by the development of leaf succulence in transgenic tobacco plants.

    Science.gov (United States)

    Han, Yansha; Wang, Wei; Sun, Jian; Ding, Mingquan; Zhao, Rui; Deng, Shurong; Wang, Feifei; Hu, Yue; Wang, Yang; Lu, Yanjun; Du, Liping; Hu, Zanmin; Diekmann, Heike; Shen, Xin; Polle, Andrea; Chen, Shaoliang

    2013-11-01

    Populus euphratica is a salt-tolerant tree species that develops leaf succulence after a prolonged period of salinity stress. In the present study, a putative xyloglucan endotransglucosylase/hydrolase gene (PeXTH) from P. euphratica was isolated and transferred to tobacco plants. PeXTH localized exclusively to the endoplasmic reticulum and cell wall. Plants overexpressing PeXTH were more salt tolerant than wild-type tobacco with respect to root and leaf growth, and survival. The increased capacity for salt tolerance was due mainly to the anatomical and physiological alterations caused by PeXTH overexpression. Compared with the wild type, PeXTH-transgenic plants contained 36% higher water content per unit area and 39% higher ratio of fresh weight to dry weight, a hallmark of leaf succulence. However, the increased water storage in the leaves in PeXTH-transgenic plants was not accompanied by greater leaf thickness but was due to highly packed palisade parenchyma cells and fewer intercellular air spaces between mesophyll cells. In addition to the salt dilution effect in response to NaCl, these anatomical changes increased leaf water-retaining capacity, which lowered the increase of salt concentration in the succulent tissues and mesophyll cells. Moreover, the increased number of mesophyll cells reduced the intercellular air space, which improved carbon economy and resulted in a 47-78% greater net photosynthesis under control and salt treatments (100-150 mM NaCl). Taken together, the results indicate that PeXTH overexpression enhanced salt tolerance by the development of succulent leaves in tobacco plants without swelling.

  20. Co-expression of NCED and ALO improves vitamin C level and tolerance to drought and chilling in transgenic tobacco and stylo plants.

    Science.gov (United States)

    Bao, Gegen; Zhuo, Chunliu; Qian, Chunmei; Xiao, Ting; Guo, Zhenfei; Lu, Shaoyun

    2016-01-01

    Abscisic acid (ABA) regulates plant adaptive responses to various environmental stresses, while L-ascorbic acid (AsA) that is also named vitamin C is an important antioxidant and involves in plant stress tolerance and the immune system in domestic animals. Transgenic tobacco (Nicotiana tabacum L.) and stylo [Stylosanthes guianensis (Aublet) Swartz], a forage legume, plants co-expressing stylo 9-cis-epoxycarotenoid dioxygenase (SgNCED1) and yeast D-arabinono-1,4-lactone oxidase (ALO) genes were generated in this study, and tolerance to drought and chilling was analysed in comparison with transgenic tobacco overexpressing SgNCED1 or ALO and the wild-type plants. Compared to the SgNCED1 or ALO transgenic plants, in which only ABA or AsA levels were increased, both ABA and AsA levels were increased in transgenic tobacco and stylo plants co-expressing SgNCED1 and ALO genes. Compared to the wild type, an enhanced drought tolerance was observed in SgNCED1 transgenic tobacco plants with induced expression of drought-responsive genes, but not in ALO plants, while an enhanced chilling tolerance was observed in ALO transgenic tobaccos with induced expression of cold-responsive genes, but not in SgNCED1 plants. Co-expression of SgNCED1 and ALO genes resulted in elevated tolerance to both drought and chilling in transgenic tobacco and stylo plants with induced expression of both drought and cold-responsive genes. Our result suggests that co-expression of SgNCED1 and ALO genes is an effective way for use in forage plant improvement for increased tolerance to drought and chilling and nutrition quality.

  1. Characterization of salt tolerance in ectoine-transformed tobacco plants (Nicotiana tabaccum): photosynthesis, osmotic adjustment, and nitrogen partitioning.

    Science.gov (United States)

    Moghaieb, R E A; Tanaka, N; Saneoka, H; Murooka, Y; Ono, H; Morikawa, H; Nakamura, A; Nguyen, N T; Suwa, R; Fujita, K

    2006-02-01

    Ectoine (1,4,5,6-tetrahydro-2-methyl-4-pyrimidinecarboxylic acid) biosynthetic genes (ect. ABC) from Halomonas elongata were introduced to tobacco plants using an Agrobacterium-mediated gene delivery system. The genes for ectoine biosynthesis were integrated in a stable manner into the tobacco genome and the corresponding transcripts were expressed. The concentration of ectoine under salt-stress conditions was higher in the roots than in leaves. A close relationship was found between stomatal conductance and the amount of transported nitrogen, suggesting that water transport through the xylem in the stem and transpiration may be involved in nitrogen transport to leaves. The data indicate that the turgor values of the ectoine transgenic lines increased with increasing salt concentration. The data revealed two ways in which ectoine enhanced salinity tolerance of tobacco plants. First, ectoine improved the maintenance of root function so that water is taken up consistently and supplied to shoots under saline conditions. Second, ectoine enhanced the nitrogen supply to leaves by increasing transpiration and by protecting Rubisco proteins from deleterious effects of salt, thereby improving the rate of photosynthesis.

  2. Cadmium resistance in transgenic tobacco plants enhanced by expressing bean heavy metal-responsive gene PvSR2

    Institute of Scientific and Technical Information of China (English)

    CHAI; Tuanyao; (柴团耀); CHEN; Qiong; (陈琼); ZHANG; Yuxiu; (张玉秀); DONG; Juan; (董娟); AN; Chengcai; (安成才)

    2003-01-01

    PvSR2 (Phaseolus vulgaris stress-related gene) has been cloned from French bean and shown to be expressed specifically upon heavy metal treatment. In order to investigate the role of PvSR2 in plant, PvSR2 gene under the control of cauliflower mosaic virus 35S promoter was introduced into tobacco mediated with Agrobacterium tumefaciens LBA4404. The regenerated plantlets were selected on medium with 100 mg/L kanamycin. PCR and Southern blot analysis showed PvSR2 gene was integrated in tobacco genome. Gus and Northern blot analysis indicated PvSR2 gene was expressed in transgenic seedling. The heavy metal resistance assay showed that the transgenic tobacco seedlings with the PvSR2 coding sequence exhibited higher tolerance to Cd compared with wild-type (WT) under Cd exposure. The Cd content accumulated in root between transgenic and WT seedlings had no obvious difference at lower Cd external concentration (0.05-0.075 mmol/L CdCl2), whereas transgenic plant showed a lower root Cd content than the control at higher external Cd concentration (0.1 mmol/L CdCl2). These results suggested that the expression of PvSR2 can enhance the Cd tolerance, and PvSR2 may be involved in Cd transportation and accumulation at the test concentration of 0.1 mmol/L Cd.

  3. Cloning of Promoter of Chinese Bean GRP 1.8 Gene and Characterization of Its Function in Transgenic Tobacco Plants

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    In order to learn the expression pattern of GRP1.8(glycine-rich-protein) gene promoter in transgenic plants and to explore its potential application in plant genetic engineering for vascular-specific expression of interested genes, GRP 1.8 promoter was amplified by PCR from Chinese bean genomic DNA. The intermediate vector was constructed by inserting vascular-specific expression promoter of GRP 1.8 gene in vector pBI 101. The regenerated tobacco plants obtained were analyzed by PCR to select the putative transgenic plants. The histochemical localization of GUS(β-D-glucosidase) activity indicates that as for that of GRP 1.8 promoter we can confer the vascular-specific expression of GUS gene.

  4. Multiple Different Defense Mechanisms Are Activated in the Young Transgenic Tobacco Plants Which Express the Full Length Genome of the Tobacco Mosaic Virus, and Are Resistant against this Virus

    Science.gov (United States)

    Jada, Balaji; Soitamo, Arto J.; Siddiqui, Shahid Aslam; Murukesan, Gayatri; Aro, Eva-Mari; Salakoski, Tapio; Lehto, Kirsi

    2014-01-01

    Previously described transgenic tobacco lines express the full length infectious Tobacco mosaic virus (TMV) genome under the 35S promoter (Siddiqui et al., 2007. Mol Plant Microbe Interact, 20: 1489–1494). Through their young stages these plants exhibit strong resistance against both the endogenously expressed and exogenously inoculated TMV, but at the age of about 7–8 weeks they break into TMV infection, with typical severe virus symptoms. Infections with some other viruses (Potato viruses Y, A, and X) induce the breaking of the TMV resistance and lead to synergistic proliferation of both viruses. To deduce the gene functions related to this early resistance, we have performed microarray analysis of the transgenic plants during the early resistant stage, and after the resistance break, and also of TMV-infected wild type tobacco plants. Comparison of these transcriptomes to those of corresponding wild type healthy plants indicated that 1362, 1150 and 550 transcripts were up-regulated in the transgenic plants before and after the resistance break, and in the TMV-infected wild type tobacco plants, respectively, and 1422, 1200 and 480 transcripts were down-regulated in these plants, respectively. These transcriptome alterations were distinctly different between the three types of plants, and it appears that several different mechanisms, such as the enhanced expression of the defense, hormone signaling and protein degradation pathways contributed to the TMV-resistance in the young transgenic plants. In addition to these alterations, we also observed a distinct and unique gene expression alteration in these plants, which was the strong suppression of the translational machinery. This may also contribute to the resistance by slowing down the synthesis of viral proteins. Viral replication potential may also be suppressed, to some extent, by the reduction of the translation initiation and elongation factors eIF-3 and eEF1A and B, which are required for the TMV

  5. Functional components of the bacterial CzcCBA efflux system reduce cadmium uptake and accumulation in transgenic tobacco plants.

    Science.gov (United States)

    Nesler, Andrea; DalCorso, Giovanni; Fasani, Elisa; Manara, Anna; Di Sansebastiano, Gian Pietro; Argese, Emanuele; Furini, Antonella

    2017-03-25

    Cadmium (Cd) is a toxic trace element released into the environment by industrial and agricultural practices, threatening the health of plants and contaminating the food/feed chain. Biotechnology can be used to develop plant varieties with a higher capacity for Cd accumulation (for use in phytoremediation programs) or a lower capacity for Cd accumulation (to reduce Cd levels in food and feed). Here we generated transgenic tobacco plants expressing components of the Pseudomonas putida CzcCBA efflux system. Plants were transformed with combinations of the CzcC, CzcB and CzcA genes, and the impact on Cd mobilization was analysed. Plants expressing PpCzcC showed no differences in Cd accumulation, whereas those expressing PpCzcB or PpCzcA accumulated less Cd in the shoots, but more Cd in the roots. Plants expressing both PpCzcB and PpCzcA accumulated less Cd in the shoots and roots compared to controls, whereas plants expressing all three genes showed a significant reduction in Cd levels only in shoots. These results show that components of the CzcCBA system can be expressed in plants and may be useful for developing plants with a reduced capacity to accumulate Cd in the shoots, potentially reducing the toxicity of food/feed crops cultivated in Cd-contaminated soils. Copyright © 2016 Elsevier B.V. All rights reserved.

  6. The alc-GR system: a modified alc gene switch designed for use in plant tissue culture.

    Science.gov (United States)

    Roberts, Gethin R; Garoosi, G Ali; Koroleva, Olga; Ito, Masaki; Laufs, Patrick; Leader, David J; Caddick, Mark X; Doonan, John H; Tomsett, A Brian

    2005-07-01

    The ALCR/alcA (alc) two-component, ethanol-inducible gene expression system provides stringent control of transgene expression in genetically modified plants. ALCR is an ethanol-activated transcription factor that can drive expression from the ALCR-responsive promoter (alcA). However, the alc system has been shown to have constitutive expression when used in plant callus or cell suspension cultures, possibly resulting from endogenous inducer produced in response to lowered oxygen availability. To widen the use of the alc system in plant cell culture conditions, the receptor domain of the rat glucocorticoid receptor (GR) was translationally fused to the C terminus of ALCR to produce ALCR-GR, which forms the basis of a glucocorticoid-inducible system (alc-GR). The alc-GR switch system was tested in tobacco (Nicotiana tabacum) Bright Yellow-2 suspension cells using a constitutively expressed ALCR-GR with four alternative alcA promoter-driven reporter genes: beta-glucuronidase, endoplasmic reticulum-targeted green fluorescent protein, haemagglutinin, and green fluorescent protein-tagged Arabidopsis (Arabidopsis thaliana) Arath;CDKA;1 cyclin-dependent kinase. Gene expression was shown to be stringently dependent on the synthetic glucocorticoid dexamethasone and, in cell suspensions, no longer required ethanol for induction. Thus, the alc-GR system allows tight control of alcA-driven genes in cell culture and complements the conventional ethanol switch used in whole plants.

  7. An acidic class III chitinase in sugar beet: induction by Cercospora beticola, characterization, and expression in transgenic tobacco plants.

    Science.gov (United States)

    Nielsen, K K; Mikkelsen, J D; Kragh, K M; Bojsen, K

    1993-01-01

    An acidic chitinase (SE) was found to accumulate in leaves of sugar beet (Beta vulgaris) during infection with Cercospora beticola. Two isoforms, SE1 and SE2, with MW of 29 kDa and pI of approximately 3.0 were purified to homogeneity. SE2 is an endochitinase that also exhibits exochitinase activity, i.e., it is capable of hydrolyzing chito-oligosaccharides, including chitobiose, into N-acetyl-glucosamine. Partial amino acid sequence data for SE2 were used to obtain a cDNA clone by polymerase chain reaction. The clone was used to isolate a cDNA clone encoding SE2. The deduced amino acid sequence for SE2 is 58-67% identical to the class III chitinases from cucumber, Arabidopsis, and tobacco. A transient induction of SE2 mRNA during the early stages of infection with C. beticola is much stronger in tolerant plants than in susceptible plants. Transgenic tobacco (Nicotiana benthamiana) plants constitutively accumulate SE2 protein in the intercellular space of their leaves. In a preliminary infection experiment, the transgenic plants did not show increase in resistance against C. nicotianae.

  8. The complete nucleotide sequence and multipartite organization of the tobacco mitochondrial genome: comparative analysis of mitochondrial genomes in higher plants.

    Science.gov (United States)

    Sugiyama, Y; Watase, Y; Nagase, M; Makita, N; Yagura, S; Hirai, A; Sugiura, M

    2005-02-01

    Tobacco is a valuable model system for investigating the origin of mitochondrial DNA (mtDNA) in amphidiploid plants and studying the genetic interaction between mitochondria and chloroplasts in the various functions of the plant cell. As a first step, we have determined the complete mtDNA sequence of Nicotiana tabacum. The mtDNA of N. tabacum can be assumed to be a master circle (MC) of 430,597 bp. Sequence comparison of a large number of clones revealed that there are four classes of boundaries derived from homologous recombination, which leads to a multipartite organization with two MCs and six subgenomic circles. The mtDNA of N. tabacum contains 36 protein-coding genes, three ribosomal RNA genes and 21 tRNA genes. Among the first class, we identified the genes rps1 and psirps14, which had previously been thought to be absent in tobacco mtDNA on the basis of Southern analysis. Tobacco mtDNA was compared with those of Arabidopsis thaliana, Beta vulgaris, Oryza sativa and Brassica napus. Since repeated sequences show no homology to each other among the five angiosperms, it can be supposed that these were independently acquired by each species during the evolution of angiosperms. The gene order and the sequences of intergenic spacers in mtDNA also differ widely among the five angiosperms, indicating multiple reorganizations of genome structure during the evolution of higher plants. Among the conserved genes, the same potential conserved nonanucleotide-motif-type promoter could only be postulated for rrn18-rrn5 in four of the dicotyledonous plants, suggesting that a coding sequence does not necessarily move with the promoter upon reorganization of the mitochondrial genome.

  9. Transgenic tobacco overexpressing Brassica juncea HMG-CoA synthase 1 shows increased plant growth, pod size and seed yield.

    Science.gov (United States)

    Liao, Pan; Wang, Hui; Wang, Mingfu; Hsiao, An-Shan; Bach, Thomas J; Chye, Mee-Len

    2014-01-01

    Seeds are very important not only in the life cycle of the plant but they represent food sources for man and animals. We report herein a mutant of 3-hydroxy-3-methylglutaryl-coenzyme A synthase (HMGS), the second enzyme in the mevalonate (MVA) pathway that can improve seed yield when overexpressed in a phylogenetically distant species. In Brassica juncea, the characterisation of four isogenes encoding HMGS has been previously reported. Enzyme kinetics on recombinant wild-type (wt) and mutant BjHMGS1 had revealed that S359A displayed a 10-fold higher enzyme activity. The overexpression of wt and mutant (S359A) BjHMGS1 in Arabidopsis had up-regulated several genes in sterol biosynthesis, increasing sterol content. To quickly assess the effects of BjHMGS1 overexpression in a phylogenetically more distant species beyond the Brassicaceae, wt and mutant (S359A) BjHMGS1 were expressed in tobacco (Nicotiana tabacum L. cv. Xanthi) of the family Solanaceae. New observations on tobacco OEs not previously reported for Arabidopsis OEs included: (i) phenotypic changes in enhanced plant growth, pod size and seed yield (more significant in OE-S359A than OE-wtBjHMGS1) in comparison to vector-transformed tobacco, (ii) higher NtSQS expression and sterol content in OE-S359A than OE-wtBjHMGS1 corresponding to greater increase in growth and seed yield, and (iii) induction of NtIPPI2 and NtGGPPS2 and downregulation of NtIPPI1, NtGGPPS1, NtGGPPS3 and NtGGPPS4. Resembling Arabidopsis HMGS-OEs, tobacco HMGS-OEs displayed an enhanced expression of NtHMGR1, NtSMT1-2, NtSMT2-1, NtSMT2-2 and NtCYP85A1. Overall, increased growth, pod size and seed yield in tobacco HMGS-OEs were attributed to the up-regulation of native NtHMGR1, NtIPPI2, NtSQS, NtSMT1-2, NtSMT2-1, NtSMT2-2 and NtCYP85A1. Hence, S359A has potential in agriculture not only in improving phytosterol content but also seed yield, which may be desirable in food crops. This work further demonstrates HMGS function in plant reproduction

  10. The expression of tga1a gene from tobacco affects the expression of exogenous gene in transgenic plant

    Institute of Scientific and Technical Information of China (English)

    路子显; 常团结; 李旭刚; 徐军望; 李慧芬; 陈宛新; 冯德江; 肖桂芳; 朱祯

    2003-01-01

    The DNA-binding protein TGA1a of tobacco can specially interact with the enhancer sequence as-1 (-83 to -63) of CaMV35S promoter and show the function of transcriptional activation. In order to study the expression of exogenous gene affected by TGA1a, a trans-actingregulation system was formed by tandem connecting tga1a under the control of the phloem-specific promoter rolC with reporter gene under the control of CaMV35S. Then, the system abovewas utilized to construct a plant expression vector. Moreover, two plant expression vectors wereconstructed with the report gene controlled by CaMV35S and rolC promoter respectively as positive controls. Tobacco leaf disc transformed by Agrobacterium-mediated method and transgenic plants were regenerated. It was proved that the reporter gene existed in the genome of transgenic plants by Southern hybridization. The results of GUS activity indicated that the expression of tga1a controlled by rolC remarkably increased the expression of the reporter gene controlled by CaMV35S. GUS activity of transgenic plants containing trans-acting regulation system was higher than that of transgenic plants containing the reporter gene under the control of CaMV35S and rolC respectively, with the highest GUS activity of about tenfolds of two positive controls. Histochemical method demonstrated that GUS staining amassed mainly in phloem tissue of transgenic plantscontaining the trans-acting regulation system. A new model for arising the expression level and tissue-specific expression of exogenous gene in transgenic plant was established in this study.

  11. A novel stress-induced sugarcane gene confers tolerance to drought, salt and oxidative stress in transgenic tobacco plants.

    Directory of Open Access Journals (Sweden)

    Kevin Begcy

    Full Text Available BACKGROUND: Drought is a major abiotic stress that affects crop productivity worldwide. Sugarcane can withstand periods of water scarcity during the final stage of culm maturation, during which sucrose accumulation occurs. Meanwhile, prolonged periods of drought can cause severe plant losses. METHODOLOGY/PRINCIPAL FINDINGS: In a previous study, we evaluated the transcriptome of drought-stressed plants to better understand sugarcane responses to drought. Among the up-regulated genes was Scdr1 (sugarcane drought-responsive 1. The aim of the research reported here was to characterize this gene. Scdr1 encodes a putative protein containing 248 amino acids with a large number of proline (19% and cysteine (13% residues. Phylogenetic analysis showed that ScDR1is in a clade with homologs from other monocotyledonous plants, separate from those of dicotyledonous plants. The expression of Scdr1 in different varieties of sugarcane plants has not shown a clear association with drought tolerance. CONCLUSIONS/SIGNIFICANCE: The overexpression of Scdr1 in transgenic tobacco plants increased their tolerance to drought, salinity and oxidative stress, as demonstrated by increased photosynthesis, water content, biomass, germination rate, chlorophyll content and reduced accumulation of ROS. Physiological parameters, such as transpiration rate (E, net photosynthesis (A, stomatal conductance (gs and internal leaf CO(2 concentration, were less affected by abiotic stresses in transgenic Scdr1 plants compared with wild-type plants. Overall, our results indicated that Scdr1 conferred tolerance to multiple abiotic stresses, highlighting the potential of this gene for biotechnological applications.

  12. A novel stress-induced sugarcane gene confers tolerance to drought, salt and oxidative stress in transgenic tobacco plants.

    Science.gov (United States)

    Begcy, Kevin; Mariano, Eduardo D; Gentile, Agustina; Lembke, Carolina G; Zingaretti, Sonia Marli; Souza, Glaucia M; Menossi, Marcelo

    2012-01-01

    Drought is a major abiotic stress that affects crop productivity worldwide. Sugarcane can withstand periods of water scarcity during the final stage of culm maturation, during which sucrose accumulation occurs. Meanwhile, prolonged periods of drought can cause severe plant losses. In a previous study, we evaluated the transcriptome of drought-stressed plants to better understand sugarcane responses to drought. Among the up-regulated genes was Scdr1 (sugarcane drought-responsive 1). The aim of the research reported here was to characterize this gene. Scdr1 encodes a putative protein containing 248 amino acids with a large number of proline (19%) and cysteine (13%) residues. Phylogenetic analysis showed that ScDR1is in a clade with homologs from other monocotyledonous plants, separate from those of dicotyledonous plants. The expression of Scdr1 in different varieties of sugarcane plants has not shown a clear association with drought tolerance. The overexpression of Scdr1 in transgenic tobacco plants increased their tolerance to drought, salinity and oxidative stress, as demonstrated by increased photosynthesis, water content, biomass, germination rate, chlorophyll content and reduced accumulation of ROS. Physiological parameters, such as transpiration rate (E), net photosynthesis (A), stomatal conductance (gs) and internal leaf CO(2) concentration, were less affected by abiotic stresses in transgenic Scdr1 plants compared with wild-type plants. Overall, our results indicated that Scdr1 conferred tolerance to multiple abiotic stresses, highlighting the potential of this gene for biotechnological applications.

  13. Plants with modified lignin content and methods for production thereof

    Science.gov (United States)

    Zhao, Qiao; Chen, Fang; Dixon, Richard A.

    2014-08-05

    The invention provides methods for decreasing lignin content and for increasing the level of fermentable carbohydrates in plants by down-regulation of the NST transcription factor. Nucleic acid constructs for down-regulation of NST are described. Transgenic plants are provided that comprise reduced lignin content. Plants described herein may be used, for example, as improved biofuel feedstock and as highly digestible forage crops. Methods for processing plant tissue and for producing ethanol by utilizing such plants are also provided.

  14. Production of hydroxylated fatty acids in genetically modified plants

    Energy Technology Data Exchange (ETDEWEB)

    Somerville, Chris (Portola Valley, CA); Broun, Pierre (Burlingame, CA); van de Loo, Frank (Weston, AU); Boddupalli, Sekhar S. (Manchester, MI)

    2011-08-23

    This invention relates to plant fatty acyl hydroxylases. Methods to use conserved amino acid or nucleotide sequences to obtain plant fatty acyl hydroxylases are described. Also described is the use of cDNA clones encoding a plant hydroxylase to produce a family of hydroxylated fatty acids in transgenic plants. In addition, the use of genes encoding fatty acid hydroxylases or desaturases to alter the level of lipid fatty acid unsaturation in transgenic plants is described.

  15. Influence of Multivalent Vaccines Against Tobacco Virus Disease on Flue-cured Tobacco Plant Traits%多价抗烟草病毒病疫苗对烤烟植物性状的影响

    Institute of Scientific and Technical Information of China (English)

    田洪彰; 刘西金; 曹光辉; 王伟; 岳秀峰; 孙昌友; 李峰; 魏代福; 裴军; 王文杰; 刘文涛

    2016-01-01

    For the situation that tobacco virus disease affected tobacco plant traits and production,set up different virus disease vaccine and control group sprayed tobacco seedlings,to explore the prevention effect of weak poison vaccine on tobacco virus disease, through the selection of terrain height consistent, balanced soil fertile, plot contiguous, surrounding crop not multifarious, plot size consistent conduct comparison test,excluded the interference of external factors, investigated and analyzed the disease in the resettling stage,prosperous stage,flat period,to conclud the effects of weak tobacco virus vaccine on tobacco disease resistance.The results showed that the control effect of TVBMV-TCP treatment was the best,could effectively reduce the occurrence of virus disease,promote tobacco normal growth,tobacco plant traits were the best,PVX-PT treatment was superior to the control, comprehensive evaluation concluded that tobacco plant spraying polyvalent virus vaccine grew well, with strong disease resistance and good plant traits.%针对烟草病毒病影响影响烟叶植物性状和产量的实际,设置不同病毒病疫苗与对照组对烟草幼苗进行喷施,探索弱毒疫苗对烟草病毒病的防治效果,通过选取地势高低一致、土质肥沃程度均衡、地块连片、周边作物不繁杂、地块大小一致的烟田进行对比试验,排除外界因素的干扰,分别在团棵期、旺长期、平顶期进行病害调查分析,得出烟草弱病毒疫苗对烟草抗病性的影响。结果表明:TVBMV-TCP处理的防治效果最好,能够有效降低病毒病的发生,促进烟叶正常生长,植物性状最好。PVX-PT处理优于对照处理。综合评价喷施多价抗病毒病疫苗的烟株大田长势良好,抗病性较强,植物性状表现优良。

  16. Improvement of pest resistance in transgenic tobacco plants expressing dsRNA of an insect-associated gene EcR.

    Directory of Open Access Journals (Sweden)

    Jin-Qi Zhu

    Full Text Available The adoption of pest-resistant transgenic plants to reduce yield loss and pesticide utilization has been successful in the past three decades. Recently, transgenic plant expressing double-stranded RNA (dsRNA targeting pest genes emerges as a promising strategy for improving pest resistance in crops. The steroid hormone, 20-hydroxyecdysone (20E, predominately controls insect molting via its nuclear receptor complex, EcR-USP. Here we report that pest resistance is improved in transgenic tobacco plants expressing dsRNA of EcR from the cotton bollworm, Helicoverpa armigera, a serious lepidopteran pest for a variety of crops. When H. armigera larvae were fed with the whole transgenic tobacco plants expressing EcR dsRNA, resistance to H. armigera was significantly improved in transgenic plants. Meanwhile, when H. armigera larvae were fed with leaves of transgenic tobacco plants expressing EcR dsRNA, its EcR mRNA level was dramatically decreased causing molting defects and larval lethality. In addition, the transgenic tobacco plants expressing H. armigera EcR dsRNA were also resistant to another lepidopteran pest, the beet armyworm, Spodoptera exigua, due to the high similarity in the nucleotide sequences of their EcR genes. This study provides additional evidence that transgenic plant expressing dsRNA targeting insect-associated genes is able to improve pest resistance.

  17. Migration of Azospirillum brasilense Yu62 from Root to Stem and Leaves Inside Rice and Tobacco Plants

    Institute of Scientific and Technical Information of China (English)

    CHIFeng; SHENShi-Hua; CHENSan-Feng; JINGYu-Xiang

    2004-01-01

    Azospirillum brasilense Tarrand, Krieg et Doebereiner is one of the important plant growthpromotion endophytes. A. brasilense Yu62 tagged with gfp gene was inoculated into roots of rice and tobacco seedlings, which were then, cultured in gnotobiotic condition. At a certain days after inoculation the different portions of the seedling were observed under laser confocal microscope, resulting in that A.brasilense Yu62 bacteria were colonized in epidermal and cortical cells, intercellular spaces and vascular system of stem and leaf tissue interiors besides in roots. Higher populations of the bacteria isolated from roots, stems and leaves indicated that A. brasilense Yu62 bacteria could ascend themselves from roots to stems and leaves of rice and tobacco. This observation lays down the foundation for ecology and cell morphology of bacterial migration inside plants, interaction between A. brasilense Yu62 bacteria and host cells as well as the plant-growth promotion, provides scientific basis for further application, and is of importance in science and practice.

  18. Activation of Pathogenesis-related Genes by the Rhizobacterium, Bacillus sp. JS, Which Induces Systemic Resistance in Tobacco Plants

    Directory of Open Access Journals (Sweden)

    Ji-Seong Kim

    2015-06-01

    Full Text Available Plant growth promoting rhizobacteria (PGPR are known to confer disease resistance to plants. Bacillus sp. JS demonstrated antifungal activities against five fungal pathogens in in vitro assays. To verify whether the volatiles of Bacillus sp. JS confer disease resistance, tobacco leaves pre-treated with the volatiles were damaged by the fungal pathogen, Rhizoctonia solani and oomycete Phytophthora nicotianae. Pre-treated tobacco leaves had smaller lesion than the control plant leaves. In pathogenesis-related (PR gene expression analysis, volatiles of Bacillus sp. JS caused the up-regulation of PR-2 encoding β-1,3-glucanase and acidic PR-3 encoding chitinase. Expression of acidic PR-4 encoding chitinase and acidic PR-9 encoding peroxidase increased gradually after exposure of the volatiles to Bacillus sp. JS. Basic PR-14 encoding lipid transfer protein was also increased. However, PR-1 genes, as markers of salicylic acid (SA induced resistance, were not expressed. These results suggested that the volatiles of Bacillus sp. JS confer disease resistance against fungal and oomycete pathogens through PR genes expression.

  19. Expression of geminiviral AC2 RNA silencing suppressor changes sugar and jasmonate responsive gene expression in transgenic tobacco plants

    Directory of Open Access Journals (Sweden)

    Soitamo Arto J

    2012-11-01

    Full Text Available Abstract Background RNA-silencing is a conserved gene regulation and surveillance machinery, which in plants, is also used as major defence mechanism against viruses. Various virus-specific dsRNA structures are recognized by the silencing machinery leading to degradation of the viral RNAs or, as in case of begomoviruses, to methylation of their DNA genomes. Viruses produce specific RNA silencing suppressor (RSS proteins to prevent these host defence mechanisms, and as these interfere with the silencing machinery they also disturb the endogenous silencing reactions. In this paper, we describe how expression of AC2 RSS, derived from African cassava mosaic geminivirus changes transcription profile in tobacco (Nicotiana tabacum leaves and in flowers. Results Expression of AC2 RSS in transgenic tobacco plants induced clear phenotypic changes both in leaves and in flowers. Transcriptomes of these plants were strongly altered, with total of 1118 and 251 differentially expressed genes in leaves and flowers, respectively. The three most up-regulated transcript groups were related to stress, cell wall modifications and signalling, whereas the three most down-regulated groups were related to translation, photosynthesis and transcription. It appears that many of the gene expression alterations appeared to be related to enhanced biosynthesis of jasmonate and ethylene, and consequent enhancement of the genes and pathways that are regulated by these hormones, or to the retrograde signalling caused by the reduced photosynthetic activity and sugar metabolism. Comparison of these results to a previous transcriptional profiling of HC-Pro RSS-expressing plants revealed that some of same genes were induced by both RSSs, but their expression levels were typically higher in AC2 than in HC-Pro RSS expressing plants. All in all, a large number of transcript alterations were found to be specific to each of the RSS expressing transgenic plants. Conclusions AC2 RSS in

  20. Expression of geminiviral AC2 RNA silencing suppressor changes sugar and jasmonate responsive gene expression in transgenic tobacco plants.

    Science.gov (United States)

    Soitamo, Arto J; Jada, Balaji; Lehto, Kirsi

    2012-11-07

    RNA-silencing is a conserved gene regulation and surveillance machinery, which in plants, is also used as major defence mechanism against viruses. Various virus-specific dsRNA structures are recognized by the silencing machinery leading to degradation of the viral RNAs or, as in case of begomoviruses, to methylation of their DNA genomes. Viruses produce specific RNA silencing suppressor (RSS) proteins to prevent these host defence mechanisms, and as these interfere with the silencing machinery they also disturb the endogenous silencing reactions. In this paper, we describe how expression of AC2 RSS, derived from African cassava mosaic geminivirus changes transcription profile in tobacco (Nicotiana tabacum) leaves and in flowers. Expression of AC2 RSS in transgenic tobacco plants induced clear phenotypic changes both in leaves and in flowers. Transcriptomes of these plants were strongly altered, with total of 1118 and 251 differentially expressed genes in leaves and flowers, respectively. The three most up-regulated transcript groups were related to stress, cell wall modifications and signalling, whereas the three most down-regulated groups were related to translation, photosynthesis and transcription. It appears that many of the gene expression alterations appeared to be related to enhanced biosynthesis of jasmonate and ethylene, and consequent enhancement of the genes and pathways that are regulated by these hormones, or to the retrograde signalling caused by the reduced photosynthetic activity and sugar metabolism. Comparison of these results to a previous transcriptional profiling of HC-Pro RSS-expressing plants revealed that some of same genes were induced by both RSSs, but their expression levels were typically higher in AC2 than in HC-Pro RSS expressing plants. All in all, a large number of transcript alterations were found to be specific to each of the RSS expressing transgenic plants. AC2 RSS in transgenic tobacco plants interferes with the silencing

  1. Use of recombinant tobacco mosaic virus to achieve RNA interference in plants against the citrus mealybug, Planococcus citri (Hemiptera: Pseudococcidae.

    Directory of Open Access Journals (Sweden)

    Arif Muhammad Khan

    Full Text Available The citrus mealybug, Planococcus citri, is an important plant pest with a very broad plant host range. P. citri is a phloem feeder and loss of plant vigor and stunting are characteristic symptoms induced on a range of host plants, but P. citri also reduces fruit quality and causes fruit drop leading to significant yield reductions. Better strategies for managing this pest are greatly needed. RNA interference (RNAi is an emerging tool for functional genomics studies and is being investigated as a practical tool for highly targeted insect control. Here we investigated whether RNAi effects can be induced in P. citri and whether candidate mRNAs could be identified as possible targets for RNAi-based P. citri control. RNAi effects were induced in P. citri, as demonstrated by specific target reductions of P. citri actin, chitin synthase 1 and V-ATPase mRNAs after injection of the corresponding specific double-stranded RNA inducers. We also used recombinant Tobacco mosaic virus (TMV to express these RNAi effectors in Nicotiana benthamiana plants. We found that P. citri showed lower fecundity and pronounced death of crawlers after feeding on recombinant TMV-infected plants. Taken together, our data show that actin, chitin synthase 1 and V-ATPase mRNAs are potential targets for RNAi against P. citri, and that recombinant TMV is an effective tool for evaluating candidate RNAi effectors in plants.

  2. Use of recombinant tobacco mosaic virus to achieve RNA interference in plants against the citrus mealybug, Planococcus citri (Hemiptera: Pseudococcidae).

    Science.gov (United States)

    Khan, Arif Muhammad; Ashfaq, Muhammad; Kiss, Zsofia; Khan, Azhar Abbas; Mansoor, Shahid; Falk, Bryce W

    2013-01-01

    The citrus mealybug, Planococcus citri, is an important plant pest with a very broad plant host range. P. citri is a phloem feeder and loss of plant vigor and stunting are characteristic symptoms induced on a range of host plants, but P. citri also reduces fruit quality and causes fruit drop leading to significant yield reductions. Better strategies for managing this pest are greatly needed. RNA interference (RNAi) is an emerging tool for functional genomics studies and is being investigated as a practical tool for highly targeted insect control. Here we investigated whether RNAi effects can be induced in P. citri and whether candidate mRNAs could be identified as possible targets for RNAi-based P. citri control. RNAi effects were induced in P. citri, as demonstrated by specific target reductions of P. citri actin, chitin synthase 1 and V-ATPase mRNAs after injection of the corresponding specific double-stranded RNA inducers. We also used recombinant Tobacco mosaic virus (TMV) to express these RNAi effectors in Nicotiana benthamiana plants. We found that P. citri showed lower fecundity and pronounced death of crawlers after feeding on recombinant TMV-infected plants. Taken together, our data show that actin, chitin synthase 1 and V-ATPase mRNAs are potential targets for RNAi against P. citri, and that recombinant TMV is an effective tool for evaluating candidate RNAi effectors in plants.

  3. Molecular phenotyping of lignin-modified tobacco reveals associated changes in cell-wall metabolism, primary metabolism, stress metabolism and photorespiration.

    Science.gov (United States)

    Dauwe, Rebecca; Morreel, Kris; Goeminne, Geert; Gielen, Birgit; Rohde, Antje; Van Beeumen, Jos; Ralph, John; Boudet, Alain-Michel; Kopka, Joachim; Rochange, Soizic F; Halpin, Claire; Messens, Eric; Boerjan, Wout

    2007-10-01

    Lignin is an important component of secondarily thickened cell walls. Cinnamoyl CoA reductase (CCR) and cinnamyl alcohol dehydrogenase (CAD) are two key enzymes that catalyse the penultimate and last steps in the biosynthesis of the monolignols. Downregulation of CCR in tobacco (Nicotiana tabacum) has been shown to reduce lignin content, whereas lignin in tobacco downregulated for CAD incorporates more aldehydes. We show that altering the expression of either or both genes in tobacco has far-reaching consequences on the transcriptome and metabolome. cDNA-amplified fragment length polymorphism-based transcript profiling, combined with HPLC and GC-MS-based metabolite profiling, revealed differential transcripts and metabolites within monolignol biosynthesis, as well as a substantial network of interactions between monolignol and other metabolic pathways. In general, in all transgenic lines, the phenylpropanoid biosynthetic pathway was downregulated, whereas starch mobilization was upregulated. CCR-downregulated lines were characterized by changes at the level of detoxification and carbohydrate metabolism, whereas the molecular phenotype of CAD-downregulated tobacco was enriched in transcript of light- and cell-wall-related genes. In addition, the transcript and metabolite data suggested photo-oxidative stress and increased photorespiration, mainly in the CCR-downregulated lines. These predicted effects on the photosynthetic apparatus were subsequently confirmed physiologically by fluorescence and gas-exchange measurements. Our data provide a molecular picture of a plant's response to altered monolignol biosynthesis.

  4. 75 FR 62096 - Agricultural Technical Advisory Committees for Trade in Tobacco, Cotton, Peanuts and Planting...

    Science.gov (United States)

    2010-10-07

    ... addresses, such as genetically modified organisms, new technologies and international negotiations, are... the United States Trade Representative (USTR), is considering modifying the existing structure of...

  5. A CBL-Interacting Protein Kinase TaCIPK2 Confers Drought Tolerance in Transgenic Tobacco Plants through Regulating the Stomatal Movement

    Science.gov (United States)

    Li, Tingting; Wang, Meng; Yang, Guangxiao; He, Guangyuan

    2016-01-01

    In plants, the CBL-CIPK signaling pathways play key roles in the response to abiotic stresses. However, functional studies of CIPKs in the important staple crop wheat are very rare. In this study, we identified a CIPK gene from wheat, designated TaCIPK2. Expression analysis results showed that TaCIPK2 could be up-regulated in wheat leaves by polyethylene glycol, abscisic acid and H2O2 treatments. Subcellular localization analyses revealed that TaCIPK2 was present in whole wheat epidermal cells. A yeast two-hybrid assay indicated that TaCIPK2 interacted with TaCBL1, 2, 3 and 4 in vitro. Transgenic tobacco plants over-expressing TaCIPK2 exhibited increased drought tolerance, indicated by a larger proportion of green cotyledons and higher survival rates under the osmotic and drought stress conditions compared with control plants. Additionally, physiological index analyses revealed that the transgenic tobacco plants had lower water loss rates and ion leakage, accumulated less malondialdehyde and H2O2, and had higher catalase and superoxide dismutase activities than the control plants. The transgenic plants also exhibited faster stomatal closure following exposure to osmotic stress conditions. The seed germination rates and stomatal aperture of TaCIPK2-overexpressing tobacco plants decreased after exogenous abscisic acid treatment was applied, implying that the transgenic tobacco plants were more sensitive to exogenous abscisic acid than the control plants. Our results indicate that TaCIPK2 plays a positive regulatory role in drought stress responses in transgenic tobacco plants. PMID:27936160

  6. The Ectopic Expression of CaRop1 Modulates the Response of Tobacco Plants to Ralstonia solanacearum and Aphids

    Directory of Open Access Journals (Sweden)

    Shuilin He

    2016-08-01

    Full Text Available In plants, Rho-related GTPases (Rops are versatile molecular switches that regulate various biological processes, although their exact roles are not fully understood. Herein, we provide evidence that the ectopic expression of a Rop derived from Capsicum annuum, designated CaRop1, in tobacco plants modulates the response of these plants to Ralstonia solanacearum or aphid attack. The deduced amino acid sequence of CaRop1 harbors a conserved Rho domain and is highly homologous to Rops of other plant species. Transient expression of a CaRop1-GFP fusion protein in Nicotiana benthamiana leaf epidermal cells revealed localization of the GFP signal to the plasma membrane, cytoplasm, and nucleus. Overexpression (OE of the wild-type CaRop1 or its dominant-negative mutant (DN-CaRop1 conferred substantial resistance to R. solanacearum infection and aphid attack, and this effect was accompanied by enhanced transcriptional expression of the hypersensitive-reaction marker gene HSR201; the jasmonic-acid (JA-responsive PR1b and LOX1; the insect resistance-associated NtPI-I, NtPI-II, and NtTPI; the ethylene (ET production-associated NtACS1; and NPK1, a mitogen-activated protein kinase kinase kinase (MAPKKK that interferes with N-, Bs2-, and Rx-mediated disease resistance. In contrast, OE of the constitutively active mutant of CaRop1(CA-CaRop1 enhanced susceptibility of the transgenic tobacco plants to R. solanacearum infection and aphid attack and downregulated or sustained the expression of HSR201, PR1b, NPK1, NtACS1, NtPI-I, NtPI-II and NtTPI. These results collectively suggest that CaRop1 acts as a signaling switch in the crosstalk between Solanaceaes’s response to R. solanacearum infection and aphid attack possibly via JA/ET-mediated signaling machinery.

  7. Analysis of the photosynthetic apparatus in transgenic tobacco plants with altered endogenous cytokinin content: a proteomic study

    Directory of Open Access Journals (Sweden)

    Valcke Roland

    2011-06-01

    Full Text Available Abstract Background Cytokinin is a plant hormone that plays a crucial role in several processes of plant growth and development. In recent years, major breakthroughs have been achieved in the elucidation of the metabolism, the signal perception and transduction, as well as the biological functions of cytokinin. An important activity of cytokinin is the involvement in chloroplast development and function. Although this biological function has already been known for 50 years, the exact mechanisms remain elusive. Results To elucidate the effects of altered endogenous cytokinin content on the structure and function of the chloroplasts, chloroplast subfractions (stroma and thylakoids from transgenic Pssu-ipt and 35S:CKX1 tobacco (Nicotiana tabacum plants with, respectively, elevated and reduced endogenous cytokinin content were analysed using two different 2-DE approaches. Firstly, thykaloids were analysed by blue-native polyacrylamide gel electrophoresis followed by SDS-PAGE (BN/SDS-PAGE. Image analysis of the gel spot pattern thus obtained from thylakoids showed no substantial differences between wild-type and transgenic tobacco plants. Secondly, a quantitative DIGE analysis of CHAPS soluble proteins derived from chloroplast subfractions indicated significant gel spot abundance differences in the stroma fraction. Upon identification by MALDI-TOF/TOF mass spectrometry, these proteins could be assigned to the Calvin-Benson cycle and photoprotective mechanisms. Conclusion Taken together, presented proteomic data reveal that the constitutively altered cytokinin status of transgenic plants does not result in any qualitative changes in either stroma proteins or protein complexes of thylakoid membranes of fully developed chloroplasts, while few but significant quantitative differences are observed in stroma proteins.

  8. Proposal for a Test Protocol for Genetically Modified Plants

    DEFF Research Database (Denmark)

    Strandberg, B.; Kjær, C.

    1999-01-01

    The report contains the proceedings from the conference Genetically Modified Organisms in Nordic Habitats - Sustainable Use or Loss of Diversity? in Helsinki, 1998......The report contains the proceedings from the conference Genetically Modified Organisms in Nordic Habitats - Sustainable Use or Loss of Diversity? in Helsinki, 1998...

  9. Analysis of Protein Amino Acids in Tobacco Using Microwave Digestion of Plant Material

    Directory of Open Access Journals (Sweden)

    Moldoveanu SC

    2014-12-01

    Full Text Available This paper describes a technique using microwave digestion and gas chromatography-mass spectrometry (GC-MS, which makes possible the analysis of protein amino acids in tobacco. The technique involves first the measurement of free amino acids, a hydrolysis using microwave digestion, and a measurement of total resulting amino acids. The content of protein amino acids is determined from the difference of total and free amino acids. The digestion is performed with aqueous 6 N HCl (with 1% phenol for two hours in a microwave at 120°C in sealed vials. The GC-MS analysis is performed after the amino acids are derivatized with N-methyl-N-(t-butyldimethylsilyltrifluoroacetamide (MTBSTFA. The technique provides reliable results with less than 10% relative standard deviation (RSD for most amino acids. Only the determination of very low level amino acids is affected by larger errors. The method provides results for free amino acids that are in very good agreement with those obtained by high performance liquid chromatography (HPLC, and also results for protein levels in tobacco in agreement with data previously reported in the literature. Results are given for several single grade tobaccos and for tobacco blends from four Kentucky reference cigarettes.

  10. Trans-activation of an artificial dTam3 transposable element in transgenic tobacco plants

    NARCIS (Netherlands)

    Haring, Michel A.; Teeuwen-de Vroomen, Marianne J.; Nijkamp, H. John J.; Hille, Jacques

    1991-01-01

    In Antirrhinum majus only autonomous Tam3 transposons have been characterized. We investigated whether an artificial dTam3 element, with a deletion in the presumptive transposase coding region, can be trans-activated in tobacco by an activator Tam3 element, which was immobilized by the deletion of o

  11. Transglutaminase activity changes during the hypersensitive reaction, a typical defense response of tobacco NN plants to TMV.

    Science.gov (United States)

    Del Duca, Stefano; Betti, Lucietta; Trebbi, Grazia; Serafini-Fracassini, Donatella; Torrigiani, Patrizia

    2007-10-01

    The occurrence of glutamyl polyamines (PAs) and changes in activity and levels of transglutaminase (TGase, EC 2.3.2.13), the enzyme responsible for their synthesis, are reported during the progression of the hypersensitive reaction (HR) of resistant NN tobacco plants (Nicotiana tabacum L. cv. Samsun) to tobacco mosaic virus (TMV). Mature leaves of tobacco were collected over 0-72 h after inoculation with TMV or phosphate buffer (mock). In vivo synthesis of polyamine glutamyl derivatives (glutamyl PAs), catalyzed by TGase activity, was evaluated after supplying labeled putrescine (Pu, a physiological substrate of TGase) to leaves. Results show that, starting from 24 h, mono-(gamma-glutamyl)-Pu and bis-(gamma-glutamyl)-Sd were recovered in TMV-inoculated samples but not in mock-inoculated ones; 2 days later, in the former, the amount of glutamyl derivatives further increased. An in vitro radiometric assay showed that, in TMV-inoculated leaves, TGase activity increased from 24 h onwards relative to mock controls. An immunoblot analysis with AtPng1p polyclonal antibody detected a 72-kDa protein whose amount increased at 72 h in TMV-inoculated leaves and in the lesion-enriched areas. A biotin-labeled cadaverine incorporation assay showed that TGase activity occurred in S1 (containing soluble proteins), S2 (proteins released by both cell walls and membranes) and S3 (membrane intrinsic proteins) fractions. In S3 fraction, where changes were the most relevant, TGase activity was enhanced in both mock-inoculated and TMV-inoculated samples, but the stimulation persisted only in the latter case. These data are discussed in the light of a possible role of TGase activity and glutamyl PAs in the defense against a viral plant pathogen.

  12. Influence of planting frame in the cultivar “Corojo 2006” of tobacco (Nicotiana tabacum L. on productive indicators

    Directory of Open Access Journals (Sweden)

    Luis Gustavo González Gómez

    2015-03-01

    Full Text Available The investigation was developed in the General CCS-F Victor Ramos Hernandez, Located in the area of the Oven, on a carbonated brown floor. The valued variety of tobacco was the Corojo 2006, with the objective of evaluating different plantation marks, for they were evaluated it four plantation distances that in turn were the valued treatments with a design of totally randomized in parcels of 20 m of long and wide The utilized materials they are characteristic of a field investigation and the cultural attentions were carried out by the Technical Instructive of the Tobacco (2011. the main indicators of the cultivation were evaluated like they were number of leaves, long and wide of the leaves, dry mass and yield, with its respective economic valuation. The data were processed with the statistical package it ESTATISTICA version 8 on Windows, through an analysis of simple variance and a test of multiple comparison for Tukey for 5% of probability of the error, The reached results indicate us that the best yields are obtained when the plants have a distance among them of 0.30 m and the lowest results when they are planted 0.25 m.

  13. An engineered pathway for glyoxylate metabolism in tobacco plants aimed to avoid the release of ammonia in photorespiration

    Directory of Open Access Journals (Sweden)

    Carvalho Josirley de FC

    2011-11-01

    Full Text Available Abstract Background The photorespiratory nitrogen cycle in C3 plants involves an extensive diversion of carbon and nitrogen away from the direct pathways of assimilation. The liberated ammonia is re-assimilated, but up to 25% of the carbon may be released into the atmosphere as CO2. Because of the loss of CO2 and high energy costs, there has been considerable interest in attempts to decrease the flux through the cycle in C3 plants. Transgenic tobacco plants were generated that contained the genes gcl and hyi from E. coli encoding glyoxylate carboligase (EC 4.1.1.47 and hydroxypyruvate isomerase (EC 5.3.1.22 respectively, targeted to the peroxisomes. It was presumed that the two enzymes could work together and compete with the aminotransferases that convert glyoxylate to glycine, thus avoiding ammonia production in the photorespiratory nitrogen cycle. Results When grown in ambient air, but not in elevated CO2, the transgenic tobacco lines had a distinctive phenotype of necrotic lesions on the leaves. Three of the six lines chosen for a detailed study contained single copies of the gcl gene, two contained single copies of both the gcl and hyi genes and one line contained multiple copies of both gcl and hyi genes. The gcl protein was detected in the five transgenic lines containing single copies of the gcl gene but hyi protein was not detected in any of the transgenic lines. The content of soluble amino acids including glycine and serine, was generally increased in the transgenic lines growing in air, when compared to the wild type. The content of soluble sugars, glucose, fructose and sucrose in the shoot was decreased in transgenic lines growing in air, consistent with decreased carbon assimilation. Conclusions Tobacco plants have been generated that produce bacterial glyoxylate carboligase but not hydroxypyruvate isomerase. The transgenic plants exhibit a stress response when exposed to air, suggesting that some glyoxylate is diverted away from

  14. The 5' flanking region of a barley B hordein gene controls tissue and developmental specific CAT expression in tobacco plants.

    Science.gov (United States)

    Marris, C; Gallois, P; Copley, J; Kreis, M

    1988-07-01

    The 549 base pairs of the 5' flanking region of a barley seed storage protein (B1 hordein) gene were linked to the reporter gene encoding chloramphenicol acetyl transferase (CAT). The chimaeric gene was transferred into tobacco plants using Agrobacterium tumefaciens. CAT enzyme activity was detected in the seeds, but not in the leaves, of the transgenic plants. Furthermore, enzyme activity was found only in the endosperm, and only from fifteen days after pollination. In contrast, the constitutive 19S promoter from cauliflower mosaic virus (CaMV) directed the expression of the CAT gene in the leaves as well as in both the endosperm and embryo and at all stages in seed development.

  15. Process for modifying the architecture and improving the yield of crop plants

    OpenAIRE

    Rojo, Enrique; Sanmartín, Maite; Sánchez Serrano, José J.

    2011-01-01

    This invention identifies the plant MINIYO (IYO) gene and the AtRTR1 gene which are essential for the initiation of cell differentiation in all plant meristems and in embryogenesis. This invention relates methods for generating transgenic plants in which expression of the IYO and/or At RTR1 genes or their orthologous genes is modified to advancing or delaying the onset of differentiation in one or more meristems of the plant.

  16. Potentiality Of Stressed Alfalfa Plants To Modify Soil Traits

    Directory of Open Access Journals (Sweden)

    Bardees M. Mickky

    2015-08-01

    Full Text Available Leguminous plants play a key role in the sustainability of agricultural systems. Nevertheless it is to somewhat scarce to find investigations about the interaction of such plants with the soil in which they grow under stress. A pot experiment was conducted to investigate the influence of graduated water regimes on the chemical features of different soil types varying in sand proportion before and after their cultivation with alfalfa Medicago sativa plants. The results indicated that soil cultivation with the studied plants whether water-satisfied or not generally increased the amount of soil ammonia nitrate peptide total soluble and total nitrogen. On contrary the amount of soil amino amide nitrite and protein nitrogen generally decreased after cultivation. Concerning soil ionic composition the post-planting values of soil sodium calcium magnesium and sodium potassium ratio were significantly lower than their pre-cultivation synonyms. Furthermore the percentages of decline caused by cultivating well-watered plants were higher than those caused by their moderately-droughted relatives which were in turn higher than those caused by severely-droughted ones. For soil chloride potassium sodium adsorption ratio and potassium adsorption ratio culturing water-unstressed plants reduced these ionic fractions while droughted plants markedly favored such values. As a general feature the maximum titers of all soil nitrogenous and ionic constituents were recorded for soil with the least sand proportion. The other chemical soil characteristics pH electric conductivity organic carbon organic matter calcium carbonate bicarbonates sulphates and total soluble salts were all fluctuated in a random fashion among the various soil types before and after planting water-stressed or control alfalfa plants. Thus the results obtained herein recommend alfalfa as a pioneer plant that can be introduced to infertile and or dry lands with a paramount efficacy to enhance soil

  17. Association of tobacco and alcohol use with earlier development of colorectal cancer: should we modify screening guidelines?

    Science.gov (United States)

    Acott, Alison A; Theus, Sue A; Marchant-Miros, Kathyrn E; Mancino, Anne T

    2008-12-01

    Current guidelines recommend initial colorectal cancer screening at age 50 years for average-risk patients. Alcohol and tobacco use can be associated with earlier onset of colorectal cancer. We hypothesized an earlier age at diagnosis and/or more advanced stage in patients with these habits. We queried our tumor registry for colorectal cancer diagnosed between January 1997 and December 2006. Data were analyzed to evaluate effects of alcohol and tobacco use. Of 335 colorectal cancer patients, 81% used tobacco, 51% used alcohol, 45% used both, and 14% used neither. Current tobacco and alcohol use were associated with younger ages at onset of colorectal cancer. Thirteen of 332 patients were diagnosed with colorectal cancer before age 50 years. All had exposure to alcohol and tobacco. Fifty-four percent (7/13) of these patients presented at stage 3/4 compared with 34% of the overall population. Modification of screening guidelines to include these habits as "high-risk" factors may be indicated.

  18. Examination of the biological effects of high anionic peroxidase production in tobacco plants grown under field conditions. I. Insect pest damage.

    Science.gov (United States)

    Dowd, Patrick F; Lagrimini, L Mark

    2006-04-01

    At least 25 wild type and high peroxidase tobacco Nicotiana tabacum L. plants were examined semiweekly over several weeks for pest insect distribution and damage in a 2 year field study. Incidence and/or severity of naturally occurring caterpillar damage (dingy cutworm (Feltia ducens Walker), black cutworm (Agrotis ipsilon (Hufnagel), tobacco hornworm (Manduca sexta L.), and false tobacco budworm (= corn earworm Helicoverpa zea (Boddie)) was significantly reduced at several sample dates for high peroxidase vs. wild type plants. These results parallel those of prior laboratory studies with caterpillars. The number of adult whiteflies (Trialeurodes vaporariorum (Westwood) per plant was significantly reduced on high peroxidase compared to wild type plants on most sample dates in both years. The number of plants with leaves containing >100 aphids (primarily Myzus persicae Sulzer) per leaf on high peroxidase plants was significantly lower that on wild type plants after an equivalent invasion period in both years. A significantly higher proportion of aphids were found dead on leaf five of high peroxidase compared to wild type plants at most sample dates in both years. These results indicate that high peroxidase plants have resistance to a wide range of insects, implicating this enzyme as a broad range resistance mechanism.

  19. Chemical modification of the inner and outer surfaces of Tobacco Mosaic Virus (TMV).

    Science.gov (United States)

    Bruckman, Michael A; Steinmetz, Nicole F

    2014-01-01

    Viral nanoparticles derived from tobacco mosaic virus (TMV) find applications in various fields. We report the purification and chemical modification of TMV which is a hollow rod-shaped plant viral nanoparticle with modifiable interior and exterior surfaces. We describe methods to isolate TMV from its tobacco plant host for spatially controlled interior and exterior chemical modification and to characterize the resulting TMV hybrid materials.

  20. Chemical Modification of the Inner and Outer Surfaces of Tobacco Mosaic Virus (TMV)

    OpenAIRE

    Bruckman, Michael A.; Steinmetz, Nicole F.

    2014-01-01

    Viral nanoparticles derive from tobacco mosaic virus (TMV) has broad applications in various fields. We report the purification and chemical engineering of TMV which is a hollow rod-shaped plant viral nanoparticle with modifiable interior and exterior surfaces. Here, we describe methods to isolate TMV from its tobacco plant host for spatially controlled interior and exterior chemical modification and to characterize the resulting TMV hybrid materials.

  1. A statistical assessment of differences and equivalences between genetically modified and reference plant varieties

    NARCIS (Netherlands)

    Voet, van der H.; Perry, J.N.; Amzal, B.; Paoletti, C.

    2011-01-01

    Background - Safety assessment of genetically modified organisms is currently often performed by comparative evaluation. However, natural variation of plant characteristics between commercial varieties is usually not considered explicitly in the statistical computations underlying the assessment. Re

  2. A statistical assessment of differences and equivalences between genetically modified and reference plant varieties

    OpenAIRE

    Amzal Billy; Perry Joe N; van der Voet Hilko; Paoletti Claudia

    2011-01-01

    Abstract Background Safety assessment of genetically modified organisms is currently often performed by comparative evaluation. However, natural variation of plant characteristics between commercial varieties is usually not considered explicitly in the statistical computations underlying the assessment. Results Statistical methods are described for the assessment of the difference between a genetically modified (GM) plant variety and a conventional non-GM counterpart, and for the assessment o...

  3. Transgenic plants protected from insect attack

    Science.gov (United States)

    Vaeck, Mark; Reynaerts, Arlette; Höfte, Herman; Jansens, Stefan; de Beuckeleer, Marc; Dean, Caroline; Zabeau, Marc; Montagu, Marc Van; Leemans, Jan

    1987-07-01

    The Gram-positive bacterium Bacillus thuringiensis produces proteins which are specifically toxic to a variety of insect species. Modified genes have been derived from bt2, a toxin gene cloned from one Bacillus strain. Transgenic tobacco plants expressing these genes synthesize insecticidal proteins which protect them from feeding damage by larvae of the tobacco hornworm.

  4. 75 FR 76921 - Tobacco Transition Payment Program; Tobacco Transition Assessments

    Science.gov (United States)

    2010-12-10

    ... Corporation 7 CFR Part 1463 RIN 0560-AH30 Tobacco Transition Payment Program; Tobacco Transition Assessments... Commodity Credit Corporation (CCC) is modifying the regulations for the Tobacco Transition Payment Program (TTPP) to clarify, consistent with current practice and as required by the Fair and Equitable...

  5. RNA-Guided Cas9-Induced Mutagenesis in Tobacco Followed by Efficient Genetic Fixation in Doubled Haploid Plants

    Science.gov (United States)

    Schedel, Sindy; Pencs, Stefanie; Hensel, Götz; Müller, Andrea; Rutten, Twan; Kumlehn, Jochen

    2017-01-01

    Customizable endonucleases are providing an effective tool for genome engineering. The resulting primary transgenic individuals (T0) are typically heterozygous and/or chimeric with respect to any mutations induced. To generate genetically fixed mutants, they are conventionally allowed to self-pollinate, a procedure which segregates individuals into mutant heterozygotes/homozygotes and wild types. The chances of recovering homozygous mutants among the progeny depend not only on meiotic segregation but also on the frequency of mutated germline cells in the chimeric mother plant. In Nicotiana species, the heritability of Cas9-induced mutations has not been demonstrated yet. RNA-guided Cas9 endonuclease-mediated mutagenesis was targeted to the green fluorescent protein (GFP) gene harbored by a transgenic tobacco line. Upon retransformation using a GFP-specific guide RNA/Cas9 construct, the T0 plants were allowed to either self-pollinate, or were propagated via regeneration from in vitro cultured embryogenic pollen which give rise to haploid/doubled haploid plants or from leaf explants that form plants vegetatively. Single or multiple mutations were detected in 80% of the T0 plants. About half of these mutations proved heritable via selfing. Regeneration from in vitro cultured embryogenic pollen allowed for homozygous mutants to be produced more efficiently than via sexual reproduction. Consequently, embryogenic pollen culture provides a convenient method to rapidly generate a variety of genetically fixed mutants following site-directed mutagenesis. The recovery of a mutation not found among sexually produced and analyzed progeny was shown to be achievable through vegetative plant propagation in vitro, which eventually resulted in heritability when the somatic clones were selfed. In addition, some in-frame mutations were associated with functional attenuation of the target gene rather than its full knock-out. The generation of mutants with compromised rather than

  6. Genetic Factors Interact With Tobacco Smoke to Modify Risk for Inflammatory Bowel Disease in Humans and Mice

    DEFF Research Database (Denmark)

    Yadav, Pankaj; Ellinghaus, David; Rémy, Gaëlle

    2017-01-01

    BACKGROUND & AIMS: The role of tobacco smoke in the etiology of inflammatory bowel disease (IBD) is unclear. We investigated interactions between genes and smoking (gene-smoking interactions) that affect risk for Crohn's disease (CD) and ulcerative colitis (UC) in a case-only study of patients an...

  7. MOLECULAR PHENOTYPING OF LIGNIN-MODIFIED TOBACCO REVEALS ASSOCIATED CHANGES IN CELL WALL METABOLISM, PRIMARY METABOLISM, STRESS METABOLISM AND PHOTORESPIRATION

    Science.gov (United States)

    Lignin is an important component of secondary thickened cell walls. Cinnamoyl CoA reductase (CCR) and cinnamyl alcohol dehydrogenase (CAD) are two key enzymes catalyzing the penultimate and last step in the biosynthesis of the monolignols. Down-regulation of CCR in tobacco has been shown to reduce l...

  8. Rhodococcus fascians infection accelerates progression of tobacco BY-2 cells into mitosis through rapid changes in plant gene expression.

    Science.gov (United States)

    Vandeputte, Olivier; Vereecke, Danny; Mol, Adeline; Lenjou, Marc; Van Bockstaele, Dirk; El Jaziri, Mondher; Baucher, Marie

    2007-01-01

    * To characterize plant cell cycle activation following Rhodococcus fascians infection, bacterial impact on cell cycle progression of tobacco BY-2 cells was investigated. * S-phase-synchronized BY-2 cells were cocultivated with R. fascians and cell cycle progression was monitored by measuring mitotic index, cell cycle gene expression and flow cytometry parameters. Cell cycle alteration was further investigated by cDNA-AFLP (amplified fragment length polymorphism). * It was shown that cell cycle progression of BY-2 cells was accelerated only upon infection with bacteria whose virulence gene expression was induced by a leafy gall extract. Thirty-eight BY-2 genes showed a differential expression within 6 h post-infection. Among these, seven were previously associated with specific plant cell cycle phases (in particular S and G2/M phases). Several genes also showed a differential expression during leafy gall formation. * R. fascians-infected BY-2 cells provide a simple model to identify plant genes related to leafy gall development. R. fascians can also be regarded as a useful biotic agent to alter cell cycle progression and, thereby, gain a better understanding of cell cycle regulation in plants.

  9. Overexpression of a Modified Plant Thionin Enhances Disease Resistance to Citrus Canker and Huanglongbing (HLB).

    Science.gov (United States)

    Hao, Guixia; Stover, Ed; Gupta, Goutam

    2016-01-01

    Huanglongbing (HLB or citrus greening disease) caused by Candidatus Liberibacter asiaticus (Las) is a great threat to the US citrus industry. There are no proven strategies to eliminate HLB disease and no cultivar has been identified with strong HLB resistance. Citrus canker is also an economically important disease associated with a bacterial pathogen (Xanthomonas citri). In this study, we characterized endogenous citrus thionins and investigated their expression in different citrus tissues. Since no HLB-resistant citrus cultivars have been identified, we attempted to develop citrus resistant to both HLB and citrus canker through overexpression of a modified plant thionin. To improve effectiveness for disease resistance, we modified and synthesized the sequence encoding a plant thionin and cloned into the binary vector pBinPlus/ARS. The construct was then introduced into Agrobacterium strain EHA105 for citrus transformation. Transgenic Carrizo plants expressing the modified plant thionin were generated by Agrobacterium-mediated transformation. Successful transformation and transgene gene expression was confirmed by molecular analysis. Transgenic Carrizo plants expressing the modified thionin gene were challenged with X. citri 3213 at a range of concentrations, and a significant reduction in canker symptoms and a decrease in bacterial growth were demonstrated compared to nontransgenic plants. Furthermore, the transgenic citrus plants were challenged with HLB via graft inoculation. Our results showed significant Las titer reduction in roots of transgenic Carrizo compared with control plants and reduced scion Las titer 12 months after graft inoculation. These data provide promise for engineering citrus disease resistance against HLB and canker.

  10. Optimization of Acidothermus Celluloyticus Endoglucanase (E1) Production in Transgenic Tobacco Plants by Transcriptional, Post-transcription and Post-Translational Modification

    Energy Technology Data Exchange (ETDEWEB)

    Dai, Ziyu; Hooker, Brian S.; Quesenberry, Ryan D.; Thomas, S. R.

    2005-10-01

    Biochemical characteristics of Acidothermus cellulolyticus endoglucanase (E1) and its physiological effects in transgenic tobacco (Nicotiana tabacum) has been studied previously. In an attempt to obtain a high level of production of intact E1 in transgenic plants, the E1 gene was expressed under the control of strong Mac promoter (a hybrid promoter of manopine synthase promoter and cauliflower mosaic virus 35S promoter enhancer region) or tomato Rubisco small subunit (RbcS-3C) promoter with different 5’ untranslated leader (UTL) sequence and targeted to different subcellular comartmentations with various transit peptides. The expression of E1 protein in transgenic tobacco plants was determined via E1 activity, protein immunobloting, and RNA gel-blotting analyses. Effects of different transit peptides on E1 protein production and its stability were examined in transgenic tobacco plants carrying one of six transgene expression vectors with the same (Mac) promoter and transcription terminator (Tmas). Transgenic tobacco plants with apoplast transit peptide (Mm-apo) had the highest average E1 activity and protein accumulation , while E1 protein was more stable in transgenic plants with no transit peptide (Mm) than others. The E1 expression under tomato RbcS-3C promoter was higher than that under Mac promoter based on the average E1 activity, E1 protein accumulation, and RNA gel-blotting. The E1 expression was increased more than two fold when the 5’-UTL of alfalfa mosaic virus RNA4 gene replaced the UTL of RbcS-3C promoter, while the UTL of alfalfa mosaic virus RNA4 gene was less effective than the UTL of Mac promoter. The optimal combination of promoter, 5’-UTL, and subcellular compartmentation (transit peptide) for E1 protein production in transgenic tobacco plants are discussed.

  11. Nicotine Concentration in Leaves of Flue-cured Tobacco Plants as Affected by Removal of the Shoot Apex and Lateral Buds

    Institute of Scientific and Technical Information of China (English)

    Shu-Sheng Wang; Qiu-Mei Shi; Wen-Qing Li; Jun-Fang Niu; Chun-Jian Li; Fu-Suo Zhang

    2008-01-01

    It is believed that the nicotine concentration in tobacco is closely correlated with the amount of nitrogen (N) supplied.On the other hand,N uptake mainly occurs at the early growth stage,whereas nicotine concentration increases at the late growth stage,especially after removing the shoot apex.To identify the causes of the increased nicotine concentration in tobacco plants,and to compare the effects of different ways of mechanical wounding on nicotine concentration,field experiments were carried out in Fuzhou,Fujian Province in 2003 and 2004.Excision of the shoot apex had almost no influence on N content in the plant;however,it caused dramatic increases in nicotine concentration in leaves,especially in the middle and upper leaves.An additional increase of the nicotine concentration was obtained by removal of axillary buds.The wounding caused by routine leaf harvests,however,did not change the leaf nicotine concentration,and neither did reducing leaf harvest times.The present results revealed no direct relationship between N supply and nicotine concentration in tobacco leaves,and indicate that not all kinds of mechanical wounding were capable of stimulating nicotine synthesis in tobacco plants.Since nicotine production is highly dependent on the removal of apical meristems and hence on the major sources of auxin in the plant,and application of 1-naphthylacetic acid onto the cut surface of the stem after removing the shoot apex markedly decreased the nicotine concentration in different leaves and the total nicotine content in the plant,the results suggest that decreased auxin supply caused by removal of the shoot apex as a kind of mechanical wounding might regulate nicotine synthesis in the roots of tobacco plants.

  12. High levels of bioplastic are produced in fertile transplastomic tobacco plants engineered with a synthetic operon for the production of polyhydroxybutyrate.

    Science.gov (United States)

    Bohmert-Tatarev, Karen; McAvoy, Susan; Daughtry, Sean; Peoples, Oliver P; Snell, Kristi D

    2011-04-01

    An optimized genetic construct for plastid transformation of tobacco (Nicotiana tabacum) for the production of the renewable, biodegradable plastic polyhydroxybutyrate (PHB) was designed using an operon extension strategy. Bacterial genes encoding the PHB pathway enzymes were selected for use in this construct based on their similarity to the codon usage and GC content of the tobacco plastome. Regulatory elements with limited homology to the host plastome yet known to yield high levels of plastidial recombinant protein production were used to enhance the expression of the transgenes. A partial transcriptional unit, containing genes of the PHB pathway and a selectable marker gene encoding spectinomycin resistance, was flanked at the 5' end by the host plant's psbA coding sequence and at the 3' end by the host plant's 3' psbA untranslated region. This design allowed insertion of the transgenes into the plastome as an extension of the psbA operon, rendering the addition of a promoter to drive the expression of the transgenes unnecessary. Transformation of the optimized construct into tobacco and subsequent spectinomycin selection of transgenic plants yielded T0 plants that were capable of producing up to 18.8% dry weight PHB in samples of leaf tissue. These plants were fertile and produced viable seed. T1 plants producing up to 17.3% dry weight PHB in samples of leaf tissue and 8.8% dry weight PHB in the total biomass of the plant were also isolated.

  13. Bioavailability of transgenic microRNAs in genetically modified plants

    Science.gov (United States)

    Transgenic expression of small RNAs is a prevalent approach in agrobiotechnology for the global enhancement of plant foods. Meanwhile, emerging studies have, on the one hand, emphasized the potential of transgenic microRNAs (miRNAs) as novel dietary therapeutics and, on the other, suggested potentia...

  14. Overexpression of plastidial thioredoxins f and m differentially alters photosynthetic activity and response to oxidative stress in tobacco plants

    Directory of Open Access Journals (Sweden)

    Pascal eREY

    2013-10-01

    Full Text Available Plants display a remarkable diversity of thioredoxins (Trxs, reductases controlling the thiol redox status of proteins. The physiological function of many of them remains elusive, particularly for plastidial Trxs f and m, which are presumed based on biochemical data to regulate photosynthetic reactions and carbon metabolism. Recent reports revealed that Trxs f and m participate in vivo in the control of starch metabolism and cyclic photosynthetic electron transfer around photosystem I, respectively. To further delineate their in planta function, we compared the photosynthetic characteristics, the level and/or activity of various Trx targets and the responses to oxidative stress in transplastomic tobacco plants overexpressing either Trx f or Trx m. We found that plants overexpressing Trx m specifically exhibit altered growth, reduced chlorophyll content, impaired photosynthetic linear electron transfer and decreased pools of glutathione and ascorbate. In both transplastomic lines, activities of two enzymes involved in carbon metabolism, NADP-malate dehydrogenase and NADP-glyceraldehyde-3-phosphate dehydrogenase are markedly and similarly altered. In contrast, plants overexpressing Trx m specifically display increased capacity for methionine sulfoxide reductases, enzymes repairing damaged proteins by regenerating methionine from oxidized methionine. Finally, we also observed that transplastomic plants exhibit distinct responses when exposed to oxidative stress conditions generated by methyl viologen or exposure to high light combined with low temperature, the plants overexpressing Trx m being notably more tolerant than Wt and those overexpressing Trx f. Altogether, these data indicate that Trxs f and m fulfill distinct physiological functions. They prompt us to propose that the m type is involved in key processes linking photosynthetic activity, redox homeostasis and antioxidant mechanisms in the chloroplast.

  15. Enhanced drought and heat stress tolerance of tobacco plants with ectopically enhanced cytokinin oxidase/dehydrogenase gene expression.

    Science.gov (United States)

    Macková, Hana; Hronková, Marie; Dobrá, Jana; Turečková, Veronika; Novák, Ondřej; Lubovská, Zuzana; Motyka, Václav; Haisel, Daniel; Hájek, Tomáš; Prášil, Ilja Tom; Gaudinová, Alena; Štorchová, Helena; Ge, Eva; Werner, Tomáš; Schmülling, Thomas; Vanková, Radomíra

    2013-07-01

    Responses to drought, heat, and combined stress were compared in tobacco (Nicotiana tabacum L.) plants ectopically expressing the cytokinin oxidase/dehydrogenase CKX1 gene of Arabidopsis thaliana L. under the control of either the predominantly root-expressed WRKY6 promoter or the constitutive 35S promoter, and in the wild type. WRKY6:CKX1 plants exhibited high CKX activity in the roots under control conditions. Under stress, the activity of the WRKY6 promoter was down-regulated and the concomitantly reduced cytokinin degradation coincided with raised bioactive cytokinin levels during the early phase of the stress response, which might contribute to enhanced stress tolerance of this genotype. Constitutive expression of CKX1 resulted in an enlarged root system, a stunted, dwarf shoot phenotype, and a low basal level of expression of the dehydration marker gene ERD10B. The high drought tolerance of this genotype was associated with a relatively moderate drop in leaf water potential and a significant decrease in leaf osmotic potential. Basal expression of the proline biosynthetic gene P5CSA was raised. Both wild-type and WRKY6:CKX1 plants responded to heat stress by transient elevation of stomatal conductance, which correlated with an enhanced abscisic acid catabolism. 35S:CKX1 transgenic plants exhibited a small and delayed stomatal response. Nevertheless, they maintained a lower leaf temperature than the other genotypes. Heat shock applied to drought-stressed plants exaggerated the negative stress effects, probably due to the additional water loss caused by a transient stimulation of transpiration. The results indicate that modulation of cytokinin levels may positively affect plant responses to abiotic stress through a variety of physiological mechanisms.

  16. Simultaneous Expression of PDH45 with EPSPS Gene Improves Salinity and Herbicide Tolerance in Transgenic Tobacco Plants

    Science.gov (United States)

    Garg, Bharti; Gill, Sarvajeet S.; Biswas, Dipul K.; Sahoo, Ranjan K.; Kunchge, Nandkumar S.; Tuteja, Renu; Tuteja, Narendra

    2017-01-01

    To cope with the problem of salinity- and weed-induced crop losses, a multi-stress tolerant trait is need of the hour but a combinatorial view of such traits is not yet explored. The overexpression of PDH45 (pea DNA helicase 45) and EPSPS (5-enoylpruvyl shikimate-3-phosphate synthase) genes have been reported to impart salinity and herbicide tolerance. Further, the understanding of mechanism and pathways utilized by PDH45 and EPSPS for salinity and herbicide tolerance will help to improve the crops of economical importance. In the present study, we have performed a comparative analysis of salinity and herbicide tolerance to check the biochemical parameters and antioxidant status of tobacco transgenic plants. Collectively, the results showed that PDH45 overexpressing transgenic lines display efficient tolerance to salinity stress, while PDH45+EPSPS transgenics showed tolerance to both the salinity and herbicide as compared to the control [wild type (WT) and vector control (VC)] plants. The activities of the components of enzymatic antioxidant machinery were observed to be higher in the transgenic plants indicating the presence of an efficient antioxidant defense system which helps to cope with the stress-induced oxidative-damages. Photosynthetic parameters also showed significant increase in PDH45 and PDH45+EPSPS overexpressing transgenic plants in comparison to WT, VC and EPSPS transgenic plants under salinity stress. Furthermore, PDH45 and PDH45+EPSPS synergistically modulate the jasmonic acid and salicylic acid mediated signaling pathways for combating salinity stress. The findings of our study suggest that pyramiding of the PDH45 gene with EPSPS gene renders host plants tolerant to salinity and herbicide by enhancing the antioxidant machinery thus photosynthesis.

  17. Transgenic tobacco plants expressing siRNA targeted against the Mungbean yellow mosaic virus transcriptional activator protein gene efficiently block the viral DNA accumulation

    OpenAIRE

    Shanmugapriya, Gnanasekaran; Das, Sudhanshu Sekhar; Veluthambi, Karuppannan

    2015-01-01

    Mungbean yellow mosaic virus (MYMV) is a bipartite begomovirus that infects many pulse crops such as blackgram, mungbean, mothbean, Frenchbean, and soybean. We tested the efficacy of the transgenically expressed intron-spliced hairpin RNA gene of the transcriptional activator protein (hpTrAP) in reducing MYMV DNA accumulation. Tobacco plants transformed with the MYMV hpTrAP gene accumulated 21–22 nt siRNA. Leaf discs of the transgenic plants, agroinoculated with the partial dimers of MYMV, di...

  18. Biochemical Studies on Curing and Fermentation Processing Periods of Different Tobacco Plant (Nicotianatabacum L. Cultivars

    Directory of Open Access Journals (Sweden)

    Szedljak I

    2014-12-01

    Full Text Available We studied the changes in the activity of the enzymes polyphenol oxidase (PPO and peroxidase (POX, the concentration of total soluble phenolic compounds and soluble protein content in different tobacco cultivars (Virginia and Burley during curing. The curing procedure was a special combination of open-air-curing and fluecuring methods and a long fermentation period. In the curing period the changes in data suggested a combination of the biochemical processes and the direct effect of oxygen. A slight increase then a decrease in the concentrations of both total soluble phenolic compounds and the soluble protein content were detected. In this period we found no correlation between the concentration of total soluble phenolic compounds, the decreasing PPO and the increasing POX activity. In the fermentation period a deactivation of the enzymes (PPO and POX and a decrease in the concentration of both total soluble phenolic compounds and soluble protein content were found, because the fermentation is not a biochemically-regulated process. Moreover, these results suggest that the end of curing period is the most favourable time for protein isolation from different tobacco cultivars.

  19. Comparison of Tobacco Host Cell Protein Removal Methods by Blanching Intact Plants or by Heat Treatment of Extracts.

    Science.gov (United States)

    Buyel, Johannes F; Hubbuch, Jürgen; Fischer, Rainer

    2016-01-01

    Plants not only provide food, feed and raw materials for humans, but have also been developed as an economical production system for biopharmaceutical proteins, such as antibodies, vaccine candidates and enzymes. These must be purified from the plant biomass but chromatography steps are hindered by the high concentrations of host cell proteins (HCPs) in plant extracts. However, most HCPs irreversibly aggregate at temperatures above 60 °C facilitating subsequent purification of the target protein. Here, three methods are presented to achieve the heat precipitation of tobacco HCPs in either intact leaves or extracts. The blanching of intact leaves can easily be incorporated into existing processes but may have a negative impact on subsequent filtration steps. The opposite is true for heat precipitation of leaf extracts in a stirred vessel, which can improve the performance of downstream operations albeit with major changes in process equipment design, such as homogenizer geometry. Finally, a heat exchanger setup is well characterized in terms of heat transfer conditions and easy to scale, but cleaning can be difficult and there may be a negative impact on filter capacity. The design-of-experiments approach can be used to identify the most relevant process parameters affecting HCP removal and product recovery. This facilitates the application of each method in other expression platforms and the identification of the most suitable method for a given purification strategy.

  20. Transgenic tobacco plants expressing synthetic Cry1Ac and Cry1le genes are more toxic to cotton bollworm than those containing one gene

    Institute of Scientific and Technical Information of China (English)

    LIAN Yun; JIA ZhiWei; HE KangLai; LIU YunJun; SONG FuPing; WANG BaoMin; WANG GuoYing

    2008-01-01

    Transgenic tobacco plants carrying Cry1Ac, Crylle or both genes were obtained. In the leaves of transgenic plants carrying both genes, the contents of CrylAc and Cry1le proteins were 0.173% and 0.131% of the total proteins, respectively, Cry1Ac protein content was 0.182 % and Cry1le protein con-tent was 0.124% of the total proteins in the leaves of transgenic plants containing only one Bt gone. Fresh leaves of transgenic tobacco and wild-type plants were used for the insect bioassay against wild-type and Cry1Ac-resistant cotton bollworm (Helicoverpa armigera). The bioassay results showed that transgenic plants carrying both genes were significantly more toxic to wild-type and Cry1Ac-resistant cotton bollworm than those carrying Cry1Ac or Cry1le alone. This study indicates that the higher toxicity of transgenic tobacco plants carrying both genes is caused by the cooperative function of both Bt proteins, thus providing a potential way to delay the development of insect resis-tance to transgenic crops.

  1. Metabolic engineering of glycine betaine synthesis: plant betaine aldehyde dehydrogenases lacking typical transit peptides are targeted to tobacco chloroplasts where they confer betaine aldehyde resistance.

    Science.gov (United States)

    Rathinasabapathi, B; McCue, K F; Gage, D A; Hanson, A D

    1994-01-01

    Certain higher plants synthesize and accumulate glycine betaine, a compound with osmoprotectant properties. Biosynthesis of glycine betaine proceeds via the pathway choline-->betaine aldehyde-->glycine betaine. Plants such as tobacco (Nicotiana tabacum L.) which do not accumulate glycine betaine lack the enzymes catalyzing both reactions. As a step towards engineering glycine betaine accumulation into a non-accumulator, spinach and sugar beet complementary-DNA sequences encoding the second enzyme of glycine-betaine synthesis (betaine aldehyde dehydrogenase, BADH, EC 1.2.1.8) were expressed in tobacco. Despite the absence of a typical transit peptide, BADH was targeted to the chloroplast in leaves of transgenic plants. Levels of extractable BADH were comparable to those in spinach and sugar beet, and the molecular weight, isoenzyme profile and Km for betaine aldehyde of the BADH enzymes from transgenic plants were the same as for native spinach or sugar beet BADH. Transgenic plants converted supplied betaine aldehyde to glycine betaine at high rates, demonstrating that they were able to transport betaine aldehyde across both the plasma membrane and the chloroplast envelope. The glycine betaine produced in this way was not further metabolized and reached concentrations similar to those in plants which accumulate glycine betaine naturally. Betaine aldehyde was toxic to non-transformed tobacco tissues whereas transgenic tissues were resistant due to detoxification of betaine aldehyde to glycine betaine. Betaine aldehyded ehydrogenase is therefore of interest as a potential selectable marker, as well as in the metabolic engineering of osmoprotectant biosynthesis.

  2. Isolation of the promoter of a cotton β-galactosidase gene (GhGal1) and its expression in transgenic tobacco plants

    Institute of Scientific and Technical Information of China (English)

    WU; Aimin; LIU; Jinyuan

    2006-01-01

    β-galactosidases (EC 3.2.1.23) constitute a widespread family of glycosyl hydrolases in plants and are thought to be involved in metabolism of cell wall polysaccharides. A cDNA of the cotton (Gossypium hirsutum) β-galactosidase gene, designated GhGal1, has previously been identified and its transcripts are highly abundant at the elongation stage of the cotton fiber. To examine the temporal and spatial control of GhGal1 expression, a transcriptional fusion of the GhGal1 promoter region (1770 bp) with the β-glucuronidase (GUS) reporter gene was introduced into tobacco plants by the Agrobacterium infection method. The resulting transgenic plants showed higher GUS activity of fruit in the transgenic plants than that in the negative and positive controls. Histochemical localization of GUS activity demonstrated that the expression of the GUS gene could be found in the meristem zones of roots, cotyledons, vascular tissues, fruit and trichomes in transgenic tobacco plants. Additionally, sequence analysis of the regulatory region also revealed several conserved motifs among which some were shared with previously reported fruit/seed-specific elements and the others were related with trichome expression. These results indicated the temporal and spatial expression characterization of the GhGal1 promoter in transgenic tobacco plants and provided an important insight into the roles of GhGal1 in cotton fiber development.

  3. Overexpression of a Grapevine Sucrose Transporter (VvSUC27 in Tobacco Improves Plant Growth Rate in the Presence of Sucrose In vitro

    Directory of Open Access Journals (Sweden)

    Yumeng Cai

    2017-06-01

    Full Text Available The import of sugar from source leaves and it further accumulation in grape berries are considerably high during ripening, and this process is mediated via sucrose transporters. In this study, a grape sucrose transporter (SUT gene, VvSUC27, located at the plasma membrane, was transferred to tobacco (Nicotiana tabacum. The transformants were more sensitive to sucrose and showed more rapid development, especially roots, when cultured on MS agar medium containing sucrose, considering that the shoot/root dry weight ratio was only half that of the control. Moreover, all transformed plants exhibited light-colored leaves throughout their development, which indicated chlorosis and an associated reduction in photosynthesis. The total sugar content in the roots and stems of transformants was higher than that in control plants. No significant difference was observed in the leaves between the transformants and control plants. The levels of growth-promoting hormones were increased, and those of stress-mediating hormones were reduced in transgenic tobacco plants. The qRT-PCR analysis revealed that the expression of VvSUC27 was 1,000 times higher than that of the autologous tobacco sucrose transporter, which suggested that the markedly increased growth rate of transformants was because of the heterogeneously expressed gene. The transgenic tobacco plants showed resistance to abiotic stresses. Strikingly, the overexpression of VvSUC27 leaded to the up regulation of most reactive oxygen species scavengers and abscisic acid-related genes that might enable transgenic plants to overcome abiotic stress. Taken together, these results revealed an important role of VvSUC27 in plant growth and response to abiotic stresses, especially in the presence of sucrose in vitro.

  4. Temperature as a modifier of plant-herbivore interaction.

    Science.gov (United States)

    Yang, Shiyong; Ruuhola, Teija; Haviola, Sanna; Rantala, Markus J

    2007-03-01

    Temperature directly affects the growth, survival, and development rates of poikilothermic insect herbivores; it may also have an important indirect impact, via the activities of plant defensive enzymes. The effects of wounding birch leaves and temperature on the growth and development rates of a Lepidopteran moth, Epirrita autumnata, were studied. We also examined the activities of a mountain birch (Betula pubescesns spp. czerepanovii) defensive enzymes, specifically the polyphenoloxidases (PPOs), in relation to temperature and wounding. The optimal temperature for early instars in terms of survival and developmental rates was between +15 and 20 degrees C. Wounding treatment had different effects on birch PPO activity depending on the temperature: at +12 degrees C, wounding decreased the activity, suggesting induced amelioration at that temperature, whereas at +25 degrees C, wounding increased the activity, suggesting induced resistance. However, larval growth was retarded slightly, but significantly, on the leaves of wounded twigs at both temperatures. Both PPO activity and larval growth rates were affected within 12 h, indicating the existence of a transcription- and translation-independent defense system in birch leaves. We suggest that underlying the increase in PPO activity and the decrease in larval growth rate may be H2O2, which has been shown to accumulate in response to wounding. Our results also provide a possible biological mechanism for the hypothesis that low temperatures promote the success of E. autumnata and other Lepidopteran larvae via decreased defensive enzyme activities of host plants at lower temperatures.

  5. Transgenic tobacco revealing altered bacterial diversity in the rhizosphere during early plant development

    NARCIS (Netherlands)

    Andreote, Fernando D.; Mendes, Rodrigo; Dini-Andreote, Francisco; Rossetto, Priscilla B.; Labate, Carlos A.; Pizzirani-Kleiner, Aline A.; van Elsas, Jan Dirck; Azevedo, Joao L.; Araujo, Welington L.

    2008-01-01

    The rhizosphere constitutes a complex niche that may be exploited by a wide variety of bacteria. Bacterium-plant interactions in this niche can be influenced by factors such as the expression of heterologous genes in the plant. The objective of this work was to describe the bacterial communities ass

  6. Transgenic tobacco revealing altered bacterial diversity in the rhizosphere during early plant development

    NARCIS (Netherlands)

    Andreote, Fernando D.; Mendes, Rodrigo; Dini-Andreote, Francisco; Rossetto, Priscilla B.; Labate, Carlos A.; Pizzirani-Kleiner, Aline A.; van Elsas, Jan Dirck; Azevedo, Joao L.; Araujo, Welington L.

    The rhizosphere constitutes a complex niche that may be exploited by a wide variety of bacteria. Bacterium-plant interactions in this niche can be influenced by factors such as the expression of heterologous genes in the plant. The objective of this work was to describe the bacterial communities

  7. Heterologous expression of plant cell wall glycosyltransferases in Pichia, pea and tobacco

    DEFF Research Database (Denmark)

    Petersen, Bent Larsen; Damager, Iben; Faber, Kirsten

    to participate in plant CW biosynthesis, has been achieved in only a few cases. We have previously reported the characterisation of two highly homologous plant-specific membrane-bound GTs, which when expressed as secreted tagged soluble proteins in the baculo virus system, catalysed the transfer of xylose from......The plant cell wall (CW) consists of numerous complex and uniqe carbohydrate polymer structures. Although the structure (sugar composition) of the various plant CW components are known in some detail, functional characterisation of of the more than 300 glycosyltransferases (GTs), that are believed...... UDP-xylose on to the monosaccharide sugar fucose. Partly based on these data, the two genes were proposed to function in the biosynthesis of pectic rhamnogalacturonan II (RG-II) and designated RhamnoGalacturonan XylosylTransferase 1 and -2 (RGXT1 and -2), accordingly (Egelund et al. 2006, The Plant...

  8. Expression of a multi-epitope DPT fusion protein in transplastomic tobacco plants retains both antigenicity and immunogenicity of all three components of the functional oligomer.

    Science.gov (United States)

    Soria-Guerra, Ruth Elena; Alpuche-Solís, Angel G; Rosales-Mendoza, Sergio; Moreno-Fierros, Leticia; Bendik, Elise M; Martínez-González, Luzmila; Korban, Schuyler S

    2009-05-01

    Expression of genes in plant chloroplasts provides an opportunity for enhanced production of target proteins. We report the introduction and expression of a fusion DPT protein containing immunoprotective exotoxin epitopes of Corynebacterium diphtheriae, Bordetella pertussis, and Clostridium tetani in tobacco chloroplasts. Using biolistic-mediated transformation, a plant-optimized synthetic DPT gene was successfully transferred to tobacco plastomes. Putative transplastomic T0 plants were identified by PCR, and Southern blot analysis confirmed homoplasmy in T1 progeny. ELISA assays demonstrated that the DPT protein retained antigenicity of the three components of the fusion protein. The highest level of expression in these transplastomic plants reached 0.8% of total soluble protein. To assess whether the functional recombinant protein expressed in tobacco plants would induce specific antibodies in test animals, a mice feeding experiment was conducted. For mice orally immunized with freeze-dried transplastomic leaves, production of IgG and IgA antibodies specific to each toxin were detected in serum and mucosal tissues.

  9. β-Ketoacyl-acyl Carrier Protein Synthase I (KASI Plays Crucial Roles in the Plant Growth and Fatty Acids Synthesis in Tobacco

    Directory of Open Access Journals (Sweden)

    Tianquan Yang

    2016-08-01

    Full Text Available Fatty acids serve many functions in plants, but the effects of some key genes involved in fatty acids biosynthesis on plants growth and development are not well understood yet. To understand the functions of 3-ketoacyl-acyl-carrier protein synthase I (KASI in tobacco, we isolated two KASI homologs, which we have designated NtKASI-1 and NtKASI-2. Expression analysis showed that these two KASI genes were transcribed constitutively in all tissues examined. Over-expression of NtKASI-1 in tobacco changed the fatty acid content in leaves, whereas over-expressed lines of NtKASI-2 exhibited distinct phenotypic features such as slightly variegated leaves and reduction of the fatty acid content in leaves, similar to the silencing plants of NtKASI-1 gene. Interestingly, the silencing of NtKASI-2 gene had no discernibly altered phenotypes compared to wild type. The double silencing plants of these two genes enhanced the phenotypic changes during vegetative and reproductive growth compared to wild type. These results uncovered that these two KASI genes had the partially functional redundancy, and that the KASI genes played a key role in regulating fatty acids synthesis and in mediating plant growth and development in tobacco.

  10. β-Ketoacyl-acyl Carrier Protein Synthase I (KASI) Plays Crucial Roles in the Plant Growth and Fatty Acids Synthesis in Tobacco.

    Science.gov (United States)

    Yang, Tianquan; Xu, Ronghua; Chen, Jianghua; Liu, Aizhong

    2016-08-08

    Fatty acids serve many functions in plants, but the effects of some key genes involved in fatty acids biosynthesis on plants growth and development are not well understood yet. To understand the functions of 3-ketoacyl-acyl-carrier protein synthase I (KASI) in tobacco, we isolated two KASI homologs, which we have designated NtKASI-1 and NtKASI-2. Expression analysis showed that these two KASI genes were transcribed constitutively in all tissues examined. Over-expression of NtKASI-1 in tobacco changed the fatty acid content in leaves, whereas over-expressed lines of NtKASI-2 exhibited distinct phenotypic features such as slightly variegated leaves and reduction of the fatty acid content in leaves, similar to the silencing plants of NtKASI-1 gene. Interestingly, the silencing of NtKASI-2 gene had no discernibly altered phenotypes compared to wild type. The double silencing plants of these two genes enhanced the phenotypic changes during vegetative and reproductive growth compared to wild type. These results uncovered that these two KASI genes had the partially functional redundancy, and that the KASI genes played a key role in regulating fatty acids synthesis and in mediating plant growth and development in tobacco.

  11. High Levels of Bioplastic Are Produced in Fertile Transplastomic Tobacco Plants Engineered with a Synthetic Operon for the Production of Polyhydroxybutyrate1[C][OA

    Science.gov (United States)

    Bohmert-Tatarev, Karen; McAvoy, Susan; Daughtry, Sean; Peoples, Oliver P.; Snell, Kristi D.

    2011-01-01

    An optimized genetic construct for plastid transformation of tobacco (Nicotiana tabacum) for the production of the renewable, biodegradable plastic polyhydroxybutyrate (PHB) was designed using an operon extension strategy. Bacterial genes encoding the PHB pathway enzymes were selected for use in this construct based on their similarity to the codon usage and GC content of the tobacco plastome. Regulatory elements with limited homology to the host plastome yet known to yield high levels of plastidial recombinant protein production were used to enhance the expression of the transgenes. A partial transcriptional unit, containing genes of the PHB pathway and a selectable marker gene encoding spectinomycin resistance, was flanked at the 5′ end by the host plant’s psbA coding sequence and at the 3′ end by the host plant’s 3′ psbA untranslated region. This design allowed insertion of the transgenes into the plastome as an extension of the psbA operon, rendering the addition of a promoter to drive the expression of the transgenes unnecessary. Transformation of the optimized construct into tobacco and subsequent spectinomycin selection of transgenic plants yielded T0 plants that were capable of producing up to 18.8% dry weight PHB in samples of leaf tissue. These plants were fertile and produced viable seed. T1 plants producing up to 17.3% dry weight PHB in samples of leaf tissue and 8.8% dry weight PHB in the total biomass of the plant were also isolated. PMID:21325565

  12. Signal Peptide of Potato PinⅡ Enhances the Expression of Cry1Ac in Transgenic Tobacco

    Institute of Scientific and Technical Information of China (English)

    Yun-Jun LIU; Yuan YUAN; Jun ZHENG; Ya-Zhong TAO; Zhi-Gang DONG; Jian-Hua WANG; Guo-Ying WANG

    2004-01-01

    The modified Cry1Ac was expressed in transgenic tobacco plants.To allow secretion of the Cry1Ac protein into the intercellular space,the signal peptide sequence of potato proteinase inhibitor Ⅱ(pinⅡ)was N-terminally fused to the CrylAc encoding region.Expression of Cry1Ac in transgenic tobacco plants was assayed with ELISA.The results showed that pinⅡ signal peptide sequence enhanced the expression of Cry 1 Ac protein and led to the secretion of the Cry 1Ac protein in transgenic tobacco plants.GFP gene was also fused to the signal peptide sequence and transformed to tobacco.The results of fluorescent detection showed that GFP had localized in the apoplast of transgenic plants.

  13. Effects of cigarette smoke, cessation and switching to a candidate modified risk tobacco product on the liver in Apoe -/- mice--a systems toxicology analysis.

    Science.gov (United States)

    Lo Sasso, Giuseppe; Titz, Bjoern; Nury, Catherine; Boué, Stéphanie; Phillips, Blaine; Belcastro, Vincenzo; Schneider, Thomas; Dijon, Sophie; Baumer, Karine; Peric, Daruisz; Dulize, Remi; Elamin, Ashraf; Guedj, Emmanuel; Buettner, Ansgar; Leroy, Patrice; Kleinhans, Samuel; Vuillaume, Gregory; Veljkovic, Emilija; Ivanov, Nikolai V; Martin, Florian; Vanscheeuwijck, Patrick; Peitsch, Manuel C; Hoeng, Julia

    2016-04-01

    The liver is one of the most important organs involved in elimination of xenobiotic and potentially toxic substances. Cigarette smoke (CS) contains more than 7000 chemicals, including those that exert biological effects and cause smoking-related diseases. Though CS is not directly hepatotoxic, a growing body of evidence suggests that it may exacerbate pre-existing chronic liver disease. In this study, we integrated toxicological endpoints with molecular measurements and computational analyses to investigate effects of exposures on the livers of Apoe(-/- )mice. Mice were exposed to 3R4F reference CS, to an aerosol from the Tobacco Heating System (THS) 2.2, a candidate modified risk tobacco product (MRTP) or to filtered air (Sham) for up to 8 months. THS2.2 takes advantage of a "heat-not-burn" technology that, by heating tobacco, avoids pyrogenesis and pyrosynthesis. After CS exposure for 2 months, some groups were either switched to the MRTP or filtered air. While no group showed clear signs of hepatotoxicity, integrative analysis of proteomics and transcriptomics data showed a CS-dependent impairment of specific biological networks. These networks included lipid and xenobiotic metabolism and iron homeostasis that likely contributed synergistically to exacerbating oxidative stress. In contrast, most proteomic and transcriptomic changes were lower in mice exposed to THS2.2 and in the cessation and switching groups compared to the CS group. Our findings elucidate the complex biological responses of the liver to CS exposure. Furthermore, they provide evidence that THS2.2 aerosol has reduced biological effects, as compared with CS, on the livers of Apoe(-/- )mice.

  14. Evaluation of a non-targeted "Omic"' approach in the safety assessment of genetically modified plants

    DEFF Research Database (Denmark)

    Metzdorff, Stine Broeng; Kok, E. J.; Knuthsen, Pia;

    2006-01-01

    Genetically modified plants must be approved before release in the European Union, and the approval is generally based upon a comparison of various characteristics between the transgenic plant and a conventional counterpart. As a case study, focusing on safety assessment of genetically modified...... plants, we here report the development and characterisation of six independently transformed Arabidopsis thaliana lines modified in the flavonoid biosynthesis. Analyses of integration events and comparative analysis for characterisation of the intended effects were performed by PCR, quantitative Real......, no unintended effects were identified. However, we found that the majority of genes showing differential expression were identified as stress-related genes and that environmental conditions had a large impact on the expression of several genes, proteins, and metabolites. We suggest that the microarray approach...

  15. The signal peptide-like segment of hpaXm is required for its association to the cell wall in transgenic tobacco plants

    Science.gov (United States)

    Li, Le; Miao, Weiguo; Liu, Wenbo; Zhang, Shujian

    2017-01-01

    Harpins, encoded by hrp (hypersensitive response and pathogenicity) genes of Gram-negative plant pathogens, are elicitors of hypersensitive response (HR). HpaXm is a novel harpin-like protein described from cotton leaf blight bacteria, Xanthomonas citri subsp. malvacearum—a synonym of X. campestris pv. malvacearum (Smith 1901–1978). A putative signal peptide (1-MNSLNTQIGANSSFL-15) of hpaXm was predicted in the nitroxyl-terminal (N-terminal)by SignalP (SignalP 3.0 server). Here, we explored the function of the N-terminal leader peptide like segment of hpaXm using transgenic tobacco (Nicotiana tabacum cv. Xanthi nc.). Transgenic tobacco lines expressing the full-length hpaXm and the signal peptide-like segment-deleted mutant hpaXmΔLP were developed using transformation mediated by Agrobacterium tumefaciens. The target genes were confirmed integrated into the tobacco genomes and expressed normally. Using immune colloidal-gold detection technique, hpaXm protein was found to be transferred to the cytoplasm, the cell membrane, and organelles such as chloroplasts, mitochondria, and nucleus, as well as the cell wall. However, the deletion mutant hpaXmΔLP expressed in transgenic tobacco was found unable to cross the membrane to reach the cell wall. Additionally, soluble proteins extracted from plants transformed with hpaXm and hpaXmΔLP were bio-active. Defensive micro-HR induced by the transgene expression of hpaXm and hpaXmΔLP were observed on transgenic tobacco leaves. Disease resistance bioassays to tobacco mosaic virus (TMV) showed that tobacco plants transformed with hpaXm and with hpaXmΔLP exhibited enhanced resistance to TMV. In summary, the N-terminal signal peptide-like segment (1–45 bp) in hpaXm sequence is not necessary for transgene expression, bioactivity of hpaXm and resistance to TMV in transgenic tobacco, but is required for the protein to be translocated to the cell wall. PMID:28141855

  16. Effects of simultaneous expression of heterologous genes involved in phytochelatin biosynthesis on thiol content and cadmium accumulation in tobacco plants.

    Science.gov (United States)

    Wawrzynski, Adam; Kopera, Edyta; Wawrzynska, Anna; Kaminska, Jolanta; Bal, Wojciech; Sirko, Agnieszka

    2006-01-01

    Transgenic tobacco (Nicotiana tabacum cv. LA Burley 21) lines expressing three genes encoding enzymes thought to be critical for the efficient production of phytochelatins, (i) serine acetyltransferase (EC 2.3.1.30) involved in the production of O-acetylserine, the cysteine precursor, (ii) gamma-glutamylcysteine synthetase (EC 6.3.2.2) involved in the production of gamma-glutamylcysteine, the precursor of glutathione, and (iii) phytochelatin synthase (EC 2.3.2.15), were obtained and analysed for non-protein thiol content and cadmium accumulation. After a 3 week exposure to 15 microM CdCl2, plants expressing transgenes (either separately or in combination) had increased cadmium concentration in roots but not in shoots compared with the wild type. Nearly all transgenic lines analysed had more non-protein thiols than the wild type. The greatest effects (about 8-fold elevation of thiols) were found in one of the lines simultaneously expressing the three transgenes. Despite the fact that a multi-transgene strategy described in this work resulted in a strong increase in the levels of several classes of non-protein thiols in transgenic plants, other factors appeared to restrict cadmium accumulation in shoots.

  17. DGP1, a drought-induced guard cell-specific promoter and its function analysis in tobacco plants

    Institute of Scientific and Technical Information of China (English)

    LI; Jun; GONG; Ximing; LIN; Huiqiong; SONG; Quanbo; CHEN; J

    2005-01-01

    The genetic regulation of stomatal movement mainly depends on an efficient control system of gene expression, and guard cell-specific promoter is becoming the best choice. Here we combined the dehydration responsive element (DRE) with guard cell specific element (GCSE) to construct a novel promoter, DGP1. Histochemical assays in transgenic tobacco carryingβ-glucuronidase (gus) gene fused to DGP1 demonstrated that GUS activity was found to be highly inducible by drought treatment and specifically restricted to guard cells. No GUS activity was detected in roots, stems or flowers after treatment. Further quantitative analysis showed that GUS activity in the epidermal strips was apparently induced by dehydration and dramatically increased with the elongation of treatment. The GUS activity after 8 h treatment was 179 times that of those without treatment. Although GUS activity in roots, stems or mesophyll increased after treatment, no great changes were observed. These results suggested that DGP1 could drive target gene expressed in guard cells when plant is subjected to drought stress. And this gets us prepared to control opening and closing of stomata through plant gene engineering.

  18. Enhanced synthesis of choline and glycine betaine in transgenic tobacco plants that overexpress phosphoethanolamine N-methyltransferase

    Science.gov (United States)

    McNeil, Scott D.; Nuccio, Michael L.; Ziemak, Michael J.; Hanson, Andrew D.

    2001-01-01

    Choline (Cho) is the precursor of the osmoprotectant glycine betaine and is itself an essential nutrient for humans. Metabolic engineering of Cho biosynthesis in plants could therefore enhance both their resistance to osmotic stresses (drought and salinity) and their nutritional value. The key enzyme of the plant Cho-synthesis pathway is phosphoethanolamine N-methyltransferase, which catalyzes all three of the methylations required to convert phosphoethanolamine to phosphocholine. We show here that overexpressing this enzyme in transgenic tobacco increased the levels of phosphocholine by 5-fold and free Cho by 50-fold without affecting phosphatidylcholine content or growth. Moreover, the expanded Cho pool led to a 30-fold increase in synthesis of glycine betaine via an engineered glycine betaine pathway. Supplying the transgenics with the Cho precursor ethanolamine (EA) further enhanced Cho levels even though the supplied EA was extensively catabolized. These latter results establish that there is further scope for improving Cho synthesis by engineering an increased endogenous supply of EA and suggest that this could be achieved by enhancing EA synthesis and/or by suppressing its degradation. PMID:11481443

  19. Comparative systems toxicology analysis of cigarette smoke and aerosol from a candidate modified risk tobacco product in organotypic human gingival epithelial cultures: A 3-day repeated exposure study.

    Science.gov (United States)

    Zanetti, Filippo; Titz, Bjoern; Sewer, Alain; Lo Sasso, Giuseppe; Scotti, Elena; Schlage, Walter K; Mathis, Carole; Leroy, Patrice; Majeed, Shoaib; Torres, Laura Ortega; Keppler, Brian R; Elamin, Ashraf; Trivedi, Keyur; Guedj, Emmanuel; Martin, Florian; Frentzel, Stefan; Ivanov, Nikolai V; Peitsch, Manuel C; Hoeng, Julia

    2017-03-01

    Smoking is one of the major lifestyle-related risk factors for periodontal diseases. Modified risk tobacco products (MRTP) offer a promising alternative in the harm reduction strategy for adult smokers unable to quit. Using a systems toxicology approach, we investigated and compared the exposure effects of a reference cigarette (3R4F) and a heat-not-burn technology-based candidate MRTP, the Tobacco Heating System (THS) 2.2. Human gingival epithelial organotypic cultures were repeatedly exposed (3 days) for 28 min at two matching concentrations of cigarette smoke (CS) or THS2.2 aerosol. Results showed only minor histopathological alterations and minimal cytotoxicity upon THS2.2 aerosol exposure compared to CS (1% for THS2.2 aerosol vs. 30% for CS, at the high concentration). Among the 14 proinflammatory mediators analyzed, only 5 exhibited significant alterations with THS2.2 exposure compared with 11 upon CS exposure. Transcriptomic and metabolomic analysis indicated a general reduction of the impact in THS2.2 aerosol-exposed samples with respect to CS (∼79% lower biological impact for the high THS2.2 aerosol concentration compared to CS, and 13 metabolites significantly perturbed for THS2.2 vs. 181 for CS). This study indicates that exposure to THS2.2 aerosol had a lower impact on the pathophysiology of human gingival organotypic cultures than CS. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

  20. Suggested Improvements for the Allergenicity Assessment of Genetically Modified Plants Used in Foods

    OpenAIRE

    Goodman, Richard E.; Tetteh, Afua O.

    2011-01-01

    Genetically modified (GM) plants are increasingly used for food production and industrial applications. As the global population has surpassed 7 billion and per capita consumption rises, food production is challenged by loss of arable land, changing weather patterns, and evolving plant pests and disease. Previous gains in quantity and quality relied on natural or artificial breeding, random mutagenesis, increased pesticide and fertilizer use, and improved farming techniques, all without a for...

  1. Overexpression of a modified plant thionin enhances disease resistance to citrus canker and Huanglongbing (HLB

    Directory of Open Access Journals (Sweden)

    Guixia Hao

    2016-07-01

    Full Text Available Huanglongbing (HLB or citrus greening disease caused by Candidatus Liberibacter asiaticus (Las is a great threat to the US citrus industry. There are no proven strategies to eliminate HLB disease and no cultivar has been identified with strong HLB resistance. Citrus canker is also an economically important disease associated with a bacterial pathogen (Xanthomonas citri. In this study, we characterized endogenous citrus thionins and investigated their expression in different citrus tissues. Since no HLB-resistant citrus cultivars have been identified, we attempted to develop citrus resistant to both HLB and citrus canker through overexpression of a modified plant thionin. To improve effectiveness for disease resistance, we modified and synthesized the sequence encoding a plant thionin and cloned into the binary vector pBinPlus/ARS. The construct was then introduced into Agrobacterium strain EHA105 for citrus transformation. Transgenic Carrizo plants expressing the modified plant thionin were generated by Agrobacterium-mediated transformation. Successful transformation and transgene gene expression was confirmed by molecular analysis. Transgenic Carrizo plants expressing the modified thionin gene were challenged with X. citri 3213 at a range of concentrations, and a significant reduction in canker symptoms and a decrease in bacterial growth were demonstrated compared to nontransgenic plants. Furthermore the transgenic citrus plants were challenged with HLB via graft inoculation. Our results showed significant Las titer reduction in roots of transgenic Carrizo compared with control plants and reduced scion Las titer twelve months after graft inoculation. These data provide promise for engineering citrus disease resistance against HLB and canker.

  2. Effect of Plant Growth Regulators on Calcium-stimulated Serine Transport into Tobacco Cells

    Science.gov (United States)

    Smith, Ivan K.

    1978-01-01

    The transport of serine into tobacco cells (Nicotiana tabacum L.) cultured in liquid medium was examined. Transport was inhibited approximately 50% by 2,4-dichlorophenoxyacetic acid, indoleacetic acid, α-naphthalene acetic acid, and kinetin at a concentration of 10 micrograms per milliliter. Transport was not inhibited by 2,6-dichlorophenoxyacetic acid and inhibited less than 25% by p-chlorophenoxyacetic acid at this concentration. Removal of 2,4-dichlorophenoxyacetic acid from the transport medium resulted in an alleviation of inhibition. Gibberellic acid at concentrations from 2 to 20 micrograms per milliliter stimulated transport. It was previously shown that inhibition of transport by La3+ was due to removal of Ca2+ from surface sites and inhibition of Ca2+ uptake by cells. None of the growth regulators tested had any significant effect on Ca2+ binding and/or transport. A contributing factor to the low transport rates in the absence of Ca2+ is the increased rate of serine efflux. None of the growth regulators tested had any significant effect on the rate of serine efflux. PMID:16660646

  3. Transgenic tobacco plants accumulating osmolytes show reduced oxidative damage under freezing stress.

    Science.gov (United States)

    Parvanova, Daniela; Ivanov, Sergei; Konstantinova, Tatyana; Karanov, Emanuil; Atanassov, Atanas; Tsvetkov, Tsvetan; Alexieva, Vera; Djilianov, Dimitar

    2004-01-01

    We studied the reaction to the oxidative component of freezing in several tobacco lines, transformed with genes coding for enzymes involved in the synthesis of osmoprotectants (proline, fructan or glycine betaine) along with their wild type. The levels of some oxidative stress markers (leakage of electrolytes, hydrogen peroxide and malondialdehyde) as well as the activity of antioxidative enzymes catalase (EC 1.11.1.6.) and guaiacol peroxidase (EC 1.11.1.7.) have been followed at acclimation, 12 and 24 h freezing and at recovery. Freezing for 24 h resulted in severe damages for the wild type. A corresponding increase of electrolyte leakage, hydrogen peroxide and malondialdehyde contents, a rise of peroxidase activity and inhibition of catalase activity occurred in the non-transformants. Similar, but significantly lower trend of the same parameters has been found for the transgenic lines. Moreover, the oxidative markers returned to their normal levels when the transformants were able to recover from freezing. It could be speculated that transfer of genes, coding for accumulation of osmoprotectants, is related to reduced intensity of freezing-induced oxidative processes. Our lines and model system could serve as a good prerequisite for additional studies to gain further insights into the complex role of osmoprotectants in freezing tolerance.

  4. How the redox state of tobacco 'Bel-W3' is modified in response to ozone and other environmental factors in a sub-tropical area?

    Energy Technology Data Exchange (ETDEWEB)

    Dias, Ana P.L.; Dafre, Marcelle; Rinaldi, Mirian C.S. [Instituto de Botanica, Caixa Postal 3005, 01061-970 Sao Paulo, SP (Brazil); Domingos, Marisa, E-mail: mmingos@superig.com.b [Instituto de Botanica, Caixa Postal 3005, 01061-970 Sao Paulo, SP (Brazil)

    2011-02-15

    This study intended to determine whether the redox state in plants of Nicotiana tabacum 'Bel-W3' fluctuates in response to the environmental factors in a sub-tropical area contaminated by ozone (Sao Paulo, SE - Brazil) and which environmental factors are related to this fluctuation, discussing their biomonitoring efficiency. We comparatively evaluated the indicators of redox state (ascorbic acid, glutathione, superoxide dismutase, ascorbate peroxidase, and glutathione reductase) and leaf injury in 17 field experiments performed in 2008. The redox state was explained by the combined effects of chronic levels of O{sub 3} and meteorological variables 4-6 days prior to the plant sampling. Moderate leaf injury was observed in most cases. The redox state of tobacco decreases few days after their placement in the sub-tropical environment, causing them to become susceptible to oxidative stress imposed by chronic doses of O{sub 3}. Its bioindicator efficiency would not be diminished in such levels of atmospheric contamination. - Research highlights: Nicotiana tabacum 'Bel-W3' is potentially a bioindicator of O{sub 3} in the sub-tropics. However, it is unknown if its redox state would affect its bioindicator performance under sub-tropical environmental conditions. This study revealed that the redox state of tobacco decreases few days after their placement in the sub-tropical environment, causing them to become susceptible to oxidative stress imposed by chronic doses of O{sub 3}. Therefore, its bioindicator efficiency would not be diminished in such levels of atmospheric contamination. However, the bioindicator efficiency N. tabacum 'Bel-W3' for biomonitoring O{sub 3} should be regionally modeled in the sub-tropics, based on both its redox state and on the flux of O{sub 3} through stomata, in response to the varying micro-meteorological conditions that govern both physiological processes. - The bioindicator efficiency of tobacco plants is not

  5. Evaluation of transgenic tobacco plants expressing a bacterial Co-Ni transporter for acquisition of cobalt.

    Science.gov (United States)

    Nair, Smitha; Joshi-Saha, Archana; Singh, Sudhir; Ramachandran, V; Singh, Surya; Thorat, Vidya; Kaushik, C P; Eapen, Susan; D'Souza, S F

    2012-11-15

    Phytoremediation is a viable strategy for management of toxic wastes in a large area/volume with low concentrations of toxic elemental pollutants. With increased industrial use of cobalt and its alloys, it has become a major metal contaminant in soils and water bodies surrounding these industries and mining sites with adverse effects on the biota. A bacterial Co-Ni permease was cloned from Rhodopseudomonas palustris and introduced into Nicotiana tabacum to explore its potential for phytoremediation and was found to be specific for cobalt and nickel. The transgenic plants accumulated more cobalt and nickel as compared to control, whereas no significant difference in accumulation of other divalent ions was observed. The transgenic plants were evaluated for cobalt content and showed increased acquisition of cobalt (up to 5 times) as compared to control. The plants were also assessed for accumulation of nickel and found to accumulate up to 2 times more nickel than control. At the same initial concentration of cobalt and nickel, transgenic plant preferentially accumulated cobalt as compared to nickel. The present study is perhaps the first attempt to develop transgenic plants expressing heterologous Co transporter with an improved capacity to uptake cobalt. Copyright © 2012 Elsevier B.V. All rights reserved.

  6. Site-specific proteolytic degradation of IgG monoclonal antibodies expressed in tobacco plants.

    Science.gov (United States)

    Hehle, Verena K; Lombardi, Raffaele; van Dolleweerd, Craig J; Paul, Mathew J; Di Micco, Patrizio; Morea, Veronica; Benvenuto, Eugenio; Donini, Marcello; Ma, Julian K-C

    2015-02-01

    Plants are promising hosts for the production of monoclonal antibodies (mAbs). However, proteolytic degradation of antibodies produced both in stable transgenic plants and using transient expression systems is still a major issue for efficient high-yield recombinant protein accumulation. In this work, we have performed a detailed study of the degradation profiles of two human IgG1 mAbs produced in plants: an anti-HIV mAb 2G12 and a tumour-targeting mAb H10. Even though they use different light chains (κ and λ, respectively), the fragmentation pattern of both antibodies was similar. The majority of Ig fragments result from proteolytic degradation, but there are only a limited number of plant proteolytic cleavage events in the immunoglobulin light and heavy chains. All of the cleavage sites identified were in the proximity of interdomain regions and occurred at each interdomain site, with the exception of the VL /CL interface in mAb H10 λ light chain. Cleavage site sequences were analysed, and residue patterns characteristic of proteolytic enzymes substrates were identified. The results of this work help to define common degradation events in plant-produced mAbs and raise the possibility of predicting antibody degradation patterns 'a priori' and designing novel stabilization strategies by site-specific mutagenesis.

  7. The Xerophyta viscosa aldose reductase (ALDRXV4) confers enhanced drought and salinity tolerance to transgenic tobacco plants by scavenging methylglyoxal and reducing the membrane damage.

    Science.gov (United States)

    Kumar, Deepak; Singh, Preeti; Yusuf, Mohd Aslam; Upadhyaya, Chandrama Prakash; Roy, Suchandra Deb; Hohn, Thomas; Sarin, Neera Bhalla

    2013-06-01

    We report the efficacy of an aldose reductase (ALDRXV4) enzyme from Xerophyta viscosa Baker in enhancing the prospects of plant's survival under abiotic stress. Transgenic tobacco plants overexpressing ALDRXV4 cDNA showed alleviation of NaCl and mannitol-induced abiotic stress. The transgenic plants survived longer periods of water deficiency and salinity stress and exhibited improved recovery after rehydration as compared to the wild type plants. The increased synthesis of aldose reductase in transgenic plants correlated with reduced methylglyoxal and malondialdehyde accumulation and an elevated level of sorbitol under stress conditions. In addition, the transgenic lines showed better photosynthetic efficiency, less electrolyte damage, greater water retention, higher proline accumulation, and favorable ionic balance under stress conditions. Together, these findings suggest the potential of engineering aldose reductase levels for better performance of crop plants growing under drought and salt stress conditions.

  8. The mungbean yellow mosaic begomovirus transcriptional activator protein transactivates the viral promoter-driven transgene and causes toxicity in transgenic tobacco plants.

    Science.gov (United States)

    Rajeswaran, Rajendran; Sunitha, Sukumaran; Shivaprasad, Padubidri V; Pooggin, Mikhail M; Hohn, Thomas; Veluthambi, Karuppannan

    2007-12-01

    The Begomovirus transcriptional activator protein (TrAP/AC2/C2) is a multifunctional protein which activates the viral late gene promoters, suppresses gene silencing, and determines pathogenicity. To study TrAP-mediated transactivation of a stably integrated gene, we generated transgenic tobacco plants with a Mungbean yellow mosaic virus (MYMV) AV1 late gene promoter-driven reporter gene and supertransformed them with the MYMV TrAP gene driven by a strong 35S promoter. We obtained a single supertransformed plant with an intact 35S-TrAP gene that activated the reporter gene 2.5-fold. However, 10 of the 11 supertransformed plants did not have the TrAP region of the T-DNA, suggesting the likely toxicity of TrAP in plants. Upon transformation of wild-type tobacco plants with the TrAP gene, six of the seven transgenic plants obtained had truncated T-DNAs which lacked TrAP. One plant, which had the intact TrAP gene, did not express TrAP. The apparent toxic effect of the TrAP transgene was abolished by mutations in its nuclear-localization signal or zinc-finger domain and by deletion of its activation domain. Therefore, all three domains of TrAP, which are required for transactivation and suppression of gene silencing, also are needed for its toxic effect.

  9. Assessing environmental impacts of genetically modified plants on non-target organisms

    NARCIS (Netherlands)

    Arpaia, Salvatore; Birch, A.N.E.; Kiss, Jozsef; Loon, van Joop J.A.; Messéan, Antoine; Nuti, Marco; Perry, Joe N.; Sweet, Jeremy B.; Tebbe, Christoph C.

    2017-01-01

    In legal frameworks worldwide, genetically modified plants (GMPs) are subjected to pre-market environmental risk assessment (ERA) with the aim of identifying potential effects on the environment. In the European Union, the EFSA Guidance Document introduces the rationale that GMPs, as well as their n

  10. Assessing environmental impacts of genetically modified plants on non-target organisms

    NARCIS (Netherlands)

    Arpaia, Salvatore; Birch, A.N.E.; Kiss, Jozsef; Loon, van Joop J.A.; Messéan, Antoine; Nuti, Marco; Perry, Joe N.; Sweet, Jeremy B.; Tebbe, Christoph C.

    2017-01-01

    In legal frameworks worldwide, genetically modified plants (GMPs) are subjected to pre-market environmental risk assessment (ERA) with the aim of identifying potential effects on the environment. In the European Union, the EFSA Guidance Document introduces the rationale that GMPs, as well as thei

  11. Biochemistry of plant class IV chitinases and fungal chitinase-modifying proteins

    Science.gov (United States)

    Plant class IV chitinases have 2 domains, a small (3 kDa) amino-terminal domain with homology to carbohydrate binding peptides, and a larger (25 kDa) catalytic domain. The biological function of these chitinases is not known. But it is known that some pathogenic fungi secrete chitinase modifying pro...

  12. Effects of genetically modified plants on microbial communities and processes in soil

    NARCIS (Netherlands)

    Bruinsma, M.; Kowalchuk, G.A.; Van Veen, J.A.

    2003-01-01

    The development and use of genetically modified plants (GMPs) has been a topic of considerable public debate in recent years. GMPs hold great promise for improving agricultural output, but the potential for unwanted effects of GMP use is still not fully understood. The majority of studies addressing

  13. Heterologous expression of plant cell wall glycosyltransferases in Pichia, pea and tobacco

    DEFF Research Database (Denmark)

    Petersen, Bent Larsen; Damager, Iben; Faber, Kirsten

    UDP-xylose on to the monosaccharide sugar fucose. Partly based on these data, the two genes were proposed to function in the biosynthesis of pectic rhamnogalacturonan II (RG-II) and designated RhamnoGalacturonan XylosylTransferase 1 and -2 (RGXT1 and -2), accordingly (Egelund et al. 2006, The Plant...

  14. Stress-inducible GmGSTU4 shapes transgenic tobacco plants metabolome towards increased salinity tolerance

    NARCIS (Netherlands)

    Kissoudis, Christos; Kalloniati, Chrissanthi; Flemetakis, Emmanouil; Madesis, Panagiotis; Labrou, Nikolaos E.; Tsaftaris, Athanasios; Nianiou-Obeidat, Irini

    2015-01-01

    The involvement of glutathione transferases (GSTs) in plant’s tolerance to abiotic stresses has been extensively studied; however, the metabolic changes occurring in the plants with altered GSTs expression have not been studied in detail. We have previously demonstrated that GmGSTU4 overexpressio

  15. Stress-inducible GmGSTU4 shapes transgenic tobacco plants metabolome towards increased salinity tolerance

    NARCIS (Netherlands)

    Kissoudis, Christos; Kalloniati, Chrissanthi; Flemetakis, Emmanouil; Madesis, Panagiotis; Labrou, Nikolaos E.; Tsaftaris, Athanasios; Nianiou-Obeidat, Irini

    2015-01-01

    The involvement of glutathione transferases (GSTs) in plant’s tolerance to abiotic stresses has been extensively studied; however, the metabolic changes occurring in the plants with altered GSTs expression have not been studied in detail. We have previously demonstrated that GmGSTU4

  16. Uptake of NO, NO 2 and O 3 by sunflower ( Helianthus annuus L.) and tobacco plants ( Nicotiana tabacum L.): dependence on stomatal conductivity

    Science.gov (United States)

    Neubert, A.; Kley, D.; Wildt, J.; Segschneider, H. J.; Förstel, H.

    The uptake of NO, NO 2 and O 3 by sunflowers ( Helianthus annuus L. var. giganteus) and tobacco plants ( Nicotiana tabacum L. var. Bel W3), using concentrations representative for moderately polluted air, has been determined by gas exchange experiments. Conductivities for these trace gases were measured at different light fluxes ranging from 820 μEm -2s -1 to darkness. The conductivities to water vapor and the trace gases are highly correlated. It is concluded that the uptake of NO, NO 2 and O 3 by sunflowers and tobacco plants is linearly dependent on stomatal opening. While the uptake of NO is limited by the mesophyll resistance, the uptake of NO 2 is only by diffusion through the stomata. Loss processes by deposition to the leaf surfaces are more pronounced for O 3 than for NO and NO 2.

  17. [Genetically modified plants and food safety. State of the art and discussion in the European Union].

    Science.gov (United States)

    Schauzu, M

    2004-09-01

    Placing genetically modified (GM) plants and derived products on the European Union's (EU) market has been regulated by a Community Directive since 1990. This directive was complemented by a regulation specific for genetically modified and other novel foods in 1997. Specific labelling requirements have been applicable for GM foods since 1998. The law requires a pre-market safety assessment for which criteria have been elaborated and continuously adapted in accordance with the state of the art by national and international bodies and organisations. Consequently, only genetically modified products that have been demonstrated to be as safe as their conventional counterparts can be commercialized. However, the poor acceptance of genetically modified foods has led to a de facto moratorium since 1998. It is based on the lack of a qualified majority of EU member states necessary for authorization to place genetically modified plants and derived foods on the market. New Community Regulations are intended to end this moratorium by providing a harmonized and transparent safety assessment, a centralised authorization procedure, extended labelling provisions and a traceability system for genetically modified organisms (GMO) and derived food and feed.

  18. Promoting flowering, lateral shoot outgrowth, leaf development, and flower abscission in tobacco plants overexpressing cotton FLOWERING LOCUS T (FT)-like gene GhFT1.

    Science.gov (United States)

    Li, Chao; Zhang, Yannan; Zhang, Kun; Guo, Danli; Cui, Baiming; Wang, Xiyin; Huang, Xianzhong

    2015-01-01

    FLOWERING LOCUS T (FT) encodes a mobile signal protein, recognized as major component of florigen, which has a central position in regulating flowering, and also plays important roles in various physiological aspects. A mode is recently emerging for the balance of indeterminate and determinate growth, which is controlled by the ratio of FT-like and TERMINAL FLOWER 1 (TFL1)-like gene activities, and has a strong influence on the floral transition and plant architecture. Orthologs of GhFT1 was previously isolated and characterized from Gossypium hirsutum. We demonstrated that ectopic overexpression of GhFT1 in tobacco, other than promoting flowering, promoted lateral shoot outgrowth at the base, induced more axillary bud at the axillae of rosette leaves, altered leaf morphology, increased chlorophyll content, had higher rate of photosynthesis and caused flowers abscission. Analysis of gene expression suggested that flower identity genes were significantly upregulated in transgenic plants. Further analysis of tobacco FT paralogs indicated that NtFT4, acting as flower inducer, was upregulated, whereas NtFT2 and NtFT3 as flower inhibitors were upregulated in transgenic plants under long-day conditions, but downregulated under short-day conditions. Our data suggests that sufficient level of transgenic cotton FT might disturb the balance of the endogenous tobacco FT paralogs of inducers and repressors and resulted in altered phenotype in transgenic tobacco, emphasizing the expanding roles of FT in regulating shoot architecture by advancing determine growth. Manipulating the ratio for indeterminate and determinate growth factors throughout FT-like and TFL1-like gene activity holds promise to improve plant architecture and enhance crop yield.

  19. Transgenic Tobacco Lines Expressing Sense or Antisense FERROCHELATASE 1 RNA Show Modified Ferrochelatase Activity in Roots and Provide Experimental Evidence for Dual Localization of Ferrochelatase 1.

    Science.gov (United States)

    Hey, Daniel; Ortega-Rodes, Patricia; Fan, Tingting; Schnurrer, Florian; Brings, Lea; Hedtke, Boris; Grimm, Bernhard

    2016-12-01

    In plants, two genes encode ferrochelatase (FC), which catalyzes iron chelation into protoporphyrin IX at the final step of heme biosynthesis. FERROCHELATASE1 (FC1) is continuously, but weakly expressed in roots and leaves, while FC2 is dominantly active in leaves. As a continuation of previous studies on the physiological consequences of FC2 inactivation in tobacco, we aimed to assign FC1 function in plant organs. While reduced FC2 expression leads to protoporphyrin IX accumulation in leaves, FC1 down-regulation and overproduction caused reduced and elevated FC activity in root tissue, respectively, but were not associated with changes in macroscopic phenotype, plant development or leaf pigmentation. In contrast to the lower heme content resulting from a deficiency of the dominant FC2 expression in leaves, a reduction of FC1 in roots and leaves does not significantly disturb heme accumulation. The FC1 overexpression was used for an additional approach to re-examine FC activity in mitochondria. Transgenic FC1 protein was immunologically shown to be present in mitochondria. Although matching only a small portion of total cellular FC activity, the mitochondrial FC activity in a FC1 overexpressor line increased 5-fold in comparison with wild-type mitochondria. Thus, it is suggested that FC1 contributes to mitochondrial heme synthesis.

  20. Impact of Raw and Bioaugmented Olive-Mill Wastewater and Olive-Mill Solid Waste on the Content of Photosynthetic Molecules in Tobacco Plants.

    Science.gov (United States)

    Parrotta, Luigi; Campani, Tommaso; Casini, Silvia; Romi, Marco; Cai, Giampiero

    2016-08-03

    Disposal and reuse of olive-mill wastes are both an economic and environmental problem, especially in countries where the cultivation of olive trees is extensive. Microorganism-based bioaugmentation can be used to reduce the pollutant capacity of wastes. In this work, bioaugmentation was used to reduce the polyphenolic content of both liquid and solid wastes. After processing, bioaugmented wastes were tested on the root development of maize seeds and on photosynthesis-related molecules of tobacco plants. In maize, we found that bioaugmentation made olive-mill wastes harmless for seed germination. In tobacco, we analyzed the content of RuBisCO (ribulose-1,5-bisphosphate carboxylase oxygenase) and of the photosynthetic pigments lutein, chlorophylls, and β-carotene. Levels of RuBisCO were negatively affected by untreated wastewater but increased if plants were treated with bioaugmented wastewater. On the contrary, levels of RuBisCO increased in the case of plants treated with raw olive-mill solid waste. Pigment levels showed dissimilar behavior because their concentration increased if plants were irrigated with raw wastewater or treated with raw olive-mill solid waste. Treatment with bioaugmented wastes restored pigment content. Findings show that untreated wastes are potentially toxic at the commencement of treatment, but plants can eventually adapt after an initial stress period. Bioaugmented wastes do not induce immediate damages, and plants rapidly recover optimal levels of photosynthetic molecules.

  1. Sympton development, X-body formation and 126-kDa-protein in plants infected with tobacco mosaic virus

    NARCIS (Netherlands)

    Wijdeveld, M.M.G.

    1990-01-01

    Upon infection with tobacco mosaic virus (TMV) sensitive tobacco varieties develop systemic mosaic symptoms in the developing leaves. These symptoms are the visible result of the interaction of the virus with its host and the nature and the severity of the symptoms are determined

  2. Infection cycle of Artichoke Italian latent virus in tobacco plants: meristem invasion and recovery from disease symptoms.

    Directory of Open Access Journals (Sweden)

    Elisa Santovito

    Full Text Available Nepoviral infections induce recovery in fully expanded leaves but persist in shoot apical meristem (SAM by a largely unknown mechanism. The dynamics of infection of a grapevine isolate of Artichoke Italian latent virus (AILV-V, genus Nepovirus in tobacco plants, including colonization of SAM, symptom induction and subsequent recovery of mature leaves from symptoms, were characterized. AILV-V moved from the inoculated leaves systemically and invaded SAM in 7 days post-inoculation (dpi, remaining detectable in SAM at least up to 40 dpi. The new top leaves recovered from viral symptoms earliest at 21 dpi. Accumulation of viral RNA to a threshold level was required to trigger the overexpression of RDR6 and DCL4. Consequently, accumulation of viral RNA decreased in the systemically infected leaves, reaching the lowest concentration in the 3rd and 4th leaves at 23 dpi, which was concomitant with recovery of the younger, upper leaves from disease symptoms. No evidence of virus replication was found in the recovered leaves, but they contained infectious virus particles and were protected against re-inoculation with AILV-V. In this study we also showed that AILV-V did not suppress initiation or maintenance of RNA silencing in transgenic plants, but was able to interfere with the cell-to-cell movement of the RNA silencing signal. Our results suggest that AILV-V entrance in SAM and activation of RNA silencing may be distinct processes since the latter is triggered in fully expanded leaves by the accumulation of viral RNA above a threshold level rather than by virus entrance in SAM.

  3. Using transgenic plants and modified plant viruses for the development of treatments for human diseases.

    Science.gov (United States)

    Loh, Hwei-San; Green, Brian J; Yusibov, Vidadi

    2017-08-08

    Production of proteins in plants for human health applications has become an attractive strategy attributed by their potentials for low-cost production, increased safety due to the lack of human or animal pathogens, scalability and ability to produce complex proteins. A major milestone for plant-based protein production for use in human health was achieved when Protalix BioTherapeutics produced taliglucerase alfa (Elelyso(®)) in suspension cultures of a transgenic carrot cell line for the treatment of patients with Gaucher's disease, was approved by the USA Food and Drug Administration in 2012. In this review, we are highlighting various approaches for plant-based production of proteins and recent progress in the development of plant-made therapeutics and biologics for the prevention and treatment of human diseases. Copyright © 2017 Elsevier B.V. All rights reserved.

  4. Transgenic tobacco plants expressing siRNA targeted against the Mungbean yellow mosaic virus transcriptional activator protein gene efficiently block the viral DNA accumulation.

    Science.gov (United States)

    Shanmugapriya, Gnanasekaran; Das, Sudhanshu Sekhar; Veluthambi, Karuppannan

    2015-06-01

    Mungbean yellow mosaic virus (MYMV) is a bipartite begomovirus that infects many pulse crops such as blackgram, mungbean, mothbean, Frenchbean, and soybean. We tested the efficacy of the transgenically expressed intron-spliced hairpin RNA gene of the transcriptional activator protein (hpTrAP) in reducing MYMV DNA accumulation. Tobacco plants transformed with the MYMV hpTrAP gene accumulated 21-22 nt siRNA. Leaf discs of the transgenic plants, agroinoculated with the partial dimers of MYMV, displayed pronounced reduction in MYMV DNA accumulation. Thus, silencing of the TrAP gene, a suppressor of gene silencing, emerged as an effective strategy to control MYMV.

  5. Normal Growth of Transgenic Tobacco Plants in the Absence of Cytosolic Pyruvate Kinase 1

    Science.gov (United States)

    Gottlob-McHugh, Sylvia G.; Sangwan, Rajender S.; Blakeley, Stephen D.; Vanlerberghe, Greg C.; Ko, Kenton; Turpin, David H.; Plaxton, William C.; Miki, Brian L.; Dennis, David T.

    1992-01-01

    The coding sequence of the cytosolic isozyme of potato tuber pyruvate kinase (PK) was attached to the transit peptide of the small subunit of pea ribulose-1,5-bisphosphate carboxylase oxygenase and placed under the control of the cauliflower mosaic virus 35S promoter. This construct was transformed into Nicotiana tabacum. Unexpectedly, two primary transformants were recovered in which PK activity in leaves was greatly reduced. The reduction in PK activity appeared to result from the complete absence of the cytosolic form of the enzyme (PKc). In addition, no PKc could be detected on western blots of leaf extracts. Metabolite analyses indicated that the levels of phosphoenolpyruvate are substantially higher in PKc-deficient leaves than in wild-type leaves, consistent with a block in glycolysis at the step catalyzed by PK. PKc deficiency in the leaves does not appear to adversely affect plant growth. Analysis of progeny indicates that PKc deficiency is a heritable trait. The leaves of PKc-deficient transformants have normal rates of photosynthetic O2 evolution and respiratory O2 consumption, indicating that these plants are using alternative pathways to bypass PK. Images Figure 2 Figure 4 PMID:16653063

  6. Rhizobacteria modify plant-aphid interactions: a case of induced systemic susceptibility.

    Science.gov (United States)

    Pineda, A; Zheng, S-J; van Loon, J J A; Dicke, M

    2012-03-01

    Beneficial microbes, such as plant growth-promoting rhizobacteria and mycorrhizal fungi, may have a plant-mediated effect on insects aboveground. The plant growth-promoting rhizobacterium Pseudomonas fluorescens can induce systemic resistance in Arabidopsis thaliana against several microbial pathogens and chewing insects. However, the plant-mediated effect of these beneficial microbes on phloem-feeding insects is not well understood. Using Arabidopsis as a model, we here report that P. fluorescens has a positive effect on the performance (weight gain and intrinsic rate of increase) of the generalist aphid Myzus persicae, while no effect was recorded on the crucifer specialist aphid Brevicoryne brassicae. Additionally, transcriptional analyses of selected marker genes revealed that in the plant-microbe interaction with M. persicae, rhizobacteria (i) prime the plant for enhanced expression of LOX2, a gene involved in the jasmonic acid (JA)-regulated defence pathway, and (ii) suppress the expression of ABA1, a gene involved in the abscisic acid (ABA) signalling pathway, at several time points. In contrast, almost no effect of the plant-microbe interaction with B. brassicae was found at the transcriptional level. This study presents the first data on rhizobacteria-induced systemic susceptibility to an herbivorous insect, supporting the pattern proposed for other belowground beneficial microbes and aboveground phloem feeders. Moreover, we provide further evidence that at the transcript level, soil-borne microbes modify plant-aphid interactions.

  7. Suggested improvements for the allergenicity assessment of genetically modified plants used in foods.

    Science.gov (United States)

    Goodman, Richard E; Tetteh, Afua O

    2011-08-01

    Genetically modified (GM) plants are increasingly used for food production and industrial applications. As the global population has surpassed 7 billion and per capita consumption rises, food production is challenged by loss of arable land, changing weather patterns, and evolving plant pests and disease. Previous gains in quantity and quality relied on natural or artificial breeding, random mutagenesis, increased pesticide and fertilizer use, and improved farming techniques, all without a formal safety evaluation. However, the direct introduction of novel genes raised questions regarding safety that are being addressed by an evaluation process that considers potential increases in the allergenicity, toxicity, and nutrient availability of foods derived from the GM plants. Opinions vary regarding the adequacy of the assessment, but there is no documented proof of an adverse effect resulting from foods produced from GM plants. This review and opinion discusses current practices and new regulatory demands related to food safety.

  8. Overexpression of a tobacco small G protein gene NtRop1 causes salt sensitivity and hydrogen peroxide production in transgenic plants

    Institute of Scientific and Technical Information of China (English)

    CAO YangRong; LI ZhiGang; CHEN Tao; ZHANG ZhiGang; ZHANG JinSong; CHEN ShouYi

    2008-01-01

    The small GTPases of Rop/Rho family is central regulators of important cellular processes in plants.Tobacco small G protein gene NtRop1 has been isolated; however, its roles in stress responses were unknown. In the present study, the genomic sequence of NtRop1 was cloned, which has seven exons and six introns, similar to the Rop gene structure from Arabidopsis. The NtRopl gene was constitutively expressed in the different organs whereas the other six Rop genes from tobacco had differential expression patterns. The expression of the NtRop1 gene was moderately induced by methyl viologen,NaCl, and ACC treatments, but slightly inhibited by ABA treatment, with no significant induction by NAA treatment. The trsnsgenic Arabidopsis plants overexpressing the NtRop1 showed increased salt sensitivity as can be seen from the reduced root growth and elevated relative electrolyte leakage. The hydrogen peroxide production was also promoted in the NtRop1-trangenic plants in comparison with wild type plants. These results imply that the NtRopl may confer salt sensitivity through activation of H2O2 production during plant response to salt stress.

  9. Overexpression of a tobacco small G protein gene NtRop1 causes salt sensitivity and hydrogen peroxide production in transgenic plants

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    The small GTPases of Rop/Rho family is central regulators of important cellular processes in plants. Tobacco small G protein gene NtRop1 has been isolated; however, its roles in stress responses were unknown. In the present study, the genomic sequence of NtRop1 was cloned, which has seven exons and six introns, similar to the Rop gene structure from Arabidopsis. The NtRop1 gene was constitutively expressed in the different organs whereas the other six Rop genes from tobacco had differential expression patterns. The expression of the NtRop1 gene was moderately induced by methyl viologen, NaCl, and ACC treatments, but slightly inhibited by ABA treatment, with no significant induction by NAA treatment. The transgenic Arabidopsis plants overexpressing the NtRop1 showed increased salt sensitivity as can be seen from the reduced root growth and elevated relative electrolyte leakage. The hydrogen peroxide production was also promoted in the NtRop1-trangenic plants in comparison with wild type plants. These results imply that the NtRop1 may confer salt sensitivity through activation of H2O2 production during plant response to salt stress.

  10. Transformation of tobacco plant (Nicotiana tabacum L. with the recombinant hepatitis B virus genes 35SHBsAg and 35SHBsAgER

    Directory of Open Access Journals (Sweden)

    Juliana Martins Ribeiro

    2010-03-01

    Full Text Available The recombinant surface antigen of hepatitis B virus (HBsAg, purified from transgenic plants, proved to be efficient when utilized for raising anti-HB antibodies for the prevention of hepatitis B. Because of the important role of the HBsAg antigen in hepatitis B prevention, the coding sequence of HBsAg antigen, with or without the addition of the carboxi-terminus sequence for protein retention in the endoplasmatic reticulum, was linked to cauliflower mosaic virus 35S promoter, tobacco mosaic virus leader sequence Ω, and the transcription terminator sequence. The aim of this work was to clone the chimeric gene 35SHBsAgER in the plant expression vector pGPTV/Kan/Asc. The resulting plasmid, called pG35SHBsAgER, and another plasmid produced previously in our laboratory called pG35SHBsAg, were transferred to Agrobacterium tumefaciens, and tobacco leaves, of the SR1 cultivar were used as explants for genetic transformation. Twenty-one fully regenerated plants were obtained (10 for the pG35SHBsAg construction and 11 for the pG35SHBsAgER construction. The genomic DNA of all plants was analyzed by PCR, and the presence of the transgene was confirmed in all plants.

  11. Induction of protective immune responses against the challenge of Actinobacillus pleuropneumoniae by the oral administration of transgenic tobacco plant expressing ApxIIA toxin from the bacteria.

    Science.gov (United States)

    Lee, Kyung-Yeol; Kim, Dong-Heon; Kang, Tae-Jin; Kim, Ju; Chung, Gook-Hyun; Yoo, Han-Sang; Arntzen, Charles J; Yang, Moon-Sik; Jang, Yong-Suk

    2006-12-01

    Actinobacillus pleuropneumoniae is the causative agent of porcine pleuropneumonia. Among the virulence factors, ApxIIA, a bacterial exotoxin, is reportedly expressed in many serotypes and is considered as a candidate for the development of a vaccine against the bacterial infection. Previously, we isolated a field strain of A. pleuropneumoniae serotype 2 in Korea and characterized its exotoxins to develop an oral vaccine. In this study, we initially confirmed the immunogenicity of ApxIIA expressed in Escherichia coli. We then developed transgenic tobacco expressing ApxIIA and tested its efficacy to induce a protective immune response against A. pleuropneumoniae infection after oral administration of the plant powder. We observed that protective immune responses were induced in mice after oral administration of the plant powder once a week for 4 weeks. Immunoassays revealed that the levels of antigen-specific immunoglobulin G against ApxIIA increased in mice that were fed a powder made from the transgenic plant, but not in mice fed a powder made from wild-type tobacco. Additionally, mice fed the transgenic plant powder were protected from an injection of a lethal dose of A. pleuropneumoniae. These results support that the transgenic plant may be a suitable candidate for an oral vaccine that could be used effectively against A. pleuropneumoniae infection.

  12. Short-term salinity stress in tobacco plants leads to the onset of animal-like PCD hallmarks in planta in contrast to long-term stress.

    Science.gov (United States)

    Andronis, Efthimios A; Roubelakis-Angelakis, Kalliopi A

    2010-01-01

    Recent results have identified mitochondria as centers of stress-induced generation of reactive oxygen species in plants. Depolarization of plant mitochondrial membrane during stress results the release of programmed cell death (PCD)-inducing factors in the cytosol in a fashion similar to the onset of animal-like PCD. Herein, we report significant similarities of animal-like PCD and salinity stress-induced plant PCD. Short-term salinity stress (3 h) led to depolarization of the mitochondrial membrane, release of cytochrome c (CYT-c), which was visualized using a contemporary molecular technique, activation of caspase-3 type proteases and the onset of PCD in wild type tobacco plants, Nicotiana tabacum cv. Petit Havana. However, PCD was not manifested during long-term salinity stress (24 h). Interestingly long-term salinity stress led to necrotic-like features, which were accompanied by collapse of respiration, reduction of key components of the respiratory chain, such as CYT-c and alternative oxidase, ATP depletion and high proteolytic activity. The results suggest that salinity stress of tobacco plants in planta leads to the onset of animal-like PCD only during the early stages post-stress, while long-term stress leads to necrotic-like features.

  13. [Analysis of tobacco regeneration plants from the protoplasts produced by electrofusion [corrected] in space].

    Science.gov (United States)

    Li, Xiu-Gen; Chen, Ai-Di; Wang, Liu-Fa; Zheng, Hui-Qiong

    2007-10-01

    Vacuolated mesophyll protoplasts of Nicotiana rustica L. were electrically fused with evacuolated protoplasts of the same genus (N. tabacum cv. 'Gexin No.1') during a 7-day space flight in the Chinese spacecraft "SZ-4". The initial cell division leading to micro-callus formation took place after landing (Fig.1). Higher plating efficiencies were observed in the flight samples than the control culture, but the frequency of plantlets regeneration reduced by about 20% of the control (Table 1). The hybrid characters were tested by chromosome counting, isozyme analysis and comparison of morphological characteristics (Figs.2-4). About 32% of the regenerates showed hybrid character. Leaf morphological modifications were found in 3 hybrids, i.e., H23, H25 and H27. After backcrossing with N. rustica, alterations in flower color and leaf shape occurred in the somatic hybrid H23 (Fig.5). These results demonstrate that the hybrids formed under microgravity condition could regenerate fertile plants.

  14. A rat pancreatic ribonuclease fused to a late cotton pollen promoter severely reduces pollen viability in tobacco plants

    Directory of Open Access Journals (Sweden)

    R.B. Bernd-Souza

    2000-06-01

    Full Text Available The effects of an animal RNase fused to the late cotton pollen-specific promoter G9 in a plant system were investigated. Expression of the chimeric genes G9-uidA and G9-RNase in tobacco plants showed that the 1.2-kb promoter fragment of the G9 gene was sufficient to maintain tissue and temporal specificity in a heterologous system. GUS (beta-glucuronidase expression was detected only in pollen from anther stage 6 through anthesis, with maximal GUS activity in pollen from stage 10 anthers. Investigating the effects of the rat RNase on pollen viability at stage 10, we found that pollen viability was reduced from 79 to 8% and from 89 to 40%, in pollen germination and fluoresceine diacetate assays, respectively, in one G9-RNase transgenic line, suggesting a lethal effect of the RNase gene. This indicates that the rat RNase produces deleterious effects in this plant system and may be useful for engineering male sterility.Foram investigados os efeitos da expressão de uma ribonuclease de origem animal em um sistema vegetal, ligando-se esta ao promotor do gene pólen-específico G9 de algodão. Examinou-se a expressão dos genes quiméricos G9-uidA e G9-RNase em plantas de tabaco e determinou-se que o fragmento de 1.2 kb do promotor do gene G9 foi suficiente para manter a especificidade temporal e espacial da expressão, em sistema heterólogo. A expressão do gene GUS foi detectada somente em pólen, do estágio 6 do desenvolvimento da antera até a antese, com atividade máxima em pólen de anteras no estágio 10. Estudos neste estágio com linhagens transgênicas contendo G9-RNase mostraram que um clone transgênico apresentava reduções na viabilidade do pólen de 79 para 8% e de 89 para 40% nos testes de germinação e coloração com diacetato de fluoresceína, respectivamente, sugerindo letalidade na expressão do gene de RNase. Estes resultados indicam que a RNase animal apresenta um efeito deletério em planta e oferece possibilidade de uso

  15. A statistical assessment of differences and equivalences between genetically modified and reference plant varieties

    Directory of Open Access Journals (Sweden)

    Amzal Billy

    2011-02-01

    Full Text Available Abstract Background Safety assessment of genetically modified organisms is currently often performed by comparative evaluation. However, natural variation of plant characteristics between commercial varieties is usually not considered explicitly in the statistical computations underlying the assessment. Results Statistical methods are described for the assessment of the difference between a genetically modified (GM plant variety and a conventional non-GM counterpart, and for the assessment of the equivalence between the GM variety and a group of reference plant varieties which have a history of safe use. It is proposed to present the results of both difference and equivalence testing for all relevant plant characteristics simultaneously in one or a few graphs, as an aid for further interpretation in safety assessment. A procedure is suggested to derive equivalence limits from the observed results for the reference plant varieties using a specific implementation of the linear mixed model. Three different equivalence tests are defined to classify any result in one of four equivalence classes. The performance of the proposed methods is investigated by a simulation study, and the methods are illustrated on compositional data from a field study on maize grain. Conclusions A clear distinction of practical relevance is shown between difference and equivalence testing. The proposed tests are shown to have appropriate performance characteristics by simulation, and the proposed simultaneous graphical representation of results was found to be helpful for the interpretation of results from a practical field trial data set.

  16. Metabolic Profiling with Gas Chromatography-Mass Spectrometry and Capillary Electrophoresis-Mass Spectrometry Reveals the Carbon-Nitrogen Status of Tobacco Leaves Across Different Planting Areas.

    Science.gov (United States)

    Zhao, Jieyu; Zhao, Yanni; Hu, Chunxiu; Zhao, Chunxia; Zhang, Junjie; Li, Lili; Zeng, Jun; Peng, Xiaojun; Lu, Xin; Xu, Guowang

    2016-02-05

    The interaction between carbon (C) and nitrogen (N) metabolism can reflect plant growth status and environmental factors. Little is known regarding the connections between C-N metabolism and growing regions under field conditions. To comprehensively investigate the relationship in mature tobacco leaves, we established metabolomics approaches based on gas chromatography-mass spectrometry (GC-MS) and capillary electrophoresis-time-of-flight-mass spectrometry (CE-TOF-MS). Approximately 240 polar metabolites were determined. Multivariate statistical analysis revealed that the growing region greatly influenced the metabolic profiles of tobacco leaves. A metabolic correlation network and related pathway maps were used to reveal the global overview of the alteration of C-N metabolism across three typical regions. In Yunnan, sugars and tricarboxylic acid (TCA) cycle intermediates were closely correlated with amino acid pools. Henan tobacco leaves showed positive correlation between the pentose phosphate pathway (PPP) intermediates and C-rich secondary metabolism. In Guizhou, the proline and asparagine had significant links with TCA cycle intermediates and urea cycle, and antioxidant accumulation was observed in response to drought. These results demonstrate that combined analytical approaches have great potential to detect polar metabolites and provide information on C-N metabolism related to planting regional characteristics.

  17. Assessing and monitoring impacts of genetically modified plants on agro-ecosystems

    DEFF Research Database (Denmark)

    Arpaia, S.; Messéan, A.; Birch, N.A.;

    2014-01-01

    -funded research project AMIGA − Assessing and monitoring Impacts of Genetically modified plants on Agro-ecosystems − aims to address this issue, by providing a framework that establishes protection goals and baselines for European agro-ecosystems, improves knowledge on the potential long term environmental...... focuses on ecological studies in different EU regions, the sustainability of GM crops is estimated by analysing the functional components of the agro-ecosystems and specific experimental protocols are being developed for this scope....

  18. A virulence factor encoded by a polydnavirus confers tolerance to transgenic tobacco plants against lepidopteran larvae, by impairing nutrient absorption.

    Directory of Open Access Journals (Sweden)

    Ilaria Di Lelio

    Full Text Available The biological control of insect pests is based on the use of natural enemies. However, the growing information on the molecular mechanisms underpinning the interactions between insects and their natural antagonists can be exploited to develop "bio-inspired" pest control strategies, mimicking suppression mechanisms shaped by long co-evolutionary processes. Here we focus on a virulence factor encoded by the polydnavirus associated with the braconid wasp Toxoneuron nigriceps (TnBV, an endophagous parasitoid of noctuid moth larvae. This virulence factor (TnBVANK1 is a member of the viral ankyrin (ANK protein family, and appears to be involved both in immunosuppression and endocrine alterations of the host. Transgenic tobacco plants expressing TnBVANK1 showed insecticide activity and caused developmental delay in Spodoptera littoralis larvae feeding on them. This effect was more evident in a transgenic line showing a higher number of transcripts of the viral gene. However, this effect was not associated with evidence of translocation into the haemocoel of the entire protein, where the receptors of TnBVANK1 are putatively located. Indeed, immunolocalization experiments evidenced the accumulation of this viral protein in the midgut, where it formed a thick layer coating the brush border of epithelial cells. In vitro transport experiments demonstrated that the presence of recombinant TnBVANK1 exerted a dose-dependent negative impact on amino acid transport. These results open new perspectives for insect control and stimulate additional research efforts to pursue the development of novel bioinsecticides, encoded by parasitoid-derived genes. However, future work will have to carefully evaluate any effect that these molecules may have on beneficial insects and on non-target organisms.

  19. A virulence factor encoded by a polydnavirus confers tolerance to transgenic tobacco plants against lepidopteran larvae, by impairing nutrient absorption.

    Science.gov (United States)

    Di Lelio, Ilaria; Caccia, Silvia; Coppola, Mariangela; Buonanno, Martina; Di Prisco, Gennaro; Varricchio, Paola; Franzetti, Eleonora; Corrado, Giandomenico; Monti, Simona M; Rao, Rosa; Casartelli, Morena; Pennacchio, Francesco

    2014-01-01

    The biological control of insect pests is based on the use of natural enemies. However, the growing information on the molecular mechanisms underpinning the interactions between insects and their natural antagonists can be exploited to develop "bio-inspired" pest control strategies, mimicking suppression mechanisms shaped by long co-evolutionary processes. Here we focus on a virulence factor encoded by the polydnavirus associated with the braconid wasp Toxoneuron nigriceps (TnBV), an endophagous parasitoid of noctuid moth larvae. This virulence factor (TnBVANK1) is a member of the viral ankyrin (ANK) protein family, and appears to be involved both in immunosuppression and endocrine alterations of the host. Transgenic tobacco plants expressing TnBVANK1 showed insecticide activity and caused developmental delay in Spodoptera littoralis larvae feeding on them. This effect was more evident in a transgenic line showing a higher number of transcripts of the viral gene. However, this effect was not associated with evidence of translocation into the haemocoel of the entire protein, where the receptors of TnBVANK1 are putatively located. Indeed, immunolocalization experiments evidenced the accumulation of this viral protein in the midgut, where it formed a thick layer coating the brush border of epithelial cells. In vitro transport experiments demonstrated that the presence of recombinant TnBVANK1 exerted a dose-dependent negative impact on amino acid transport. These results open new perspectives for insect control and stimulate additional research efforts to pursue the development of novel bioinsecticides, encoded by parasitoid-derived genes. However, future work will have to carefully evaluate any effect that these molecules may have on beneficial insects and on non-target organisms.

  20. From gene to harvest: insights into upstream process development for the GMP production of a monoclonal antibody in transgenic tobacco plants.

    Science.gov (United States)

    Sack, Markus; Rademacher, Thomas; Spiegel, Holger; Boes, Alexander; Hellwig, Stephan; Drossard, Juergen; Stoger, Eva; Fischer, Rainer

    2015-10-01

    The EU Sixth Framework Programme Integrated Project 'Pharma-Planta' developed an approved manufacturing process for recombinant plant-made pharmaceutical proteins (PMPs) using the human HIV-neutralizing monoclonal antibody 2G12 as a case study. In contrast to the well-established Chinese hamster ovary platform, which has been used for the production of therapeutic antibodies for nearly 30 years, only draft regulations were initially available covering the production of recombinant proteins in transgenic tobacco plants. Whereas recombinant proteins produced in animal cells are secreted into the culture medium during fermentation in bioreactors, intact plants grown under nonsterile conditions in a glasshouse environment provide various 'plant-specific' regulatory and technical challenges for the development of a process suitable for the acquisition of a manufacturing licence for clinical phase I trials. During upstream process development, several generic steps were addressed (e.g. plant transformation and screening, seed bank generation, genetic stability, host plant uniformity) as well as product-specific aspects (e.g. product quantity). This report summarizes the efforts undertaken to analyse and define the procedures for the GMP/GACP-compliant upstream production of 2G12 in transgenic tobacco plants from gene to harvest, including the design of expression constructs, plant transformation, the generation of production lines, master and working seed banks and the detailed investigation of cultivation and harvesting parameters and their impact on biomass, product yield and intra/interbatch variability. The resulting procedures were successfully translated into a prototypic manufacturing process that has been approved by the German competent authority.

  1. Overexpression of carnation S-adenosylmethionine decarboxylase gene generates a broad-spectrum tolerance to abiotic stresses in transgenic tobacco plants.

    Science.gov (United States)

    Wi, Soo Jin; Kim, Woo Taek; Park, Ky Young

    2006-10-01

    Polyamines (PAs), such as putrescine, spermidine, and spermine, are present in all living organism and implicate in a wide range of cellular physiological processes. We have used transgenic technology in an attempt to evaluate their potential for mitigating the adverse effects of several abiotic stresses in plants. Sense construct of full-length cDNA for S-adenosylmethionine decarboxylase (SAMDC), a key enzyme in PA biosynthesis, from carnation (Dianthus caryophyllus L.) flower was introduced into tobacco (Nicotiana tabacum L.) by Agrobacterium tumefaciens-mediated transformation. Several transgenic lines overexpressing SAMDC gene under the control of cauliflower mosaic virus 35S promoter accumulated soluble total PAs by 2.2 (S16-S-4) to 3.1 (S16-S-1) times than wild-type plants. The transgenic tobacco did not show any difference in organ phenotype compared to the wild-type. The number and weight of seeds increased, and net photosynthetic rate also increased in transgenic plants. Stress-induced damage was attenuated in these transgenic plants, in the symptom of visible yellowing and chlorophyll degradation after all experienced stresses such as salt stress, cold stress, acidic stress, and abscisic acid treatment. H2O2-induced damage was attenuated by spermidine treatment. Transcripts for antioxidant enzymes (ascorbate peroxidase, manganase superoxide dismutase, and glutathione S-transferase) in transgenic plants and GUS activity transformed with SAMDC promoter::GUS fusion were induced more significantly by stress treatment, compared to control. These results that the transgenic plants with sense SAMDC cDNA are more tolerant to abiotic stresses than wild-type plants suggest that PAs may play an important role in contributing stress tolerance in plants.

  2. A modified method for diffusive monitoring of 3-ethenylpyridine as a specific marker of environmental tobacco smoke

    Science.gov (United States)

    Kuusimäki, Leea; Peltonen, Kimmo; Vainiotalo, Sinikka

    A previously introduced method for monitoring environmental tobacco smoke (ETS) was further validated. The method is based on diffusive sampling of a vapour-phase marker, 3-ethenylpyridine (3-EP), with 3 M passive monitors (type 3500). Experiments were done in a dynamic chamber to assess diffusive sampling in comparison with active sampling in charcoal tubes or XAD-4 tubes. The sampling rate for 3-EP collected on the diffusive sampler was 23.1±0.6 mL min -1. The relative standard deviation for parallel samples ( n=6) ranged from 4% to 14% among experiments ( n=9). No marked reverse diffusion of 3-EP was detected nor any significant effect of relative humidity at 20%, 50% or 80%. The diffusive sampling of 3-EP was validated in field measurements in 15 restaurants in comparison with 3-EP and nicotine measurements using active sampling. The 3-EP concentration in restaurants ranged from 0.01 to 9.8 μg m -3, and the uptake rate for 3-EP based on 92 parallel samples was 24.0±0.4 mL min -1. A linear correlation ( r=0.98) was observed between 3-EP and nicotine concentrations, the average ratio of 3-EP to nicotine being 1:8. Active sampling of 3-EP and nicotine in charcoal tubes provided more reliable results than sampling in XAD-4 tubes. All samples were analysed using gas chromatography-mass spectrometry after elution with a 15% solution of pyridine in toluene. For nicotine, the limit of quantification of the charcoal tube method was 4 ng per sample, corresponding to 0.04 μg m -3 for an air sample of 96 L. For 3-EP, the limit of quantification of the diffusive method was 0.5-1.0 ng per sample, corresponding to 0.04-0.09 μg m -3 for 8 h sampling. The diffusive method proved suitable for ETS monitoring, even at low levels of ETS.

  3. Patterns of phenylpropanoids in non-inoculated and potato virus Y-inoculated leaves of transgenic tobacco plants expressing yeast-derived invertase.

    Science.gov (United States)

    Baumert, A; Mock, H P; Schmidt, J; Herbers, K; Sonnewald, U; Strack, D

    2001-03-01

    The patterns of secondary metabolites in leaves of yeast invertase-transgenic tobacco plants (Nicotiana tabacum L. cv. Samsun NN) were analyzed. Plants expressing cytosolic yeast-derived invertase (cytInv) or apoplastic (cell wall associated) yeast invertase (cwInv) showed a characteristic phytochemical phenotype compared to untransformed controls (wild-type plants). The level of phenylpropanoids decreased in the cytInv plants but increased in the cwInv plants, which showed an induced de novo synthesis of a caffeic acid amide, i.e. N-caffeoylputrescine. In addition, the level of the coumarin glucoside scopolin was markedly enhanced. Increased accumulation of scopolin in the cwInv plants is possibly correlated with the induction of defense reactions and the appearance of necrotic lesions similar to the hypersensitive response caused by avirulent pathogens. This is consistent with results from potato virus Y-infected plants. Whereas there was no additional increase in the coumarins in leaves following infection in cwInv plants, wild-type plants showed a slight increase and cytInc a marked increase.

  4. Tobacco plants transformed with the bean. alpha. ai gene express an inhibitor of insect. alpha. -amylase in their seeds. [Nicotiana tabacum; Tenebrio molitor

    Energy Technology Data Exchange (ETDEWEB)

    Altabella, T.; Chrispeels, M.J. (Univ. of California, San Diego, La Jolla (USA))

    1990-06-01

    Bean (Phaseolus vulgaris L.) seeds contain a putative plant defense protein that inhibits insect and mammalian but not plant {alpha}-amylases. We recently presented strong circumstantial evidence that this {alpha}-amylase inhibitor ({alpha}Al) is encoded by an already-identified lectin gene whose product is referred to as lectin-like-protein (LLP). We have now made a chimeric gene consisting of the coding sequence of the lectin gene that encodes LLP and the 5{prime} and 3{prime} flanking sequences of the lectin gene that encodes phytohemagglutinin-L. When this chimeric gene was expressed in transgenic tobacco (Nicotiana tabacum), we observed in the seeds a series of polypeptides (M{sub r} 10,000-18,000) that cross-react with antibodies to the bean {alpha}-amylase inhibitor. Most of these polypeptides bind to a pig pancreas {alpha}-amylase affinity column. An extract of the seeds of the transformed tobacco plants inhibits pig pancreas {alpha}-amylase activity as well as the {alpha}-amylase present in the midgut of Tenebrio molitor. We suggest that introduction of this lectin gene (to be called {alpha}ai) into other leguminous plants may be a strategy to protect the seeds from the seed-eating larvae of Coleoptera.

  5. Promoting Flowering, Lateral Shoot Outgrowth, Leaf Development, and Flower Abscission in Tobacco Plants Overexpressing Cotton FLOWERING LOCUS T (FT-Like Gene GhFT1

    Directory of Open Access Journals (Sweden)

    Chao eLi

    2015-06-01

    Full Text Available FLOWERING LOCUS T (FT encodes a mobile signal protein, recognized as major component of florigen, which has a central position in regulating flowering, and also plays important roles in various physiological aspects. A mode is recently emerging for the balance of indeterminate and determinate growth, which is controlled by the ratio of FT-like and TERMINAL FLOWER 1 (TFL1-like gene activities, and has a strong influence on the floral transition and plant architecture. Orthologs of GhFT1 was previously isolated and characterized from Gossypium hirsutum. We demonstrated that ectopic overexpression of GhFT1 in tobacco, other than promoting flowering, promoted lateral shoot outgrowth at the base, induced more axillary bud at the axillae of rosette leaves, altered leaf morphology, increased chlorophyll content, had higher photosynthesis and caused flowers abscission. Analysis of gene expression suggested that flower identity genes were significantly upregulated in transgenic plants. Further analysis of tobacco FT paralogs indicated that NtFT4, acting as flower inducer, was upregulated, whereas NtFT2 and NtFT3 as flower inhibitors were upregulated in transgenic plants under long-day conditions, but downregulated under short-day conditions. Our data suggested that sufficient level of foreign FT might disturb the balance of the endogenous FT paralogs of inducers and repressors and resulted in altered phenotype in transgenic tobacco, emphasizing the expanding roles of FT in regulating shoot architecture by advancing determine growth. Manipulating the ratio for indeterminate and determinate growth factors throughout FT-like and TFL1-like gene activity holds promise to improve plant architecture and enhance crop yield.

  6. Screening of plants containing Naja naja siamensis cobra venom inhibitory activity using modified ELISA technique.

    Science.gov (United States)

    Daduang, Sakda; Sattayasai, Nison; Sattayasai, Jintana; Tophrom, Pattara; Thammathaworn, Achra; Chaveerach, Arunrat; Konkchaiyaphum, Monruedee

    2005-06-15

    Enzyme-linked immunosorbent assay (ELISA) has been modified for screening plants with antagonistic activity to Naja naja siamensis cobra venom. Aqueous extracts from plants were investigated for their inhibitory effects on the binding of anti-cobra venom antibody to antigen, cobra venom, fixed onto 96-well microtiter plates. Ingredients in extracts were allowed to react with immobilized venom before the subsequent addition of antivenom antibody. Venom components affected by exposure to the extracts, unable to interact with their specific antibody, were predicted to be unable to bind to their native destinations or natural receptors. Curcuma cf. zedoaria, an old Thai medicinal plant, showed clear inhibitory activity in the ELISA test. Neurotoxin and protein degradative enzymes, major components in venom, were identified as targets of this extract in Western immunoblotting analysis. Ingredients in the extract showed high affinity to the toxin in competition assay by immunoprecipitation. The extract attenuated toxin activity by extending contraction time of diaphragm muscle after envenomation and had a potency to protect cellular proteins from venom degradative enzymes. Curcuma parviflora, with less activity in ELISA, exhibited acceptable results in two experiments but negative results in two experiments, whereas Curcuma longa, having low activity in the ELISA test, never showed any favorable results. Screening of 36 samples could classify plants into an inhibition range of 0 to 86%. This modified ELISA is recommended as a preliminary screening method for inhibitors with a large number of samples.

  7. Genetic basis and detection of unintended effects in genetically modified crop plants.

    Science.gov (United States)

    Ladics, Gregory S; Bartholomaeus, Andrew; Bregitzer, Phil; Doerrer, Nancy G; Gray, Alan; Holzhauser, Thomas; Jordan, Mark; Keese, Paul; Kok, Esther; Macdonald, Phil; Parrott, Wayne; Privalle, Laura; Raybould, Alan; Rhee, Seung Yon; Rice, Elena; Romeis, Jörg; Vaughn, Justin; Wal, Jean-Michel; Glenn, Kevin

    2015-08-01

    In January 2014, an international meeting sponsored by the International Life Sciences Institute/Health and Environmental Sciences Institute and the Canadian Food Inspection Agency titled "Genetic Basis of Unintended Effects in Modified Plants" was held in Ottawa, Canada, bringing together over 75 scientists from academia, government, and the agro-biotech industry. The objectives of the meeting were to explore current knowledge and identify areas requiring further study on unintended effects in plants and to discuss how this information can inform and improve genetically modified (GM) crop risk assessments. The meeting featured presentations on the molecular basis of plant genome variability in general, unintended changes at the molecular and phenotypic levels, and the development and use of hypothesis-driven evaluations of unintended effects in assessing conventional and GM crops. The development and role of emerging "omics" technologies in the assessment of unintended effects was also discussed. Several themes recurred in a number of talks; for example, a common observation was that no system for genetic modification, including conventional methods of plant breeding, is without unintended effects. Another common observation was that "unintended" does not necessarily mean "harmful". This paper summarizes key points from the information presented at the meeting to provide readers with current viewpoints on these topics.

  8. Biological impact of cigarette smoke compared to an aerosol produced from a prototypic modified risk tobacco product on normal human bronchial epithelial cells.

    Science.gov (United States)

    Kogel, U; Gonzalez Suarez, I; Xiang, Y; Dossin, E; Guy, P A; Mathis, C; Marescotti, D; Goedertier, D; Martin, F; Peitsch, M C; Hoeng, J

    2015-12-01

    Cigarette smoking causes serious and fatal diseases. The best way for smokers to avoid health risks is to quit smoking. Using modified risk tobacco products (MRTPs) may be an alternative to reduce the harm caused for those who are unwilling to quit smoking, but little is known about the toxic effects of MRTPs, nor were the molecular mechanisms of toxicity investigated in detail. The toxicity of an MRTP and the potential molecular mechanisms involved were investigated in high-content screening tests and whole genome transcriptomics analyses using human bronchial epithelial cells. The prototypic (p)MRTP that was tested had less impact than reference cigarette 3R4F on the cellular oxidative stress response and cell death pathways. Higher pMRTP aerosol extract concentrations had impact on pathways associated with the detoxification of xenobiotics and the reduction of oxidative damage. A pMRTP aerosol concentration up to 18 times higher than the 3R4F caused similar perturbation effects in biological networks and led to the perturbation of networks related to cell stress, and proliferation biology. These results may further facilitate the development of a systems toxicology-based impact assessment for use in future risk assessments in line with the 21st century toxicology paradigm, as shown here for an MRTP. Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.

  9. A cold-induced myo-inositol transporter-like gene confers tolerance to multiple abiotic stresses in transgenic tobacco plants.

    Science.gov (United States)

    Sambe, Mame Abdou Nahr; He, Xueying; Tu, Qinghua; Guo, Zhenfei

    2015-03-01

    A full length cDNA encoding a myo-inositol transporter-like protein, named as MfINT-like, was cloned from Medicago sativa subsp. falcata (herein falcata), a species with greater cold tolerance than alfalfa (M. sativa subsp. sativa). MfINT-like is located on plasma membranes. MfINT-like transcript was induced 2-4 h after exogenous myo-inositol treatment, 24-96 h with cold, and 96 h by salinity. Given that myo-inositol accumulates higher in falcata after 24 h of cold treatment, myo-inositol is proposed to be involved in cold-induced expression of MfINT-like. Higher levels of myo-inositol was observed in leaves of transgenic tobacco plants overexpressing MfINT-like than the wild-type but not in the roots of plants grown on myo-inositol containing medium, suggesting that transgenic plants had higher myo-inositol transport activity than the wild-type. Transgenic plants survived better to freezing temperature, and had lower ion leakage and higher maximal photochemical efficiency of photosystem II (Fv /Fm ) after chilling treatment. In addition, greater plant fresh weight was observed in transgenic plants as compared with the wild-type when plants were grown under drought or salinity stress. The results suggest that MfINT-like mediated transport of myo-inositol is associated with plant tolerance to abiotic stresses.

  10. Recombinant protein expression plasmids optimized for industrial E. coli fermentation and plant systems produce biologically active human insulin-like growth factor-1 in transgenic rice and tobacco plants.

    Science.gov (United States)

    Panahi, Mitra; Alli, Zaman; Cheng, Xiongying; Belbaraka, Loubaba; Belgoudi, Jaafar; Sardana, Ravinder; Phipps, Jenny; Altosaar, Illimar

    2004-06-01

    Human insulin-like growth factor-1 (hIGF-1) is a growth factor with clinical significance in medicine. The therapeutic potential of recombinant hIGF-1 (rthIGF-1) stems from the fact that hIGF-1 resembles insulin in many aspects of physiology. The expression of hIGF-1 in transgenic tobacco and rice plants using different expression cassettes is reported here. In the present study, two coding sequences were tested, one with the original human sequence, but partially optimized for expression in E. coli and the other with a plant-codon-optimized sequence that was expected to give a higher level of expression in plant systems. Three different hIGF-1 recombinant expression constructs were generated. All expression constructs utilized the maize ubiquitin 1 promoter with or without a signal sequence. Analyses conducted using a hIGF-1 specific ELISA kit showed all transgenic plants produced hIGF-1 and the accumulated hIGF-1 increased from the E. coli codon bias to higher levels when the hIGF-1 coding sequence was codon-optimized to match that of the maize zeamatin protein--the most transcribed gene in maize endosperm suspension cells. Further analyses that compared the functionality of the bacterial signal peptide Lam B in plants showed that this leader peptide led to lower expression levels when compared to transgenic plants that did not contain this sequence. This indicated that this expression construct was functional without removal of the bacterial signal sequence. The maize ubiquitin 1 promoter was found to be more active in rice plants than tobacco plants indicating that in this case, there was a class preference that was biased towards a monocot host. Biological analyses conducted using protein extracts from transgenic plants showed that the rthIGF-1 was effective in stimulating the in vitro growth and proliferation of human SH-SY5Y neuroblastoma cells. This indicated that the plant-produced rthIGF-1 was stable and biologically active. As some plants have been

  11. Production by Tobacco Transplastomic Plants of Recombinant Fungal and Bacterial Cell-Wall Degrading Enzymes to Be Used for Cellulosic Biomass Saccharification.

    Science.gov (United States)

    Longoni, Paolo; Leelavathi, Sadhu; Doria, Enrico; Reddy, Vanga Siva; Cella, Rino

    2015-01-01

    Biofuels from renewable plant biomass are gaining momentum due to climate change related to atmospheric CO2 increase. However, the production cost of enzymes required for cellulosic biomass saccharification is a major limiting step in this process. Low-cost production of large amounts of recombinant enzymes by transgenic plants was proposed as an alternative to the conventional microbial based fermentation. A number of studies have shown that chloroplast-based gene expression offers several advantages over nuclear transformation due to efficient transcription and translation systems and high copy number of the transgene. In this study, we expressed in tobacco chloroplasts microbial genes encoding five cellulases and a polygalacturonase. Leaf extracts containing the recombinant enzymes showed the ability to degrade various cell-wall components under different conditions, singly and in combinations. In addition, our group also tested a previously described thermostable xylanase in combination with a cellulase and a polygalacturonase to study the cumulative effect on the depolymerization of a complex plant substrate. Our results demonstrate the feasibility of using transplastomic tobacco leaf extracts to convert cell-wall polysaccharides into reducing sugars, fulfilling a major prerequisite of large scale availability of a variety of cell-wall degrading enzymes for biofuel industry.

  12. Production by Tobacco Transplastomic Plants of Recombinant Fungal and Bacterial Cell-Wall Degrading Enzymes to Be Used for Cellulosic Biomass Saccharification

    Directory of Open Access Journals (Sweden)

    Paolo Longoni

    2015-01-01

    Full Text Available Biofuels from renewable plant biomass are gaining momentum due to climate change related to atmospheric CO2 increase. However, the production cost of enzymes required for cellulosic biomass saccharification is a major limiting step in this process. Low-cost production of large amounts of recombinant enzymes by transgenic plants was proposed as an alternative to the conventional microbial based fermentation. A number of studies have shown that chloroplast-based gene expression offers several advantages over nuclear transformation due to efficient transcription and translation systems and high copy number of the transgene. In this study, we expressed in tobacco chloroplasts microbial genes encoding five cellulases and a polygalacturonase. Leaf extracts containing the recombinant enzymes showed the ability to degrade various cell-wall components under different conditions, singly and in combinations. In addition, our group also tested a previously described thermostable xylanase in combination with a cellulase and a polygalacturonase to study the cumulative effect on the depolymerization of a complex plant substrate. Our results demonstrate the feasibility of using transplastomic tobacco leaf extracts to convert cell-wall polysaccharides into reducing sugars, fulfilling a major prerequisite of large scale availability of a variety of cell-wall degrading enzymes for biofuel industry.

  13. Characterization of the Tomato Prosystemin Promoter: Organ-specific Expression, Hormone Specificity and Methyl Jasmonate Responsiveness by Deletion Analysis in Transgenic Tobacco Plants(F)

    Institute of Scientific and Technical Information of China (English)

    Hamlet Avilés-Arnaut; John Paul Délano-Frier

    2012-01-01

    Tomato systemin is a bioactive peptide that regulates the systemic activation of wound-responsive genes.It is released from its 200 amino acid precursor called prosystemin.Initial tissue-localization and hormone-induced expression assays indicated that the tomato prosystemin gene (SIPS) accumulates mainly in floral tissues and in response to exogenous abscisic acid and methyl jasmonate (MeJA)treatments,respectively.Later,the promoter regions of the PS gene in tomato (Solanum lycopersicum L.cv.Castlemart),pepper (Capsicum annuum) and potato (Solanum tuberosum) were isolated and an in silico analysis of the SIPS promoter revealed an over-representation of stress- and MeJA-responsive motifs.A subsequent 5' deletion analysis of the SIPS promoter fused to theβ-glucuronidase reporter (GUS) gene showed that the -221 to +40 bp proximal SIPS promoter region was sufficient to direct the stigma,vascular bundle-specific and MeJA-responsive expression of GUS in transgenic tobacco plants.Important vascular-tissue-specific,light- and MeJA-responsive cis-elements were also present in this region.These findings provide relevant information regarding the transcriptional regulation mechanisms of the SIPS promoter operating in transgenic tobacco plants.They also suggest that its tissue-specificity and inducible nature could have wide applicability in plant biotechnology.

  14. The capacity of modified nickel catalysts derived from discharged catalyst of fertilizer plants for NOx treatment

    Science.gov (United States)

    Ha, T. M. P.; Luong, N. T.; Le, P. N.

    2016-11-01

    In Vietnam for recent years, a large amount of hazardous waste containing nickel (Ni) derived from discharged catalyst of fertilizer plants has caused environmental problems in landfill overloading and the risk of soil or surface water sources pollution. Taking advantage of discharged catalyst, recycling Ni components and then synthesizing new catalysts apply for mono-nitrogen oxides (NOx) treatments is an approach to bring about both economic and environmental benefits. This study was carried out with the main objective: Evaluate the performance of modified catalysts (using recovered Ni from the discharged RKS-2-7H catalyst of Phu My Fertilizer Plant) on NOx treatment. The catalysts was synthesized and modified with active phases consist of recovered Ni and commercial Barium oxide (BaO), Manganese dioxide (MnO2) / Cerium (IV) oxide (CeO2) on the support Aluminium oxide (γ-Al2O3). The results show that the modified catalysts with Ni, Ba, Ce was not more beneficial for NOx removal than which with Ni, Ba, Mn. 98% NOx removal at 350°C with the start temperature at 115°C and the T60 value at 307°C can be obtained with 10Ni10Ba10Mn/Al catalyst.

  15. Optimization of CRISPR/Cas9 genome editing to modify abiotic stress responses in plants

    Science.gov (United States)

    Osakabe, Yuriko; Watanabe, Takahito; Sugano, Shigeo S; Ueta, Risa; Ishihara, Ryosuke; Shinozaki, Kazuo; Osakabe, Keishi

    2016-01-01

    Genome editing using the CRISPR/Cas9 system can be used to modify plant genomes, however, improvements in specificity and applicability are still needed in order for the editing technique to be useful in various plant species. Here, using genome editing mediated by a truncated gRNA (tru-gRNA)/Cas9 combination, we generated new alleles for OST2, a proton pump in Arabidopsis, with no off-target effects. By following expression of Cas9 and the tru-gRNAs, newly generated mutations in CRIPSR/Cas9 transgenic plants were detected with high average mutation rates of up to 32.8% and no off-target effects using constitutive promoter. Reducing nuclear localization signals in Cas9 decreased the mutation rate. In contrast, tru-gRNA Cas9 cassettes driven by meristematic- and reproductive-tissue-specific promoters increased the heritable mutation rate in Arabidopsis, showing that high expression in the germ line can produce bi-allelic mutations. Finally, the new mutant alleles obtained for OST2 exhibited altered stomatal closing in response to environmental conditions. These results suggest further applications in molecular breeding to improve plant function using optimized plant CRISPR/Cas9 systems. PMID:27226176

  16. Optimization of CRISPR/Cas9 genome editing to modify abiotic stress responses in plants.

    Science.gov (United States)

    Osakabe, Yuriko; Watanabe, Takahito; Sugano, Shigeo S; Ueta, Risa; Ishihara, Ryosuke; Shinozaki, Kazuo; Osakabe, Keishi

    2016-05-26

    Genome editing using the CRISPR/Cas9 system can be used to modify plant genomes, however, improvements in specificity and applicability are still needed in order for the editing technique to be useful in various plant species. Here, using genome editing mediated by a truncated gRNA (tru-gRNA)/Cas9 combination, we generated new alleles for OST2, a proton pump in Arabidopsis, with no off-target effects. By following expression of Cas9 and the tru-gRNAs, newly generated mutations in CRIPSR/Cas9 transgenic plants were detected with high average mutation rates of up to 32.8% and no off-target effects using constitutive promoter. Reducing nuclear localization signals in Cas9 decreased the mutation rate. In contrast, tru-gRNA Cas9 cassettes driven by meristematic- and reproductive-tissue-specific promoters increased the heritable mutation rate in Arabidopsis, showing that high expression in the germ line can produce bi-allelic mutations. Finally, the new mutant alleles obtained for OST2 exhibited altered stomatal closing in response to environmental conditions. These results suggest further applications in molecular breeding to improve plant function using optimized plant CRISPR/Cas9 systems.

  17. RNAi-mediated mortality of the whitefly through transgenic expression of double-stranded RNA homologous to acetylcholinesterase and ecdysone receptor in tobacco plants

    Science.gov (United States)

    Malik, Hassan Jamil; Raza, Amir; Amin, Imran; Scheffler, Jodi A.; Scheffler, Brian E.; Brown, Judith K.; Mansoor, Shahid

    2016-01-01

    The whitefly Bemisia tabaci (Genn.) is a pest and vector of plant viruses to crop and ornamental plants worldwide. Using RNA interference (RNAi) to down regulate whitefly genes by expressing their homologous double stranded RNAs in plants has great potential for management of whiteflies to reduce plant virus disease spread. Using a Tobacco rattle virus-derived plasmid for in planta transient expression of double stranded RNA (dsRNA) homologous to the acetylcholinesterase (AChE) and ecdysone receptor (EcR) genes of B. tabaci, resulted in significant adult whitefly mortality. Nicotiana tabacum L. plants expressing dsRNA homologous to B. tabaci AChE and EcR were constructed by fusing sequences derived from both genes. Mortality of adult whiteflies exposed to dsRNA by feeding on N. tabacum plants, compared to non-dsRNA expressing plants, recorded at 24-hr intervals post-ingestion for three days, was >90% and 10%, respectively. Analysis of gene expression by real time quantitative PCR indicated that whitefly mortality was attributable to the down-regulation of both target genes by RNAi. Results indicated that knock down of whitefly genes involved in neuronal transmission and transcriptional activation of developmental genes, has potential as a bio-pesticide to reduce whitefly population size and thereby decrease virus spread. PMID:27929123

  18. RNAi-mediated mortality of the whitefly through transgenic expression of double-stranded RNA homologous to acetylcholinesterase and ecdysone receptor in tobacco plants.

    Science.gov (United States)

    Malik, Hassan Jamil; Raza, Amir; Amin, Imran; Scheffler, Jodi A; Scheffler, Brian E; Brown, Judith K; Mansoor, Shahid

    2016-12-08

    The whitefly Bemisia tabaci (Genn.) is a pest and vector of plant viruses to crop and ornamental plants worldwide. Using RNA interference (RNAi) to down regulate whitefly genes by expressing their homologous double stranded RNAs in plants has great potential for management of whiteflies to reduce plant virus disease spread. Using a Tobacco rattle virus-derived plasmid for in planta transient expression of double stranded RNA (dsRNA) homologous to the acetylcholinesterase (AChE) and ecdysone receptor (EcR) genes of B. tabaci, resulted in significant adult whitefly mortality. Nicotiana tabacum L. plants expressing dsRNA homologous to B. tabaci AChE and EcR were constructed by fusing sequences derived from both genes. Mortality of adult whiteflies exposed to dsRNA by feeding on N. tabacum plants, compared to non-dsRNA expressing plants, recorded at 24-hr intervals post-ingestion for three days, was >90% and 10%, respectively. Analysis of gene expression by real time quantitative PCR indicated that whitefly mortality was attributable to the down-regulation of both target genes by RNAi. Results indicated that knock down of whitefly genes involved in neuronal transmission and transcriptional activation of developmental genes, has potential as a bio-pesticide to reduce whitefly population size and thereby decrease virus spread.

  19. Increased production of wax esters in transgenic tobacco plants by expression of a fatty acid reductase:wax synthase gene fusion.

    Science.gov (United States)

    Aslan, Selcuk; Hofvander, Per; Dutta, Paresh; Sun, Chuanxin; Sitbon, Folke

    2015-12-01

    Wax esters are hydrophobic lipids consisting of a fatty acid moiety linked to a fatty alcohol with an ester bond. Plant-derived wax esters are today of particular concern for their potential as cost-effective and sustainable sources of lubricants. However, this aspect is hampered by the fact that the level of wax esters in plants generally is too low to allow commercial exploitation. To investigate whether wax ester biosynthesis can be increased in plants using transgenic approaches, we have here exploited a fusion between two bacterial genes together encoding a single wax ester-forming enzyme, and targeted the resulting protein to chloroplasts in stably transformed tobacco (Nicotiana benthamiana) plants. Compared to wild-type controls, transgenic plants showed both in leaves and stems a significant increase in the total level of wax esters, being eight-fold at the whole plant level. The profiles of fatty acid methyl ester and fatty alcohol in wax esters were related, and C16 and C18 molecules constituted predominant forms. Strong transformants displayed certain developmental aberrations, such as stunted growth and chlorotic leaves and stems. These negative effects were associated with an accumulation of fatty alcohols, suggesting that an adequate balance between formation and esterification of fatty alcohols is crucial for a high wax ester production. The results show that wax ester engineering in transgenic plants is feasible, and suggest that higher yields may become achieved in the near future.

  20. Suppression of Reactive Oxygen Species Accumulation in Chloroplasts Prevents Leaf Damage but Not Growth Arrest in Salt-Stressed Tobacco Plants.

    Directory of Open Access Journals (Sweden)

    Anabella F Lodeyro

    Full Text Available Crop yield reduction due to salinity is a growing agronomical concern in many regions. Increased production of reactive oxygen species (ROS in plant cells accompanies many abiotic stresses including salinity, acting as toxic and signaling molecules during plant stress responses. While ROS are generated in various cellular compartments, chloroplasts represent a main source in the light, and plastid ROS synthesis and/or elimination have been manipulated to improve stress tolerance. Transgenic tobacco plants expressing a plastid-targeted cyanobacterial flavodoxin, a flavoprotein that prevents ROS accumulation specifically in chloroplasts, displayed increased tolerance to many environmental stresses, including drought, excess irradiation, extreme temperatures and iron starvation. Surprisingly, flavodoxin expression failed to protect transgenic plants against NaCl toxicity. However, when high salt was directly applied to leaf discs, flavodoxin did increase tolerance, as reflected by preservation of chlorophylls, carotenoids and photosynthetic activities. Flavodoxin decreased salt-dependent ROS accumulation in leaf tissue from discs and whole plants, but this decline did not improve tolerance at the whole plant level. NaCl accumulation in roots, as well as increased osmotic pressure and salt-induced root damage, were not prevented by flavodoxin expression. The results indicate that ROS formed in chloroplasts have a marginal effect on plant responses during salt stress, and that sensitive targets are present in roots which are not protected by flavodoxin.

  1. Thai ethnomedicinal plants as resistant modifying agents for combating Acinetobacter baumannii infections

    Directory of Open Access Journals (Sweden)

    Phatthalung Pinanong

    2012-04-01

    Full Text Available Abstracts Background Acinetobacter baumannii is well-recognized as an important nosocomial pathogen, however, due to their intrinsic resistance to several antibiotics, treatment options are limited. Synergistic effects between antibiotics and medicinal plants, particularly their active components, have intensively been studied as alternative approaches. Methods Fifty-one ethanol extracts obtained from 44 different selected medicinal plant species were tested for resistance modifying agents (RMAs of novobiocin against A. baumannii using growth inhibition assay. Results At 250 μg/ml, Holarrhena antidysenterica, Punica granatum, Quisqualis indica, Terminalia bellirica, Terminalia chebula, and Terminalia sp. that possessed low intrinsic antibacterial activity significantly enhanced the activity of novobiocin at 1 μg/ml (1/8xminimum inhibitory concentration against this pathogen. Holarrhena antidysenterica at 7.8 μg/ml demonstrated remarkable resistant modifying ability against A. baumannii in combination with novobiocin. The phytochemical study revealed that constituents of this medicinal plant contain alkaloids, condensed tannins, and triterpenoids. Conclusion The use of Holarrhena antidysenterica in combination with novobiocin provides an effective alternative treatment for multidrug resistant A. baumannii infections.

  2. Summary of Investigations of the Use of Modified Turbine Inlet Conditions in a Binary Power Plant

    Energy Technology Data Exchange (ETDEWEB)

    Mines, Gregory Lee

    2000-09-01

    Investigators at the Idaho National Engineering and Environmental Laboratory (INEEL) are developing technologies that will enhance the feasibility of generating electrical power from a hydrothermal resource. One of the concepts investigated is the use of modified inlet conditions in geothermal binary power plant turbines to increase the power generation. An inlet condition of interest allows the expanding vapor to enter the two-phase region, a mode of operation typically avoided because of concern that condensate would form and damage the turbine, degrading performance. INEEL investigators postulated that initially a supersaturated vapor would be supported, and that no turbine damage would occur. This paper summarizes the investigation of these expansions that began with testing of their condensation behavior, and culminated with the incorporation of these expansions into the operation of several commercial binary plant turbines.

  3. Endogenous allergens and compositional analysis in the allergenicity assessment of genetically modified plants.

    Science.gov (United States)

    Fernandez, A; Mills, E N C; Lovik, M; Spoek, A; Germini, A; Mikalsen, A; Wal, J M

    2013-12-01

    Allergenicity assessment of genetically modified (GM) plants is one of the key pillars in the safety assessment process of these products. As part of this evaluation, one of the concerns is to assess that unintended effects (e.g. over-expression of endogenous allergens) relevant for the food safety have not occurred due to the genetic modification. Novel technologies are now available and could be used as complementary and/or alternative methods to those based on human sera for the assessment of endogenous allergenicity. In view of these developments and as a step forward in the allergenicity assessment of GM plants, it is recommended that known endogenous allergens are included in the compositional analysis as additional parameters to be measured.

  4. Plants with stacked genetically modified events: to assess or not to assess?

    DEFF Research Database (Denmark)

    Kok, Esther J.; Pedersen, Jan W.; Onori, Roberta

    2014-01-01

    of these differences relates to the so-called ‘stacked GM events’, that is, GMOs, plants so far, where new traits are combined by conventional crossing of different GM plants. This paper advocates rethinking the current food/feed safety assessment of stacked GM events in Europe based on an analysis of different......The principles for the safety assessment of genetically modified (GM) organisms (GMOs) are harmonised worldwide to a large extent. There are, however, still differences between the European GMO regulations and the GMO regulations as they have been formulated in other parts of the world. One...... aspects that currently form the rationale for the safety assessment of stacked GM events....

  5. 种植密度和施肥量对烟叶生长和质量的影响%Effects of Planting Density and Fertilization on the Growth and Quality of Tobacco

    Institute of Scientific and Technical Information of China (English)

    夏玉珍; 鲁绍坤; 王毅; 杨宇虹; 李正风

    2015-01-01

    为研究种植密度和施肥量对烟叶生长和质量的影响,在云南玉溪选取试验点,对烤烟‘K326’品种进行种植密度和施肥量2因素3水平的裂区试验。结果表明,烤烟种植密度及施肥量对烟叶质量与产量均有显著影响,其中种植密度对烤烟产量的影响更大;施肥量对烤烟内在化学成分的影响更大。%In order to study the effects of planting density and fertilizer application on the growth and quality of tobacco leaves, the experimental points were selected in Yuxi Yunnan to conduct the split plot experiment of different planting densities and fertilizer rates of two factors and three levels of flue-cured tobacco varieties‘K326’. The results showed that the planting density and fertilization of flue-cured tobacco were significantly affected tobacco quality and yield, planting density had more effect on the yield and fertilization had more effect on intrinsic chemical composition of flue-cured tobacco.

  6. Determinants of Plant Growth-promoting Ochrobactrum lupini KUDC1013 Involved in Induction of Systemic Resistance against Pectobacterium carotovorum subsp. carotovorum in Tobacco Leaves

    Directory of Open Access Journals (Sweden)

    Marilyn Sumayo

    2013-06-01

    Full Text Available The plant growth-promoting rhizobacterium Ochrobactrum lupini KUDC1013 elicited induced systemic resistance (ISR in tobacco against soft rot disease caused by Pectobacterium carotovorum subsp. carotovorum. We investigated of its factors involved in ISR elicitation. To characterize the ISR determinants, KUDC1013 cell suspension, heat-treated cells, supernatant from a culture medium, crude bacterial lipopolysaccharide (LPS and flagella were tested for their ISR activities. Both LPS and flagella from KUDC1013 were effective in ISR elicitation. Crude cell free supernatant elicited ISR and factors with the highest ISR activity were retained in the n-butanol fraction. Analysis of the ISR-active fraction revealed the metabolites, phenylacetic acid (PAA, 1-hexadecene and linoleic acid (LA, as elicitors of ISR. Treatment of tobacco with these compounds significantly decreased the soft rot disease symptoms. This is the first report on the ISR determinants by plant growth-promoting rhizobacteria (PGPR KUDC1013 and identifying PAA, 1-hexadecene and LA as ISR-related compounds. This study shows that KUDC1013 has a great potential as biological control agent because of its multiple factors involved in induction of systemic resistance against phytopathogens.

  7. The use of statistical tools in field testing of putative effects of genetically modified plants on non-target organisms

    NARCIS (Netherlands)

    Semenov, Alexander V.; van Elsas, Jan Dirk; Glandorf, Debora C. M.; Schilthuizen, Menno; de Boer, Willem F.

    2013-01-01

    To fulfill existing guidelines, applicants that aim to place their genetically modified (GM) insect-resistant crop plants on the market are required to provide data from field experiments that address the potential impacts of the GM plants on nontarget organisms (NTO's). Such data may be based on va

  8. The use of statistical tools in field testing of putative effects of genetically modified plants on nontarget organisms

    NARCIS (Netherlands)

    Semenov, A.V.; Elsas, van J.D.; Glandorf, D.C.M.; Schilthuizen, M.; Boer, de W.F.

    2013-01-01

    To fulfill existing guidelines, applicants that aim to place their genetically modified (GM) insect-resistant crop plants on the market are required to provide data from field experiments that address the potential impacts of the GM plants on nontarget organisms (NTO's). Such data may be based on

  9. The use of statistical tools in field testing of putative effects of genetically modified plants on non-target organisms

    NARCIS (Netherlands)

    Semenov, Alexander V.; van Elsas, Jan Dirk; Glandorf, Debora C. M.; Schilthuizen, Menno; de Boer, Willem F.

    To fulfill existing guidelines, applicants that aim to place their genetically modified (GM) insect-resistant crop plants on the market are required to provide data from field experiments that address the potential impacts of the GM plants on nontarget organisms (NTO's). Such data may be based on

  10. A statistical simulation model for fiels testing of non-target organisms in environmental risk assessment of genetically modified plants

    NARCIS (Netherlands)

    Goedhart, P.W.; Voet, van der H.; Baldacchino, F.; Arpaia, S.

    2014-01-01

    Genetic modification of plants may result in unintended effects causing potentially adverse effects on the environment. A comparative safety assessment is therefore required by authorities, such as the European Food Safety Authority, in which the genetically modified plant is compared with its conve

  11. Overexpression of a cytosolic abiotic stress responsive universal stress protein (SbUSP mitigates salt and osmotic stress in transgenic tobacco plants

    Directory of Open Access Journals (Sweden)

    Pushpika eUdawat

    2016-04-01

    Full Text Available The Universal Stress Protein (USP is a ubiquitous protein and plays an indispensable role in plant abiotic stress tolerance. The genome of Salicornia brachiata contains two homologues of intron less SbUSP gene which encodes for salt and osmotic responsive universal stress protein. In vivo localization reveals that SbUSP is a membrane bound cytosolic protein. The role of the gene was functionally validated by developing transgenic tobacco and compared with control (wild type and vector control plants under different abiotic stress condition. Transgenic lines (T1 exhibited higher chlorophyll, relative water, proline, total sugar, reducing sugar, free amino acids, polyphenol contents, osmotic potential, membrane stability and lower electrolyte leakage and lipid peroxidation (malondialdehyde content under stress treatments than control (WT and VC plants. Lower accumulation of H2O2 and O2- radicals was also detected in transgenic lines compared to control plants under stress conditions. Present study confers that overexpression of the SbUSP gene enhances plant growth, alleviates ROS buildup, maintains ion homeostasis and improves the physiological status of the plant under salt and osmotic stresses. Principal component analysis (PCA exhibited a statistical distinction of plant response to salinity stress, and a significant response was observed for transgenic lines under stress, which provides stress endurance to the plant. A possible signaling role is proposed that some downstream genes may get activated by abiotic stress responsive cytosolic SbUSP, which leads to the protection of cell from oxidative damages. The study unveils that ectopic expression of the gene mitigates salt or osmotic stress by scavenging ROS and modulating the physiological process of the plant.

  12. Analysis of Comprehensive Efficiency of Traditional and Organic Tobacco Planting Method%有机与常规种植方式烤烟综合效益分析

    Institute of Scientific and Technical Information of China (English)

    张光煦; 郭怡卿; 马剑雄; 欧阳文; 阮天伟; 陈云松; 张艳波; 安经明

    2012-01-01

    随着人们对环境与食品安全的关注,有机产品的消费需求应运而生,有机烟草生产也开始在国内出现.为了探索有机烤烟种植方式的可行性,作者于2008年至2010年在大理州宾川县拉乌乡开展了有机烟叶生产试验示范,从种植技术、烤烟农艺性状、病害发生、常规化学成分、产量效益等,综合分析了有机种植与常规种植的差异,为有机烟叶规模化生产提供指导.结果表明,种植密度对烟叶中后期病害的发生影响较大,有机种植应适当降低种植密度;施肥量不仅影响烤烟生长、产量与品质,而且与烟叶中后期病害程度直接相关;有机种植的初烤烟叶较常规种植的产量平均减少18.7%,因此发展有机烤烟种植,主管部门应出台相应的补贴政策.%With the increase of concern on security of environment and food, organic products became popular and organic tobacco production also rise in China. In order to explore the possibility of organic tobacco production, experiments and extension were conducted in Lawu County, Dali of Yunnan Province. Planting technique, agricultural characters, diseases,yield and quality of tobacco were investigated to analyze the difference between organic and traditional planting method. The results showed that planting density significantly affected the disease of tobacco leave in the later period, so it was better to decrease the planting density in organic tobacco planting method. The rate of fertilize was not only closely related to plant growth, yield and quality, but also to the disease of tobacco leave in the later period. After curing, the yield of organic tobacco leaf was decreased 18.7% than that of traditional method. Therefore, for organic tobacco production development, Tobacco Company should set up composition policy to encourage the production.

  13. Smoked Tobacco Products

    Science.gov (United States)

    ... United States from India and other Southeast Asian countries. They are tobacco wrapped in a tendu or temburni leaf—plants native to Asia—that may be tied with colorful string at one or both ends. Bidis can be flavored—such as chocolate, cherry, or mango—or unflavored. Flavored bidis, however, ...

  14. Growth phase-dependent expression of proteins with decreased plant-specific N-glycans and immunogenicity in tobacco BY2 cells

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    Plants possess some desirable characteristics to synthesize recombinant glycoproteins for pharma-ceutical application.However,the mammalian glycoproteins produced in plants are somewhat different from their natural counterparts in terms of N-glycoforms.The immunogenicity of plant-specific glyco-epitopes is the major concern in human therapy.Here,the distribution of N-glycans in different growth phases of tobacco BY2 cells and their immunogenicity in mice were determined.It was ob-served that the percentage of β1,2-xylose and α1,3-fucose in proteins of growing cells increased and the corresponding protein extracts caused accelerated immune response in mice.Based on this observation,the recombinant erythropoietin in BY2 cells was expressed and characterized,and Western blot analysis showed that the recombinant erythropoietin contained a relatively small amount of plant-specific glyco-epitopes in the early phase of culture growth.This study may provide a simple but effective strategy for the production of therapeutic glycoproteins with human-like N-glycan structures in plant hosts to avoid a great allergenic risk.

  15. Altered apoplastic ascorbate redox state in tobacco plants via ascorbate oxidase overexpression results in delayed dark-induced senescence in detached leaves.

    Science.gov (United States)

    Fotopoulos, Vasileios; Kanellis, Angelos K

    2013-12-01

    Ascorbate oxidase (AO) is an apoplastic enzyme that uses oxygen to catalyse the oxidation of ascorbate (AA) to dehydroascorbate (DHA) via the unstable radical monodehydroascorbate (MDHA). Here, we report that transgenic tobacco plants (Nicotiana tabacum L. cv. Xanthi) with an in vivo lowered apoplastic AA redox state through increased AO expression demonstrate signs of delayed dark-induced senescence compared with wild-type plants, as shown by chlorophyll loss assay. In situ localization of hydrogen peroxide (H2O2) suggests that, although transgenic plants have higher constitutive levels of H2O2 under normal growth conditions, imposed dark-induced senescence results in smaller induction levels of H2O2, an observation which correlates with increased antioxidant enzyme activities and an induction in the expression of AA recycling genes compared with that in wild-type plants. Our current findings, combined with previous studies which showed the contribution of AO in the regulation of AA redox state, suggest that the reduction in AA redox state in the leaf apoplast of these transgenic plants results in an increase in the endogenous levels of H2O2, which provides a form of 'acquired tolerance' to oxidative stress imposed by dark-induced senescence.

  16. Expression of an alfalfa (Medicago sativa L.) ethylene response factor gene MsERF8 in tobacco plants enhances resistance to salinity.

    Science.gov (United States)

    Chen, Tingting; Yang, Qingchuan; Gruber, Margaret; Kang, Junmei; Sun, Yan; Ding, Wang; Zhang, Tiejun; Zhang, Xinquan

    2012-05-01

    Ethylene response factors (ERF) play crucial roles in plant development and response to stresses. Here, a novel cDNA fragment (MsERF8) encoding an ERF protein with an AP2 domain was isolated and characterized from alfalfa. The MsERF8 cDNA has an open reading frame of 603 bp and encodes a nuclear protein of 201 amino acids. Q-RT-PCR analysis revealed that MsERF8 was strongly enriched in roots and leaves compared with stems, flower buds and flowers of mature alfalfa plants. Bioinformatic analysis of the MsERF8 promoter indicated a number of elements associated with stress-related responses, and MsERF8 transcripts in alfalfa seedlings were induced by NaCl, PEG6000, Al(2)(SO(4))(3) and five different hormones. Expression of MsERF8 in transgenic tobacco plants resulted in higher tolerance to salinity than with non-transgenic plants. This data shows that MsERF8 is a gene which prevents or alleviates salinity damage and has strong potential to impart salt tolerance to other crop plants.

  17. Genetically modified plants and food hypersensitivity diseases: usage and implications of experimental models for risk assessment.

    Science.gov (United States)

    Prescott, Vanessa E; Hogan, Simon P

    2006-08-01

    The recent advances in biotechnology in the plant industry have led to increasing crop production and yield that in turn has increased the usage of genetically modified (GM) food in the human food chain. The usage of GM foods for human consumption has raised a number of fundamental questions including the ability of GM foods to elicit potentially harmful immunological responses, including allergic hypersensitivity. To assess the safety of foods derived from GM plants including allergenic potential, the US FDA, Food and Agriculture Organization of the United Nations (FAO)/World Health Organization (WHO), and the EU have developed approaches for evaluation assessment. One assessment approach that has been a very active area of research and debate is the development and usage of animal models to assess the potential allergenicity of GM foods. A number of specific animal models employing rodents, pigs, and dogs have been developed for allergenicity assessment. However, validation of these models is needed and consideration of the criteria for an appropriate animal model for the assessment of allergenicity in GM plants is required. We have recently employed a BALB/c mouse model to assess the potential allergenicity of GM plants. We have been able to demonstrate that this model is able to detect differences in antigenicity and identify aspects of protein post-translational modifications that can alter antigenicity. Furthermore, this model has also enabled us to examine the usage of GM plants as a therapeutic approach for the treatment of allergic diseases. This review discusses the current approaches to assess the allergenic potential of GM food and particularly focusing on the usage of animal models to determine the potential allergenicity of GM foods and gives an overview of our recent findings and implications of these studies.

  18. A modified Gateway cloning strategy for overexpressing tagged proteins in plants

    Directory of Open Access Journals (Sweden)

    Bowler Chris

    2008-01-01

    Full Text Available Abstract Background Recent developments, including the sequencing of a number of plant genomes, have greatly increased the amount of data available to scientists and has enabled high throughput investigations where many genes are investigated simultaneously. To perform these studies, recombinational cloning methods such as the Gateway system have been adapted to plant transformation vectors to facilitate the creation of overexpression, tagging and silencing vectors on a large scale. Results Here we present a hybrid cloning strategy which combines advantages of both recombinational and traditional cloning and which is particularly amenable to low-to-medium throughput investigations of protein function using techniques of molecular biochemistry and cell biology. The system consists of a series of twelve Gateway Entry cassettes into which a gene of interest can be inserted using traditional cloning methods to generate either N- or C-terminal fusions to epitope tags and fluorescent proteins. The resulting gene-tag fusions can then be recombined into Gateway-based Destination vectors, thus providing a wide choice of resistance marker, promoter and expression system. The advantage of this modified Gateway cloning strategy is that the entire open reading frame encoding the tagged protein of interest is contained within the Entry vectors so that after recombination no additional linker sequences are added between the tag and the protein that could interfere with protein function and expression. We demonstrate the utility of this system for both transient and stable Agrobacterium-mediated plant transformations. Conclusion This modified Gateway cloning strategy is complementary to more conventional Gateway-based systems because it expands the choice of tags and higher orders of combinations, and permits more control over the linker sequence lying between a protein of interest and an epitope tag, which can be particularly important for studies of protein

  19. Specificity of expression of the GUS reporter gene (uidA) driven by the tobacco ASA2 promoter in soybean plants and tissue cultures.

    Science.gov (United States)

    Inaba, Yoshimi; Zhong, Wei Qun; Zhang, Xing-Hai; Widholm, Jack M

    2007-07-01

    Twelve independent lines were transformed by particle bombardment of soybean embryogenic suspension cultures with the tobacco anthranilate synthase (ASA2) promoter driving the uidA (beta-glucuronidase, GUS) reporter gene. ASA2 appears to be expressed in a tissue culture specific manner in tobacco (Song H-S, Brotherton JE, Gonzales RA, Widholm JM. Tissue culture specific expression of a naturally occurring tobacco feedback-insensitive anthranilate synthase. Plant Physiol 1998;117:533-43). The transgenic lines also contained the hygromycin phosphotransferase (hpt) gene and were selected using hygromycin. All the selected cultures or the embryos that were induced from these cultures expressed GUS measured histochemically. However, no histochemical GUS expression could be found in leaves, stems, roots, pods and root nodules of the plants formed from the embryos and their progeny. Pollen from some of the plants and immature and mature seeds and embryogenic cultures initiated from immature cotyledons did show GUS activity. Quantitative 4-methylumbelliferyl-glucuronide (MUG) assays of the GUS activity in various tissues showed that all with observable histochemical GUS activity contained easily measurable activities and leaves and stems that showed no observable histochemical GUS staining did contain very low but measurable MUG activity above that of the untransformed control but orders of magnitude lower than the constitutive 35S-uidA controls used. Low but clearly above background levels of boiling sensitive GUS activity could be observed in the untransformed control immature seeds and embryogenic cultures using the MUG assay. Thus in soybean the ASA2 promoter drives readily observable GUS expression in tissue cultures, pollen and seeds, with only extremely low levels seen in vegetative tissues of the plants. The ASA2 driven expression seen in mature seed was, however, much lower than that seen with the constitutive 35S promoter; less than 2% in seed coats and less than

  20. Regulatory approval and a first-in-human phase I clinical trial of a monoclonal antibody produced in transgenic tobacco plants.

    Science.gov (United States)

    Ma, Julian K-C; Drossard, Jürgen; Lewis, David; Altmann, Friedrich; Boyle, Julia; Christou, Paul; Cole, Tom; Dale, Philip; van Dolleweerd, Craig J; Isitt, Valerie; Katinger, Dietmar; Lobedan, Martin; Mertens, Hubert; Paul, Mathew J; Rademacher, Thomas; Sack, Markus; Hundleby, Penelope A C; Stiegler, Gabriela; Stoger, Eva; Twyman, Richard M; Vcelar, Brigitta; Fischer, Rainer

    2015-10-01

    Although plant biotechnology has been widely investigated for the production of clinical-grade monoclonal antibodies, no antibody products derived from transgenic plants have yet been approved by pharmaceutical regulators for clinical testing. In the Pharma-Planta project, the HIV-neutralizing human monoclonal antibody 2G12 was expressed in transgenic tobacco (Nicotiana tabacum). The scientific, technical and regulatory demands of good manufacturing practice (GMP) were addressed by comprehensive molecular characterization of the transgene locus, confirmation of genetic and phenotypic stability over several generations of transgenic plants, and by establishing standard operating procedures for the creation of a master seed bank, plant cultivation, harvest, initial processing, downstream processing and purification. The project developed specifications for the plant-derived antibody (P2G12) as an active pharmaceutical ingredient (API) based on (i) the guidelines for the manufacture of monoclonal antibodies in cell culture systems; (ii) the draft European Medicines Agency Points to Consider document on quality requirements for APIs produced in transgenic plants; and (iii) de novo guidelines developed with European national regulators. From the resulting process, a GMP manufacturing authorization was issued by the competent authority in Germany for transgenic plant-derived monoclonal antibodies for use in a phase I clinical evaluation. Following preclinical evaluation and ethical approval, a clinical trial application was accepted by the UK national pharmaceutical regulator. A first-in-human, double-blind, placebo-controlled, randomized, dose-escalation phase I safety study of a single vaginal administration of P2G12 was carried out in healthy female subjects. The successful completion of the clinical trial marks a significant milestone in the commercial development of plant-derived pharmaceutical proteins. © 2015 Society for Experimental Biology, Association of

  1. Assessment of endogenous allergenicity of genetically modified plants exemplified by soybean - Where do we stand?

    Science.gov (United States)

    Selb, R; Wal, J M; Moreno, F J; Lovik, M; Mills, C; Hoffmann-Sommergruber, K; Fernandez, A

    2017-03-01

    According to EU regulation, genetically modified (GM) plants considered to be allergenic have to be assessed concerning their endogenous allergens before placement on the EU market, in line with the international standards described in Codex Alimentarius. Under such premises, a quantitative relevant increase in allergens might occur in GM plants as an unintended effect compared with conventionally produced crops, which could pose a risk to consumers. Currently, data showing a connection between dose and allergic sensitisation are scarce since the pathophysiological mechanisms of sensitisation are insufficiently understood. In contrast, data on population dose-distribution relationships acquired by oral food challenge are available showing a connection between quantity of allergenic protein consumed and the population of allergic individuals experiencing reactions. Soybean is currently the only recognised allergenic GM food by law for which EFSA has received applications and was therefore taken as an example for defining an assessment strategy. Identification of potential allergens, methodology for quantification as well as risk assessment considerations, are discussed. A strategy is proposed for the identification, assessment and evaluation of potential hazards/risks concerning endogenous allergenicity in food derived from plants developed by biotechnology. This approach could be expanded to other allergenic foods in the future, whenever required. Copyright © 2017 Elsevier Ltd. All rights reserved.

  2. Emerging Importance of Helicases in Plant Stress Tolerance: Characterization of Oryza sativa Repair Helicase XPB2 Promoter and Its Functional Validation in Tobacco under Multiple Stresses.

    Science.gov (United States)

    Raikwar, Shailendra; Srivastava, Vineet K; Gill, Sarvajeet S; Tuteja, Renu; Tuteja, Narendra

    2015-01-01

    Genetic material always remains at the risk of spontaneous or induced damage which challenges the normal functioning of DNA molecule, thus, DNA repair is vital to protect the organisms against genetic damage. Helicases, the unique molecular motors, are emerged as prospective molecules to engineer stress tolerance in plants and are involved in nucleic acid metabolism including DNA repair. The repair helicase, XPB is an evolutionary conserved protein present in different organisms, including plants. Availability of few efficient promoters for gene expression in plants provoked us to study the promoter of XPB for better understanding of gene regulation under stress conditions. Here, we report the in silico analysis of novel stress inducible promoter of Oryza sativa XPB2 (OsXPB2). The in vivo validation of functionality/activity of OsXPB2 promoter under abiotic and hormonal stress conditions was performed by Agrobacterium-mediated transient assay in tobacco leaves using OsXPB2::GUS chimeric construct. The present research revealed that OsXPB2 promoter contains cis-elements accounting for various abiotic stresses (salt, dehydration, or cold) and hormone (Auxin, ABA, or MeJA) induced GUS expression/activity in the promoter-reporter assay. The promoter region of OsXPB2 contains CACG, GTAACG, CACGTG, CGTCA CCGCCGCGCT cis acting-elements which are reported to be salt, dehydration, cold, MeJA, or ABA responsive, respectively. Functional analysis was done by Agrobacterium-mediated transient assay using agroinfiltration in tobacco leaves, followed by GUS staining and fluorescence quantitative analyses. The results revealed high induction of GUS activity under multiple abiotic stresses as compared to mock treated control. The present findings suggest that OsXPB2 promoter is a multi-stress inducible promoter and has potential applications in sustainable crop production under abiotic stresses by regulating desirable pattern of gene expression.

  3. The N-homologue LRR domain adopts a folding which explains the TMV-Cg-induced HR-like response in sensitive tobacco plants.

    Science.gov (United States)

    Stange, Claudia; Matus, José Tomás; Domínguez, Calixto; Perez-Acle, Tomás; Arce-Johnson, Patricio

    2008-01-01

    Following leaf infection with the tobacco mosaic virus (TMV), Nicotiana species that carry the disease resistance N gene develop a hypersensitive response (HR) that blocks the systemic movement of the virus. TMV-sensitive tobacco plants that lack the N gene develop classical disease symptoms following infection with most of the tobamoviruses. However, upon infection with TMV-Cg, these plants display a HR-like response that is unable to limit viral spread. We previously identified the NH gene in sensitive plants; this gene is homologous to the resistance N gene and both belong to the TIR/NBS/LRR family. Isolation and analysis of the NH transcript enabled the prediction of the amino acid sequence in which we detected a leucine-rich repeat domain, proposed to be involved in pathogen recognition. This domain is found in four of five classes of pathogen resistant proteins, in which sequence and structural changes may generate different specificities. In order to study the possible functional role of the LRR domain in the HR-like response, we developed a comparative three-dimensional model for the NH and N gene products, by means of functional and structural domains recognition, secondary structure prediction, domain assignment through profile Hidden Markov Models (HMM) and molecular dynamics (MD) simulations. Based on our results we postulate that the NH protein could adopt a LRR fold with a functional role in the HR-like response. Our two reliable LRR three-dimensional models (N-LRR, NH-LRR) can be used as structural frameworks for future experiments in which the structure-function relationships regarding the protein-protein interaction process may be revealed. Evolutionary aspects of the N and NH genes in Nicotiana species are also discussed.

  4. TOBACCO TIGHTROPE

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    China's monopoly tobacco industry is trying to maintain revenue levels while adjusting to stricter policies aimed at curbing smoking While China is increasingly opening the doors to its booming economy, reducing the number of state-owned enterprises and welcoming foreign businesses, when it comes to tobacco, the government is still screening out the smoke. A major source of government tax rev-

  5. Transformation and analysis of tobacco plant var Petit havana with T-urf13 gene under anther-specific TA29 promoter.

    Science.gov (United States)

    Arun, V; Kuriakose, Boney; Sridhar, Vaniyambadi V; Thomas, George

    2011-09-01

    T-urf13, a well-documented cms-associated gene from maize, has been shown to render methomyl sensitivity to heterologous systems like rice, yeast and bacteria when expressed constitutively. Since these transgenic plants were fertile, it was hypothesized that T-urf13 gene if expressed in anthers may result in male sterility that could be used for hybrid seed production. Hence, this work was aimed at analysing whether T-urf13 gene when expressed in anthers can result in male sterile plants or requires methomyl treatment to cause male sterility (controllable). This is the first report of transformation of tobacco with T-urf13 gene under anther-specific promoter (TA29) with or without mitochondrial targeting sequence. Most of the transgenic plants obtained were fertile; this was surprising as many male sterile plants were expected as T-urf13 gene is a cms associated gene. Our results suggest that it may not be possible to obtain male sterility by expressing URF13 in the anther by itself or by methomyl application.

  6. Utilization of a tobacco rattle virus vector to clone an Nicotiana benthamiana cDNA library for VIGS

    Science.gov (United States)

    Virus-induced gene silencing (VIGS) is an efficient and rapid method to identify plant gene functions. One of the most widely used VIGS vectors is Tobacco rattle virus (TRV) which has been used successfully for RNA interference (RNAi) in N. benthamiana and tomato. We previously modified a TRV VIGS v...

  7. Crop systems and plant roots can modify the soil water holding capacity

    Science.gov (United States)

    Doussan, Claude; Cousin, Isabelle; Berard, Annette; Chabbi, Abad; Legendre, Laurent; Czarnes, Sonia; Toussaint, Bruce; Ruy, Stéphane

    2015-04-01

    At the interface between atmosphere and deep sub-soil, the root zone plays a major role in regulating the flow of water between major compartments: groundwater / surface / atmosphere (drainage, runoff, evapotranspiration). This role of soil as regulator/control of water fluxes, but also as a supporting medium to plant growth, is strongly dependent on the hydric properties of the soil. In turn, the plant roots growing in the soil can change its structure; both in the plow layer and in the deeper horizons and, therefore, could change the soil properties, particularly hydric properties. Such root-related alteration of soil properties can be linked to direct effect of roots such as soil perforation during growth, aggregation of soil particles or indirect effects such as the release of exudates by roots that could modify the properties of water or of soil particles. On an another hand, the rhizosphere, the zone around roots influenced by the activity of root and associated microorganisms, could have a high influence on hydric properties, particularly the water retention. To test if crops and plant roots rhizosphere may have a significant effect on water retention, we conducted various experiment from laboratory to field scales. In the lab, we tested different soil and species for rhizospheric effect on soil water retention. Variation in available water content (AWC) between bulk and rhizospheric soil varied from non-significant to a significant increase (to about 16% increase) depending on plant species and soil type. In the field, the alteration of water retention by root systems was tested in different pedological settings for a Maize crop inoculated or not with the bacteria Azospirillum spp., known to alter root structure, growth and morphology. Again, a range of variation in AWC was evidenced, with significant increase (~30%) in some soil types, but more linked to innoculated/non-innoculated plants rather than to a difference between rhizospheric and bulk soil

  8. General recommendations for soil ecotoxicological tests suitable for the environmental risk assessment of genetically modified plants.

    Science.gov (United States)

    Römbke, Jörg; Jänsch, Stephan; Meier, Matthias; Hilbeck, Angelika; Teichmann, Hanka; Tappeser, Beatrix

    2010-04-01

    Before a genetically modified plant (GMP) can be placed on the market in the European Union (EU), an environmental risk assessment has to be conducted according to EU-Directive 2001/18/EC or Regulation (EC) No. 1829/2003 of the European Parliament and of the Council. However, no harmonized concept for ecotoxicological testing is available today that considers the characteristics of GMPs as a whole. In fact, to date, mainly ecotoxicological tests originally developed and standardized for pesticides are used for this purpose. Frequently in these tests, not the whole GMP is tested but only specific transgene products (mainly toxins). In this contribution, ecotoxicological methods developed for the testing of pesticides are evaluated for whether they are suitable for risk assessment of GMPs as well. In total, 105 test methods covering a wide range of terrestrial invertebrates, microbes, and plants (laboratory, semifield, and field levels) were assessed. Only 7 of them had already been used with GMPs, and in about 20 studies the existing tests methods were modified, mostly in a way such that nonstandard species were used. In the laboratory, few earthworm and nontarget arthropod (NTA) species as well as collembolans and isopods were tested, and, in the field, only the litter-bag test was used. Clearly, more species than these few standard organisms currently in use have to be selected for testing purposes. A more detailed analysis of GMP tests with soil invertebrates published in the literature revealed that some of the relevant GMP exposure routes, such as via bulk soil, soil porewater, and litter from GMPs, are well covered. However, studies addressing either consumption of GMPs themselves or secondary exposure after GMPs have been taken up by invertebrates that feed on living or dead GMPs are underrepresented. (c) 2009 SETAC.

  9. Tobacco Research and Its Relevance to Science, Medicine and Industry

    Directory of Open Access Journals (Sweden)

    Tso TC

    2014-12-01

    Full Text Available This article is a historical review and a vision for the future of tobacco plant research. This is the perspective of an experienced tobacco scientist who devoted his total professional career to tobacco research. From the very beginning, pioneering tobacco research was the foundation of plant science at the dawn of modern development, in such areas as light, nutrition, genetics, growth control, disorders and metabolism. Tobacco research led to current advancements in plant biotechnology. In addition, tobacco plant research contributed significantly to public health research in radioactive elements, mycotoxins, and air pollutants. However, public support for tobacco research has today greatly declined to almost total elimination because of a sense of political correctness. This author points out that tobacco is one of the most valuable research tools, and is a most abundant source of scientific information. Research with tobacco plants will contribute far beyond the frontiers of agricultural science: tobacco can be a source of food supply with nutrition value similar to that of milk; tobacco can be a source of health supplies including medical chemicals and various vaccines; tobacco can be a source of biofuel. All we need is to treat tobacco with respect; the use of tobacco is only in its initial stages.

  10. From miracle fruit to transgenic tomato: mass production of the taste-modifying protein miraculin in transgenic plants.

    Science.gov (United States)

    Hiwasa-Tanase, Kyoko; Hirai, Tadayoshi; Kato, Kazuhisa; Duhita, Narendra; Ezura, Hiroshi

    2012-03-01

    The utility of plants as biofactories has progressed in recent years. Some recombinant plant-derived pharmaceutical products have already reached the marketplace. However, with the exception of drugs and vaccines, a strong effort has not yet been made to bring recombinant products to market, as cost-effectiveness is critically important for commercialization. Sweet-tasting proteins and taste-modifying proteins have a great deal of potential in industry as substitutes for sugars and as artificial sweeteners. The taste-modifying protein, miraculin, functions to change the perception of a sour taste to a sweet one. This taste-modifying function can potentially be used not only as a low-calorie sweetener but also as a new seasoning that could be the basis of a new dietary lifestyle. However, miraculin is far from inexpensive, and its potential as a marketable product has not yet been fully developed. For the last several years, biotechnological production of this taste-modifying protein has progressed extensively. In this review, the characteristics of miraculin and recent advances in its production using transgenic plants are summarized, focusing on such topics as the suitability of plant species as expression hosts, the cultivation method for transgenic plants, the method of purifying miraculin and future advances required to achieve industrial use.

  11. 烟草甲植物源驱避剂的筛选%Screening of plant-derived repellents against tobacco beetle, Lasioderma serricorne (Fabricius)

    Institute of Scientific and Technical Information of China (English)

    李为争; 范荫荫; 安靖靖; 王琼; 郭线茹; 罗梅浩; 原国辉

    2014-01-01

    为筛选烟草甲植物源高效驱避剂,对12种芳香植物材料进行了筛选,比较了肉桂醛及其2种结构类似物(肉桂酸甲酯和肉桂酸乙酯)的驱避活性,测定了肉桂醛琼脂胶包结物的驱避效果。Y形嗅觉仪测定结果表明,肉桂皮粉和丁香粉对烟草甲具有极显著的驱避活性,3种化合物的驱避活性次序为:肉桂醛>肉桂酸甲酯>肉桂酸乙酯。盆形嗅觉仪测定结果表明,肉桂醛琼脂胶包结物在4 d内对烟草甲的驱避反应指数可达100%。肉桂醛是烟用香精的成分,可制作琼脂胶包结驱避剂用于烟仓中烟草甲的防治。%Twelve aroma plant species were chosen to screen effective plant repellent to tobacco beetle, Lasioderma serricorne. Repellent activity of cinnamaldehyde was compared against its two structural analogs (methyl cinnamate and ethyl cinnamate), and repellent effect of cinnamaldehyde formulated in agaro-pectin was also determined. Results in Y-typed olfactometer showed that the Cinnamomun cassia bark powder and Syringa oblata bud powder had extremely significant repellent effect, and repellent activity of three tested compounds was expressed in a declining order of cinnamaldehyde, methyl cinnamate and ethyl cinnamate. Results in basin-shaped olfactometer showed that repellent index of cinnamaldehyde formulated in agaro-pectin almost reached 100%against Lasioderma serricorne during a four-day test period. Thus cinnamaldehyde as component of tobacco flavor could be used to make agaro-pectin repellent against Lasioderma serricorne in tobacco warehouse.

  12. Identification and Characterization of CYP9A40 from the Tobacco Cutworm Moth (Spodoptera litura), a Cytochrome P450 Gene Induced by Plant Allelochemicals and Insecticides

    Science.gov (United States)

    Wang, Rui-Long; Staehelin, Christian; Xia, Qing-Qing; Su, Yi-Juan; Zeng, Ren-Sen

    2015-01-01

    Cytochrome P450 monooxygenases (P450s) of insects play crucial roles in the metabolism of endogenous and dietary compounds. Tobacco cutworm moth (Spodoptera litura), an important agricultural pest, causes severe yield losses in many crops. In this study, we identified CYP9A40, a novel P450 gene of S. litura, and investigated its expression profile and potential role in detoxification of plant allelochemicals and insecticides. The cDNA contains an open reading frame encoding 529 amino acid residues. CYP9A40 transcripts were found to be accumulated during various development stages of S. litura and were highest in fifth and sixth instar larvae. CYP9A40 was mainly expressed in the midgut and fat body. Larval consumption of xenobiotics, namely plant allelochemicals (quercetin and cinnamic acid) and insecticides (deltamethrin and methoxyfenozide) induced accumulation of CYP9A40 transcripts in the midgut and fat body. Injection of dsCYP9A40 (silencing of CYP9A40 by RNA interference) significantly increased the susceptibility of S. litura larvae to the tested plant allelochemicals and insecticides. These results indicate that CYP9A40 expression in S. litura is related to consumption of xenobiotics and suggest that CYP9A40 is involved in detoxification of these compounds. PMID:26393579

  13. Identification and Characterization of CYP9A40 from the Tobacco Cutworm Moth (Spodoptera litura), a Cytochrome P450 Gene Induced by Plant Allelochemicals and Insecticides.

    Science.gov (United States)

    Wang, Rui-Long; Staehelin, Christian; Xia, Qing-Qing; Su, Yi-Juan; Zeng, Ren-Sen

    2015-09-18

    Cytochrome P450 monooxygenases (P450s) of insects play crucial roles in the metabolism of endogenous and dietary compounds. Tobacco cutworm moth (Spodoptera litura), an important agricultural pest, causes severe yield losses in many crops. In this study, we identified CYP9A40, a novel P450 gene of S. litura, and investigated its expression profile and potential role in detoxification of plant allelochemicals and insecticides. The cDNA contains an open reading frame encoding 529 amino acid residues. CYP9A40 transcripts were found to be accumulated during various development stages of S. litura and were highest in fifth and sixth instar larvae. CYP9A40 was mainly expressed in the midgut and fat body. Larval consumption of xenobiotics, namely plant allelochemicals (quercetin and cinnamic acid) and insecticides (deltamethrin and methoxyfenozide) induced accumulation of CYP9A40 transcripts in the midgut and fat body. Injection of dsCYP9A40 (silencing of CYP9A40 by RNA interference) significantly increased the susceptibility of S. litura larvae to the tested plant allelochemicals and insecticides. These results indicate that CYP9A40 expression in S. litura is related to consumption of xenobiotics and suggest that CYP9A40 is involved in detoxification of these compounds.

  14. Identification and Characterization of CYP9A40 from the Tobacco Cutworm Moth (Spodoptera litura, a Cytochrome P450 Gene Induced by Plant Allelochemicals and Insecticides

    Directory of Open Access Journals (Sweden)

    Rui-Long Wang

    2015-09-01

    Full Text Available Cytochrome P450 monooxygenases (P450s of insects play crucial roles in the metabolism of endogenous and dietary compounds. Tobacco cutworm moth (Spodoptera litura, an important agricultural pest, causes severe yield losses in many crops. In this study, we identified CYP9A40, a novel P450 gene of S. litura, and investigated its expression profile and potential role in detoxification of plant allelochemicals and insecticides. The cDNA contains an open reading frame encoding 529 amino acid residues. CYP9A40 transcripts were found to be accumulated during various development stages of S. litura and were highest in fifth and sixth instar larvae. CYP9A40 was mainly expressed in the midgut and fat body. Larval consumption of xenobiotics, namely plant allelochemicals (quercetin and cinnamic acid and insecticides (deltamethrin and methoxyfenozide induced accumulation of CYP9A40 transcripts in the midgut and fat body. Injection of dsCYP9A40 (silencing of CYP9A40 by RNA interference significantly increased the susceptibility of S. litura larvae to the tested plant allelochemicals and insecticides. These results indicate that CYP9A40 expression in S. litura is related to consumption of xenobiotics and suggest that CYP9A40 is involved in detoxification of these compounds.

  15. A school-based programme for tobacco and alcohol prevention in special education: effectiveness of the modified 'healthy school and drugs' intervention and moderation by school subtype

    NARCIS (Netherlands)

    Turhan, Abdullah; Onrust, Simone; ten Klooster, Peter M.; Pieterse, Marcel E.

    2017-01-01

    AIMS: To test the effectiveness of the Healthy School and Drugs (HSD) programme on tobacco and alcohol use in Dutch secondary special education (SE) schools, and whether this depends upon subtypes of SE schools and the level of implementation. DESIGN: In a quasi-experimental design with baseline and

  16. Modulation of Quorum Sensing in Acylhomoserine Lactone-Producing or -Degrading Tobacco Plants Leads to Alteration of Induced Systemic Resistance Elicited by the Rhizobacterium Serratia marcescens 90-166.

    Science.gov (United States)

    Ryu, Choong-Min; Choi, Hye Kyung; Lee, Chi-Ho; Murphy, John F; Lee, Jung-Kee; Kloepper, Joseph W

    2013-06-01

    Numerous root-associated bacteria (rhizobacteria) are known to elicit induced systemic resistance (ISR) in plants. Bacterial cell-density-dependent quorum sensing (QS) is thought to be important for ISR. Here, we investigated the role of QS in the ISR elicited by the rhizobacterium, Serratia marcescens strain 90-166, in tobacco. Since S. marcescens 90-166 produces at least three QS signals, QS-mediated ISR in strain 90-166 has been difficult to understand. Therefore, we investigated the ISR capacity of two transgenic tobacco (Nicotiana tabacum) plants that contained either bacterial acylhomoserine lactone-producing (AHL) or -degrading (AiiA) genes in conjunction with S. marcescens 90-166 to induce resistance against bacterial and viral pathogens. Root application of S. marcescens 90-166 increased ISR to the bacterial pathogens, Pectobacterium carotovorum subsp. carotovorum and Pseudomonas syringae pv. tabaci, in AHL plants and decreased ISR in AiiA plants. In contrast, ISR to Cucumber mosaic virus was reduced in AHL plants treated with S. marcescens 90-166 but enhanced in AiiA plants. Taken together, these data indicate that QS-dependent ISR is elicited by S. marcescens 90-166 in a pathogen-dependent manner. This study provides insight into QS-dependent ISR in tobacco elicited by S. marcescens 90-166.

  17. Modulation of Quorum Sensing in Acylhomoserine Lactone-Producing or -Degrading Tobacco Plants Leads to Alteration of Induced Systemic Resistance Elicited by the Rhizobacterium Serratia marcescens 90-166

    Directory of Open Access Journals (Sweden)

    Choong-Min Ryu

    2013-06-01

    Full Text Available Numerous root-associated bacteria (rhizobacteria are known to elicit induced systemic resistance (ISR in plants. Bacterial cell-density-dependent quorum sensing (QS is thought to be important for ISR. Here, we investigated the role of QS in the ISR elicited by the rhizobacterium, Serratia marcescens strain 90–166, in tobacco. Since S. marcescens 90–166 produces at least three QS signals, QS-mediated ISR in strain 90–166 has been difficult to understand. Therefore, we investigated the ISR capacity of two transgenic tobacco (Nicotiana tabacum plants that contained either bacterial acylhomoserine lactone-producing (AHL or -degrading (AiiA genes in conjunction with S. marcescens 90–166 to induce resistance against bacterial and viral pathogens. Root application of S. marcescens 90–166 increased ISR to the bacterial pathogens, Pectobacterium carotovorum subsp. carotovorum and Pseudomonas syringae pv. tabaci, in AHL plants and decreased ISR in AiiA plants. In contrast, ISR to Cucumber mosaic virus was reduced in AHL plants treated with S. marcescens 90–166 but enhanced in AiiA plants. Taken together, these data indicate that QS-dependent ISR is elicited by S. marcescens 90–166 in a pathogen-dependent manner. This study provides insight into QS-dependent ISR in tobacco elicited by S. marcescens 90–166.

  18. Smokeless Tobacco

    Science.gov (United States)

    ... toothpick-sized sticks. Some of these also contain sweeteners or flavoring and look a lot like candy. ... Still, tobacco companies often market these products as alternatives to smoking in places where smoking isn’t ...

  19. Tobacco Dilemma

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    Although a major fiscal revenue source, the tobacco industry is always under a watchful eye while many industries continue to suffer negative growth, even with economic recovery efforts in full swing, profits from Chinese tobacco companies allowed the industry to pay 513.11 billion yuan ($75.13 billion) in taxes in 2009, a year-on-year increase of 12.2 percent.

  20. Deliberate release of genetically modified plants into the environment in Slovenia

    Directory of Open Access Journals (Sweden)

    Zlata LUTHAR

    2015-11-01

    Full Text Available Deliberate release of genetically modified higher plants (GMHPs into the environment in Slovenia is regulated by the Law on the Management of Genetically Modified Organisms (ZRGSO Ur. l. RS 23/2005 and 21/2010, III chapter. For each deliberate release of GMPs into the environment a license issued by the Ministry of Environment and Spatial Planning (MESP must be acquired. The application or notification should contain a very accurate and complex description of the GMP, of the field where it will be released and of wider surroundings or environment. The application consists of Annex 2 with accessories: 1. Part A (technical data for the authorization of deliberate GMP release into the environment; 2. Part B (environmental risk assessment; 3. Application summary in Slovenian and English language for the release of GMP into environment, which is transmitted to Brussels by MESP; 4. Extract from the Land Cadastre of the field to which the GMP will be released. The release procedure runs (till here under the above mentioned Law, which has been in place for several years and which clearly defines that it is possible to release GMP in Slovenia. In the case of GM rice in 2011, the law applied till the site selection of the experiment. Here, the law was not sufficiently taken into account. It was prevailed by the regulation of Farmland and Forest Fund of the Republic of Slovenia and municipal decision, which was stronger than the national law and prevented the cultivation of GM rice in an area that is legally suitable for release of GMO into the environment. Rice is not grown in Slovenia and does not have wild ancestors or close relatives with whom it might mate. Nearest area of cultivation is in neighboring Italy, which is from potentially selected location in Slovenia more than 70 km away.

  1. A Member of the 14-3-3 Gene Family in Brachypodium distachyon, BdGF14d, Confers Salt Tolerance in Transgenic Tobacco Plants

    Science.gov (United States)

    He, Yuan; Zhang, Yang; Chen, Lihong; Wu, Chunlai; Luo, Qingchen; Zhang, Fan; Wei, Qiuhui; Li, Kexiu; Chang, Junli; Yang, Guangxiao; He, Guangyuan

    2017-01-01

    Plant 14-3-3 proteins are involved in diverse biological processes, but for the model monocotyledonous species, Brachypodium distachyon, their roles in abiotic stress tolerance are not well understood. In this study, a total of eight Bd14-3-3 genes were identified from B. distachyon and these were designated respectively as BdGF14a–BdGF14g. The qRT-PCR analyses of 3-month-old plants of B. distachyon showed that these genes were all expressed in the stems, leaves, and spikelets. By contrast, most of the plants had relatively lower transcriptional levels in their roots, except for the BdGF14g gene. The different expression profiles of the Bd14-3-3s under various stress treatments, and the diverse interaction patterns between Bd14-3-3s and BdAREB/ABFs, suggested that these gene products probably had a range of functions in the stress responses. The NaCl-induced Bd14-3-3 gene, BdGF14d, was selected for overexpression in tobacco. BdGF14d was found to be localized throughout the cell and it conferred enhanced tolerance to salt in the transgenic plants. Lowered contents of malondialdehyde, H2O2, and Na+, and lower relative electronic conductance (Rec%), yet greater activities of catalase and peroxidase, were observed in the overexpressing plants. Higher photosynthetic rate, transpiration rate, stomatal conductance, and water use efficiency were measured in the transgenic lines. Following abscisic acid (ABA) or NaCl treatment, stomatal aperture in leaves of the BdGF14d-overexpression plants was significantly lower than in leaves of the wild type (WT) controls. The stress-related marker genes involved in the ABA signaling pathway, the reactive oxygen species (ROS)-scavenging system, and the ion transporters were all up-regulated in the BdGF14d-overexpressing plants as compared with WT. Taken together, these results demonstrate that the Bd14-3-3 genes play important roles in abiotic stress tolerance. The ABA signaling pathway, the ROS-scavenging system, and ion

  2. Expression of the yeast trehalose-6-phosphate synthase gene in transgenic tobacco plants: pleiotropic phenotypes include drought tolerance.

    Science.gov (United States)

    Romero, C; Bellés, J M; Vayá, J L; Serrano, R; Culiáñez-Macià, F A

    1997-03-01

    The yeast trehalose-6-phosphate synthase gene (TPS1) was engineered under the control of the cauliflower mosaic virus regulatory sequences (CaMV35S) for expression in plants. Using Agrobacterium-mediated transfer, the gene was incorporated into the genomic DNA and constitutively expressed in Nicotiana tabacum L. plants. Trehalose was determined in the transformants, by anion-exchange chromatography coupled to pulsed amperometric detection. The non-reducing disaccharide accumulated up to 0.17 mg per g fresh weight in leaf extracts of transgenic plants. Trehaloseaccumulating plants exhibited multiple phenotypic alterations, including stunted growth, lancet-shaped leaves, reduced sucrose content and improved drought tolerance. These pleiotropic effects, and the fact that water loss from detached leaves was not significantly affected by trehalose accumulation, suggest that synthesis of this sugar, rather than leading to an osmoprotectant effect, had altered sugar metabolism and regulatory pathways affecting plant development and stress tolerance.

  3. Variance Analysis and Integrated Evaluation on Sensory Quality of Flue-cured Tobacco in Tobacco planting Areas of Liaoning Province%辽宁烟区烤烟感官质量的差异分析和综合评价

    Institute of Scientific and Technical Information of China (English)

    焦敬华; 陈晓波

    2011-01-01

    [Objective] The study aimed to analyze the difference on the sensory quality of flue-cured tobacco in different tobacco planting areas of Liaoning Province. [ Method] With 9 tobacco planting counties as the objects, 86 samples of upper, middle and lower tobacco leaves were collected to make the variance analysis and comprehensive evaluation on the sensory quality of flue-cured tobacco in tobacco-planting areas of Liaoning Province. [ Result] Among the various tobacco planting areas, the comprehensive sensory quality score of upper leaves in Beipi-ao County was the highest, while that in Kaiyuan and Fengcheng counties were relatively lower. The total sensory assessment and comprehensive evaluation score of middle leaves in Beipiao and Kuandian counties were better and the total sensory assessment and usability and comprehensive evaluation score of lower leaves in Xifeng, Changtu, Beipiao and Jianping counties were better. Smoking quality comprehensive evaluation scores of middle leaves showed a same trend among different tobacco planting areas in 2007, 2008 and 2009 and had a increasing trend annually. The average smoking quality scores of middle leaves in all around the province had a little increase with the year. [ Conclusion] On the total, in different tobacco planting areas of Liaoning Province, the sensory quality of the flue-cured tobacco produced in Beipiao County was better and that in Fengcheng County was poor. The smoking quality of middle leaves showed a trend of increasing year by year.%[目的]分析辽宁各烟区烤烟感官质量的差异,为当地烟草生产提供理论依据.[方法]以辽宁省9个植烟县为对象,采集了烟株上、中、下部叶烟叶样品共86个,对辽宁烟区烤烟感官质量进行差异分析和综合评价.[结果]辽宁各烟区烤烟上部叶感官质量综合评价得分以北票最高,开原和凤城的综合评价得分均较低;中部叶以北票和宽甸总体感官评定和综合评价得分表现较好

  4. Validation of PCR methods for quantitation of genetically modified plants in food.

    Science.gov (United States)

    Hübner, P; Waiblinger, H U; Pietsch, K; Brodmann, P

    2001-01-01

    For enforcement of the recently introduced labeling threshold for genetically modified organisms (GMOs) in food ingredients, quantitative detection methods such as quantitative competitive (QC-PCR) and real-time PCR are applied by official food control laboratories. The experiences of 3 European food control laboratories in validating such methods were compared to describe realistic performance characteristics of quantitative PCR detection methods. The limit of quantitation (LOQ) of GMO-specific, real-time PCR was experimentally determined to reach 30-50 target molecules, which is close to theoretical prediction. Starting PCR with 200 ng genomic plant DNA, the LOQ depends primarily on the genome size of the target plant and ranges from 0.02% for rice to 0.7% for wheat. The precision of quantitative PCR detection methods, expressed as relative standard deviation (RSD), varied from 10 to 30%. Using Bt176 corn containing test samples and applying Bt176 specific QC-PCR, mean values deviated from true values by -7to 18%, with an average of 2+/-10%. Ruggedness of real-time PCR detection methods was assessed in an interlaboratory study analyzing commercial, homogeneous food samples. Roundup Ready soybean DNA contents were determined in the range of 0.3 to 36%, relative to soybean DNA, with RSDs of about 25%. Taking the precision of quantitative PCR detection methods into account, suitable sample plans and sample sizes for GMO analysis are suggested. Because quantitative GMO detection methods measure GMO contents of samples in relation to reference material (calibrants), high priority must be given to international agreements and standardization on certified reference materials.

  5. Assays of dioxins and dioxin-like compounds in actually contaminated soils using transgenic tobacco plants carrying a recombinant mouse aryl hydrocarbon receptor-mediated β-glucuronidase reporter gene expression system.

    Science.gov (United States)

    Inui, Hideyuki; Gion, Keiko; Utani, Yasushi; Wakai, Taketo; Kodama, Susumu; Eun, Heesoo; Kim, Yun-Seok; Ohkawa, Hideo

    2012-01-01

    The transgenic tobacco plant XD4V-26 carrying the recombinant mouse aryl hydrocarbon receptor XD4V-mediated β-glucuronidase (GUS) reporter gene expression system was used for assay of dioxins and dioxin-like compounds consisting of polychlorinated dibenzeno-p-dioxins, polychlorinated dibenzofurans, and coplanar polychlorinated biphenyls (Co-PCBs) in actually contaminated soils. The transgenic tobacco plant XD4V-26 showed a significant dose-dependent induced GUS activity when cultured on MS medium containing PCB126 [toxic equivalency factor (TEF) = 0.1]. In contrast, PCB169 and PCB180, which have 0.03 of TEF and unassigned TEF values, respectively, did not significantly induce GUS activity under the same conditions as with PCB126. When the tobacco plants were cultivated for up to 5 weeks on actually contaminated soils with dioxins and dioxin-like compounds collected from the periphery of an incinerator used for disposal of residential and industrial wastes, GUS activity in the leaves was dose-dependently increased. The plants clearly detected 360 pg-TEQ g(-1) of dioxins and dioxin-like compounds in this assay. There was a positive correlation between GUS activity and TEQ value of dioxins and dioxin-like compounds in the plants. This assay does not require any extraction and purification processes for the actually contaminated soil samples.

  6. Transformation of tobacco and Arabidopsis plants with Stellaria media genes encoding novel hevein-like peptides increases their resistance to fungal pathogens.

    Science.gov (United States)

    R Shukurov, Rahim; D Voblikova, Vera; Nikonorova, Alexandra K; Komakhin, Roman A; V Komakhina, Vera; A Egorov, Tsezi; V Grishin, Eugene; V Babakov, Alexey

    2012-04-01

    Two novel antifungal hevein-like peptides, SmAMP1.1a and SmAMP2.2a, were previously isolated from seeds of Stellaria media. It has been established that these peptides accumulate in this weed as a result of proteolysis of two propeptides, pro-SmAMP1 and pro-SmAMP2. The primary structure of these propeptides is unique; in addition to having a signal peptide and negatively charged C-terminus, each of these structures consists of two hevein-like peptides of different length separated by a space rather than a single peptide. In this work, we demonstrated that the expression of the pro-SmAMP1 and pro-SmAMP2 genes was tissue-specific and increased substantially under exposure to fungal infection. To elucidate whether S. media has any advantages in defending against phytopathogens due to its unusual structure of pro-SmAMP1 and pro-SmAMP2, on the basis of the pro-SmAMP1 gene, we created three genetic constructs. Arabidopsis and tobacco plants were subsequently transformed with these constructs. Transgenic plants bearing the full-length pro-SmAMP1 gene exhibited the best resistance to the phytopathogens Bipolaris sorokiniana and Thielaviopsis basicola. The resistance of S. media plants to phytopathogenic fungi was likely due to the fungal-inducible expression of pro-SmAMP1 and pro-SmAMP2 genes, and due to the specific features of the primary structure of the corresponding propeptides. As a result of the processing of these propeptides, two different antimicrobial peptides were released simultaneously. Based on our results, we conclude that the genes for antimicrobial peptides from S. media may be promising genetic tools for the improvement of plant resistance to fungal diseases.

  7. Pronounced phenotypic changes in transgenic tobacco plants overexpressing sucrose synthase may reveal a novel sugar signaling pathway

    Directory of Open Access Journals (Sweden)

    Quynh Anh eNguyen

    2016-01-01

    Full Text Available Soluble sugars not only serve as nutrients, but also act as signals for plant growth and development, but how sugar signals are perceived and translated into physiological responses in plants remains unclear. We manipulated sugar levels in transgenic plants by overexpressing sucrose synthase (SuSy, which is a key enzyme believed to have reversible sucrose synthesis and sucrose degradation functions. The ectopically expressed SuSy protein exhibited sucrose-degrading activity, which may change the flux of sucrose demand from photosynthetic to non-photosynthetic cells, and trigger an unknown sucrose signaling pathway that lead to increased sucrose content in the transgenic plants. An experiment on the transition from heterotrophic to autotrophic growth demonstrated the existence of a novel sucrose signaling pathway, which stimulated photosynthesis, and enhanced photosynthetic synthesis of sucrose, which was the direct cause or the sucrose increase. In addition, a light/dark time treatment experiment, using different day length ranges for photosynthesis/respiration showed the carbohydrate pattern within a 24-hour day and consolidated the role of sucrose signaling pathway as a way to maintain sucrose demand, and indicated the relationships between increased sucrose and upregulation of genes controlling development of the shoot apical meristem (SAM. As a result, transgenic plants featured a higher biomass and a shorter time required to switch to reproduction compared to those of control plants, indicating altered phylotaxis and more rapid advancement of developmental stages in the transgenic plants.

  8. Dynamic area-partition evaluation of flue-cured tobacco planting climates in Yunnan based on GIS%基于GIS的云南烤烟种植气候动态分区评估

    Institute of Scientific and Technical Information of China (English)

    黄中艳

    2011-01-01

    基于气象、烟叶化学成分含量的大样本数据,通过完善烤烟种植气候适宜性定量评价指标体系,构建气象条件对烟叶品质影响的定量评判标准,在精细化模拟推算基本气象要素地理空间分布的基础上,应用GIS技术实现了烤烟种植气候的动态分区评估。以云南省两个典型气候年份为例,在GIS平台下将统计模型推算与空间插值技术方法有机结合,尝试突破传统"空间静态性"作物气候区划,实现"空间+时间动态性"作物种植气候精细化分区评估;直观形象地展现了区域地理"面"气候年际波动变化及其对烤烟种植适宜性和烟叶品质的影响。两种动态分区评估相结合,%The research mainly focuses on improving the effect of crop-climate regionalization and promoting perfect meteorological service.Based on a large sample of metorological data and chemical composition contents of tobacco leaf,the dynamic climate zoning evaluation of tobacco planting was accomplished by improving quantificational estimate criterion about climatic suitability in tobacco planting,constituting qualificationally-graded evaluation criteria about the effects of meteorological condition on tobacco ingredients,subtly simulating and calculating the geographical distribution of essential meteorological elements with the support of GIS software. With a view to speaking concisely and clearly,two typical years with different climates were taken as examples to demonstrate the two refined dynamic regionalization estimates considering mainly the temporal and spatial changes of meteorological conditions.On one hand,the paper carried out climate-suitability zoning evaluation,in which the proportion of each suitability-grade subarea for the two typical years was compared with the zoning results of the climatic mean in Yunnan.On the other hand,the zoning maps of the relative change grades for two chemical ingredients of tobacco between the two

  9. Chloroplast Redox Status Modulates Genome-Wide Plant Responses during the Non-host Interaction of Tobacco with the Hemibiotrophic Bacterium Xanthomonas campestris pv. vesicatoria

    Directory of Open Access Journals (Sweden)

    Juan J. Pierella Karlusich

    2017-07-01

    Full Text Available Non-host resistance is the most ample and durable form of plant resistance against pathogen infection. It includes induction of defense-associated genes, massive metabolic reprogramming, and in many instances, a form of localized cell death (LCD at the site of infection, purportedly designed to limit the spread of biotrophic and hemibiotrophic microorganisms. Reactive oxygen species (ROS have been proposed to act as signals for LCD orchestration. They are produced in various cellular compartments including chloroplasts, mitochondria and apoplast. We have previously reported that down-regulation of ROS build-up in chloroplasts by expression of a plastid-targeted flavodoxin (Fld suppressed LCD in tobacco leaves inoculated with the non-host bacterium Xanthomonas campestris pv. vesicatoria (Xcv, while other defensive responses were unaffected, suggesting that chloroplast ROS and/or redox status play a major role in the progress of LCD. To better understand these effects, we compare here the transcriptomic alterations caused by Xcv inoculation on leaves of Fld-expressing tobacco plants and their wild-type siblings. About 29% of leaf-expressed genes were affected by Xcv and/or Fld. Surprisingly, 5.8% of them (1,111 genes were regulated by Fld in the absence of infection, presumably representing pathways responsive to chloroplast ROS production and/or redox status during normal growth conditions. While the majority (∼75% of pathogen-responsive genes were not affected by Fld, many Xcv responses were exacerbated, attenuated, or regulated in opposite direction by expression of this protein. Particularly interesting was a group of 384 genes displaying Xcv responses that were already triggered by Fld in the absence of infection, suggesting that the transgenic plants had a larger and more diversified suite of constitutive defenses against the attacking microorganism compared to the wild type. Fld modulated many genes involved in pathogenesis, signal

  10. Type I J-domain NbMIP1 proteins are required for both Tobacco mosaic virus infection and plant innate immunity.

    Directory of Open Access Journals (Sweden)

    Yumei Du

    Full Text Available Tm-2² is a coiled coil-nucleotide binding-leucine rich repeat resistance protein that confers durable extreme resistance against Tomato mosaic virus (ToMV and Tobacco mosaic virus (TMV by recognizing the viral movement protein (MP. Here we report that the Nicotiana benthamiana J-domain MIP1 proteins (NbMIP1s associate with tobamovirus MP, Tm-2² and SGT1. Silencing of NbMIP1s reduced TMV movement and compromised Tm-2²-mediated resistance against TMV and ToMV. Furthermore, silencing of NbMIP1s reduced the steady-state protein levels of ToMV MP and Tm-2². Moreover, NbMIP1s are required for plant resistance induced by other R genes and the nonhost pathogen Pseudomonas syringae pv. tomato (Pst DC3000. In addition, we found that SGT1 associates with Tm-2² and is required for Tm-2²-mediated resistance against TMV. These results suggest that NbMIP1s function as co-chaperones during virus infection and plant immunity.

  11. Type I J-Domain NbMIP1 Proteins Are Required for Both Tobacco Mosaic Virus Infection and Plant Innate Immunity

    Science.gov (United States)

    Liu, Qi; Zhang, Haili; Wang, Yan; Hong, Yiguo; Xiao, Fangming; Zhang, Ling; Shen, Qianhua; Liu, Yule

    2013-01-01

    Tm-22 is a coiled coil-nucleotide binding-leucine rich repeat resistance protein that confers durable extreme resistance against Tomato mosaic virus (ToMV) and Tobacco mosaic virus (TMV) by recognizing the viral movement protein (MP). Here we report that the Nicotiana benthamiana J-domain MIP1 proteins (NbMIP1s) associate with tobamovirus MP, Tm-22 and SGT1. Silencing of NbMIP1s reduced TMV movement and compromised Tm-22-mediated resistance against TMV and ToMV. Furthermore, silencing of NbMIP1s reduced the steady-state protein levels of ToMV MP and Tm-22. Moreover, NbMIP1s are required for plant resistance induced by other R genes and the nonhost pathogen Pseudomonas syringae pv. tomato (Pst) DC3000. In addition, we found that SGT1 associates with Tm-22 and is required for Tm-22-mediated resistance against TMV. These results suggest that NbMIP1s function as co-chaperones during virus infection and plant immunity. PMID:24098120

  12. 百色烟区烤烟合理种植密度及施肥量研究%Studies on Planting Density and Fertilizer Application Rate of Flue-cured Tobacco in Baise

    Institute of Scientific and Technical Information of China (English)

    周文亮; 赖洪敏; 黄瑾; 林北森; 高华军

    2012-01-01

    [目的]为了掌握百色烟区合理的种植密度及施肥量,为烤烟生产提供科学指导和应用依据.[方法]研究了不同种植密度和施肥水平对烟株生长、烟叶产量和质量的影响.[结果]烤烟生育前期受种植密度和施肥量影响不大,但腰叶、顶叶成熟的时间随着施氮量的增大而变晚.不同种植密度和施肥量对烤烟农艺性状的影响较大,在种植密度相同时,随施肥量的增加,烟株株高、茎围、节距、有效叶片数和叶片长度呈递增趋势;减小种植密度有利于顶叶的开片生长及有效叶片数的增加.综合各经济性状及烟叶主要内在化学成分和评吸质量,A2B3处理(行株距120 cm×50 cm,施氮量112.5 kg/hm2,氮、磷、钾比例1∶1∶3)能够得到较好的经济效益和烟叶质量.[结论]在百色特定的气候、中等土壤肥力条件下,烤烟种植密度以行距120 cm,株距50 cm,纯氮用量112.5 kg/hm2(氮、磷、钾比例1∶1∶3)为宜.%[Objective] To explore the suitable planting density and fertilizer application rate of flue-cured tobacco and to provide the scientific basis and reference applications for guiding its production in Baise. [Method] The effects of different planting density and fertilizer application rate on the growth of tobacco plants, the yield and quality of tobacco leaves were studied. [ Result ] Planting density and fertilizer application rate had a little effects on the growth period of tobacco plants at early growth stage, but the mature stage of central and top leaves delayed with the increasing of fertilizer application rate. Different planting density and fertilizer application rate had significant effect on the agronomic characters, with the increasing fertilizer application rate, the plant height, stem diameter, node distance, effecttive number of leaves and length of leaf increased at the same planting density; when planting density was decreased, top leaves advantageous to growing and

  13. Influences of clinoptilolite and surfactant-modified clinoptilolite zeolite on nitrate leaching and plant growth.

    Science.gov (United States)

    Malekian, Raheleh; Abedi-Koupai, Jahangir; Eslamian, Sayed Saeid

    2011-01-30

    The increasing demands for environmental protection and sustainable food production require an increase in the use of natural and non-toxic materials for agriculture. In this study, the feasibility of using surfactant-modified zeolite (SMZ) in comparison with zeolite clinoptilolite (Cp) application to reduce nitrate leaching and enhance crop growth was investigated. The effects of size (millimeter and nanometer) and application rate (20 g kg(-1) and 60 g kg(-1)) of Cp and SMZ on nitrate leaching and crop response were also evaluated. Using soil lysimeters, it was determined that the maximum and mean nitrate concentration in the leachate of SMZ-amended soil were significantly (p<0.05) lower than those of Cp-amended soil. The amount of NO(3)-N leached from SMZ- and Cp-amended lysimeters at the higher application rate (60 g kg(-1)) was approximately 26% and 22% lower, respectively, than that from the control system. The mean grain yield, grain nitrogen content, stover dry matter, and N uptake were significantly greater in Cp-amended than SMZ-amended lysimeters. There was no significant effect due to the particle size of the two soil amendments. The results implicitly suggest that plants may have a better response if Cp is used as a fertilizer carrier rather than SMZ when applied at a rate of 60 g kg(-1).

  14. Influences of clinoptilolite and surfactant-modified clinoptilolite zeolite on nitrate leaching and plant growth

    Energy Technology Data Exchange (ETDEWEB)

    Malekian, Raheleh [Isfahan University of Technology, Isfahan, 84156-83111 (Iran, Islamic Republic of); Abedi-Koupai, Jahangir, E-mail: koupai@cc.iut.ac.ir [Isfahan University of Technology, Isfahan, 84156-83111 (Iran, Islamic Republic of); Eslamian, Sayed Saeid [Isfahan University of Technology, Isfahan, 84156-83111 (Iran, Islamic Republic of)

    2011-01-30

    The increasing demands for environmental protection and sustainable food production require an increase in the use of natural and non-toxic materials for agriculture. In this study, the feasibility of using surfactant-modified zeolite (SMZ) in comparison with zeolite clinoptilolite (Cp) application to reduce nitrate leaching and enhance crop growth was investigated. The effects of size (millimeter and nanometer) and application rate (20 g kg{sup -1} and 60 g kg{sup -1}) of Cp and SMZ on nitrate leaching and crop response were also evaluated. Using soil lysimeters, it was determined that the maximum and mean nitrate concentration in the leachate of SMZ-amended soil were significantly (p < 0.05) lower than those of Cp-amended soil. The amount of NO{sub 3}-N leached from SMZ- and Cp-amended lysimeters at the higher application rate (60 g kg{sup -1}) was approximately 26% and 22% lower, respectively, than that from the control system. The mean grain yield, grain nitrogen content, stover dry matter, and N uptake were significantly greater in Cp-amended than SMZ-amended lysimeters. There was no significant effect due to the particle size of the two soil amendments. The results implicitly suggest that plants may have a better response if Cp is used as a fertilizer carrier rather than SMZ when applied at a rate of 60 g kg{sup -1}.

  15. Modifying rainfall patterns in a Mediterranean shrubland: system design, plant responses, and experimental burning

    Science.gov (United States)

    Parra, Antonio; Ramírez, David A.; Resco, Víctor; Velasco, Ángel; Moreno, José M.

    2012-11-01

    Global warming is projected to increase the frequency and intensity of droughts in the Mediterranean region, as well as the occurrence of large fires. Understanding the interactions between drought, fire and plant responses is therefore important. In this study, we present an experiment in which rainfall patterns were modified to simulate various levels of drought in a Mediterranean shrubland of central Spain dominated by Cistus ladanifer, Erica arborea and Phillyrea angustifolia. A system composed of automatic rainout shelters with an irrigation facility was used. It was designed to be applied in vegetation 2 m tall, treat relatively large areas (36 m2), and be quickly dismantled to perform experimental burning and reassembled back again. Twenty plots were subjected to four rainfall treatments from early spring: natural rainfall, long-term average rainfall (2 months drought), moderate drought (25% reduction from long-term rainfall, 5 months drought) and severe drought (45% reduction, 7 months drought). The plots were burned in late summer, without interfering with rainfall manipulations. Results indicated that rainfall manipulations caused differences in soil moisture among treatments, leading to reduced water availability and growth of C. ladanifer and E. arborea in the drought treatments. However, P. angustifolia was not affected by the manipulations. Rainout shelters had a negligible impact on plot microenvironment. Experimental burns were of high fire intensity, without differences among treatments. Our system provides a tool to study the combined effects of drought and fire on vegetation, which is important to assess the threats posed by climate change in Mediterranean environments.

  16. Using genetically modified tomato crop plants with purple leaves for absolute weed/crop classification.

    Science.gov (United States)

    Lati, Ran N; Filin, Sagi; Aly, Radi; Lande, Tal; Levin, Ilan; Eizenberg, Hanan

    2014-07-01

    Weed/crop classification is considered the main problem in developing precise weed-management methodologies, because both crops and weeds share similar hues. Great effort has been invested in the development of classification models, most based on expensive sensors and complicated algorithms. However, satisfactory results are not consistently obtained due to imaging conditions in the field. We report on an innovative approach that combines advances in genetic engineering and robust image-processing methods to detect weeds and distinguish them from crop plants by manipulating the crop's leaf color. We demonstrate this on genetically modified tomato (germplasm AN-113) which expresses a purple leaf color. An autonomous weed/crop classification is performed using an invariant-hue transformation that is applied to images acquired by a standard consumer camera (visible wavelength) and handles variations in illumination intensities. The integration of these methodologies is simple and effective, and classification results were accurate and stable under a wide range of imaging conditions. Using this approach, we simplify the most complicated stage in image-based weed/crop classification models. © 2013 Society of Chemical Industry.

  17. [Smokeless tobacco].

    Science.gov (United States)

    Underner, M; Perriot, J

    2011-10-01

    Use of smokeless tobacco (ST) (chewing tobacco and snuff) can lead to a number of consequences detrimental to health. ST rapidly delivers high doses of nicotine, which can lead to dependence and is also a source of carcinogenic nitrosamines. Changes usually develop in the mouth area where the ST is most often placed. Non-malignant oral lesions include leuko-oedema, hyperkeratotic lesions of the oral mucosa and localised periodontal disease. Oral premalignant lesions are leukoplakia, erythroplakia, submucosal fibrosis and lichen planus. Betel chewing, with or without tobacco, may increase the incidence of oral cancer. There is conflicting evidence with regard to snuff users about the risk of oral and gastro-oesophageal cancer. ST use is a risk factor for pancreatic cancer and may increase the risk of fatal myocardial infarction and ischemic stroke. During pregnancy, ST is associated with an increase in pre-eclampsia, preterm delivery and stillbirth. Nicotine replacement therapy and bupropion reduce withdrawal symptoms and tobacco craving during ST cessation. However, they have not been shown to help long-term abstinence. Information concerning the potential hazards of ST products should be incorporated into educational programmes to discourage its use and to help users to quit. Smokeless tobacco is not recommended to help smoking cessation.

  18. Consumo precoz de tabaco y alcohol como factores modificadores del riesgo de uso de marihuana Early tobacco and alcohol consumption as modifying risk factors on marijuana use

    Directory of Open Access Journals (Sweden)

    Verónica Iglesias

    2007-08-01

    idade de início de consumo de tabaco e/ou álcool e a intensidade de uso de tabaco. Uso de maconha e idade de início do consumo foram as variáveis de desfecho. Para a análise dos dados se utilizou regressão de Poisson e regressão de Weibull. RESULTADOS: Os escolares apresentaram alta prevalência de consumo de tabaco, álcool maconha (77%, 79% e 23%, respectivamente. O consumo de tabaco na presença de consumo de álcool foi um fator de risco de uso de maconha (RP=10,4; IC 95%: 8,9;12,2. O início tardio de consumo de tabaco (HR=0,85; IC 95%: 0,84;0,86 e álcool (HR=0,90; IC 95%: 0,89;0,91 resultou ser um fator protetor do uso de maconha. A probabilidade de consumo de maconha foi maior naqueles que fumavam diariamente ou quase, em relação àqueles que fumavam somente aos fins de semana (RP=3,11; IC 95%: 2,96;3,26 vs. RP=1,70; IC 95%: 1,58;1,83. CONCLUSÕES: O risco de consumo de maconha se associou significativamente à idade de início de consumo de tabaco, à freqüência de consumo de tabaco e ao consumo simultâneo de álcool. As estratégias de prevenção deveriam proteger os escolares do consumo precoce de tabaco.OBJECTIVE: To assess the relationship between early tobacco and alcohol use and the risk of marijuana consumption among schoolchildren. METHODS: A cross-sectional study comprising data from the Fourth National Study on Drug Use in the Chilean School Population (2001. There were studied 54,001 schoolchildren aged between eight and 20 years. Predictors were self-reported tobacco and alcohol use (ever users, age at initiation of tobacco and/or alcohol use, and intensity of tobacco use. The study outcome was self-reported marijuana use (ever users versus never users and age at initiation of marijuana use. Poisson regression and Weibull regression were used for data analysis. RESULTS: Lifetime prevalence of tobacco use was high: 77%, alcohol 79%, and marijuana 23%. Tobacco consumption increased the likelihood of marijuana use (PR=10.4; 95% CI: 8

  19. Analysis of Correlation between Physiochemical Characteristics of Tobacco Planting Soil and Chemical Compositions of Tobacco Leaf in Youyang County of Chongqing Province%重庆酉阳植烟土壤理化性状与烟叶化学成分的相关性分析

    Institute of Scientific and Technical Information of China (English)

    王勇

    2014-01-01

    Using the methods of correlation analysis and canonical correlation analysis, the relationships between physiochemical characteristics of tobacco planting soil and chemical compositions of tobacco leaf in Youyang Cangling Base of China Tobacco Hunan Industrial Co., Ltd. were studied. The results showed that the pH value of tobacco planting soil in the base was generally appropriate, the available phosphor was relatively lacking, the contents of available potassium, available nitrogen and organic matter were at middle level, and the content of water-soluble chloride was relatively low. The correlation analysis results showed that the soil pH value was positively correlated with the content of reducing sugar in tobacco leaf, but negatively correlated with the content of total nitrogen in tobacco leaf;the content of available phosphorus in soil was negatively correlated with the nicotine content in tobacco leaf; the contents of available phosphorus and available nitrogen in soil were significantly positively correlated with the ratio of nitrogen to nicotine in tobacco leaf. Canonical correlation analysis results showed that the effects of soil physiochemical characteristics on chemical compositions of tobacco leaf was in a order of pH value> available phosphor> organic matters> available potassium> available nitrogen> water-soluble chlorine; the contents of available potassium and organic matters mainly influenced the sugar content in tobacco leaf, in which the content of available potassium in soil was beneficial to accumulation of reducing sugar in tobacco leaf, but disbenefit for accumulation of total sugar; the effects of the content of organic matters in soil on the sugar content in tobacco leaf were opposite to the effects of the content of available potassium on that.%采用相关分析、典型相关分析研究了湖南中烟重庆酉阳苍岭基地植烟土壤理化性状对烟叶化学成分的影响。烟区植烟土壤pH值总体适宜,速效磷相对

  20. A Review on Material Management for Planting Tobacco Based on RFID Technology%基于射频识别技术的烟用物资管理研究文献综述

    Institute of Scientific and Technical Information of China (English)

    刘骏; 李家俊; 何轶; 焦剑; 田必文; 金祥

    2014-01-01

    The material for planting tobacco (i.e. tobacco material) is one of the most important produce material in the to-bacco industry. Under the background of the"agriculture informatization", how to make use of RFID technology to efficient use of materials has become a problem to be solved. This article from the view application in material management for plan-ting tobacco as the breakthrough point, combing the related literature at home and abroad, to provide the reference for rese-archers of cigarette material management.%烤烟生产烟用物资(简称烟用物资)是烟草业重要的生产资料,在当前“农业生产信息化”背景下,如何利用射频识别技术规范、高效地使用烟用物资已经成为亟待解决的问题。本文从射频识别在烟用物资管理中的应用为切入点,梳理了国内外有关文献,为众多烟用物资管理研究者提供参考。

  1. FUELS IN TOBACCO PRODUCTION

    Directory of Open Access Journals (Sweden)

    M. Čavlek

    2008-09-01

    Full Text Available Energy production from biomass can reduce „greenhouse effect” and contribute to solving energy security especially in the agricultural households which rely on energy from fossil fuels. In Croatia fuel-cured tobacco is produced on about 5000 ha. Gross income for the whole production is about 180 000 000 kn/year. Flue-cured tobacco is a high energy consuming crop. There are two parts of energy consumption, for mechanization used for the field production (11% and, energy for bulk-curing (89%. In each case, presently used fuels of fossil origin need to be substituted by an alternative energy source of organic origin. Hereafter attention is paid to finding a more economic and ecologically acceptable fuel for curing tobacco. Curing flue-cured tobacco is done by heated air in curing burns. Various sources of heat have been used; wood, coal, oil and gas. In each case different burning facilities of different efficiency have been used. This has had an impact on curing costs and ecology. Recently, mostly used fuel has been natural gas. However, gas is getting expensive. Consequently, an alternative fuel for curing tobacco is sought for. According to literature, agricultural crops suitable for the latter purpose could be wheat, barley, maize, sorghum, sugar beet and some other annual and perennial plant species. Wooden pellets (by-products are suitable for combustion too. Ligno-cellulose fuels have been used for heating since long time. However, not sufficient research has been done from an applied point of view (Venturi and Venturi, 2003. Fuel combustion is getting more efficient with developing technological innovations. The curing barn manufacturers are offering technology for combusting wooden pellets (by-products for curing tobacco. The pellets are available on domestic market. The same technology can be used for combustion of maize grain. Within “Hrvatski duhani” research on suitability of using wooden pellets and maize grain and whole

  2. TB & Tobacco

    Directory of Open Access Journals (Sweden)

    Tjandra Y. Aditama

    2003-03-01

    Full Text Available One third of the world population is infected with tuberculosis, and over 8 millions people were developing each year. On the other hand tobacco is responsible for 3 millions death in the world. For Indonesia, our country has the third biggest TB cases in the world. Whereas Indonesia is ranked as having the fourth largest number of smokers in the world. A relationship between smoking and TB has been suspected for a long time, even though the epidemiological evidence has not been convincing so far, as well as the pathophysiology and the biomolecullar changes. At present time there are more and more epidemiological data to suggest relationship between TB and tobacco. Further research should be done to get more indepth relationship as well as avoiding the confounder factor. To be able to perform TB control as well as tobacco control successfully there should be emphasize on synergistic public health approaches. Tuberculosis –which Indonesia got 3rd rank in the world- as well as smoking problem –which Indonesia got 4th rank in the world- are two important public health problem for the country. If there are relationship between tobacco and tuberculosis, health problem faced by Indonesian even become bigger. Knowledge about tuberculosis as well as tobacco among Indonesian population is very essential to improve the public health situation. Tuberculosis control programme as well as smoking control programme are essential tools for the well being of Indonesian people. (Med J Indones 2003; 12: 48-52 Keywords: tobacco, tuberculosis, epidemiological data

  3. A systems toxicology approach for comparative assessment: Biological impact of an aerosol from a candidate modified-risk tobacco product and cigarette smoke on human organotypic bronchial epithelial cultures.

    Science.gov (United States)

    Iskandar, Anita R; Mathis, Carole; Schlage, Walter K; Frentzel, Stefan; Leroy, Patrice; Xiang, Yang; Sewer, Alain; Majeed, Shoaib; Ortega-Torres, Laura; Johne, Stephanie; Guedj, Emmanuel; Trivedi, Keyur; Kratzer, Gilles; Merg, Celine; Elamin, Ashraf; Martin, Florian; Ivanov, Nikolai V; Peitsch, Manuel C; Hoeng, Julia

    2017-03-01

    This study reports a comparative assessment of the biological impact of a heated tobacco aerosol from the tobacco heating system (THS) 2.2 and smoke from a combustible 3R4F cigarette. Human organotypic bronchial epithelial cultures were exposed to an aerosol from THS2.2 (a candidate modified-risk tobacco product) or 3R4F smoke at similar nicotine concentrations. A systems toxicology approach was applied to enable a comprehensive exposure impact assessment. Culture histology, cytotoxicity, secreted pro-inflammatory mediators, ciliary beating, and genome-wide mRNA/miRNA profiles were assessed at various time points post-exposure. Series of experimental repetitions were conducted to increase the robustness of the assessment. At similar nicotine concentrations, THS2.2 aerosol elicited lower cytotoxicity compared with 3R4F smoke. No morphological change was observed following exposure to THS2.2 aerosol, even at nicotine concentration three times that of 3R4F smoke. Lower levels of secreted mediators and fewer miRNA alterations were observed following exposure to THS2.2 aerosol than following 3R4F smoke. Based on the computational analysis of the gene expression changes, 3R4F (0.13 mg nicotine/L) elicited the highest biological impact (100%) in the context of Cell Fate, Cell Proliferation, Cell Stress, and Inflammatory Network Models at 4 h post-exposure. Whereas, the corresponding impact of THS2.2 (0.14 mg nicotine/L) was 7.6%. Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.

  4. 翻耕与改良剂施用对土壤植烟适应性的影响%Effects of Tillage and Amendment Application on the Suitability of Soils for Tobacco Cultivation in Tobacco-Planting Areas

    Institute of Scientific and Technical Information of China (English)

    蒋珍茂; 赵秀兰; 魏世强; 张永华; 罗畅; 徐畅; 陈益银

    2015-01-01

    Effects of soil turnover depth and application of amendments on the physico‐chemical properties of tobacco‐planting soils were studied through a two‐year field experiment , conducted in Pengshui , Chongqing from April ,2012 to September ,2013 ,and the suitability of soils for tobacco cultivation after consolidation was evaluated .The results indicated that turnover of soil ameliorated its physical properties :soil bulk density was reduced by 1.38% -15.17% and soil porosity increased by 1.68% -18.68% ;how‐ever ,it decreased the contents of soil organic matter and reduced the availabilities of nitrogen ,phosphor‐us ,iron ,manganese and copper .These negative effects increased with the increase in soil plough depth . Lime ,biochar and compost showed different roles in ameliorating soil physico‐chemical properties :liming increased soil pH but reduced the availabilities of most nutrients ,biochar promoted the accumulation of soil organic matter and enhanced the role of lime in acid soil improvement ,while compost had positive roles both in improvement of soil physical properties and balanced supply of nutrients .Combined applica‐tion of the soil amendments tested showed a synergistic effect for improving soil physico‐chemical proper‐ties ,resulting in better suitability for tobacco cultivation .Topsoil application of amendments showed bet‐ter effects than application to the w hole soil profile .%在重庆市典型烟区进行了2a时间的田间试验,研究了土壤翻耕深度、土壤改良剂类型及其施用方式等土地整理因子对植烟土壤质量的影响,评估了整理后土壤的植烟适宜性.结果表明,土壤翻耕可改善土壤物理性质,使得土壤容质量下降1.38%~15.17%,孔隙度增加1.68%~18.68%,但同时也降低了土壤的有机质、有效氮、磷、铁、锰、铜等含量,且随土地整理深度增加,降低幅度增大.石灰、生物质炭、堆肥等土壤改良剂在改

  5. The role of chloroplast NAD(P)H dehydrogenase in protection of tobacco plant against heat stress

    Institute of Scientific and Technical Information of China (English)

    WANG Peng; YE Jiyu; SHEN Yungang; MI Hualing

    2006-01-01

    After incubation at 42℃ for more than 48 h, brown damages occurred on the stems of tobacco (Nicotiana tabacum L.) ndhC-ndhK-ndhJ deletion mutant (ΔndhCKJ), followed by wilt of the leaves, while less the phenotype was found in its wild type (WT). Analysis of the kinetics of post-illumination rise in chlorophyll fluorescence indicated that the PSI cyclic electron flow and the chlororespiration mediated by NAD(P)H dehydrogenase (NDH) was significantly enhanced in WT under the high temperature. After leaf disks were treated with methyl viologen (MV), photosynthetic apparatus of ΔndhCKJ exhibited more severe photo-oxidative damage, even bleaching of chlorophyll. Analysis of P700 oxidation and reduction showed that the NDH mediated cyclic electron flow probably functioned as an electron competitor with Mehler reaction, to reduce the accumulation of reactive oxygen species (ROS). When leaf disks were heat stressed at 42℃ for 6 h, the photochemical activity declined more markedly in ΔndhCKJ than in WT, accompanied with more evident decrease in the amount of soluble Rubisco activase. In addition, the slow phase of millisecond-delayed light emission (ms-DLE) of chlorophyll fluorescence indicated that NDH was involved in the building-up of transthylakoid proton gradient (ΔpH), while the consumption of ΔpH was highly inhibited in ΔndhCKJ after heat stress. Based on the results, we supposed that the cyclic electron flow mediated by NDH could be stimulated under the heat stressed conditions, to divert excess electrons via chlororespiration pathway, and sustain CO2 assimilation by providing extra ΔpH, thus reducing the photooxidative damage.

  6. Experimental tests on a new harvesting system for Burley tobacco

    Directory of Open Access Journals (Sweden)

    Salvatore Faugno

    2013-09-01

    Full Text Available The globalization of the tobacco production has led to a drop in competitiveness of the Italian tobacco on the world market. Burley is the main variety of tobacco cropped in Campania region of Southern Italy. Its leaves have to be sewn, in the curing phase. Aim of this work is to show the results of the implementation of a new harvest machine prototype. Basically, the machine used for Bright tobacco, totally mechanical harvested, which doesn’t need to be sewn because it requires an indirect-fire treatment into the curing furnaces. The machine was modified in order to mechanize harvesting of Burley tobacco, and tested on four cultivars of Burley tobacco under three different planting layouts. The Burley tobacco leaves can be harvested mechanically by pulling individual leaves off the stalk; leaves are then sorted and tied in bundles prior to sewing. A mechanical burley tobacco harvesting system was evaluated. This machine consists in realizing a leaves orientation system based on the different weight between the leaf blade and the stalk enhanced by an air flow. The measurements taken were harvest timing, work capacity, and quality standards of the work carried out. The results, in terms of user time, range from 6.67 h/ha to 7.80 h/ha while in terms of operational efficiency are between 88% and 89%. The average user capacity recorded for the four cultivars is equal to 0.14 ha/h, a value far from the one recorded for the same harvesting machine used for Bright tobacco (0.25 ha/h. The harvest timing capacity, range from 0.51 t/h to 0.99 t/h. The work productivity goes from 0.17 t to 0.33 t per hour of human unit respectively. The average number of detached leaves, depending on the cultivar, has been between 523 and 744. Concerning the leaf orientation, a general percentage of 73% was achieved.

  7. Comparative Analysis on Physicochemical Characteristic of Tobacco Planted in Rice Paddy and Mountain Area%稻田烟叶与山地烟叶理化特性比较分析

    Institute of Scientific and Technical Information of China (English)

    李平; 杨虹琦; 赵松义; 杨红武; 管维; 曹晓玉

    2012-01-01

    对湖南省稻田烟与山地烟理化特性进行了分析,结果表明稻田烟叶与山地烟叶理化指标大多存在显著或极显著差异,山地烟叶含梗率高于稻田烟叶,去梗干物质、单位叶面积重比稻田烟叶小;稻田烟叶的物理特性比山地烟叶相对稳定;稻田烟叶总糖、总氮、钾、氯、两糖差显著高于山地烟叶,而烟碱、钾氯比低于山地烟叶;稻田烟叶的β-胡萝卜素和芸香苷含量显著高于山地烟叶.叶黄素含量显著低于山地烟叶,而绿原酸含量差异不显著.%The study mainly analyzed the physicochemical characteristics of tobacco planted in rice paddy and mountain area in Hunan. The results showed the physicochemical indicators of tobacco in the soil of rice paddy and mountain reached the significant differences and very significant differences. Among the results, the indicator of containing terrier of the tobacco planted in mountain area was higher than that of the rice paddies, but the indicators of dry matter removed stems and the leaf weight of elemental area were lower than those of the rice paddies; The physicochemical characteristics of tobacco planted in rice paddy were more steady than that of the mountain area, the total sugar, total nitrogen, potassium , chloride, the content of total sugar and reducing sugar of tobacco in the soil of the rice paddy were obviously higher than those of the mountain area, but nicotine, potassium chlorine ratio of the tobacco in the soil of the rice paddy were lower; There was a significant difference in the indicators, including the β - carotene, lutein , rutin content, but the difference was not significant in the content of chlorogenic acid, the content of the β - carotene and rutin in the tobacco of the soil of rice paddy was higher,while, the content of lutein was lower.

  8. Antibacterial and antifungal activity of a snakin-defensin hybrid protein expressed in tobacco and potato plants

    Science.gov (United States)

    In this study, for the first time, functionally active recombinant cysteine-rich plant proteins snakin-1 (SN1) and defensin (PTH1) were successfully expressed and purified using a prokaryotic (bacterial) expression system. The overall level of antimicrobial activities of SN1 and PTH1 produced in Esc...

  9. Analysis of Relationship between Different Types of Tobacco Planting Soil and Tobacco Leaf Yield, Quality in Malong County%马龙县不同类型植烟土壤与烟叶产量·品质的关系

    Institute of Scientific and Technical Information of China (English)

    杨丽平; 许树德; 王小兵; 王斌

    2012-01-01

    [Objective]The relationship between" the different types of tobacco planting soil in Malong County and tobacco leaf yield, quality was analyzed. [Method]357 soil samples were collected in eight villages and towns, Malong County, Qujing City, Yunnan Province. In addition, grading and leaf sampling method were used to collect the corresponding soil sample. A set of tobacco leaf samples was collected and the soil and tobacco leaf chemical composition was analyzed. [Result] In 30 -90 d after transplanting, agronomic traits of tobacco plant in rice soil were better than those in red soil and purple soil. In comparison with purple soil and red soil, total nitrogen and nicotine content in tobacco leaves in rice soil were higher, and total sugar and reducing sugar content were low obviously. Based on the sugar/alkali and N/alkali ratio of flue-cured tobacco leaves, the quality of tobacco leaves in purple soil was closer to that of the high quality tobacco leaves. The order was purple soil > red soil > rice soil. In 3 types of soil, lutein, β-carotene, coffee tannin polyphenols in tobacco in purple soil were the highest, those in red soil were lower , and those in paddy soil were the lowest. The order of other carotenoids, flavonoids, rutin content of polyphenols was red soil > purple soil > rice soil, and the order of its smoking results was purple soil > red soil > rice soil. [Conclusion] In order to strive for high yield and good quality in flue-cured tobacco production,fertilization amount should be determined based on regional soil types.%[目的]研究云南曲靖市马龙县不同类型植烟土壤与烟叶产量、品质之间的关系.[方法]在云南省曲靖市马龙县8个乡镇避雨采集土壤样品357个.同时,在对应采集土样的田块,采用定等级、定叶位取样法取烟叶样品1套,分析土壤、烟叶中的化学成分.[结果]在烟株移栽后30~90 d,水稻土烟株的农艺性状指标明显优于红壤土、紫色土.与紫色土

  10. Tobacco Dilemma

    Institute of Scientific and Technical Information of China (English)

    LAN XINZHEN

    2010-01-01

    @@ While many industries continue to suffer negative growth, even with economic recovery efforts in full swing, profits from Chinese tobacco companies allowed the industry to pay 513.11 billion yuan ($75.13 billion) in taxes in 2009, a year-on-year increase of 12.2 percent.

  11. Removal of polycyclic aromatic hydrocarbons from aqueous solution by raw and modified plant residue materials as biosorbents.

    Science.gov (United States)

    Xi, Zemin; Chen, Baoliang

    2014-04-01

    Removal of polycyclic aromatic hydrocarbons (PAHs), e.g., naphthalene, acenaphthene, phenanthrene and pyrene, from aqueous solution by raw and modified plant residues was investigated to develop low cost biosorbents for organic pollutant abatement. Bamboo wood, pine wood, pine needles and pine bark were selected as plant residues, and acid hydrolysis was used as an easily modification method. The raw and modified biosorbents were characterized by elemental analysis, Fourier transform infrared spectroscopy and scanning electron microscopy. The sorption isotherms of PAHs to raw biosorbents were apparently linear, and were dominated by a partitioning process. In comparison, the isotherms of the hydrolyzed biosorbents displayed nonlinearity, which was controlled by partitioning and the specific interaction mechanism. The sorption kinetic curves of PAHs to the raw and modified plant residues fit well with the pseudo second-order kinetics model. The sorption rates were faster for the raw biosorbents than the corresponding hydrolyzed biosorbents, which was attributed to the latter having more condensed domains (i.e., exposed aromatic core). By the consumption of the amorphous cellulose component under acid hydrolysis, the sorption capability of the hydrolyzed biosorbents was notably enhanced, i.e., 6-18 fold for phenanthrene, 6-8 fold for naphthalene and pyrene and 5-8 fold for acenaphthene. The sorption coefficients (Kd) were negatively correlated with the polarity index [(O+N)/C], and positively correlated with the aromaticity of the biosorbents. For a given biosorbent, a positive linear correlation between logKoc and logKow for different PAHs was observed. Interestingly, the linear plots of logKoc-logKow were parallel for different biosorbents. These observations suggest that the raw and modified plant residues have great potential as biosorbents to remove PAHs from wastewater. Copyright © 2014 The Research Centre for Eco-Environmental Sciences, Chinese Academy of

  12. In Vitro Antibacterial and Antibiotic Resistance Modifying Effect of Bioactive Plant Extracts on Methicillin-Resistant Staphylococcus epidermidis

    OpenAIRE

    Romana Chovanová; Mária Mikulášová; Štefánia Vaverková

    2013-01-01

    The crude extracts of plants from Asteraceae and Lamiaceae family and essential oils from Salvia officinalis and Salvia sclarea were studied for their antibacterial as well as antibiotic resistance modifying activity. Using disc diffusion and broth microdilution assays we determined higher antibacterial effect of three Salvia spp. and by evaluating the leakage of 260 nm absorbing material we detected effect of extracts and, namely, of essential oils on the disruption of cytoplasmic membrane. ...

  13. Photorespiration plays an important role in the regulation of photosynthetic electron flow under fluctuating light in tobacco plants grown under full sunlight

    Directory of Open Access Journals (Sweden)

    Wei eHuang

    2015-08-01

    Full Text Available Plants usually experience dynamic fluctuations of light intensities under natural conditions. However, the responses of mesophyll conductance, CO2 assimilation, and photorespiration to light fluctuation are not well understood. To address this question, we measured photosynthetic parameters of gas exchange and chlorophyll fluorescence in tobacco leaves at 2-min intervals while irradiance levels alternated between 100 and 1200 μmol photons m-2 s-1. Compared with leaves exposed to a constant light of 1200 μmol photons m-2 s-1, both stomatal and mesophyll conductances were significantly restricted in leaves treated with fluctuating light condition. Meanwhile, CO2 assimilation rate and electron flow devoted to RuBP carboxylation at 1200 μmol photons m-2 s-1 under fluctuating light were limited by the low chloroplast CO2 concentration. Analysis based on the C3 photosynthesis model indicated that, at 1200 μmol photons m-2 s-1 under fluctuating light, the CO2 assimilation rate was limited by RuBP carboxylation. Electron flow devoted to RuBP oxygenation at 1200 μmol photons m-2 s-1 under fluctuating light remained at nearly the maximum level throughout the experimental period. We conclude that fluctuating light restricts CO2 assimilation by decreasing both stomatal and mesophyll conductances. Under such conditions, photorespiration plays an important role in the regulation of photosynthetic electron flow.

  14. Functional analysis of the stress-inducible soybean calmodulin isoform-4 (GmCaM-4) promoter in transgenic tobacco plants.

    Science.gov (United States)

    Park, Hyeong Cheol; Kim, Man Lyang; Kang, Yun Hwan; Jeong, Jae Cheol; Cheong, Mi Sun; Choi, Wonkyun; Lee, Sang Yeol; Cho, Moo Je; Kim, Min Chul; Chung, Woo Sik; Yun, Dae-Jin

    2009-04-30

    The transcription of soybean (Glycine max) calmodulin isoform-4 (GmCaM-4) is dramatically induced within 0.5 h of exposure to pathogen or NaCl. Core cis-acting elements that regulate the expression of the GmCaM-4 gene in response to pathogen and salt stress were previously identified, between -1,207 and -1,128 bp, and between -858 and -728 bp, in the GmCaM-4 promoter. Here, we characterized the properties of the DNA-binding complexes that form at the two core cis-acting elements of the GmCaM-4 promoter in pathogen-treated nuclear extracts. We generated GUS reporter constructs harboring various deletions of approximately 1.3-kb GmCaM-4 promoter, and analyzed GUS expression in tobacco plants transformed with these constructs. The GUS expression analysis suggested that the two previously identified core regions are involved in inducing GmCaM-4 expression in the heterologous system. Finally, a transient expression assay of Arabidopsis protoplasts showed that the GmCaM-4 promoter produced greater levels of GUS activity than did the CaMV35S promoter after pathogen or NaCl treatments, suggesting that the GmCaM-4 promoter may be useful in the production of conditional gene expression systems.

  15. Photorespiration plays an important role in the regulation of photosynthetic electron flow under fluctuating light in tobacco plants grown under full sunlight.

    Science.gov (United States)

    Huang, Wei; Hu, Hong; Zhang, Shi-Bao

    2015-01-01

    Plants usually experience dynamic fluctuations of light intensities under natural conditions. However, the responses of mesophyll conductance, CO2 assimilation, and photorespiration to light fluctuation are not well understood. To address this question, we measured photosynthetic parameters of gas exchange and chlorophyll fluorescence in tobacco leaves at 2-min intervals while irradiance levels alternated between 100 and 1200 μmol photons m(-2) s(-1). Compared with leaves exposed to a constant light of 1200 μmol photons m(-2) s(-1), both stomatal and mesophyll conductances were significantly restricted in leaves treated with fluctuating light condition. Meanwhile, CO2 assimilation rate and electron flow devoted to RuBP carboxylation at 1200 μmol photons m(-2) s(-1) under fluctuating light were limited by the low chloroplast CO2 concentration. Analysis based on the C3 photosynthesis model indicated that, at 1200 μmol photons m(-2) s(-1) under fluctuating light, the CO2 assimilation rate was limited by RuBP carboxylation. Electron flow devoted to RuBP oxygenation at 1200 μmol photons m(-2) s(-1) under fluctuating light remained at nearly the maximum level throughout the experimental period. We conclude that fluctuating light restricts CO2 assimilation by decreasing both stomatal and mesophyll conductances. Under such conditions, photorespiration plays an important role in the regulation of photosynthetic electron flow.

  16. DEVELOPMENT OF A MULTI-TIERED INSECT RESISTANCE MANAGEMENT PROGRAM FOR GENETICALLY MODIFIED CORN HYBRIDS EXPRESSING THE PLANT INCORPORATED PROTECTANT, BACILLUS THURINGIENSIS

    Science.gov (United States)

    A significant increase in genetically modified corn planting driven by biofuel demand is expected for the 2007 growing season with future planted acreages approaching 80% of total corn plantings anticipated by 2009. As demand increases, incidence of farmer non-compliance with ma...

  17. Metabolic Engineering of Plant-derived (E)-β-farnesene Synthase Genes for a Novel Type of Aphid-resistant Genetically Modified Crop Plants

    Institute of Scientific and Technical Information of China (English)

    Xiu-Dao Yu; John Pickett; You-Zhi Ma; Toby Bruce; Johnathan Napier; Huw D.Jones; Lan-Qin Xia

    2012-01-01

    Aphids are major agricultural pests that cause significant yield losses of crop plants each year.Excessive dependence on insecticides for long-term aphid control is undesirable because of the development of insecticide resistance,the potential negative effects on non-target organisms and environmental pollution.Transgenic crops engineered for resistance to aphids via a non-toxic mode of action could be an efficient alternative strategy.(E)-β-Farnesene (EβF) synthases catalyze the formation of EβF,which for many pest aphids is the main component of the alarm pheromone involved in the chemical communication within these species.EβF can also be synthesized by certain plants but is then normally contaminated with inhibitory compounds.Engineering of crop plants capable of synthesizing and emitting EβF could cause repulsion of aphids and also the attraction of natural enemies that use EβF as a foraging cue,thus minimizing aphid infestation.In this review,the effects of aphids on host plants,plants' defenses against aphid herbivory and the recruitment of natural enemies for aphid control in an agricultural setting are briefly introduced.Furthermore,the plant-derived EβF synthase genes cloned to date along with their potential roles in generating novel aphid resistance via genetically modified approaches are discussed.

  18. A Study on the Properties and Chloride Resistance of Modified Sulfur Concrete for Nuclear Power Plant and Marine Structures

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Soon Myun; Chang, Hyun Young; Park, Heung Bae [KEPCO EnC, Seongnam (Korea, Republic of)

    2015-05-15

    The mechanical, physical and chemical properties of concrete with modified sulfur have been compared and assessed against ordinary concrete. As its excellent chloride resistance and extended service life have been verified, the technology to apply modified sulfur to the construction of nuclear power plant and marine structures has been developed and secured. Recently, modified sulfur concrete has been applied for road pavement and repair works in more than 20 sites including highway and airport in Korea. Also, in the U.S., Federal Highway Administration and Virginia Department of Transportation are implementing tests to apply modified sulfur to bridge road pavement, and the modified sulfur concrete has been recognized for its good performance. Based on these cases, this study carried out tests on physical, mechanical and chemical properties of concrete after adding modified sulfur by building concrete specimens based on the concrete mix design employed to construct the Shin-Kori Units 3 and 4 containment building. Multiple tests were performed particularly for chemical resistance, a factor directly related to concrete service life. As a result, it has been verified that concrete with 5% modified sulfur content relative to cement weight has equal mechanical properties (compressive strength, tensile strength, etc.) and much better workability (slump change) and chemical resistance (resistance to chloride ion penetration, concrete carbonation) compared with ordinary concrete. Based on this, it has been concluded that an addition of modified sulfur can double the service life of concrete. In general, studies demonstrate that a significant amount of slag should be mixed into concrete to raise chemical resistance (but with decreasing mechanical properties). Considering this, this study is unparalleled.

  19. Assessing and monitoring impacts of genetically modified plants on agro-ecosystems: the approach of AMIGA project

    Directory of Open Access Journals (Sweden)

    S. Arpaia

    2014-06-01

    Full Text Available The environmental impacts of genetically modified crops is still a controversial issue in Europe. The overall risk assessment framework has recently been reinforced by the European Food Safety Authority(EFSA and its implementation requires harmonized and efficient methodologies. The EU-funded research project AMIGA − Assessing and monitoring Impacts of Genetically modified plants on Agro-ecosystems − aims to address this issue, by providing a framework that establishes protection goals and baselines for European agro-ecosystems, improves knowledge on the potential long term environmental effects of genetically modified (GM plants, tests the efficacy of the EFSA Guidance Document for the Environmental Risk Assessment, explores new strategies for post market monitoring, and provides a systematic analysis of economic aspects of Genetically Modified crops cultivation in the EU. Research focuses on ecological studies in different EU regions, the sustainability of GM crops is estimated by analysing the functional components of the agro-ecosystems and specific experimental protocols are being developed for this scope.

  20. Interaction of the tobacco lectin with histone proteins

    OpenAIRE

    Schouppe, Dieter; Ghesquière, Bart; Menschaert, Gerben; De Vos, Winnok; Bourque, Stéphane; Trooskens, Geert; Proost, Paul; Gevaert, Kris; Van Damme, Els

    2011-01-01

    The tobacco (Nicotiana tabacum) agglutinin or Nictaba is a member of a novel class of plant lectins residing in the nucleus and the cytoplasm of tobacco cells. Since tobacco lectin expression is only observed after the plant has been subjected to stress situations such as jasmonate treatment or insect attack, Nictaba is believed to act as a signaling protein involved in the stress physiology of the plant. In this paper, a nuclear proteomics approach was followed to identify the binding partne...

  1. Study on Agricultural Weather Forecast for Tobacco Planting in South Anhui Province%皖南烟叶生产农用天气预报服务研究

    Institute of Scientific and Technical Information of China (English)

    李龙; 凤宝文; 张民蓓; 孙秀邦

    2012-01-01

    [Objective] To study the agricultural weather forecast for tobacco planting in South Anhui Province. [Method] According to the regulations of farming activities in tobacco field and the relationships between tobacco production and climatic conditions, Xuancheng agrome-teorological experiment station analyzed the actual needs and contents of agricultural weather forecast in lobarco planting. [Result] Seven main forecast objects were given, including severe snow forecast, suitable transplanting period forecast, the weather forecast for pests and diseases controlling, chilling and drainage forecast, irrigation forecast, and the forecast for water logging controlling and harvesting. An agricultural weather forecast model was constructed based on the tobacco growth-weather relationships, and a meteorological services program was formulated to explore the tobacco production layout, track its growth dynamics, bring forward meteorological services timely and propose practical measures to guarantee excellent tobacco quality. [Conclusion] The study provides meteorological service and technology support for producing high-quality tobacco in South Anhui Province.%[目的]研究皖南烟叶生产农用天气预报服务.[方法]宣城市农业气象试验站根据皖南烟叶田间管理规程确定的农事活动,通过皖南烟叶生产与天气关系研究,分析农用天气预报的内容及实际需求.[结果]总结归纳出7种主要预报对象,分别是大棚防雪灾预报、适宜移栽期预报、病虫害防治预报、清沟排渍预报、灌溉预报、排涝及适宜收获期预报;提出用皖南烟叶生长天气关系模型制作农用天气预报预报,并编制了皖南烟叶田间生产周年气象服务方案,通过掌握烟叶生产布局和追踪烟叶生长动态,及时开展服务,提出保障烟叶优质生产可行的农事措施.[结论]该研究为皖南烟叶优质适产提供了农业气象科技支撑.

  2. DNA-free genome editing in plants with preassembled CRISPR-Cas9 ribonucleoproteins.

    Science.gov (United States)

    Woo, Je Wook; Kim, Jungeun; Kwon, Soon Il; Corvalán, Claudia; Cho, Seung Woo; Kim, Hyeran; Kim, Sang-Gyu; Kim, Sang-Tae; Choe, Sunghwa; Kim, Jin-Soo

    2015-11-01

    Editing plant genomes without introducing foreign DNA into cells may alleviate regulatory concerns related to genetically modified plants. We transfected preassembled complexes of purified Cas9 protein and guide RNA into plant protoplasts of Arabidopsis thaliana, tobacco, lettuce and rice and achieved targeted mutagenesis in regenerated plants at frequencies of up to 46%. The targeted sites contained germline-transmissible small insertions or deletions that are indistinguishable from naturally occurring genetic variation.

  3. Grazing-induced effects on soil properties modify plant competitive interactions in semi-natural mountain grasslands.

    Science.gov (United States)

    Medina-Roldán, Eduardo; Paz-Ferreiro, Jorge; Bardgett, Richard D

    2012-09-01

    Plant-soil feedbacks are widely recognized as playing a significant role in structuring plant communities through their effects on plant-plant interactions. However, the question of whether plant-soil feedbacks can be indirectly driven by other ecological agents, such as large herbivores, which are known to strongly modify plant community structure and soil properties, remains poorly explored. We tested in a glasshouse experiment how changes in soil properties resulting from long-term sheep grazing affect competitive interactions (intra- and inter-specific) of two graminoid species: Nardus stricta, which is typically abundant under high sheep grazing pressure in British mountain grasslands; and Eriophorum vaginatum, whose abundance is typically diminished under grazing. Both species were grown in monocultures and mixtures at different densities in soils taken from adjacent grazed and ungrazed mountain grassland in the Yorkshire Dales, northern England. Nardus stricta performed better (shoot and root biomass) when grown in grazing-conditioned soil, independent of whether or not it grew under inter-specific competition. Eriophorum vaginatum also grew better when planted in soil from the grazed site, but this occurred only when it did not experience inter-specific competition with N. stricta. This indicates that plant-soil feedback for E. vaginatum is dependent on the presence of an inter-specific competitor. A yield density model showed that indirect effects of grazing increased the intensity of intra-specific competition in both species in comparison with ungrazed-conditioned soil. However, indirect effects of grazing on the intensity of inter-specific competition were species-specific favouring N. stricta. We explain these asymmetric grazing-induced effects on competition on the basis of traits of the superior competitor and grazing effects on soil nutrients. Finally, we discuss the relevance of our findings for plant community dynamics in grazed, semi

  4. EFSA guidance on the submission of applications for authorisation of genetically modified plants under Regulation (EC No 1829/2003

    Directory of Open Access Journals (Sweden)

    European Food Safety Authority

    2013-12-01

    Full Text Available This document provides guidance to applicants for submitting an application for authorisation of genetically modified (GM plants for food and feed uses, import and processing, and/or cultivation in the European Union under Regulation (EC No 1829/2003. The EFSA submission guidance describes the community procedures in the European Union for handling GM plant applications, and provides instructions to applicants on how to prepare and present data in an application. It is supplemented with seven appendices providing templates of data presentation to be followed by applicants, including a completeness checklist. The earlier versions are now updated to account for requirements outlined in Implementing Regulation (EU No 503/2013. Instructions for submission described in this EFSA guidance are applicable to all GM plant applications submitted under Articles 5, 11, 17 and 23 of Regulation (EC No 1829/2003.

  5. In Vitro Antibacterial and Antibiotic Resistance Modifying Effect of Bioactive Plant Extracts on Methicillin-Resistant Staphylococcus epidermidis

    Directory of Open Access Journals (Sweden)

    Romana Chovanová

    2013-01-01

    Full Text Available The crude extracts of plants from Asteraceae and Lamiaceae family and essential oils from Salvia officinalis and Salvia sclarea were studied for their antibacterial as well as antibiotic resistance modifying activity. Using disc diffusion and broth microdilution assays we determined higher antibacterial effect of three Salvia spp. and by evaluating the leakage of 260 nm absorbing material we detected effect of extracts and, namely, of essential oils on the disruption of cytoplasmic membrane. The evaluation of in vitro interactions between plant extracts and oxacillin described in terms of fractional inhibitory concentration (FIC indices revealed synergistic or additive effects of plant extracts and clearly synergistic effects of essential oil from Salvia officinalis with oxacillin in methicillin-resistant Staphylococcus epidermidis.

  6. Control Efficacy on Tobacco Virus Diseases of 6% Oligosaccharins・ Plant Activation Protein WP%6%寡糖・链蛋白可湿性粉剂对烟草病毒病的防治效果

    Institute of Scientific and Technical Information of China (English)

    徐传涛; 王李芳; 赵锦超; 谢强; 张永辉; 彭勇; 谢云波; 夏建华; 顾勇

    2016-01-01

    [目的]明确6%寡糖・链蛋白可湿性粉剂对泸州烟区烟草病毒病的防治效果。[方法]在四川省泸州市叙永县麻城乡采用田间试验研究6%寡糖・链蛋白可湿性粉剂1000倍稀释液对烟草病毒病的防治效果。[结果]通过苗期喷施1次,大田期喷施2次6%寡糖・链蛋白可湿性粉剂1000倍稀释液,对烟草病毒病的防治效果达65.44%。[结论]6%寡糖・链蛋白可湿性粉剂可用于防治泸州烟区烟草病毒病。%Objective] The aim was to clarify control effect on tobacco virus diseases of 6% oligosaccharins・ plant activation protein WP in Luzhou.[ Method] The field plot trials were conducted to assess the control efficacy on tobacco virus diseases in Luzhou .[ Result] The control efficacy on tobacco virus diseases were 65.44%, when 1 000-fold diluted of 6% oligosaccharins・ plant activation protein WP was applied dur-ing the period of seeding and field stage.[Conclusions] Tobacco virus diseases could be prevented and controlled effectively by applying 6%oli-gosaccharins・ plant activation protein WP.

  7. Can tobacco have a potentially beneficial effect to our health?

    Science.gov (United States)

    Macek, Tomas; Surá, Martina; Pavliková, Daniela; Francová, Katerina; Scouten, William H; Szekeres, Miklos; Sylvestre, Michel; Macková, Martina

    2005-01-01

    With urgent pressure to clean up the contaminated environment, new approaches are needed. Phyto- and rhizoremediation using plants and related bacteria is a promising approach, but has its inborn limitations. To overcome the slow performance of the process, transgenic plants have been prepared specifically tailored for phytoremediation purposes. Our projects addressed a group of widespread synthetic organic xenobiotics, polychlorinated biphenyls (PCBs), and heavy metals as representatives of inorganic contaminants. Beside basic research studies in the field of phyto/rhizoremediation of the mentioned toxicants we focused on genetically modified plants as a highly promising tool for these purposes. We tried to prepare tobacco plants expressing the bacterial enzyme responsible for cleaving PCBs, coded by the gene bphC from the bacterial biphenyl operon. The expression of bphC product in fusion with the green fluorescent protein is described together with evaluation of the twice increased resistance of transgenic seeds towards PCBs. The other model is addressing improvement of cadmium accumulation by preparing plants bearing fused transgenes of metal binding protein (yeast metallothionein) with an introduced additional metal binding domain--polyhistidine anchor with high affinity to metals. The genetically modified plants exhibit 190% Cd accumulation of the control in harvestable parts, higher resistance and lower Cd content in roots. The performance of the plants in real contaminated soil is also evaluated.

  8. A contribution to identification of novel regulators of plant response to sulfur deficiency: characteristics of a tobacco gene UP9C, its protein product and the effects of UP9C silencing.

    Science.gov (United States)

    Lewandowska, Malgorzata; Wawrzynska, Anna; Moniuszko, Grzegorz; Lukomska, Jolanta; Zientara, Katarzyna; Piecho, Marta; Hodurek, Pawel; Zhukov, Igor; Liszewska, Frantz; Nikiforova, Victoria; Sirko, Agnieszka

    2010-03-01

    Extensive changes in plant transcriptome and metabolome have been observed by numerous research groups after transferring plants from optimal conditions to sulfur (S) deficiency. Despite intensive studies and recent important achievements, like identification of SLIM1/EIL3 as a major transcriptional regulator of the response to S-deficiency, many questions concerning other elements of the regulatory network remain unanswered. Investigations of genes with expression regulated by S-deficiency stress encoding proteins of unknown function might help to clarify these problems. This study is focused on the UP9C gene and the UP9-like family in tobacco. Homologs of these genes exist in other plant species, including a family of four genes of unknown function in Arabidopsis thaliana (LSU1-4), of which two were reported as strongly induced by S-deficit and to a lesser extent by salt stress and nitrate limitation. Conservation of the predicted structural features, such as coiled coil region or nuclear localization signal, suggests that these proteins might have important functions possibly mediated by interactions with other proteins. Analysis of transgenic tobacco plants with silenced expression of UP9-like genes strongly argues for their significant role in regulation of plant response to S-deficit. Although our study shows that the UP9-like proteins are important components of such response and they might be also required during other stresses, their molecular functions remain a mystery.

  9. [Smokeless tobacco].

    Science.gov (United States)

    Underner, Michel; Perriot, Jean; Peiffer, Gérard

    2012-01-01

    The use of snus (smokeless tobacco) can be detrimental to health. Containing carcinogenic nitrosamines (Swedish snus do not contain nitrosamine). Snus delivers rapidly high doses of nicotine which can lead to dependence. It do not induce bronchial carcinoma differently smoked tobacco. Lesions usually develop in the area of the mouth where the snus is placed. Non-malignant oral lesions include leukoedema, hyperkeratotic lesions of the oral mucosa and localised periodontal disease. The most frequently occurring premalignant lesion is leukoplakia. Studies reveal conflicting evidence about the risk of oral and gastroesophageal cancer with regard to snus users. However, the use of snus has proved to be a risk factor in developing pancreatic cancer and increases the risk of fatal myocardial infarction and ischemic stroke. During pregnancy, snus is associated with an increased risk of pre-eclampsia and premature delivery. Nicotine substitution therapy and bupropion and varenicline reduce withdrawal symptoms and tobacco craving during snus cessation. However, they have not been shown to assist in long-term abstinence. Information concerning potential hazards of using snus products must be incorporated into health educational programmes in order to discourage its use. Snus is not a recommended product to help in stopping to smoke.

  10. Green Tobacco Sickness among Thai Traditional Tobacco Farmers, Thailand

    Directory of Open Access Journals (Sweden)

    T Saleeon

    2015-07-01

    Full Text Available Background: Traditional Thai tobacco (Nicotiana abacus L. is known as a non-Virginia type whose mature leaf contains three to four times more nicotine than that of a Virginia type. As such, the process of Thai traditional tobacco production may lead to adverse health effects such as green tobacco sickness (GTS.Objective: To investigate the prevalence of GTS and risk factors related to GTS among Thai traditional tobacco farmers in Nan province, northern Thailand.Methods: 473 Thai traditional tobacco farmers from rural areas in Nan province were randomly selected and interviewed in person by means of questionnaires and environmental survey. Statistical analyses were used to identify potential risk factors for GTS.Results: The prevalence of GTS was 22.6% (95% CI 19.1% to 26.6%. Multivariate analysis showed various risk factors associated with GTS including gender of the farmer (ORadj 0.44, 95% CI 0.26 to 0.73, smoking (ORadj 4.36, 95% CI 1.41 to 13.47, skin rash (ORadj 0.36, 95% CI 0.19 to 0.68, wearing a wet suit (ORadj 1.91, 95% CI 1.12 to 3.23, process of curing tobacco leaves (ORadj 0.06, 95% CI 0.02 to 0.16, and watering tobacco plants (ORadj 0.42, 95% CI 0.25 to 0.72.Conclusion: The process of traditional Thai tobacco production can result in increased dermal exposure and can be considered a major risk factor for GTS. Body soaking during watering may further increase adverse health effects related to GTS.

  11. Regulation of galactan synthase expression to modify galactan content in plants

    Energy Technology Data Exchange (ETDEWEB)

    None

    2017-08-22

    The disclosure provides methods of engineering plants to modulate galactan content. Specifically, the disclosure provides methods for engineering a plant to increase the galactan content in a plant tissue by inducing expression of beta-1,4-galactan synthase (GALS), modulated by a heterologous promoter. Further disclosed are the methods of modulating expression level of GALS under the regulation of a transcription factor, as well as overexpression of UDP-galactose epimerse in the same plant tissue. Tissue specific promoters and transcription factors can be used in the methods are also provided.

  12. Green tobacco sickness in children and adolescents.

    Science.gov (United States)

    McKnight, Robert H; Spiller, Henry A

    2005-01-01

    Tobacco (Nicotiana tabacum) is cultivated in more than 100 countries, and in 2004, some 5.73 million metric tons dry weight of tobacco were grown worldwide. The top five tobacco producers forecast for 2004 are China (2.01 million metric tons; 35.1%), Brazil (757 thousand metric tons; 13.2%), India (598 thousand metric tons; 10.4%), United States (358 thousand metric tons; 6.2%), and Malawi (138 thousand metric tons; 2.4%). Together, these five countries account for two-thirds of worldwide tobacco production. Tobacco farming presents several hazards to those who cultivate and harvest the plant. Although some of these hazards, such as pesticide exposure and musculoskeletal trauma, are faced by workers in other types of agricultural production, tobacco production presents some unique hazards, most notably acute nicotine poisoning, a condition also known as green tobacco sickness (GTS). GTS is an occupational poisoning that can affect workers who cultivate and harvest tobacco. It occurs when workers absorb nicotine through the skin as they come into contact with leaves of the mature tobacco plant. GTS is characterized largely by nausea, vomiting, headache, muscle weakness, and dizziness. Historically, children have played a role in agricultural production in the United States, and they continue to do so today. This includes tobacco farming. The North American Guidelines for Children's Agricultural Tasks, a set of injury prevention guidelines prepared by the National Children's Center for Rural and Agricultural Health and Safety, lists GTS as one of several hazards children face when working on tobacco farms. Children 17 years of age and younger who work on U.S. tobacco farms come from three main groups: members of farm families, migrant youth laborers (primarily Latinos), and other hired local children. All three groups are at risk for GTS. Beyond the U.S., tobacco production using child labor is an emerging topic of concern in developing nations. An international

  13. Effects of Different Fertilization Ways on Growth and Development of Early Planting Flue-cured Tobacco%不同施肥方式对早植烟生长发育的影响

    Institute of Scientific and Technical Information of China (English)

    辉树光; 彭坚强; 江皓; 杨晓; 马阳红; 秦蔚

    2014-01-01

    In order to improve the technical level of early planting tobacco transplanting fertilizer, through the field plot comparative test on three kinds of fertilization methods, as bulking farmyard manure application, banding middle fertilizer, aroundding middle fertilizer. The results showed that layer of fertilization in circulation application could make flue-cured tobacco growth peak in advance, the whole growth stages could extended about 10 d, agronomic characters, economic characters and the control group had significant difference. It had certain guiding significance for promoting the level of tobacco planting.%为提高早植烟移栽施肥技术水平,通过对早植烟移栽时散施农家肥、条施中层肥、环施中层肥3种施肥方式进行大田小区对比试验。结果表明:环施中层肥能使早植烟生长高峰期提前,整个生育周期延长约10 d,农艺性状、经济性状与对照组差异显著,环施中层肥对提升早植烟种植水平有一定指导意义。

  14. Rapid Py-GC/MS assessment of the structural alterations of lignins in genetically modified plants

    NARCIS (Netherlands)

    Rencoret, Jorge; Del Río, José Carlos; Nierop, Klaas G J|info:eu-repo/dai/nl/182329895; Gutiérrez, Ana; Ralph, John

    Genetic modifications for perturbing the lignin pathway in three different species of angiosperm plants, including non-woody (Arabidopsis and alfalfa) and woody (poplar) plants, were readily evaluated by analytical pyrolysis coupled to gas chromatography-mass spectrometry (Py-GC/MS). Pyrolysis

  15. IL13 gene polymorphisms modify the effect of exposure to tobacco smoke on persistent wheeze and asthma in childhood, a longitudinal study

    Directory of Open Access Journals (Sweden)

    Kurukulaaratchy Ramesh

    2008-01-01

    Full Text Available Abstract Background Tobacco smoke and genetic susceptibility are risk factors for asthma and wheezing. The aim of this study was to investigate whether there is a combined effect of interleukin-13 gene (IL13 polymorphisms and tobacco smoke on persistent childhood wheezing and asthma. Methods In the Isle of Wight birth cohort (UK, 1989–1999, five IL13 single nucleotide polymorphisms (SNPs: rs1800925 (-1112C/T, rs2066960, rs1295686, rs20541 (R130Q and rs1295685 were genotyped. Parents were asked whether their children had wheezed in the last 12 months at ages 1, 2, 4 and 10 years. Children who reported wheeze in the first 4 years of life and also had wheezing at age 10 were classified as early-onset persistent wheeze phenotype; non-wheezers never wheezed up to age 10. Persistent asthma was defined as having a diagnosis of asthma both during the first four years of life and at age 10. Logistic regression methods were used to analyze data on 791 children with complete information. Potential confounders were gender, birth weight, duration of breast feeding, and household cat or dog present during pregnancy. Results Maternal smoking during pregnancy was associated with early-onset persistent wheeze (OR 2.93, p IL13 were not (OR 1.15, p = 0.60 for the common haplotype pair. However, the effect of maternal smoking during pregnancy was stronger in children with the common IL13 haplotype pair compared to those without it (OR 5.58 and OR 1.29, respectively; p for interaction = 0.014. Single SNP analysis revealed a similar statistical significance for rs20541 (p for interaction = 0.02. Comparable results were observed for persistent childhood asthma (p for interaction = 0.03. Conclusion This is the first report that shows a combined effect of in utero exposure to smoking and IL13 on asthma phenotypes in childhood. The results emphasize that genetic studies need to take environmental exposures into account, since they may explain contradictory findings.

  16. A school-based programme for tobacco and alcohol prevention in special education: effectiveness of the modified 'healthy school and drugs' intervention and moderation by school subtype.

    Science.gov (United States)

    Turhan, Abdullah; Onrust, Simone A; Ten Klooster, Peter M; Pieterse, Marcel E

    2017-03-01

    To test the effectiveness of the Healthy School and Drugs (HSD) programme on tobacco and alcohol use in Dutch secondary special education (SE) schools, and whether this depends upon subtypes of SE schools and the level of implementation. In a quasi-experimental design with baseline and post-treatment follow-up, 363 students were allocated arbitrarily or depending on teacher motivation to either intervention condition (n = 205) or usual curriculum (n = 158). Thirteen secondary SE schools spread throughout the Netherlands. Participants were recruited during the autumn of 2013 from three school subtypes: SE for adolescents with intellectual/physical disabilities (SEI; n = 13), behavioural/emotional difficulties (SEB; n = 136) and learning disabilities/developmental disorders (SEL; n = 214). Self-reported life-time smoking prevalence and life-time drinking frequency as outcomes, and school subtype (SEL/SEB) and implementation fidelity (high/low) as moderators. No significant differences were found at follow-up in life-time smoking [odds ratio (OR) = 1.52; 95% confidence interval (CI) = 0.74-3.12] and drinking frequency (d = 0.01; 95% CI = -0.16 to 0.18). Interaction analyses revealed adverse effects in SEB students for alcohol use (d = 0.43; 95% CI = 0.16-0.69). Effect on tobacco refusal self-efficacy was moderated positively by implementation fidelity (d = 0.35; 95% CI = 0.07-0.63). The Healthy School and Drugs programme adapted for secondary special education in the Netherlands lacked clear evidence for effects on all outcomes. This pilot study suggests further that, within special education, substance use interventions may need to be targeted at school subtypes, as these may have harmful effects among students with behavioural difficulties. Finally, limited evidence was found that programme effectiveness may depend upon implementation fidelity. © 2016 Society for the Study of Addiction.

  17. Assessing environmental impacts of genetically modified plants on non-target organisms: The relevance of in planta studies.

    Science.gov (United States)

    Arpaia, Salvatore; Birch, A Nicholas E; Kiss, Jozsef; van Loon, Joop J A; Messéan, Antoine; Nuti, Marco; Perry, Joe N; Sweet, Jeremy B; Tebbe, Christoph C

    2017-04-01

    In legal frameworks worldwide, genetically modified plants (GMPs) are subjected to pre-market environmental risk assessment (ERA) with the aim of identifying potential effects on the environment. In the European Union, the EFSA Guidance Document introduces the rationale that GMPs, as well as their newly produced metabolites, represent the potential stressor to be evaluated during ERA. As a consequence, during several phases of ERA for cultivation purposes, it is considered necessary to use whole plants or plant parts in experimental protocols. The importance of in planta studies as a strategy to address impacts of GMPs on non-target organisms is demonstrated, to evaluate both effects due to the intended modification in plant phenotype (e.g. expression of Cry proteins) and effects due to unintended modifications in plant phenotype resulting from the transformation process (e.g. due to somaclonal variations or pleiotropic effects). In planta tests are also necessary for GMPs in which newly expressed metabolites cannot easily be studied in vitro. This paper reviews the scientific literature supporting the choice of in planta studies as a fundamental tool in ERA of GMPs in cultivation dossiers; the evidence indicates they can realistically mimic the ecological relationships occurring in their receiving environments and provide important insights into the biology and sustainable management of GMPs.

  18. Overexpression of ARGOS Genes Modifies Plant Sensitivity to Ethylene, Leading to Improved Drought Tolerance in Both Arabidopsis and Maize.

    Science.gov (United States)

    Shi, Jinrui; Habben, Jeffrey E; Archibald, Rayeann L; Drummond, Bruce J; Chamberlin, Mark A; Williams, Robert W; Lafitte, H Renee; Weers, Ben P

    2015-09-01

    Lack of sufficient water is a major limiting factor to crop production worldwide, and the development of drought-tolerant germplasm is needed to improve crop productivity. The phytohormone ethylene modulates plant growth and development as well as plant response to abiotic stress. Recent research has shown that modifying ethylene biosynthesis and signaling can enhance plant drought tolerance. Here, we report novel negative regulators of ethylene signal transduction in Arabidopsis (Arabidopsis thaliana) and maize (Zea mays). These regulators are encoded by the ARGOS gene family. In Arabidopsis, overexpression of maize ARGOS1 (ZmARGOS1), ZmARGOS8, Arabidopsis ARGOS homolog ORGAN SIZE RELATED1 (AtOSR1), and AtOSR2 reduced plant sensitivity to ethylene, leading to enhanced drought tolerance. RNA profiling and genetic analysis suggested that the ZmARGOS1 transgene acts between an ethylene receptor and CONSTITUTIVE TRIPLE RESPONSE1 in the ethylene signaling pathway, affecting ethylene perception or the early stages of ethylene signaling. Overexpressed ZmARGOS1 is localized to the endoplasmic reticulum and Golgi membrane, where the ethylene receptors and the ethylene signaling protein ETHYLENE-INSENSITIVE2 and REVERSION-TO-ETHYLENE SENSITIVITY1 reside. In transgenic maize plants, overexpression of ARGOS genes also reduces ethylene sensitivity. Moreover, field testing showed that UBIQUITIN1:ZmARGOS8 maize events had a greater grain yield than nontransgenic controls under both drought stress and well-watered conditions.

  19. [Research progress in chemical communication among insect-resistant genetically modified plants, insect pests and natural enemies].

    Science.gov (United States)

    Liu, Qing-Song; Li, Yun-He; Chen, Xiu-Ping; Peng, Yu-Fa

    2014-08-01

    Semiochemicals released by plants or insects play an important role in the communication among plants, phytophagous insects and their natural enemies. They thus form a chemical information network which regulates intra- and inter-specific behaviors and sustains the composition and structure of plant and insect communities. The application of insect-resistant genetically modified (IRGM) crops may affect the chemical communication within and among the tritrophic levels, and thus cause disturbances to the biotic community structure and the stability of the farmland ecosystem. This has raised concerns about the environmental safety of IRGM crops and triggered research worldwide. In the current article we provided a brief summary of the chemical communication among plants, herbivores and natural enemies; analyzed the potential of IRGM crops to affect the chemical communication between plants and arthropods and the related mechanisms; and discussed the current research progress and the future prospects in this field. We hope that this will promote the research in this field by Chinese scientists and increase our understanding of the potential effects of growing of IRGM crops on the arthropod community structure.

  20. Effects of specific expression of iaaL gene in tobacco tapetum on pollen embryogenesis

    Institute of Scientific and Technical Information of China (English)

    杨洪全; 卫志明; 许智宏

    1997-01-01

    The indoleacetic-acid-lysine synthetase (iaaL) gene from Pseudomonas syringae subsp. savastanoi was fused to tobacco tapetum-specific expression promoter TA29, and introduced into tobacco. The expression pattern of this chimeric gene was studied, and the endogenous indoleacetic acid (IAA) levels in different organs were assayed. The results demonstrated that TA29 promoter was only able to direct the specific expression of iaaL gene in transgenic tobacco anther, and resulted in the decrease of endogenous IAA levels in transgenic tobacco anther. No significant phe-notype variation was observed among the transgenic plants at the whole plant level. However, the percentage of pollen embryogenesis was reduced to 11 % when anthers of the transgenic plants were cultured on the modified hormone-free Nistch H (NH) medium, while those of both CK1 and CK2 (see sec. 1.2.2) were more than 50% ; when the an-thers were cultured on NH medium supplemented with 0. 2 mg/L IAA, the percentage of pollen embryogenesis re-stor

  1. Ectopic localization of auxin and cytokinin in tobacco seedlings by the plant-oncogenic AK-6b gene of Agrobacterium tumefaciens AKE10.

    Science.gov (United States)

    Takahashi, Sachiko; Sato, Rui; Takahashi, Miho; Hashiba, Noriko; Ogawa, Atsushi; Toyofuku, Kyoko; Sawata, Taiki; Ohsawa, Yuki; Ueda, Kenji; Wabiko, Hiroetsu

    2013-10-01

    The oncogenic 6b gene of Agrobacterium tumefaciens induces a number of morphological and metabolic alterations in plants. Although molecular functions associated with the 6b genes have been proposed, including auxin transport, sugar transport, transcriptional regulation, and miRNA metabolism, so far an unequivocal conclusion has not been obtained. We investigated the association between auxin accumulation and tumor development of the tobacco seedlings expressing the AK-6b gene under the control of the dexamethasone-inducible promoter. Indole-3-acetic acid (IAA) localization was examined by immunochemical staining with monoclonal antibody against IAA and by histochemical analysis using the IAA-specific induced construct, DR5::GUS (β-glucuronidase). Both procedures indicated that IAA preferentially accumulated in the tumorous protrusions as well as in newly developing vascular bundles in the tumors. Furthermore, true leaves also showed abaxial IAA localization, leading to altered leaves in which the adaxial and abaxial identities were no longer evident. Co-localization of cytokinin and auxin in the abaxial tumors was verified by immunochemical staining with an antibody against cytokinin. Treatment of AK-6b-seedlings with N-1-naphthylphthalamic acid, an inhibitor of polar auxin transport, promoted the morphological severity of phenotypes, whereas 1-naphthoxyacetic acid, a specific auxin influx carrier inhibitor, induced tumor regression on cotyledons and new tumorous proliferations on hypocotyls. Prominent accumulation of both auxin and cytokinin was observed in both regressed and newly developing tumors. We suggest from these results that modulation of auxin/cytokinin localization as a result of AK-6b gene expression is responsible for the tumorous proliferation.

  2. Impact of salinity-tolerant MCM6 transgenic tobacco on soil enzymatic activities and the functional diversity of rhizosphere microbial communities.

    Science.gov (United States)

    Chaudhry, Vasvi; Dang, Hung Quang; Tran, Ngoc Quang; Mishra, Aradhana; Chauhan, Puneet Singh; Gill, Sarvajeet Singh; Nautiyal, Chandra Shekhar; Tuteja, Narendra

    2012-01-01

    The development of genetically modified plants for agriculture has provided numerous economic benefits, but has also raised concern over the potential impact of transgenic plants upon the environment. The rhizosphere is the soil compartment that is directly under the influence of living roots; it constitutes a complex niche likely to be exploited by a wide variety of bacteria potentially influenced by the introduction of transgenes in genetically modified plants. In the present study, the impact of overexpression of the salinity stress-tolerant minichromosome maintenance complex subunit 6 (MCM6) gene upon functional diversity and soil enzymatic activity in the rhizosphere of transgenic tobacco in the presence and absence of salt stress was examined. The diversity of culturable bacterial communities and soil enzymes, namely, dehydrogenases and acid phosphatases, was assessed and revealed no significant (or only minor) alterations due to transgenes in the rhizosphere soil of tobacco plants. Patterns in principal components analysis showed clustering of transgenic and non-transgenic tobacco plants according to the fingerprint of their associated bacterial communities. However, the presence of MCM6 tobacco did not cause changes in microbial populations, soil enzymatic activities or the functional diversity of the rhizosphere soil microbial community.

  3. Large grazers modify effects of aboveground–belowground interactions on small-scale plant community composition

    NARCIS (Netherlands)

    Veen, G.F.; Geuverink, E.; Olff, H.

    2012-01-01

    Aboveground and belowground organisms influence plant community composition by local interactions, and their scale of impact may vary from millimeters belowground to kilometers aboveground. However, it still poorly understood how large grazers that select their forage on large spatial scales

  4. Large grazers modify effects of aboveground-belowground interactions on small-scale plant community composition

    NARCIS (Netherlands)

    Veen, G. F. (Ciska); Geuverink, Elzemiek; Olff, Han; Schmid, Bernhard

    Aboveground and belowground organisms influence plant community composition by local interactions, and their scale of impact may vary from millimeters belowground to kilometers aboveground. However, it still poorly understood how large grazers that select their forage on large spatial scales

  5. Evaluation of buffers toxicity in tobacco cells: Homopiperazine-1,4-bis (2-ethanesulfonic acid) is a suitable buffer for plant cells studies at low pH.

    Science.gov (United States)

    Borgo, Lucélia

    2017-06-01

    Low pH is an important environmental stressor of plant root cells. Understanding the mechanisms of stress and tolerance to acidity is critical; however, there is no widely accepted pH buffer for studies of plant cells at low pH. Such a buffer might also benefit studies of Al toxicity, in which buffering at low pH is also important. The challenge is to find a buffer with minimal cellular effects. We examined the cytotoxicity and possible metabolic disturbances of four buffers that have adequate pKa values and potential use for studies in the pH range of 4.0-5.0. These were homopipes (homopiperazine-1,4-bis (2-ethanesulfonic acid); pKa1 4.4), 3,3-dimethylglutaric acid (pKa1 3.73), β-alanine (pKa1 3.70) and potassium biphthalate (pKa1 2.95; pKa2 5.41). First, tobacco BY-2 cells were grown in a rich medium containing 10 mM of each buffer or MES (2-(N-morpholino) ethanesulfonic acid) as a control, with the pH initially adjusted to 5.7. β-alanine was clearly toxic and dimethylgluturate and biphthalate were found to be cytostatic, in which no culture growth occurred but cell viability was either unaffected or decreased only after 5 days. Only homopipes allowed normal culture growth and cell viability. Homopipes (10 mM) was then tested in cell cultures with an initial pH of 4.3 ± 0.17 in minimal medium to examine whether its undissociated species (H2A) displayed any cellular effects and no cytotoxic effects were observed. It is possible to conclude that among tested buffers, homopipes is the most suitable for studies at low pH, and may be especially useful for aluminum toxicity experiments. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

  6. Moderating mycorrhizas: arbuscular mycorrhizas modify rhizosphere chemistry and maintain plant phosphorus status within narrow boundaries.

    Science.gov (United States)

    Nazeri, Nazanin K; Lambers, Hans; Tibbett, Mark; Ryan, Megan H

    2014-04-01

    Pastures often experience a pulse of phosphorus (P) when fertilized. We examined the role of arbuscular mycorrhizal fungi (AMF) in the uptake of P from a pulse. Five legumes (Kennedia prostrata, Cullen australasicum, Bituminaria bituminosa, Medicago sativa and Trifolium subterraneum) were grown in a moderate P, sterilized field soil, either with (+AMF) or without (-AMF) addition of unsterilized field soil. After 9-10 weeks, half the pots received 15 mg P kg(-1) of soil. One week later, we measured: shoot and root dry weights; percentage of root length colonized by AMF; plant P, nitrogen and manganese (Mn) concentrations; and rhizosphere carboxylates, pH and plant-available P. The P pulse raised root P concentration by a similar amount in uncolonized and colonized plants, but shoot P concentration increased by 143% in uncolonized plants and 53% in colonized plants. Inoculation with AMF decreased the amount of rhizosphere carboxylates by 52%, raised rhizosphere pH by ∼0.2-0.7 pH units and lowered shoot Mn concentration by 38%. We conclude that AMF are not simply a means for plants to enhance P uptake when P is limiting, but also act to maintain shoot P within narrow boundaries and can affect nutrient uptake through their influence on rhizosphere chemistry. © 2013 John Wiley & Sons Ltd.

  7. Tobacco packaging design for preventing tobacco uptake

    OpenAIRE

    McNeill, Ann; Bauld, Linda; Birken, Mary; Hammond, David; Moodie, Crawford; Stead, Martine; Hitchman, Sara; Hartmann-Boyce, Jamie

    2016-01-01

    This is the protocol for a review and there is no abstract. The objectives are as follows: To assess the effectiveness of standardised tobacco packaging in preventing initiation into, or regular use of, tobacco.

  8. A statistical simulation model for field testing of non-target organisms in environmental risk assessment of genetically modified plants.

    Science.gov (United States)

    Goedhart, Paul W; van der Voet, Hilko; Baldacchino, Ferdinando; Arpaia, Salvatore

    2014-04-01

    Genetic modification of plants may result in unintended effects causing potentially adverse effects on the environment. A comparative safety assessment is therefore required by authorities, such as the European Food Safety Authority, in which the genetically modified plant is compared with its conventional counterpart. Part of the environmental risk assessment is a comparative field experiment in which the effect on non-target organisms is compared. Statistical analysis of such trials come in two flavors: difference testing and equivalence testing. It is important to know the statistical properties of these, for example, the power to detect environmental change of a given magnitude, before the start of an experiment. Such prospective power analysis can best be studied by means of a statistical simulation model. This paper describes a general framework for simulating data typically encountered in environmental risk assessment of genetically modified plants. The simulation model, available as Supplementary Material, can be used to generate count data having different statistical distributions possibly with excess-zeros. In addition the model employs completely randomized or randomized block experiments, can be used to simulate single or multiple trials across environments, enables genotype by environment interaction by adding random variety effects, and finally includes repeated measures in time following a constant, linear or quadratic pattern in time possibly with some form of autocorrelation. The model also allows to add a set of reference varieties to the GM plants and its comparator to assess the natural variation which can then be used to set limits of concern for equivalence testing. The different count distributions are described in some detail and some examples of how to use the simulation model to study various aspects, including a prospective power analysis, are provided.

  9. Genetic basis and detection of unintended effects in genetically modified crop plants

    NARCIS (Netherlands)

    Ladics, G.S.; Bartholomaeus, A.; Bregitzer, P.; Doerrer, N.G.; Gray, A.; Holzhauzer, T.; Jordan, M.; Keese, P.; Kok, E.J.; Macdonald, P.; Parrott, W.; Privalle, L.; Raybould, A.; Rhee, S.Y.; Rice, E.; Romeis, J.; Vaughn, J.; Wal, J.M.; Glenn, K.

    2015-01-01

    In January 2014, an international meeting sponsored by the International Life Sciences Institute/Health and Environmental Sciences Institute and the Canadian Food Inspection Agency titled “Genetic Basis of Unintended Effects in Modified Plants” was held in Ottawa, Canada, bringing together over 75 s

  10. Opinion Building on a Socio-Scientific Issue: The Case of Genetically Modified Plants

    Science.gov (United States)

    Ekborg, Margareta

    2008-01-01

    This paper presents results from a study with the following research questions: (a) are pupils' opinions on genetically modified organisms (GMOs) influenced by biology teaching; and (b) what is important for the opinion pupils hold and how does knowledge work together with other parameters such as values? 64 pupils in an upper secondary school…

  11. Plants with stacked genetically modified events: to assess or not to assess?

    NARCIS (Netherlands)

    Kok, E.J.; Pedersen, J.; Onori, R.; Sowa, S.; Schauzu, M.; Schrijver, De A.; Teeri, T.H.

    2014-01-01

    The principles for the safety assessment of genetically modified (GM) organisms (GMOs) are harmonised worldwide to a large extent. There are, however, still differences between the European GMO regulations and the GMO regulations as they have been formulated in other parts of the world. One of these

  12. Quantitative proteomics analysis using 2D-PAGE to investigate the effects of cigarette smoke and aerosol of a prototypic modified risk tobacco product on the lung proteome in C57BL/6 mice.

    Science.gov (United States)

    Elamin, Ashraf; Titz, Bjoern; Dijon, Sophie; Merg, Celine; Geertz, Marcel; Schneider, Thomas; Martin, Florian; Schlage, Walter K; Frentzel, Stefan; Talamo, Fabio; Phillips, Blaine; Veljkovic, Emilija; Ivanov, Nikolai V; Vanscheeuwijck, Patrick; Peitsch, Manuel C; Hoeng, Julia

    2016-08-11

    Smoking is associated with several serious diseases, such as lung cancer and chronic obstructive pulmonary disease (COPD). Within our systems toxicology framework, we are assessing whether potential modified risk tobacco products (MRTP) can reduce smoking-related health risks compared to conventional cigarettes. In this article, we evaluated to what extent 2D-PAGE/MALDI MS/MS (2D-PAGE) can complement the iTRAQ LC-MS/MS results from a previously reported mouse inhalation study, in which we assessed a prototypic MRTP (pMRTP). Selected differentially expressed proteins identified by both LC-MS/MS and 2D-PAGE approaches were further verified using reverse-phase protein microarrays. LC-MS/MS captured the effects of cigarette smoke (CS) on the lung proteome more comprehensively than 2D-PAGE. However, an integrated analysis of both proteomics data sets showed that 2D-PAGE data complement the LC-MS/MS results by supporting the overall trend of lower effects of pMRTP aerosol than CS on the lung proteome. Biological effects of CS exposure supported by both methods included increases in immune-related, surfactant metabolism, proteasome, and actin cytoskeleton protein clusters. Overall, while 2D-PAGE has its value, especially as a complementary method for the analysis of effects on intact proteins, LC-MS/MS approaches will likely be the method of choice for proteome analysis in systems toxicology investigations. Quantitative proteomics is anticipated to play a growing role within systems toxicology assessment frameworks in the future. To further understand how different proteomics technologies can contribute to toxicity assessment, we conducted a quantitative proteomics analysis using 2D-PAGE and isobaric tag-based LC-MS/MS approaches and compared the results produced from the 2 approaches. Using a prototypic modified risk tobacco product (pMRTP) as our test item, we show compared with cigarette smoke, how 2D-PAGE results can complement and support LC-MS/MS data, demonstrating

  13. Genetically modified plants in phytoremediation of heavy metal and metalloid soil and sediment pollution.

    Science.gov (United States)

    Kotrba, Pavel; Najmanova, Jitka; Macek, Tomas; Ruml, Tomas; Mackova, Martina

    2009-01-01

    Phytoremediation to clean up metal- and metalloid-contaminated soil or sediments has gained increasing attention as environmental friendly and cost effective. Achievements of the last decade suggest that genetic engineering of plants can be instrumental in improving phytoremediation. Transgenic approaches successfully employed to promote phytoextraction of metals (mainly Cd, Pb, Cu) and metalloids (As, Se) from soil by their accumulation in the aboveground biomass involved mainly implementation of metal transporters, improved production of enzymes of sulphur metabolism and production of metal-detoxifying chelators - metallothioneins and phytochelatins. Plants producing bacterial mercuric reductase and organomercurial lyase can covert the toxic ion or organomercury to metallic Hg volatized from the leaf surface. Phytovolatization of selenium compounds was promoted in plants overexpressing genes encoding enzymes involved in production of gas methylselenide species. This paper provides a broad overview of the evidence supporting suitability and prospects of transgenic research in phytoremediation of heavy metals and metalloids.

  14. Modified cellulose synthase gene from Arabidopsis thaliana confers herbicide resistance to plants

    Science.gov (United States)

    Somerville, Chris R.; Scheible, Wolf

    2007-07-10

    Cellulose synthase ("CS"), a key enzyme in the biosynthesis of cellulose in plants is inhibited by herbicides comprising thiazolidinones such as 5-tert-butyl-carbamoyloxy-3-(3-trifluromethyl)phenyl-4-thiazolidinone (TZ), isoxaben and 2,6-dichlorobenzonitrile (DCB). Two mutant genes encoding isoxaben and TZ-resistant cellulose synthase have been isolated from isoxaben and TZ-resistant Arabidopsis thaliana mutants. When compared with the gene coding for isoxaben or TZ-sensitive cellulose synthase, one of the resistant CS genes contains a point mutation, wherein glycine residue 998 is replaced by an aspartic acid. The other resistant mutation is due to a threonine to isoleucine change at amino acid residue 942. The mutant CS gene can be used to impart herbicide resistance to a plant; thereby permitting the utilization of the herbicide as a single application at a concentration which ensures the complete or substantially complete killing of weeds, while leaving the transgenic crop plant essentially undamaged.

  15. Biochemical Changes in β-Cryptogein-Elicited Tobacco: A possible Basis of Acquired Resistance

    Directory of Open Access Journals (Sweden)

    Edreva A

    2014-12-01

    Full Text Available β-Cryptogein, a proteinaceous elicitor from the phytopathogenic fungus Phytophthoracryptogea, is known to induce leaf necrosis in tobacco and non-specific resistance (expressed in the perinecrotic leaf area against a wide range of tobacco pathogens. To reveal mechanisms underlying the acquired resistance, biochemical changes in leaves of β-cryptogein-elicited tobacco were followed three, five and ten days after elicitation. The activities of peroxidase, β-1,3-glucanase and β-glucosidase, as well as the patterns of acidic pathogenesis-related (PR-proteins were determined. The protected part (perinecrotic area and the non-protected part (distant extra-perinecrotic area of leaves of β-cryptogein-stem treated tobacco (cv. Xanthin.c. were analyzed. Leaves of water-stem treated tobacco served as controls. It was shown that in the protected leaf part β-cryptogein caused significant metabolic shifts early after elicitation, persisting during the whole period studied. An important increase of peroxidase and β-1,3-glucanase activity was recorded. PR-protein components appeared that were absent in the controls. There were negligible changes in β-glucosidase activity. In the non-protected leaf part late and non-significant changes occurred. Taking into account the antimicrobial, regulatory and structure-modifying properties of the biochemical components studied, it may be admitted that β-cryptogein elicited the development of a hostile environment, i.e. a potential for plant resistance against subsequent pathogen invasion.

  16. DECOMPOSTION OF GENETICALLY ENGINEERED TOBACCO UNDER FIELD CONDITIONS: PERSISTENCE OF THE PROTEINASE INHIBITOR I PRODUCT AND EFFECTS OF SOIL MICROBIAL RESPIRATION AND PROTOZOA, NEMATODE AND MICROARTHR

    Science.gov (United States)

    1. To evaluate the potential effects of genetically engineered (transgenic) plants on soil ecosystems, litterbags containing leaves of non-engineered (parental) and transgenic tobacco plants were buried in field plots. The transgenic tobacco plants were genetically engineered to ...

  17. Growth at elevated CO2 concentrations leads to modified profiles of secondary metabolites in tobacco cv. SamsunNN and to increased resistance against infection with potato virus Y.

    Science.gov (United States)

    Matros, Andrea; Amme, Steffen; Kettig, Barbara; Buck-Sorlin, Gerhard H; Sonnewald, Uwe; Mock, Hans-Peter

    2006-01-01

    The effect of elevated CO2 concentrations on the levels of secondary metabolites was investigated in tobacco plants grown under two nitrogen supply (5 and 8 mM NH4NO3) and CO2 conditions (350 and 1000 p.p.m.) each. High CO2 resulted in a dramatic increase of phenylpropanoids in the leaves, including the major carbon-rich compound chlorogenic acid (CGA) and the coumarins scopolin and scopoletin at both nitrogen fertilizations. This was accompanied by increased PAL activity in leaves and roots, which was even higher at the lower nitrogen supply. Hardly any change was observed for the structural phenolic polymer lignin and the sesquiterpenoid capsidiol. In contrast, elevated CO2 led to clearly decreased levels of the main nitrogen-rich constituent nicotine at the lower N-supply (5 mM NH4NO3) but not when plants were grown at the higher N-supply (8 mM NH4NO3). Inoculation experiments with potato virus Y (PVY) were used to evaluate possible ecological consequences of elevated CO2. The titre of viral coat-protein was markedly reduced in leaves under these conditions at both nitrogen levels. Since PR-gene expression and free salicylic acid (SA) levels remained unchanged at elevated CO2, we suggest that the accumulation of phenylpropanoids, for example, the major compound CGA and the coumarins scopolin and scopoletin may result in an earlier confinement of the virus at high CO2. Based on our results two final conclusions emerge. First, elevated CO2 leads to a shift in secondary metabolite composition that is dependent on the availability of nitrogen. Second, changes in the pool of secondary metabolites have important consequences for plant-pathogen interactions as shown for PVY as a test organism.

  18. DNA staining with the fluorochromes EtBr, DAPI and YOYO-1 in the comet assay with tobacco plants after treatment with ethyl methanesulphonate, hyperthermia and DNase-I.

    Science.gov (United States)

    Gichner, Tomás; Mukherjee, Anita; Velemínský, Jirí

    2006-06-16

    We applied the alkaline version of the single-cell gel electrophoresis (comet) assay to roots and leaves of tobacco (Nicotiana tabacum var. xanthi) seedlings or isolated leaf nuclei treated with: (1) the alkylating agent ethyl methanesulphonate, (2) necrotic heat treatments at 50 degrees C, and (3) DNase-I. All three treatments induced a dose-dependent increase in DNA migration, expressed as percentage of tail DNA. A comparison of the fluorochrome DNA dyes ethidium bromide, DAPI and YOYO-1 demonstrated that for the alkaline version of the comet assay in plants, the commonly used fluorescent dye ethidium bromide can be used with the same efficiency as DAPI or YOYO-1.

  19. Effects of Salicylic Acid to Early Growth and Quick Tillering of Transplanted Tobacco (Nicotiana tabacum L.) Plants%水杨酸对移栽后烟株早生快发的影响

    Institute of Scientific and Technical Information of China (English)

    赵静; 徐照丽; 段胜智; 王丽特; 杨利云; 盛业龙; 龚明

    2013-01-01

      为促进移栽后烟株的早生快发,以K326为试验材料,采用温室大棚漂浮育苗,在幼苗长至3~4叶1心时,叶面分别喷施0、0.5、1.0、1.5 mmol/L的水杨酸,成苗后移栽至大田。结果表明,水杨酸通过增强抗氧化系统的活性,及时有效地清除活性氧,为移栽后烟株快速适应变化的环境提供有利的条件,从而促进烟株根系的快速发育、增强根系活力和根冠比,最终缩短烟株的还苗期,促进早生快发,为后续的快速生长和干物质积累奠定基础。水杨酸浓度不同,作用效果不同,1.0、1.5 mmol/L浓度的作用效果好于0.5 mmol/L。%In order to accelerate early growth and quick tillering of transplanted tobacco plants, experimental material K326 seedlings were floated in greenhouse. Spraying 0, 0.5, 1.0, 1.5 mmol/L salicylic acid on foliar respectively when the seedlings with 3-4 leaves. Then the grown seedlings were transplanted into field. The results showed that transplanted tobacco plants could adapt to the changed environment quickly after treated by salicylic acid because it could increase the activity of antioxidant system and then remove reactive oxygen species ROS timely and effectively. So salicylic acid could promote the development of root system rapidly, strengthening the root vigor and increasing root-shoot ratio. The result was rosette stage was shorted; early growth and quick tillering were accelerated of transplanted tobacco plants. All of those were beneficial to tobacco plants subsequently grow and accumulate dry matter rapidly. Effects of 4 different salicylic acid concentrations were different. The effects of 1.0 and 1.5 mmol/L concentration were better than 0.5 mmol/L.

  20. 植物诱导抗病剂组合应用对烟草花叶病的防控效果%Control Effects of Combination of Plant Induced Resistant Agents against Tobacco Mosaic Virus (TMV)

    Institute of Scientific and Technical Information of China (English)

    徐兴阳; 董家红; 端永明; 张廷金; 杨龙祥

    2011-01-01

    [Objective] The paper was explore the control effects of combination of plant induced resistant agents against tobacco mosaic virus (TMV). [ Method] The control effects of 6 different combinations of plant induced resistant agents against TMV of flue-cured tobacco cultivar HongDa were studied under the environment of simulated disease nursery. [ Result] The combination of two induced agents polypeptide-agents and 3-acetonyl-3-hydroxyoxindole (AHO) had good control effect against TMV, which could obviously delay the incidence time of TMV in infected tobacco plants. With water and Duxiao as control, their average control effects against TMV of tobacco plants during field period reached 69.64% and 43.25% after transplanting for 70 d. They also showed obvious superiority in the aspects of plant height and leaf number, and the growth and development condition of leaves was good. Tobacco seedlings carrying TMV virus had no direct correlation with whether the symptoms performed, the seedlings carrying virus would perform symptom only when the incidence condition was suitable. The peak period for the incidence of TMV in seedlings carrying virus was during 19 d after transplanting. Spraying effective agents during nursery stage and field period, as well as promoting early growth and quick tiller of tobacco seedlings after transplanting were the key measures to control its incidence. [Conclusion] The study provided theoretical basis for preparing the control measures against TMV.%[目的]探讨植物诱导抗病剂组合对烟草花叶病的防控效果.[方法]在模拟病圃环境下,研究了6种植物诱导抗病剂的不同组合对烤烟栽培品种红花大金元的烟草花叶病的防控效果.[结果]多肽保与AHO 2种诱导剂的组合应用对控制TMV有良好效果,能明显推迟带毒隐症烟株TMV的发病时间,移栽后70d对以清水和东旺毒消为对照的大田期烟株TMV的平均防效分别达到69.64%和43.25%,且

  1. Regression Analysis of Nutrient Elements between Soil and Tobacco Leaves and Their Influences on Aroma Quality of Flue-cured Tobacco

    Institute of Scientific and Technical Information of China (English)

    Xueqin XU; Xiaolan LI; Zhiyan CHEN; Defen ZENG; Aifei XU

    2014-01-01

    This study aimed to analyze the relationship between nutrient elements K, Ca, Mg, Cu, Mn, Zn and Fe in tobacco-planting soils and tobacco leaves from six main tobacco-producing areas, and to investigate the influences of these elements on chemical composition and aroma components in tobacco leaves. Results showed that there were certain relationship between contents of nutrient elements in tobac-co-planting soils and contents of corresponding elements in tobacco leaves; various elements exhibited different influences on the aroma quality of flue-cured tobacco. Based on the actual situation of nutrient contents in soils from different tobacco-producing areas, contents of various elements in tobacco leaves should be regulat-ed by soil fertilization and foliar spraying, thereby improving the aroma quality of flue-cured tobacco.

  2. Genetic engineering of flavonoid pigments to modify flower color in floricultural plants.

    Science.gov (United States)

    Nishihara, Masahiro; Nakatsuka, Takashi

    2011-03-01

    Recent advances in genetic transformation techniques enable the production of desirable and novel flower colors in some important floricultural plants. Genetic engineering of novel flower colors is now a practical technology as typified by commercialization of a transgenic blue rose and blue carnation. Many researchers exploit knowledge of flavonoid biosynthesis effectively to obtain unique flower colors. So far, the main pigments targeted for flower color modification are anthocyanins that contribute to a variety of colors such as red, pink and blue, but recent studies have also utilized colorless or faint-colored compounds. For example, chalcones and aurones have been successfully engineered to produce yellow flowers, and flavones and flavonols used to change flower color hues. In this review, we summarize examples of successful flower color modification in floricultural plants focusing on recent advances in techniques.

  3. Prevention of autoimmune hypothyroidism by modifying iodine intake and the use of tobacco and alcohol is manoeuvring between Scylla and Charybdis.

    Science.gov (United States)

    Laurberg, Peter; Andersen, Stig; Pedersen, Inge Bülow; Knudsen, Nils; Carlé, Allan

    2013-01-01

    Although autoimmune hypothyroidism has generally been considered to be a disease that mainly develops because of genetic aberrations and for which adjustment of environment would bring about but slight risk modification, this understanding is increasingly appearing to be incorrect. We describe how iodine intake, smoking cessation and alcohol intake are all strong modifiers of risk that, combined, may influence risk by a factor of up to 30. Unfortunately, promotion of an environment leading to substantial lowering of the risk of autoimmune hypothyroidism (i.e. improvement of dietary iodine deficiency, decrease or cessation of smoking, and moderate alcohol intake) is not incorporated within current public health promoting programs. Nevertheless, it is increasingly becoming evident that knowledge of the importance of these factors for disease development is likely to assist in the planning of health promotion programs, while it will surely also be of value in the care of individual patients.

  4. Prevention of autoimmune hypothyroidism by modifying iodine intake and the use of tobacco and alcohol is manoeuvring between Scylla and Charybdis

    DEFF Research Database (Denmark)

    Laurberg, Peter; Andersen, Stig; Pedersen, Inge Bülow

    2012-01-01

    iodine intake, smoking cessation and alcohol intake are all strong modifiers of risk that, combined, may influence risk by a factor of up to 30. Unfortunately, promotion of an environment leading to substantial lowering of the risk of autoimmune hypothyroidism (i.e. improvement of dietary iodine...... deficiency, decrease or cessation of smoking, and moderate alcohol intake) is not incorporated within current public health promoting programs. Nevertheless, it is increasingly becoming evident that knowledge of the importance of these factors for disease development is likely to assist in the planning......Although autoimmune hypothyroidism has generally been considered to be a disease that mainly develops because of genetic aberrations and for which adjustment of environment would bring about but slight risk modification, this understanding is increasingly appearing to be incorrect. We describe how...

  5. Plant functional group composition modifies the effects of precipitation change on grassland ecosystem function.

    Science.gov (United States)

    Fry, Ellen L; Manning, Pete; Allen, David G P; Hurst, Alex; Everwand, Georg; Rimmler, Martin; Power, Sally A

    2013-01-01

    Temperate grassland ecosystems face a future of precipitation change, which can alter community composition and ecosystem functions through reduced soil moisture and waterlogging. There is evidence that functionally diverse plant communities contain a wider range of water use and resource capture strategies, resulting in greater resistance of ecosystem function to precipitation change. To investigate this interaction between composition and precipitation change we performed a field experiment for three years in successional grassland in southern England. This consisted of two treatments. The first, precipitation change, simulated end of century predictions, and consisted of a summer drought phase alongside winter rainfall addition. The second, functional group identity, divided the plant community into three groups based on their functional traits- broadly described as perennials, caespitose grasses and annuals- and removed these groups in a factorial design. Ecosystem functions related to C, N and water cycling were measured regularly. Effects of functional groupidentity were apparent, with the dominant trend being that process rates were higher under control conditions where a range of perennial species were present. E.g. litter decomposition rates were significantly higher in plots containing several perennial species, the group with the highest average leaf N content. Process rates were also very strongly affected by the precipitation change treatmentwhen perennial plant species were dominant, but not where the community contained a high abundance of annual species and caespitose grasses. This contrasting response could be attributable to differing rooting patterns (shallower structures under annual plants, and deeper roots under perennials) and faster nutrient uptake in annuals compared to perennials. Our results indicate that precipitation change will have a smaller effect on key process rates in grasslandscontaining a range of perennial and annual species

  6. Plant functional group composition modifies the effects of precipitation change on grassland ecosystem function.

    Directory of Open Access Journals (Sweden)

    Ellen L Fry

    Full Text Available Temperate grassland ecosystems face a future of precipitation change, which can alter community composition and ecosystem functions through reduced soil moisture and waterlogging. There is evidence that functionally diverse plant communities contain a wider range of water use and resource capture strategies, resulting in greater resistance of ecosystem function to precipitation change. To investigate this interaction between composition and precipitation change we performed a field experiment for three years in successional grassland in southern England. This consisted of two treatments. The first, precipitation change, simulated end of century predictions, and consisted of a summer drought phase alongside winter rainfall addition. The second, functional group identity, divided the plant community into three groups based on their functional traits- broadly described as perennials, caespitose grasses and annuals- and removed these groups in a factorial design. Ecosystem functions related to C, N and water cycling were measured regularly. Effects of functional groupidentity were apparent, with the dominant trend being that process rates were higher under control conditions where a range of perennial species were present. E.g. litter decomposition rates were significantly higher in plots containing several perennial species, the group with the highest average leaf N content. Process rates were also very strongly affected by the precipitation change treatmentwhen perennial plant species were dominant, but not where the community contained a high abundance of annual species and caespitose grasses. This contrasting response could be attributable to differing rooting patterns (shallower structures under annual plants, and deeper roots under perennials and faster nutrient uptake in annuals compared to perennials. Our results indicate that precipitation change will have a smaller effect on key process rates in grasslandscontaining a range of perennial

  7. Modified cellulose synthase gene from 'Arabidopsis thaliana' confers herbicide resistance to plants

    Energy Technology Data Exchange (ETDEWEB)

    Somerville, Chris R.; Scieble, Wolf

    2000-10-11

    Cellulose synthase ('CS'), a key enzyme in the biosynthesis of cellulose in plants is inhibited by herbicides comprising thiazolidinones such as 5-tert-butyl-carbamoyloxy-3-(3-trifluromethyl) phenyl-4-thiazolidinone (TZ), isoxaben and 2,6-dichlorobenzonitrile (DCB). Two mutant genes encoding isoxaben and TZ-resistant cellulose synthase have been isolated from isoxaben and TZ-resistant Arabidopsis thaliana mutants. When compared with the gene coding for isoxaben or TZ-sensitive cellulose synthase, one of the resistant CS genes contains a point mutation, wherein glycine residue 998 is replaced by an aspartic acid. The other resistant mutation is due to a threonine to isoleucine change at amino acid residue 942. The mutant CS gene can be used to impart herbicide resistance to a plant; thereby permitting the utilization of the herbicide as a single application at a concentration which ensures the complete or substantially complete killing of weeds, while leaving the transgenic crop plant essentially undamaged.

  8. Lusus naturae:climate and invasions of plant pathogens modify agricultural and forest lands

    Directory of Open Access Journals (Sweden)

    Alessandro Ragazzi

    2009-10-01

    Full Text Available The ecological and economic sustainability of agricultural and forest systems of many advanced and underdeveloped Countries are strongly threatened by the increasing introduction of exotic plant pathogens. This article provides an overview of the main causes behind these invasions. Some important diseases caused by non native phytopathogens, whose arrival in the past century had a disastrous impact on the environment and economy of vast rural areas of our Country are reported. Some dangerous, emerging pathogens, which are literally destroying whole territories in various parts of the Planet, with severe damage to agricultural crops, landscape, economy and local tourism are also reported. Action strategies to prevent immigration of unwanted pathogens, and mitigation strategies, aimed at the development of various measures to mitigate the negative effects of plant parasites already established in the territory are then discussed. Finally, it is highlighted how such a far-reaching problem can be properly tackled only with the active contribution of governments, institutions responsible for plant health monitoring (warning services, research, and agricultural, tourism and transport operators.

  9. Impact of ubiquitous inhibitors on the GUS gene reporter system: evidence from the model plants Arabidopsis, tobacco and rice and correction methods for quantitative assays of transgenic and endogenous GUS

    Directory of Open Access Journals (Sweden)

    Gerola Paolo D

    2009-12-01

    Full Text Available Abstract Background The β-glucuronidase (GUS gene reporter system is one of the most effective and employed techniques in the study of gene regulation in plant molecular biology. Improving protocols for GUS assays have rendered the original method described by Jefferson amenable to various requirements and conditions, but the serious limitation caused by inhibitors of the enzyme activity in plant tissues has thus far been underestimated. Results We report that inhibitors of GUS activity are ubiquitous in organ tissues of Arabidopsis, tobacco and rice, and significantly bias quantitative assessment of GUS activity in plant transformation experiments. Combined with previous literature reports on non-model species, our findings suggest that inhibitors may be common components of plant cells, with variable affinity towards the E. coli enzyme. The reduced inhibitory capacity towards the plant endogenous GUS discredits the hypothesis of a regulatory role of these compounds in plant cells, and their effect on the bacterial enzyme is better interpreted as a side effect due to their interaction with GUS during the assay. This is likely to have a bearing also on histochemical analyses, leading to inaccurate evaluations of GUS expression. Conclusions In order to achieve reliable results, inhibitor activity should be routinely tested during quantitative GUS assays. Two separate methods to correct the measured activity of the transgenic and endogenous GUS are presented.

  10. Durability testing modified compression ignition engines fueled with straight plant oil

    Energy Technology Data Exchange (ETDEWEB)

    Basinger, M.; Lackner, K.S. [Earth and Environmental Engineering, Columbia University, New York City 10027 (United States); Reding, T. [Mechanical Engineering, Manhattan College, New York City (United States); Rodriguez-Sanchez, F.S. [Mali Biocarburant, Bamako (Mali); Modi, V. [Mechanical Engineering, Columbia University, New York City 10027 (United States)

    2010-08-15

    Many short-run studies point to the potential for direct fueling of compression ignition engines with plant oil fuels. There is a much smaller body of work that examines the potential for these fuels in long-run tests that illuminate engine endurance and longevity issues. Generally, longevity studies involving direct fueling of engines with straight plant oils have shown significant impact to the life of the engine, though test results vary widely depending on the oil, engine type, test conditions, and measurement approach. This study utilizes a previously designed modification kit to investigate the longevity implications of directly fueling straight plant oil in an indirect injection (IDI) listeroid type, slow speed stationary engine common in agro-processing applications in developing countries. Specifically this study focuses on the lubrication oil by developing a model to characterize the engine wear and estimate lube oil change frequency. The model is extended to an analysis of the piston rings. Cylinder liner wear, emissions, engine performance, and a visual investigation of several critical engine components are also studied. The 500 hour test with waste vegetable oil fuel resulted in several important findings. The engine break-in period was identified as taking between 200 and 300 h. Emissions analysis supported the break-in definition as smoke opacity and carbon monoxide values fell from 9% and 600 ppm (respectively) during the first few hundred hours, to 5% and 400 ppm in the final 200 h. Lubrication oil viscosity was found to be the limiting degradation factor in the lube oil, requiring oil to be changed every 110 h. Piston ring mass loss was found to correlate very closely with chromium buildup in the lubrication oil and the mathematical model that was developed was used to estimate that piston ring inspection and replacement should occur after 1000 h. Cylinder ovalisation was found to be most sever at top dead center (TDC) at 53 microns of averaged

  11. A modified MS2 bacteriophage plaque reduction assay for the rapid screening of antiviral plant extracts

    Directory of Open Access Journals (Sweden)

    Ian Cock

    2010-01-01

    Full Text Available Introduction: Traditional methods of screening plant extracts and purified components for antiviral activity require up to a week to perform, prompting the need to develop more rapid quantitative methods to measure the ability of plant based preparations to block viral replication. We describe an adaption of an MS2 plaque reduction assay for use in S. aureus. Results: MS2 bacteriophage was capable of infecting and replicating in B. cereus, S. aureus and F+ E. coli but not F- E. coli. Indeed, both B. cereus and S. aureus were more sensitive to MS2 induced lysis than F+ E. coli. When MS2 bacteriophage was mixed with Camellia sinensis extract (1 mg/ml, Scaevola spinescens extract (1 mg/ml or Aloe barbadensis juice and the mixtures inoculated into S. aureus, the formation of plaques was reduced to 8.9 ± 3.8%, 5.4 ± 2.4% and 72.7 ± 20.9% of the untreated MS2 control values respectively. Conclusions: The ability of the MS2 plaque reduction assay to detect antiviral activity in these known antiviral plant preparations indicates its suitability as an antiviral screening tool. An advantage of this assay compared with traditionally used cytopathic effect reduction assays and replicon based assays is the more rapid acquisition of results. Antiviral activity was detected within 24 h of the start of testing. The MS2 assay is also inexpensive and non-pathogenic to humans making it ideal for initial screening studies or as a simulant for pathogenic viruses.

  12. An intergenic region shared by At4g35985 and At4g35987 in Arabidopsis thaliana is a tissue specific and stress inducible bidirectional promoter analyzed in transgenic arabidopsis and tobacco plants.

    Directory of Open Access Journals (Sweden)

    Joydeep Banerjee

    Full Text Available On chromosome 4 in the Arabidopsis genome, two neighboring genes (calmodulin methyl transferase At4g35987 and senescence associated gene At4g35985 are located in a head-to-head divergent orientation sharing a putative bidirectional promoter. This 1258 bp intergenic region contains a number of environmental stress responsive and tissue specific cis-regulatory elements. Transcript analysis of At4g35985 and At4g35987 genes by quantitative real time PCR showed tissue specific and stress inducible expression profiles. We tested the bidirectional promoter-function of the intergenic region shared by the divergent genes At4g35985 and At4g35987 using two reporter genes (GFP and GUS in both orientations in transient tobacco protoplast and Agro-infiltration assays, as well as in stably transformed transgenic Arabidopsis and tobacco plants. In transient assays with GFP and GUS reporter genes the At4g35985 promoter (P85 showed stronger expression (about 3.5 fold compared to the At4g35987 promoter (P87. The tissue specific as well as stress responsive functional nature of the bidirectional promoter was evaluated in independent transgenic Arabidopsis and tobacco lines. Expression of P85 activity was detected in the midrib of leaves, leaf trichomes, apical meristemic regions, throughout the root, lateral roots and flowers. The expression of P87 was observed in leaf-tip, hydathodes, apical meristem, root tips, emerging lateral root tips, root stele region and in floral tissues. The bidirectional promoter in both orientations shows differential up-regulation (2.5 to 3 fold under salt stress. Use of such regulatory elements of bidirectional promoters showing spatial and stress inducible promoter-functions in heterologous system might be an important tool for plant biotechnology and gene stacking applications.

  13. Debating the future of genetically modified plants - bridging knowledge dimensions. A technology foresight study

    DEFF Research Database (Denmark)

    Borch, Kristian; Rasmussen, Birgitte

    2003-01-01

    to offer a coordinating method for developing and strengthening those linkages. To test this, a technological foresight study was performed on genetically modified (GM) crop technology in the Danish context. Thebackground to the study was the conflict and intense debate in Denmark over applications of gene...... technology, and especially over the deliberate release of GM crops. However, the current debate characteristically involves sharply opposed fronts. In it,stakeholders and experts on both side of the conflict advocate widely differing opinions. Without a proper, generally intelligible dialogue, the broader...... public audience finds it hard to comprehend this type of debate. The study pursues the notion thatpublic dialogue can act as a driver of future applications in the technological domain, specifically GM crops. The study concluded with a stakeholder workshop that revealed three key issues that might...

  14. Scientific Opinion on the assessment of potential impacts of genetically modified plants on non-target organisms

    DEFF Research Database (Denmark)

    Arpaia, Salvatore; Bartsch, Detlef; Delos, Marc

    , for which assessment and measurement endpoints shall be developed; the need to initiate the scientific risk assessment by setting testable hypotheses; criteria for appropriate selection of test species and ecological functional groups; appropriate laboratory and field studies to collect relevant NTO data......The European Food Safety Authority (EFSA) asked the Panel on Genetically Modified Organisms to establish a self-tasking Working Group with the aim of (1) producing a scientific review of the current guidance of the GMO Panel for Environmental Risk Assessment (ERA), focusing on the potential impacts...... and individual species within a tiered approach. The present scientific opinion provides guidance to risk assessors for assessing potential effects of GM plants on NTOs, together with rationale for data requirements in order to complete a comprehensive ERA for NTOs. In this respect, guidance to applicants...

  15. Herbicide resistance and biodiversity: agronomic and environmental aspects of genetically modified herbicide-resistant plants.

    Science.gov (United States)

    Schütte, Gesine; Eckerstorfer, Michael; Rastelli, Valentina; Reichenbecher, Wolfram; Restrepo-Vassalli, Sara; Ruohonen-Lehto, Marja; Saucy, Anne-Gabrielle Wuest; Mertens, Martha

    2017-01-01

    Farmland biodiversity is an important characteristic when assessing sustainability of agricultural practices and is of major international concern. Scientific data indicate that agricultural intensification and pesticide use are among the main drivers of biodiversity loss. The analysed data and experiences do not support statements that herbicide-resistant crops provide consistently better yields than conventional crops or reduce herbicide amounts. They rather show that the adoption of herbicide-resistant crops impacts agronomy, agricultural practice, and weed management and contributes to biodiversity loss in several ways: (i) many studies show that glyphosate-based herbicides, which were commonly regarded as less harmful, are toxic to a range of aquatic organisms and adversely affect the soil and intestinal microflora and plant disease resistance; the increased use of 2,4-D or dicamba, linked to new herbicide-resistant crops, causes special concerns. (ii) The adoption of herbicide-resistant crops has reduced crop rotation and favoured weed management that is solely based on the use of herbicides. (iii) Continuous herbicide resistance cropping and the intensive use of glyphosate over the last 20 years have led to the appearance of at least 34 glyphosate-resistant weed species worldwide. Although recommended for many years, farmers did not counter resistance development in weeds by integrated weed management, but continued to rely on herbicides as sole measure. Despite occurrence of widespread resistance in weeds to other herbicides, industry rather develops transgenic crops with additional herbicide resistance genes. (iv) Agricultural management based on broad-spectrum herbicides as in herbicide-resistant crops further decreases diversity and abundance of wild plants and impacts arthropod fauna and other farmland animals. Taken together, adverse impacts of herbicide-resistant crops on biodiversity, when widely adopted, should be expected and are indeed very hard

  16. Mesoporous Silicon with Modified Surface for Plant Viruses and Their Protein Particle Sensing

    Directory of Open Access Journals (Sweden)

    Kae Dal Kwack

    2008-10-01

    Full Text Available Changes in electric parameters of a mesoporous silicon treated by a plasma chemical etching with fluorine and hydrogen ions, under the adsorption of NEPO (Nematodetransmitted Polyhedral plant viruses such as TORSV (Tomato Ringspot Virus, GFLV (Grapevine Fan Leaf Virus and protein macromolecule from TORSV particles are described. The current response to the applied voltage is measured for each virus particle to investigate the material parameters which are sensitive to the adsorbed particles. The peculiar behaviors of the response are modeled by the current-voltage relationship in a MOSFET. This model explains the behavior well and the double gate model of the MOSFET informs that the mesoporous silicon is a highly sensitive means of detecting the viruses in the size range less than 50 nm.

  17. Monitoring the persistence of genes deriving from genetically modified plants in the soil environment.

    Science.gov (United States)

    Degand, I; Laporte, J; Pussemier, L

    2002-01-01

    To study the gene persistence in the soil environment, soil samples were collected from sugar beet (Beta vulgaris) and chicory (Cichorium intybus) experimental fields just before harvest. They were homogenized, mixed and stored at constant humidity in a non-heated room. Sub-samples of soils were subsequently collected at regular intervals, dried and sieved through a 1.8-mm mesh before DNA was extracted. Specific primers were then used for the detection of plant DNA by hot start PCR. Results reveal that, under laboratory conditions, transgenic and non-transgenic sugar beet DNA was still detected after 25 days incubation in the soil taken from a sugar beet experimental plot while detection of chicory DNA was still possible after 50 days incubation in soil taken from the chicory experimental plot. This might be in correlation with the stronger resistance of chicory radicles to decomposition as compared to radicles from sugar beets.

  18. DAILY SCHEDULING OF SMALL HYDRO POWER PLANTS DISPATCH WITH MODIFIED PARTICLES SWARM OPTIMIZATION

    Directory of Open Access Journals (Sweden)

    Sinvaldo Rodrigues Moreno

    2015-04-01

    Full Text Available This paper presents a new approach for short-term hydro power scheduling of reservoirs using an algorithm-based Particle Swarm Optimization (PSO. PSO is a population-based algorithm designed to find good solutions to optimization problems, its characteristics have encouraged its adoption to tackle a variety of problems in different fields. In this paper the authors consider an optimization problem related to a daily scheduling of small hydro power dispatch. The goal is construct a feasible solution that maximize the cascade electricity production, following the environmental constraints and water balance. The paper proposes an improved Particle Swarm Optimization (PSO algorithm, which takes advantage of simplicity and facility of implementation. The algorithm was successfully applied to the optimization of the daily schedule strategies of small hydro power plants, considering maximum water utilization and all constraints related to simultaneous water uses. Extensive computational tests and comparisons with other heuristics methods showed the effectiveness of the proposed approach.

  19. Modifying woody plants for efficient conversion to liquid and gaseous fuels

    Energy Technology Data Exchange (ETDEWEB)

    Dinus, R.J.; Dimmel, D.R.; Feirer, R.P.; Johnson, M.A.; Malcolm, E.W. (Institute of Paper Science and Technology, Atlanta, GA (USA))

    1990-07-01

    The Short Rotation Woody Crop Program (SRWCP), Department of Energy, is developing woody plant species as sources of renewable energy. Much progress has been made in identifying useful species, and testing site adaptability, stand densities, coppicing abilities, rotation lengths, and harvesting systems. Conventional plant breeding and intensive cultural practices have been used to increase above-ground biomass yields. Given these and foreseeable accomplishments, program leaders are now shifting attention to prospects for altering biomass physical and chemical characteristics, and to ways for improving the efficiency with which biomass can be converted to gaseous and liquid fuels. This report provides a review and synthesis of literature concerning the quantity and quality of such characteristics and constituents, and opportunities for manipulating them via conventional selection and breeding and/or molecular biology. Species now used by SRWCP are emphasized, with supporting information drawn from others as needed. Little information was found on silver maple (Acer saccharinum), but general comparisons (Isenberg 1981) suggest composition and behavior similar to those of the other species. Where possible, conclusions concerning means for and feasibility of manipulation are given, along with expected impacts on conversion efficiency. Information is also provided on relationships to other traits, genotype X environment interactions, and potential trade-offs or limitations. Biomass productivity per se is not addressed, except in terms of effects that may by caused by changes in constituent quality and/or quantity. Such effects are noted to the extent they are known or can be estimated. Likely impacts of changes, however effected, on suitability or other uses, e.g., pulp and paper manufacture, are notes. 311 refs., 4 figs., 9 tabs.

  20. The Complexity of Tobacco and Tobacco Smoke

    Directory of Open Access Journals (Sweden)

    Perfetti TA

    2014-12-01

    Full Text Available Tobacco and tobacco smoke are both complex mixtures. We previously reported 8430 unique chemical components identified in these complex mixtures but two years later our updated number was 8889. Addition of unlisted isomers raised these numbers to 8622 and 9081, respectively. Our previous number of 4994 identified tobacco components is now 5229; our previous number of 5315 identified tobacco smoke components is now 5685. An operational definition of a complex mixture is as follows: A complex mixture is a characterizable substance containing many chemical components (perhaps thousands in inexact proportions.

  1. Spatial Variability and Comprehensive Suitability Evaluation of Soil Fertility Factors in Xiangcheng Tobacco Planting Region%河南襄城植烟田土壤肥力空间变异性与适宜性综合评价

    Institute of Scientific and Technical Information of China (English)

    陈海生; 裴孝钟; 魏跃伟; 刘国顺

    2011-01-01

    It is essential to understand spital variability of soil properties in tobacco planting regions, for the right decision can be made as to how many fertilizer rates are likely to be appropriate for the high quality tobacco production. In this paper, a total of 159 soil samples were taken from surface soil(0 ~20) of Xiangcheng tobacco planting fields, Henan province in April 2007 for examing the concentration of soil organic matter, pH, available N, available P and available K . Spatial variability of above mentioned was evaluated using geostatistics and geographic information system( GIS) analyses. The objective of this study was to provide a scientific basis for field management targeting at improving soil quality in this tobacco planting region. The results showed that among those five soil fertility factors, the variation coefficient of soil pH is the smallest, while that of available P is the greatest which demands more samples under the same precisions, which are resulted from different cultivation methods and uneven fertilization. Analysis of the isotropic variogram indicated that the soil pH, organic matter and available P semivariogram were well described with spherical models,with the distance of spatial dependence being 21 020,19 150 and 8 460 m respectively,while the available N and available K semivariogram were well described with exponential model, with the distance of spatial dependence being ranged from 7 484 to 25 320 m. Soil pH value was strongly spatiallydependence with CO/sill being 0. 193 5 , while other four soil fertility factors were moderately spatial dependent with CO/sill being range from 0. 352 8 to 0. 526 0. With the kriging analysids, the spatial distribution maps of soil properties in Xiangcheng tobacco planting regions was drawn. In the paper, the index system of soil fertility suitability of tobacco was established. S-or parabola-type dependence functions were developed according to their effect on tobacco crop and the function value

  2. Silencing of potato virus X coat protein gene in transgenic tobaccos by codon replacement that confers resistance to PVX infection

    Institute of Scientific and Technical Information of China (English)

    FENG Dejiang; LIU Xiang; MENG Kun; LIAO Lili; WEI Xiaoli; XU Honglin; ZHU Zhen

    2003-01-01

    To understand the effect of rare codon on the silencing ratio of foreign gene, some preferred codon in potato virus X (PVX) coat protein gene (cp) were substituted with synonymous rare codons. The modified PVX coat protein gene (cpm) and wild-type cp gene (cpw) were inserted into binary vector under the control of CaMV35S promoter, and these two plant expression constructs were transferred into tobacco (Nicotiana tabacum cv. Xanthi) genomes via Agrobacterium mediated method and transgenic plants were generated. Northern blot analysis of RNA isolated from these plants showed that the silencing ratio of cpm gene in transgenic tobaccos was higher than that of cpw (35% and 6.25% respectively). Run on results indicate that the silencing of cp gene happened at post-transcriptional level. The resistance of transgenic tobaccos carrying cpm genes to PVX is increased compared with that of transformants carrying cpw genes. These results suggest that the resistance of transgenic tobacco to PVX can be enhanced by codon replacement.

  3. Larval host plant origin modifies the adult oviposition preference of the female European grapevine moth Lobesia botrana

    Science.gov (United States)

    Moreau, J.; Rahme, J.; Benrey, B.; Thiery, D.

    2008-04-01

    According to the ‘natal habitat preference induction’ (NHPI) hypothesis, phytophagous insect females should prefer to lay their eggs on the host species on which they developed as larvae. We tested whether this hypothesis applies to the breeding behaviour of polyphagous European grapevine moth, Lobesia botrana, an important pest in European vineyards. We previously found that different grape cultivars affect several life history traits of the moth. Because the different cultivars of grapes are suspected to provide different plant quality, we tested the NHPI hypothesis by examining oviposition choice of L. botrana among three Vitis vinifera cultivars (Pinot, Chasselas and Chardonnay). In a choice situation, females of L. botrana that had never experienced grapes were able to discriminate between different grape cultivars and preferentially selected Pinot as an oviposition substrate. This ‘naive’ preference of oviposition could be modified by larval environment: Females raised on grapes as larvae preferred to lay eggs on the cultivar that they had experienced. Furthermore, experience of the host plant during adult emergence could be excluded because when pupae originating from our synthetic diet were exposed to grapes, the emerging adults did not show preference for the cultivar from which they emerged. The NHPI hypothesis that includes the two sub-hypothesis “Hopkins host selection principle” and “chemical legacy” may thus be relevant in this system.

  4. Enzymatic glucose sensor based on Au nanoparticle and plant-like ZnO film modified electrode

    Energy Technology Data Exchange (ETDEWEB)

    Tian, Kun [Nanostructured Materials Research Laboratory, Department of Materials Science and Engineering, University of Utah, Salt Lake City, UT 84112 (United States); Alex, Saji [Nanostructured Materials Research Laboratory, Department of Materials Science and Engineering, University of Utah, Salt Lake City, UT 84112 (United States); Department of Chemistry, Government College for Women, Thiruvananthapuram, Kerala 695014 (India); Siegel, Gene [Nanostructured Materials Research Laboratory, Department of Materials Science and Engineering, University of Utah, Salt Lake City, UT 84112 (United States); Tiwari, Ashutosh, E-mail: tiwari@eng.utah.edu [Nanostructured Materials Research Laboratory, Department of Materials Science and Engineering, University of Utah, Salt Lake City, UT 84112 (United States)

    2015-01-01

    A novel electrochemical glucose sensor was developed by employing a composite film of plant-like Zinc oxide (ZnO) and chitosan stabilized spherical gold nanoparticles (AuNPs) on which Glucose oxidaze (GOx) was immobilized. The ZnO was deposited on an indium tin oxide (ITO) coated glass and the AuNPs of average diameter of 23 nm were loaded on ZnO as the second layer. The prepared ITO/ZnO/AuNPs/GOx bioelectrode exhibited a low value of Michaelis–Menten constant of 1.70 mM indicating a good bio-matrix for GOx. The studies of electrochemical properties of the electrode using cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) showed that, the presence of AuNPs provides significant enhancement of the electron transfer rate during redox reactions. The linear sweep voltammetry (LSV) shows that the ITO/ZnO/AuNPs/GOx based sensor has a high sensitivity of 3.12 μA·mM{sup −1}·cm{sup −2} in the range of 50 mg/dL to 400 mg/dL glucose concentration. The results show promising application of the gold nanoparticle modified plant-like ZnO composite bioelectrode for electrochemical sensing of glucose.

  5. Molecular characterization of denitrifying bacteria isolated from the anoxic reactor of a modified DEPHANOX plant performing enhanced biological phosphorus removal.

    Science.gov (United States)

    Zafiriadis, Ilias; Ntougias, Spyridon; Mirelis, Paraskevi; Kapagiannidis, Anastasios G; Aivasidis, Alexander

    2012-06-01

    Enhanced Biological Phosphorus Removal (EBPR) under anoxic conditions was achieved using a Biological Nutrient Removal (BNR) system based on a modification of the DEPHANOX configuration. Double-probe Fluorescence in Situ Hybridization (FISH) revealed that Polyphosphate Accumulating Organisms (PAOs) comprised 12.3 +/- 3.2% of the total bacterial population in the modified DEPHANOX plant. The growing bacterial population on blood agar and Casitone Glycerol Yeast Autolysate agar (CGYA) medium was 16.7 +/- 0.9 x 10(5) and 3.0 +/- 0.6 x 10(5) colony forming units (cfu) mL(-1) activated sludge, respectively. A total of 121 bacterial isolates were characterized according to their denitrification ability, with 26 bacterial strains being capable of reducing nitrate to gas. All denitrifying isolates were placed within the alpha-, beta-, and gamma-subdivisions of Proteobacteria and the family Flavobacteriaceae. Furthermore, a novel denitrifying bacterium within the genus Pseudomonas was identified. This is the first report on the isolation and molecular characterization of denitrifying bacteria from EBPR sludge using a DEPHANOX-type plant.

  6. Youth and tobacco.

    Science.gov (United States)

    Tanski, S E; Prokhorov, A V; Klein, J D

    2004-12-01

    Youth around the world take up smoking and use tobacco products at high rates. Young people may not grasp the long-term consequences of tobacco use, although tobacco consumption and exposure has been shown to have significant negative health effects. Youth use a variety of tobacco products that are smoked, chewed, or sniffed, including machine-manufactured cigarettes, cigars, bidis, kreteks, sticks, and snuff. Prevention efforts have focused on countering those aspects that are believed to contribute to smoking uptake, such as tobacco industry advertising and promotion, and access to tobacco. There are many aspects of tobacco promotion through the media that have been more difficult to control, however, such as product placement within popular cinema movies. Once a youth has taken up tobacco, he or she is more likely than an adult to become addicted and should be offered treatment for tobacco cessation. Although there is not yet sufficient evidence to prove efficacy, the same treatments are suggested for youth as are recommended for adults, including nicotine replacement products. Given the severity of the tobacco epidemic worldwide and the devastating health effects on an individual and population basis, there are currently many efforts to curtail the tobacco problem, including the World Health Organization (WHO) sponsored Framework Convention on Tobacco Control. It is through comprehensive and collaborative efforts such as this that the global hazard of tobacco is most likely to be overcome.

  7. Overexpression of Arabidopsis MiR396 Enhances Drought Tolerance in Transgenic Tobacco Plants%高表达拟南芥miR396提高烟草抗旱性

    Institute of Scientific and Technical Information of China (English)

    杨凤玺; 余迪求

    2009-01-01

    MiR396是一个由21个核苷酸组成的单链非编码RNA小分子.烟草内的miR396受干旱诱导说明其可能参与烟草的干旱应答.在35S强启动子作用下我们将miR396转入到烟草体内获得高表达转基因植株,生理学测试表明高表达miR396的转基因烟草耐旱性增强,同时叶片表现出比野生型较低的失水率和较高的相对含水量,进一步分析表明转基因植株除了叶片变得更为窄小外,其气孔密度和气孔系数都比野生型降低,这些都表明miR396作为一个正调节因子参与烟草的干旱胁迫应答.%MiR396 was a single-stranded noncoding small RNA with 21 nucleotides, and the expression of MiR369 in leaves was strongly induced in water deficit condition in tobacco, which suggested a possible role of miR396 in drought stress response. Under the control of 35S promoter, MIR396 was introduced into tobacco mediated by Agrobacterium tumefaciens. Physiological tests indicated that the elevated levels of miR396 increased drought tolerance in tobacco accompanying with lower water loss rate and higher relative water content. Further more miR396-overexpressing plants exhibited visible reductions both in stomatal density and stomatal index as well as a narrow and small leaf phenotype in comparison with wild-type plants. The present study indicated that miR396 was a positive regulator in response to drought stress in tobacco.

  8. Trans-α-xylosidase and trans-β-galactosidase activities, widespread in plants, modify and stabilize xyloglucan structures.

    Science.gov (United States)

    Franková, Lenka; Fry, Stephen C

    2012-07-01

    Cell-wall components are hydrolysed by numerous plant glycosidase and glycanase activities. We investigated whether plant enzymes also modify xyloglucan structures by transglycosidase activities. Diverse angiosperm extracts exhibited transglycosidase activities that progressively transferred single sugar residues between xyloglucan heptasaccharide (XXXG or its reduced form, XXXGol) molecules, at 16 μM and above, creating octa- to decasaccharides plus smaller products. We measured remarkably high transglycosylation:hydrolysis ratios under optimized conditions. To identify the transferred monosaccharide(s), we devised a dual-labelling strategy in which a neutral radiolabelled oligosaccharide (donor substrate) reacted with an amino-labelled non-radioactive oligosaccharide (acceptor substrate), generating radioactive cationic products. For example, 37 μM [Xyl-³H]XXXG plus 1 mM XXLG-NH₂ generated ³H-labelled cations, demonstrating xylosyl transfer, which exceeded xylosyl hydrolysis 1.6- to 7.3-fold, implying the presence of enzymes that favour transglycosylation. The transferred xylose residues remained α-linked but were relatively resistant to hydrolysis by plant enzymes. Driselase digestion of the products released a trisaccharide (α-[³H]xylosyl-isoprimeverose), indicating that a new xyloglucan repeat unit had been formed. In similar assays, [Gal-³H]XXLG and [Gal-³H]XLLG (but not [Fuc-³H]XXFG) yielded radioactive cations. Thus plants exhibit trans-α-xylosidase and trans-β-galactosidase (but not trans-α-fucosidase) activities that graft sugar residues from one xyloglucan oligosaccharide to another. Reconstructing xyloglucan oligosaccharides in this way may alter oligosaccharin activities or increase their longevity in vivo. Trans-α-xylosidase activity also transferred xylose residues from xyloglucan oligosaccharides to long-chain hemicelluloses (xyloglucan, water-soluble cellulose acetate, mixed-linkage β-glucan, glucomannan and arabinoxylan). With

  9. Smokeless Tobacco Consumption by Mexican-American University Students.

    Science.gov (United States)

    Lopez, Linda C.

    A modified version of the Illinois Department of Public Health Tobacco Use Survey was used to assess smokeless tobacco consumption among students attending a state university in New Mexico. Respondents included 65 male and 83 female Mexican-Americans, as well as 59 male and 118 female Anglo-Americans. Ages ranged from 16 to 67; subgroup median…

  10. The use of statistical tools in field testing of putative effects of genetically modified plants on nontarget organisms.

    Science.gov (United States)

    Semenov, Alexander V; Elsas, Jan Dirk; Glandorf, Debora C M; Schilthuizen, Menno; Boer, Willem F

    2013-08-01

    To fulfill existing guidelines, applicants that aim to place their genetically modified (GM) insect-resistant crop plants on the market are required to provide data from field experiments that address the potential impacts of the GM plants on nontarget organisms (NTO's). Such data may be based on varied experimental designs. The recent EFSA guidance document for environmental risk assessment (2010) does not provide clear and structured suggestions that address the statistics of field trials on effects on NTO's. This review examines existing practices in GM plant field testing such as the way of randomization, replication, and pseudoreplication. Emphasis is placed on the importance of design features used for the field trials in which effects on NTO's are assessed. The importance of statistical power and the positive and negative aspects of various statistical models are discussed. Equivalence and difference testing are compared, and the importance of checking the distribution of experimental data is stressed to decide on the selection of the proper statistical model. While for continuous data (e.g., pH and temperature) classical statistical approaches - for example, analysis of variance (ANOVA) - are appropriate, for discontinuous data (counts) only generalized linear models (GLM) are shown to be efficient. There is no golden rule as to which statistical test is the most appropriate for any experimental situation. In particular, in experiments in which block designs are used and covariates play a role GLMs should be used. Generic advice is offered that will help in both the setting up of field testing and the interpretation and data analysis of the data obtained in this testing. The combination of decision trees and a checklist for field trials, which are provided, will help in the interpretation of the statistical analyses of field trials and to assess whether such analyses were correctly applied. We offer generic advice to risk assessors and applicants that will

  11. Expression of Recombinant Tryptophan Decarboxylase in Different Subcellular Compartments in Tobacco Plant%重组色氨酸脱羧酶在烟草不同亚细胞区室的表达

    Institute of Scientific and Technical Information of China (English)

    王淼; 李秋荣; Stefano Di DIORE

    2002-01-01

    The gene encoded for tryptophan decarboxylase (TDC), which is the key enzyme in terpenoil indole alkaloids pathway, was targeted to different subcellular compartments and stably expressed in transgenic tobacco (Nicotiana tabacum L.) plants at the levels detected by Western blot and tryptamine accumulation analysis. It was shown that the TDC was located in subcellular compartments, the chloroplasts and cytosol. The recombinant TDC targeted to chloroplasts and cytosol in tobacco plants was effectively expressed as soluble protein by Western blot analysis and enzymatic assay. The level of tryptamine accumulation in chloroplast was higher than that in cytosol and very low in vacuole and endoplasmic reticulum (ER) to be hardly detected by Western blot analysis. It was indicated that the highest amount of tryptamine was in chloroplasts, lower in endoplasmic reticula and the lowest in vacuoles as compared to those in wild type plants. The TDC targeted to different subcellular compartments of tobacco plants and its expression level were studied by different nucleotide sequences coding signal peptides at 5′-end of tdc gene in order to know the effects of the TDC in compartmentation on its functionality.%将萜烯类吲哚生物碱代谢关键酶--色氨酸脱羧酶(TDC)的编码基因转到烟草(Nicotiana tabacum L.)植物体内,标定在不同的亚细胞区室表达。通过蛋白免疫印迹法和色胺在植物体内的累积量测定分析,对转基因植物进行筛选。结果表明,TDC在叶绿体和胞液中高效表达,TDC在叶绿体中的表达水平最高,高于在胞液中的表达,在内质网和液泡中表达水平很低,用蛋白免疫印迹法未检出。

  12. Default cycle phases determined after modifying discrete DNA sequences in plant cells

    Energy Technology Data Exchange (ETDEWEB)

    Sans, J.; Leyton, C. [Universidad de Chile, Santiago (Chile). Facultad de Medicina; Gimenez-Abian, M.I.; Gimenez-Abian, J.F.; Aller, P.; De La Torre, C. [Centro de Investigaciones Biologicas, CSIC, Madrid (Spain)

    1997-02-01

    After bromosubstituting DNA sequences replicated in the first, second, or third part of the S phase, in Allium cepa L. meristematic cells, radiation at 313 nm wavelength under anoxia allowed ascription of different sequences to both the positive and negative regulation of some cycle phase transitions. The present report shows that the radiation forced cells in late G{sub 1} phase to advance into S, while those in G{sub 2} remained in G{sub 2} and cells in prophase returned to G{sub 2} when both sets of sequences involved in the positive and negative controls were bromosubstituted and later irradiated. In this way, not only G{sub 2} but also the S phase behaved as cycle phases where cells accumulated by default when signals of different sign functionally cancelled out. The treatment did not halt the rates of replication or transcription of plant bromosubstituted DNA. The irradiation under hypoxia apparently prevents the binding of regulatory proteins to Br-DNA. (author).

  13. Earthworms and Plant Residues Modify Nematodes in Tropical Cropping Soils (Madagascar: A Mesocosm Experiment

    Directory of Open Access Journals (Sweden)

    Cécile Villenave

    2010-01-01

    Full Text Available Free-living nematodes present several characteristics that have led to their use as bioindicators of soil quality. Analyzing the structure of nematofauna is a pertinent way to understand soil biological processes. Earthworms play an important role in soil biological functioning and organic matter dynamics. Their effects on soil nematofauna have seldom been studied. We studied the effect of the tropical endogeic earthworm, Pontoscolex corethrurus, on nematode community structure in a 5-month field mesocosm experiment conducted in Madagascar. Ten different treatments with or without earthworms and with or without organic residues (rice, soybean were compared. Organic residues were applied on the soil surface or mixed with the soil. The abundance of nematodes (bacterial and fungal feeders was higher in presence of P. corethrurus than in their absence. The type of plant residues as well as their localisation had significant effects on the abundance and composition of soil nematodes. The analysis of nematode community structure showed that earthworm activity led to an overall activation of the microbial compartment without specific stimulation of the bacterial or fungal compartment.

  14. Overexpressing tagged proteins in plants using a modified gateway cloning strategy.

    Science.gov (United States)

    Dubin, Manu J; Bowler, Chris; Benvenuto, Giovanna

    2010-03-01

    In recent years, sequence-specific recombination cloning methods such as the Gateway system have become increasingly popular for (over)expressing tagged proteins in high-throughput investigations in many different organisms, including plants. Because of their versatility and ease of use, these methods have gained favor in low- and medium-throughput investigations as well. However, due to the recombination step, the resulting fusion proteins contain long and often highly charged polylinker sequences that can interfere with their physiological function. Furthermore, in some cases the gene of interest must be cloned twice (once with and once without a stop codon) for N- and C-terminal tagging. Here, we present a hybrid combinatorial cloning strategy that overcomes many of these limitations. In the first step, the gene of interest is cloned into an entry vector containing standardized cloning sites with the desired N- or C-terminal tag and an optimized polylinker sequence. A Gateway recombination reaction is used to transfer the protein-tag fusion from the entry clone to a Gateway destination vector with the desired promoter and selectable marker for the organism of interest. As experimental requirements evolve, constructs for expressing the protein of interest with the desired tag, promoter, and selectable marker or other features can rapidly and easily be created.

  15. 27 CFR 19.914 - Medium plants.

    Science.gov (United States)

    2010-04-01

    ... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Medium plants. 19.914 Section 19.914 Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE BUREAU, DEPARTMENT... Medium plants. Any person wishing to establish a medium plant shall make application for and obtain in...

  16. A gene for plant protection: expression of a bean polygalacturonase inhibitor in tobacco confers a strong resistance against Rhizoctonia solani and two oomycetes.

    Directory of Open Access Journals (Sweden)

    Orlando eBorras-Hidalgo

    2012-12-01

    Full Text Available We have tested whether a gene encoding a polygalacturonase-inhibiting protein (PGIP protects tobacco against a fungal pathogen (Rhizoctonia solani and two oomycetes (Phytophthora parasitica var. nicotianae and Peronospora hyoscyami f. sp. tabacina. The trials were performed in greenhouse conditions for R. solani and P. parasitica and in the field for P. hyoscyami. Our results show that expression of PGIP is a powerful way of engineering a broad-spectrum disease resistance.

  17. The politics of tobacco.

    Science.gov (United States)

    Mackay, J L

    1998-12-01

    Smoking prevalence and tobacco-attributable mortality will increase substantially in the Asia- Pacific region well into the next century, due to population expansion, ageing populations, and the fact that more women are smoking. The economic costs of tobacco, already substantial, will rise. Of particular concern is the penetration of the region by the transnational tobacco companies, which deny the health evidence of the harm from tobacco, use sophisticated promotions, and lobby to oppose tobacco control measures. There is an urgent need for robust tobacco control action to be taken by every country, but many governments have little experience in combatting this new epidemic or in countering the tobacco companies. They are needlessly concerned that tobacco control action will harm their economy, leading to loss of tax revenue and jobs. Arguments to convince governments must be shaped to address economic issues and illustrate that such action is not only good for a nation's health, but also good for its economy.

  18. Risks of tobacco

    Science.gov (United States)

    Skip navigation U.S. National Library of Medicine The navigation menu has been collapsed. Menu ... tobacco URL of this page: //medlineplus.gov/ency/article/002032.htm Risks of tobacco To use the sharing features ...

  19. You(th) & Tobacco

    Science.gov (United States)

    ... that don’t sell tobacco to kids. Frequent restaurants and other places that are tobacco-free. Be ... reduces the oxygen available for muscles used in sports. Smokers suffer from shortness of breath almost 3 ...

  20. Allegheny County Tobacco Vendors

    Data.gov (United States)

    Allegheny County / City of Pittsburgh / Western PA Regional Data Center — The tobacco vendor information provides the location of all tobacco vendors in Allegheny County in 2015. Data was compiled from administrative records managed by...

  1. Youth and Tobacco

    Science.gov (United States)

    ... with emails, text messages, RSS Feeds, content syndication, social media and more to learn about the latest federal tobacco regulations. Preventing Tobacco Use Among Youth and Young Adults: A Report of the Surgeon ...

  2. RNAi-mediated Mortality of the Whitefly through Transgenic Expression of Double-stranded RNA Homologous to Acetylcholinesterase and Ecdysone Receptor in Tobacco Plants

    Science.gov (United States)

    The whitefly Bemisia tabaci (Genn.) is a pest and vector of plant viruses affecting plants worldwide. Using RNA interference (RNAi) to downregulate whitefly genes by expressing their homologous double stranded RNAs in plants has great potential for management of whiteflies to reduce plant virus dise...

  3. Low plasma 25-hydroxyvitamin d and risk of tobacco-related cancer

    DEFF Research Database (Denmark)

    Afzal, Shoaib; Bojesen, Stig E; Nordestgaard, Børge G

    2013-01-01

    Tobacco smoke chemicals may influence vitamin D metabolism and function, and conversely vitamin D may modify the carcinogenicity of tobacco smoke chemicals. We tested the hypothesis that lower plasma 25-hydroxyvitamin D [25(OH)D] is associated with a higher risk of tobacco-related cancer...

  4. Assessment of the carcinogenic N-nitrosodiethanolamine in tobacco products and tobacco smoke

    Energy Technology Data Exchange (ETDEWEB)

    Brunnemann, K.D.; Hoffmann, D.

    1981-01-01

    A simple, reproducible gas chromatography-thermal energy analyzer (g.c.-TEA) method has been developed for the analysis of N-nitrosodiethanolamine (NDELA) in tobacco and tobacco smoke. The extract of tobacco or the trapped particulates of tobacco smoke are chromatographed on silica gel. The NDELA containing fractions are concentrated, silylated and analyzed with a modified g.c.-TEA system. (/sup 14/C)NDELA serves as internal standard for the quantitative analysis. Experimental cigarettes made from tobaccos which were treated with the sucker growth inhibitor maleic hydrazidediethanolamine (MH-DELA) contained 115--420 p.p.b. of NDELA and their smoke contained 20--290 ng/cigarette, whereas hand-suckered tobacco and its smoke were free of NDELA. The tobacco of US smoking products contained 115--420 p.p.b. of NDELA and the mainstream smoke from such products yielded 10--68 ng/cigar or cigarette. NDELA levels in chewing tobacco ranged from 220--280 p.p.b. and in two commercial snuff products were 3,200 and 6,800 p.p.b. Although the five analyzed MH-DELA preparations contained between 0.6--1.9 p.p.m. NDELA it is evident that the major portion of NDELA in tobacco is formed from the DELA residue during the tobacco processing. Based on bioassay data from various laboratories which have shown that NDELA is a relatively strong carcinogen and based on the results of this study the use of MH-DELA for the cultivation of tobacco is questioned.

  5. DNA methylation polymorphism in flue-cured tobacco and candidate markers for tobacco mosaic virus resistance

    Institute of Scientific and Technical Information of China (English)

    Jie-hong ZHAO; Ji-shun ZHANG; Yi WANG; Ren-gang WANG; Chun WU; Long-jiang FAN; Xue-liang REN

    2011-01-01

    DNA methylation plays an important role in the epigenetic regulation of gene expression during plant growth,development,and polyploidization.However,there is still no distinct evidence in tobacco regarding the distribution of the methylation pattern and whether it contributes to qualitative characteristics.We studied the levels and patterns of methylation polymorphism at CCGG sites in 48 accessions of allotetraploid flue-cured tobacco,Nicotiana tabacum,using a methylation-sensitive amplified polymorphism (MSAP) technique.The results showed that methylation existed at a high level among tobacco accessions,among which 49.3% sites were methylated and 69.9% allelic sites were polymorphic.A cluster analysis revealed distinct patterns of geography-specific groups.In addition,three polymorphic sites significantly related to tobacco mosaic virus (TMV) resistance were explored.This suggests that tobacco breeders should pay more attention to epigenetic traits.

  6. Tobacco and Pregnancy

    Science.gov (United States)

    This paper will review the epidemiology of the impact of cigarette smoking and other forms of tobacco exposure on human development. Sources of exposure described include cigarettes and other forms of smoked tobacco, secondhand (environmental) tobacco smoke, several forms of smok...

  7. Cancer and Tobacco Use

    Science.gov (United States)

    ... CDC-recommended levels. www.cdc.gov/tobacco/stateandcommunity/best_practices Make tobacco cessation treatments more available to people ... what cancer screening tests are needed and are best for them. Make sure their ... their practice. Everyone can Quit using tobacco or never start. ...

  8. North Carolina Tobacco Farmers' Changing Perceptions of Tobacco Control and Tobacco Manufacturers

    Science.gov (United States)

    Crankshaw, Erik C.; Beach, Robert H.; Austin, W. David; Altman, David G.; Jones, Alison Snow

    2009-01-01

    Purpose: To examine tobacco farmers' attitudes toward tobacco control, public health, and tobacco manufacturers in order to determine the extent to which rapidly changing economic conditions have influenced North Carolina tobacco farmer attitudes in ways that may provide tobacco control advocates with new opportunities to promote tobacco control…

  9. A tannin-blocking agent does not modify the preference of sheep towards tannin-containing plants.

    Science.gov (United States)

    Hernández-Orduño, G; Torres-Acosta, J F J; Sandoval-Castro, C A; Capetillo-Leal, C M; Aguilar-Caballero, A J; Alonso-Díaz, M A

    2015-06-01

    Sheep have been suggested to use their senses to perceive plant properties and associate their intake with consequences after ingestion. However, sheep with browsing experience do not seem to select against tannin-rich browsing materials in cafeteria trials. Thus, the objective of the present study was to evaluate the relationship between the chemical composition, selectivity index (SI), preference and intake rate (IR) of tannin-containing forage trees offered to sheep in cafeteria experiments. Four trees were selected for their condensed tannin content and their varying biological activities. Havardia albicans (high biological activity), Leucaena leucocephala (medium biological activity), Acacia gaumeri (low biological activity) and Brosimum alicastrum (very low biological activity) were used in this study. Ten hair sheep (23.7kg±1.43LW) with eight months of browsing experience in native vegetation were used in this study. Polyethylene glycol (PEG 3600MW) was administered to five sheep during all experiments. In experiment 1, fresh foliage from all trees was offered ad libitum for 4h. In experiment 2, B. alicastrum was withdrawn and the preference was determined again. The forage preference in experiment 1 was A. gaumeri (14.77gDM/kgLW)>B. alicastrum (11.77gDM/kgLW)>H. albicans (3.71gDM/kgLW)=L. leucocephala (1.87gDM/kgLW) (PH. albicans=L. leucocephala. PEG administration had no effect on the preference or IR. The intake rate seemed to have been affected by the plant density. Moreover, fiber compounds were found to be better predictors of DM intake than polyphenolic compounds at levels typically found in the evaluated forages. It was concluded that tannins and PEG did not modify the preferences of sheep in cafeteria trials. Thus, tannins are not involved in the preference regulation of animals with browsing experience.

  10. Effects of bio-organic fertilizer's application on flue-cured tobacco planted on continuous cropping soil%生物有机肥改良连作土壤及烤烟生长发育的效应

    Institute of Scientific and Technical Information of China (English)

    邵孝侯; 刘旭; 周永波; 于静

    2011-01-01

    The effects of compost mixed with earthworm fertilizer and effective microorganism on continuous cropping soil was studied. It discussed the effects of bio-organic fertilizer and chemical fertilizer's mixing application by different ratios on fluecured planting and the soil chemical properties with pot experiment. The result indicated that: the fertilization significantly promoted the growth of flue-cured tobacco, the best proportion of bio-organic fertilizer was fifty percent, followed by seventy percent, while the yield of flue-cured tobacco was the highest when the proportion was fifty percent, the growth and yield of flue-cured tobacco of pure bio-organic fertilizer treatment was better than pure fertilizer treatment, the bio-organic fertilizer could promote the nitrogen and potassium absorption when the proportion was between thirty percent and fifty percent, and was more propitious to the production of high quality tobacco, the application of organic fertilizer could significantly increase the cropping soil's pH, organic matter content and available nitrogen, phosphorus and potassium content.%以蚯蚓肥和有效微生物制剂堆置发酵制成生物有机肥,采用盆栽试验研究了生物有机肥和化肥按不同配比混合施用后对烤烟生长发育及植烟土壤理化性质的影响.结果表明,施肥显著促进烤烟的生长发育,以生物有机肥比例为50%时最佳,70%次之,同时生物有机肥比例为50%时烤烟产量最高;纯生物有机肥施用后烤烟的生长指标和产量均优于纯化肥处理;生物有机肥比例为30%~50%时可以促进烤烟对氮素、钾素的吸收,更有利于优质烟叶的生产;施用生物有机肥可显著提高连作土壤的pH值、有机质含量和碱解氮、速效磷及速效钾的含量.

  11. Coat protein mutations in an attenuated Cucumber mosaic virus encoding mutant 2b protein that lacks RNA silencing suppressor activity induces chlorosis with photosynthesis gene repression and chloroplast abnormalities in infected tobacco plants.

    Science.gov (United States)

    Mochizuki, Tomofumi; Yamazaki, Ryota; Wada, Tomoya; Ohki, Satoshi T

    2014-05-01

    In tobacco plants, the Cucumber mosaic virus (CMV) pepo strain induces mosaic symptoms, including pale green chlorosis and malformed tissues. Here, we characterized the involvement of 2b protein and coat protein (CP) in the development of mosaic symptoms. A 2b mutant (R46C) that lacks viral suppressor of RNA silencing (VSR) activity showed an asymptomatic phenotype with low levels of virus accumulation. Tomato spotted wilt virus NSs protein did not complement the virulence of the R46C, although it did restore high-level virus accumulation. However, R46C mutants expressing mutated CP in which the amino acid P129 was mutated to A, E, C, Q, or S induced chlorosis that was associated with reduced expression of chloroplast and photosynthesis related genes (CPRGs) and abnormal chloroplasts with fewer thylakoid membranes. These results suggest that the CP of the CMV pepo strain acquires virulence by amino acid mutations, which causes CPRG repression and chloroplast abnormalities.

  12. 关于提升烟叶规模化种植水平的思考--以放马岭模式为例%Think about Level Promotion of Scale Planting for Tobacco:Taking Fangmaling Mode of Example

    Institute of Scientific and Technical Information of China (English)

    王苜华; 陈秀斋

    2013-01-01

    Land circulation is an important means to enhance the level of scale planting.The Common tobacco field land circulation mode and its disadvantage was introduced.The advantage and development direction of Fangmaling land circulation mode was analyzed,and reference meaning was indicated to guide land circulation.%开展土地流转是提升规模化种植水平的重要手段,介绍常见烟田的土地流转模式及其弊端,分析放马岭模式的优点和发展方向,指出其借鉴意义,以指导烟田的土地流转。

  13. Meteorological Risk Assessment and Tobacco Planting Regionalization in Western Hubei under the Background of Climate Change%气候变化背景下鄂西烟草种植气象风险评价与区划

    Institute of Scientific and Technical Information of China (English)

    孟丹; 陈正洪; 李建平; 阳威; 何飞; 陈振国

    2015-01-01

    为了确定湖北西部植烟区的气象灾害风险程度,根据灾害学系统理论,从致灾因子、孕灾环境、承灾体和防灾减灾能力4个子系统出发,综合考虑气候条件、地理地形、社会经济状况等多个因素,利用GIS平台对鄂西烟草种植气象灾害风险进行评价与区划。结果表明,鄂西北地区气象灾害危险性东高西低,南北向无明显规律,而鄂西南地区是南高北低;鄂西南巴东和恩施烟草易损性较小,而鄂西北的枣阳和丹江口市易损性较大;鄂西南北部地区的气象风险低于南部地区,而鄂西北南部地区气象风险较小,东部气象风险最大;神农架局部地区和鹤峰、丹江口市东部等地的气象风险最严重。总体而言,鄂西地区适宜种植烟草,尽管存在风险,但是大部分地区风险等级在中等及中等以下,这对指导当地合理规划烟草种植和生产具有重要意义。%Based on previous disaster science researches, we assessed and regionalized meteorological disasters via GIS in order to identify the degree of meteorological disaster risk in tobacco-planting areas of western Hubei. The assessment process was composed of four subsystems including disaster inducing factors, disaster-forming environment, disaster-bearing body and the capability of natural disaster prevention and reduction. The results showed that the risk of meteorological disaster in the eastern areas is higher than the western areas in northwest Hubei. In southwest Hubei meteorological disaster risk is high in the south but low in the north. The vulnerability of tobacco planting is lower in southwest Hubei Badong and Enshi city and higher in northwest Hubei Zaoyang and Danjiangkou city. The meteorological risk in the north is lower than in the south of southwest Hubei. In northwest Hubei, meteorological risk is lower in southern regions and higher in eastern regions. Meteorological risk is the highest

  14. Les plantes génétiquement modifiées en Afrique : enjeux et recherches

    Directory of Open Access Journals (Sweden)

    De Lattre-Gasquet Marie

    2002-11-01

    Full Text Available En ce qui concerne la politique sur plantes génétiquement modifiées, l’Afrique est un terrain de luttes d’influence et de débats. Les enjeux sont à la fois économiques, politiques et écologiques. Dans cet article, nous montrons que les biotechnologies végétales sont l’une des techniques, à combiner avec d’autres, qui pourraient permettre d’assurer la sécurité alimentaire et sanitaire de l’Afrique, et que les risques qu’entraîneront l’utilisation de cette technique doivent être soigneusement analysés. Nous présenterons ensuite la position des différents pays africains vis-à-vis des conventions internationales (propriété intellectuelle, biosécurité et commerce et nous caractériserons leurs politiques nationales et de recherche vis-à-vis des OGM.

  15. Streptococcus parauberis associated with modified atmosphere packaged broiler meat products and air samples from a poultry meat processing plant.

    Science.gov (United States)

    Koort, Joanna; Coenye, Tom; Vandamme, Peter; Björkroth, Johanna

    2006-02-15

    Lactic acid bacteria (LAB) isolated from marinated or non-marinated, modified atmosphere packaged (MAP) broiler leg products and air samples of a large-scale broiler meat processing plant were identified and analyzed for their phenotypic properties. Previously, these strains had been found to be coccal LAB. However, the use of a 16 and 23S rRNA gene RFLP database had not resulted in species identification because none of the typically meat-associated LAB type strains had clustered together with these strains in the numerical analysis of the RFLP patterns. To establish the taxonomic position of these isolates, 16S rRNA gene sequence analysis, numerical analysis of ribopatterns, and DNA-DNA hybridization experiments were done. The 16S rRNA gene sequences of three isolates possessed the highest similarities (over 99%) with the sequence of S. parauberis type strain. However, in the numerical analysis of HindIII ribopatterns, the type strain did not cluster together with these isolates. Reassociation values between S. parauberis type or reference strain and the strains studied varied from 82 to 97%, confirming that these strains belong to S. parauberis. Unexpectedly, most of the broiler meat-originating strains studied for their phenotypical properties did not utilize lactose at all and the same strains fermented also galactose very weakly, properties considered atypical for S. parauberis. This is, to our knowledge, the first report of lactose negative S. parauberis strains and also the first report associating S. parauberis with broiler slaughter and meat products.

  16. Application of β-cyclodextrin-modified, carbon nanotube-reinforced hollow fiber to solid-phase microextraction of plant hormones.

    Science.gov (United States)

    Song, Xin-Yue; Ha, Wei; Chen, Juan; Shi, Yan-Ping

    2014-12-29

    A new, efficient, and environmental friendly solid-phase microextraction (SPME) medium based on β-cyclodextrin (β-CD)-modified carbon nanotubes (CNTs) and a hollow fiber (HF) was prepared. Functionalized β-CD was covalently linked to the surface of the carboxylic CNTs and then the obtained nanocomposite was immobilized into the wall pores of HFs under ultrasonic-assisted effect. The scanning electron microscope was used to inspect surface characteristics of fibers, demonstrating the presence of nanocomposites in their wall pores. The reinforced HF was employed in SPME, and its extraction performance was evaluated by analyzing 1-naphthaleneacetic acid (NAA) and 2-naphthoxyacetic acid (2-NOA) in vegetables. Without any tedious clean-up procedure, analytes were extracted from the sample to the adsorbent and organic solvent immobilized in HFs and then desorbed in acetonitrile prior to chromatographic analysis. Under the optimized extraction conditions, the method provided 275- and 283-fold enrichment factors of NAA and 2-NOA, low limits of detection and quantification (at an ngg(-1) level), satisfactory spiked recoveries, good inter-fiber repeatability, and batch-to-batch reproducibility. The selectivity of the developed fiber was investigated to three structurally similar compounds and two reference compounds with recognition coefficients up to 3.18. The obtained results indicate that the newly developed fiber is a feasible, selective, green, and cost-effective microextraction medium and could be successfully applied for extraction and determination of naphthalene-derived plant hormones in complex matrices.

  17. Ecotoxicity of natural insecticide based on tobacco plant extract and hematological effects on the Nile tilapia (Oreochromis niloticus. Ecotoxicity and hematological effects of a natural insecticide based on tobacco (Nicotiana tabacum extract on Nile tilapia (Oreochromis niloticus - doi: 10.4025/actascibiolsci.v35i2.14131

    Directory of Open Access Journals (Sweden)

    Marisa Narciso Fernandes

    2013-05-01

    Full Text Available Natural insecticides derived from plant extracts have been used as an alternative to synthetic products in order to reduce environmental contamination. The present study aimed to examine the effects of Fumydro®, a natural insecticide based in the tobacco plant Nicotiana tabacum, on the Nile tilapia (Oreochromis niloticus by determining the 48-h LC50 and evaluating their effects on hematological variables. Adult specimens of O. niloticus were exposed to four Fumydro® concentrations (200, 300, 400 and 500 μL L-1. The 48-h LC50 of Fumydro® was determined as 370 ± 50 μL L-1. Surviving fish showed increasing in the red blood cells, hemoglobin concentration, mean corpuscular hemoglobin and mean corpuscular hemoglobin concentration. The thrombocytes did not change but the percentage of neutrophils increased. These results indicated that the insecticide Fumydro® is toxic to Nile tilapia and the changes of the erythrocyte variables suggested hypoxemia induction with low effect on the immune system.Natural insecticides from plant extracts represent an alternative to the highly toxic synthetic products in order to reduce environmental contamination; however some might also be toxic for non-target organisms. The present study determined the 50% lethal concentration (48h; LC50 for adults Nile tilapia (Oreochromis niloticus exposed to the natural insecticide Fumydro®, based on the tobacco plant (Nicotiana tabacum, and evaluated its effect on hematological variables. After preliminary tests, adult specimens of O. niloticus were exposed to four Fumydro® concentrations (200, 300, 400 and 500 μL L-1. The 48h; LC50 of Fumydro® was determined at 370 ± 50 μL L-1. The surviving fish after exposure to Fumydro® showed an increase in the number of red blood cells, hemoglobin concentration, mean corpuscular hemoglobin and mean corpuscular hemoglobin concentration. The number of thrombocytes and leukocytes has not changed, unlike the differential leukocyte

  18. 抑制烟草花叶病毒侵染的植物提取物的筛选%Selecting materials of antiviral activity to tobacco mosaic virus infection from some plant extracts

    Institute of Scientific and Technical Information of China (English)

    马学萍; 孔宝华; 叶敏; 段云晖

    2007-01-01

    Ethanol extracts from thirty-three kinds of plant were tested for inhibiting Tobacco mosaic virus (TMV) infection by using half leaf technique on Nicotiana glutinosa.The results showed that the extracts of Cedrus deodara (Roxb.) G.Don and Eucalyptus maideni F.V.Muell performed better inhibitory activities to TMV infection comparing other plant species.The inhibition percentage were 89.7%and 78.3%,respectively.The extracts of Cuscuta chinensis Lam.,Rosa moyesii Hemsl.et wils,Morus alba L.,Callistemon rigidus R.Br,R.simsii planch,Osmanthus fragrans (Thunb)Lour,Dianthus chinensis L.,Ehretia coryltfolia C.H.Wight,Brassica juncea(L.) Czern.et coss,Aloe vera(L.) Burm.f.showed inhibitory effect as well.The inhibition percentage all of them were over 50%.%用半叶枯斑法,在心叶烟上测试了33种植物乙醇提取物水溶液对烟草花叶病毒(Tobacco mosaic virus,TMV)侵染的抑制效果.结果表明,雪松、桉树提取物能较好地抑制病毒(TMV)侵染, 抑制率达到89.7%,78.3%;此外,菟丝子、红花蔷薇、桑、红千层、杜鹃、桂花、香石竹、滇厚朴、荠菜、芦荟等植物提取物也具有抑制烟草花叶病毒(TMV)侵染的效果,抑制率均在50%以上.

  19. A Fire in the Global Village: Teaching Ethical Reasoning and Stakeholder Interests Utilizing Tobacco

    Science.gov (United States)

    Dhooge, Lucien J.

    2012-01-01

    Tobacco has been an agricultural staple from the time of the first recorded European encounter with the plant in the fifteenth century. The pervasive nature of its cultivation and consumption has made tobacco one of the most profitable crops in world agricultural history. This case study examines the role of tobacco in the global marketplace with…

  20. Cannabis and tobacco smoke are not equally carcinogenic

    Directory of Open Access Journals (Sweden)

    Melamede Robert

    2005-10-01

    Full Text Available Abstract More people are using the cannabis plant as modern basic and clinical science reaffirms and extends its medicinal uses. Concomitantly, concern and opposition to smoked medicine has occurred, in part due to the known carcinogenic consequences of smoking tobacco. Are these reactions justified? While chemically very similar, there are fundamental differences in the pharmacological properties between cannabis and tobacco smoke. Cannabis smoke contains cannabinoids whereas tobacco smoke contains nicotine. Available scientific data, that examines the carcinogenic properties of inhaling smoke and its biological consequences, suggests reasons why tobacco smoke, but not cannabis smoke, may result in lung cancer.

  1. [Tobacco--a source of biofuels].

    Science.gov (United States)

    Budzianowska, Anna; Budzianowski, Jaromir

    2012-01-01

    One of the concepts of global protection of environment is to reduce greenhouse gas emissions, mainly carbon dioxide, into the atmosphere by replacing fossil fuels by the so-called biofuels, which can be obtained from cultivated plants or any plant waste biomass. Currently applied industrial technologies allow the production of biofuels to receive ethanol, mostly from the reserve carbohydrates of sugar cane and corn as well as biodiesel from oil, mainly from rapeseed or oil palm. Tobacco, which provides a high biomass, can be used to produce biogas, bioethanol and biodiesel. The latter derived from oil from seeds and leaves of tobacco has proved useful for driving cars. Modest oil content in tobacco leaves can be increased by the expression of foreign genes encoding its biosynthesis. Promising future source of biofuels is a waste plant biomass consisting mainly of cell walls, which can be subjected to the degradation to produce sugars suitable for fermentation and the production of bioethanol. A number of enzymes needed for efficient degradation of plant cell walls can be produced using recombinant DNA technology in a variety of plants, particularly in chloroplasts of tobacco.

  2. Vacuum and Co-cultivation Agroinfiltration of (Germinated) Seeds Results in Tobacco Rattle Virus (TRV) Mediated Whole-Plant Virus-Induced Gene Silencing (VIGS) in Wheat and Maize

    Science.gov (United States)

    Zhang, Ju; Yu, Deshui; Zhang, Yi; Liu, Kun; Xu, Kedong; Zhang, Fuli; Wang, Jian; Tan, Guangxuan; Nie, Xianhui; Ji, Qiaohua; Zhao, Lu; Li, Chengwei

    2017-01-01

    Tobacco rattle virus (TRV)-mediated virus-induced gene silencing (VIGS) has been frequently used in dicots. Here we show that it can also be used in monocots, by presenting a system involving use of a novel infiltration solution (containing acetosyringone, cysteine, and Tween 20) that enables whole-plant level VIGS of (germinated) seeds in wheat and maize. Using the established system, phytoene desaturase (PDS) genes were successfully silenced, resulting in typical photo-bleaching symptoms in the leaves of treated wheat and maize. In addition, three wheat homoeoalleles of MLO, a key gene repressing defense responses to powdery mildew in wheat, were simultaneously silenced in susceptible wheat with this system, resulting in it becoming resistant to powdery mildew. The system has the advantages generally associated with TRV-mediated VIGS systems (e.g., high-efficiency, mild virus infection symptoms, and effectiveness in different organs). However, it also has the following further advantages: (germinated) seed-stage agroinfiltration; greater rapidity and convenience; whole-plant level gene silencing; adequately stable transformation; and suitability for studying functions of genes involved in seed germination and early plant development stages.

  3. Nicotine Levels and Presence of Selected Tobacco-Derived Toxins in Tobacco Flavoured Electronic Cigarette Refill Liquids

    Directory of Open Access Journals (Sweden)

    Konstantinos E. Farsalinos

    2015-03-01

    Full Text Available Background. Some electronic cigarette (EC liquids of tobacco flavour contain extracts of cured tobacco leaves produced by a process of solvent extraction and steeping. These are commonly called Natural Extract of Tobacco (NET liquids. The purpose of the study was to evaluate nicotine levels and the presence of tobacco-derived toxins in tobacco-flavoured conventional and NET liquids. Methods. Twenty-one samples (10 conventional and 11 NET liquids were obtained from the US and Greek market. Nicotine levels were measured and compared with labelled values. The levels of tobacco-derived chemicals were compared with literature data on tobacco products. Results. Twelve samples had nicotine levels within 10% of the labelled value. Inconsistency ranged from −21% to 22.1%, with no difference observed between conventional and NET liquids. Tobacco-specific nitrosamines (TSNAs were present in all samples at ng/mL levels. Nitrates were present almost exclusively in NET liquids. Acetaldehyde was present predominantly in conventional liquids while formaldehyde was detected in almost all EC liquids at trace levels. Phenols were present in trace amounts, mostly in NET liquids. Total TSNAs and nitrate, which are derived from the tobacco plant, were present at levels 200–300 times lower in 1 mL of NET liquids compared to 1 gram of tobacco products. Conclusions. NET liquids contained higher levels of phenols and nitrates, but lower levels of acetaldehyde compared to conventional EC liquids. The lower levels of tobacco-derived toxins found in NET liquids compared to tobacco products indicate that the extraction process used to make these products did not transfer a significant amount of toxins to the NET. Overall, all EC liquids contained far lower (by 2–3 orders of magnitude levels of the tobacco-derived toxins compared to tobacco products.

  4. Nicotine levels and presence of selected tobacco-derived toxins in tobacco flavoured electronic cigarette refill liquids.

    Science.gov (United States)

    Farsalinos, Konstantinos E; Gillman, I Gene; Melvin, Matt S; Paolantonio, Amelia R; Gardow, Wendy J; Humphries, Kathy E; Brown, Sherri E; Poulas, Konstantinos; Voudris, Vassilis

    2015-03-24

    Some electronic cigarette (EC) liquids of tobacco flavour contain extracts of cured tobacco leaves produced by a process of solvent extraction and steeping. These are commonly called Natural Extract of Tobacco (NET) liquids. The purpose of the study was to evaluate nicotine levels and the presence of tobacco-derived toxins in tobacco-flavoured conventional and NET liquids. Twenty-one samples (10 conventional and 11 NET liquids) were obtained from the US and Greek market. Nicotine levels were measured and compared with labelled values. The levels of tobacco-derived chemicals were compared with literature data on tobacco products. Twelve samples had nicotine levels within 10% of the labelled value. Inconsistency ranged from -21% to 22.1%, with no difference observed between conventional and NET liquids. Tobacco-specific nitrosamines (TSNAs) were present in all samples at ng/mL levels. Nitrates were present almost exclusively in NET liquids. Acetaldehyde was present predominantly in conventional liquids while formaldehyde was detected in almost all EC liquids at trace levels. Phenols were present in trace amounts, mostly in NET liquids. Total TSNAs and nitrate, which are derived from the tobacco plant, were present at levels 200-300 times lower in 1 mL of NET liquids compared to 1 gram of tobacco products. NET liquids contained higher levels of phenols and nitrates, but lower levels of acetaldehyde compared to conventional EC liquids. The lower levels of tobacco-derived toxins found in NET liquids compared to tobacco products indicate that the extraction process used to make these products did not transfer a significant amount of toxins to the NET. Overall, all EC liquids contained far lower (by 2-3 orders of magnitude) levels of the tobacco-derived toxins compared to tobacco products.

  5. Tobacco control in Asia.

    Science.gov (United States)

    Mackay, Judith; Ritthiphakdee, Bungon; Reddy, K Srinath

    2013-05-04

    For the purpose of this article, Asia refers to WHO's combined South-East Asia and Western Pacific regions and thus includes Australia and New Zealand. Asia has the highest number of tobacco users and is the prime target of transnational tobacco companies. The future of global tobacco control rests in this region and the challenges are clear. China, India, and Indonesia are key markets and Asia is a frontrunner in tobacco control measures, such as plain packaging of cigarettes. Some countries in Asia have a long history of tobacco control activities beginning in the 1970s, and WHO's Western Pacific Region is still the only region where all countries have ratified WHO's Framework Convention on Tobacco Control. We reviewed the history, research, epidemiology, tobacco control action, obstacles, and potential responses and solutions to the tobacco epidemic in this region. Levels of development, systems of government, and population size are very different between countries, with population size ranging from 1500 to 1·3 billion, but similarities exist in aspects of the tobacco epidemic, harms caused, obstacles faced, and tobacco control actions needed. Copyright © 2013 Elsevier Ltd. All rights reserved.

  6. Application of omics technologies for environmental risk assessment of genetically modified plants : arabidopsis and modified defence mechanisms as a model study

    NARCIS (Netherlands)

    Houshyani Hassanzadeh, B.

    2012-01-01

    As a result of rapid biotechnological developments in the past century, genetically modified (GM) crops were developed and introduced for field application. Despite the advantages of these crops and the professional marketing policies, people also started questioning the safety of GM products for

  7. Application of omics technologies for environmental risk assessment of genetically modified plants : arabidopsis and modified defence mechanisms as a model study

    NARCIS (Netherlands)

    Houshyani Hassanzadeh, B.

    2012-01-01

    As a result of rapid biotechnological developments in the past century, genetically modified (GM) crops were developed and introduced for field application. Despite the advantages of these crops and the professional marketing policies, people also started questioning the safety of GM products for hu