WorldWideScience

Sample records for plant tissues

  1. Plant tissue culture techniques

    Directory of Open Access Journals (Sweden)

    Rolf Dieter Illg

    1991-01-01

    Full Text Available Plant cell and tissue culture in a simple fashion refers to techniques which utilize either single plant cells, groups of unorganized cells (callus or organized tissues or organs put in culture, under controlled sterile conditions.

  2. Plant tissue culture techniques

    OpenAIRE

    Rolf Dieter Illg

    1991-01-01

    Plant cell and tissue culture in a simple fashion refers to techniques which utilize either single plant cells, groups of unorganized cells (callus) or organized tissues or organs put in culture, under controlled sterile conditions.

  3. Plant Tissue Culture Studies.

    Science.gov (United States)

    Smith, Robert Alan

    Plant tissue culture has developed into a valid botanical discipline and is considered a key area of biotechnology, but it has not been a key component of the science curriculum because of the expensive and technical nature of research in this area. This manual presents a number of activities that are relatively easy to prepare and perform. The…

  4. Epigenetics in plant tissue culture

    NARCIS (Netherlands)

    Smulders, M.J.M.; Klerk, de G.J.M.

    2011-01-01

    Plants produced vegetatively in tissue culture may differ from the plants from which they have been derived. Two major classes of off-types occur: genetic ones and epigenetic ones. This review is about epigenetic aberrations. We discuss recent studies that have uncovered epigenetic modifications at

  5. History of plant tissue culture.

    Science.gov (United States)

    Thorpe, Trevor

    2012-01-01

    Plant tissue culture, or the aseptic culture of cells, tissues, organs, and their components under defined physical and chemical conditions in vitro, is an important tool in both basic and applied studies as well as in commercial application. It owes its origin to the ideas of the German scientist, Haberlandt, at the beginning of the twentieth century. The early studies led to root cultures, embryo cultures, and the first true callus/tissue cultures. The period between the 1940s and the 1960s was marked by the development of new techniques and the improvement of those that were already in use. It was the availability of these techniques that led to the application of tissue culture to five broad areas, namely, cell behavior (including cytology, nutrition, metabolism, morphogenesis, embryogenesis, and pathology), plant modification and improvement, pathogen-free plants and germplasm storage, clonal propagation, and product (mainly secondary metabolite) formation, starting in the mid-1960s. The 1990s saw continued expansion in the application of the in vitro technologies to an increasing number of plant species. Cell cultures have remained an important tool in the study of basic areas of plant biology and biochemistry and have assumed major significance in studies in molecular biology and agricultural biotechnology in the twenty-first century. The historical development of these in vitro technologies and their applications is the focus of this chapter.

  6. Plant Tissue Culture in a Bag.

    Science.gov (United States)

    Beck, Mike

    2000-01-01

    Describes the use of an oven bag as a sterile chamber for culture initiation and tissue transfer. Plant tissue culture is an ideal tool for introducing students to plants, cloning, and experimental design. Includes materials, methods, discussion, and conclusion sections. (SAH)

  7. Elicitors in Plant Tissue Culture

    Directory of Open Access Journals (Sweden)

    R. Krishnamurthy

    2013-07-01

    Full Text Available Plants or Plant cells in vitro, show physiological and morphological response to microbial, physical or chemical factors which are known as ‘elicitors’. Elicitation is a process of induced or enhanced synthesis of secondary metabolites by the plants to ensure their survival persistence and competitiveness. The application of elicitors, which is currently the focus of research, has been considered as one of the most effective methods to improve the synthesis of secondary metabolites in medicinal plants. Plant secondary metabolites are unique sources for pharmaceuticals, food additives, flavours and other industrial materials. Accumulation of such metabolites often occurs in plants subjected to stresses including various elicitors or signal molecules. Commonly tested chemical elicitors are salicylic acid, methyl salicylate, bezoic acid, chitosan and so forth which affect production of phenolic compounds and activation of various defense-related enzymes in plants. Plants are challenged by a variety of biotic stresses like fungal, bacterial or viral infections. This lead to the great loss to a plant yield. Here we discuss the classification of elicitors, mechanism of elicitor, the use of elicitors and the different features of elicitors.

  8. A continuous growth model for plant tissue

    Science.gov (United States)

    Bozorg, Behruz; Krupinski, Pawel; Jönsson, Henrik

    2016-12-01

    Morphogenesis in plants and animals involves large irreversible deformations. In plants, the response of the cell wall material to internal and external forces is determined by its mechanical properties. An appropriate model for plant tissue growth must include key features such as anisotropic and heterogeneous elasticity and cell dependent evaluation of mechanical variables such as turgor pressure, stress and strain. In addition, a growth model needs to cope with cell divisions as a necessary part of the growth process. Here we develop such a growth model, which is capable of employing not only mechanical signals but also morphogen signals for regulating growth. The model is based on a continuous equation for updating the resting configuration of the tissue. Simultaneously, material properties can be updated at a different time scale. We test the stability of our model by measuring convergence of growth results for a tissue under the same mechanical and material conditions but with different spatial discretization. The model is able to maintain a strain field in the tissue during re-meshing, which is of particular importance for modeling cell division. We confirm the accuracy of our estimations in two and three-dimensional simulations, and show that residual stresses are less prominent if strain or stress is included as input signal to growth. The approach results in a model implementation that can be used to compare different growth hypotheses, while keeping residual stresses and other mechanical variables updated and available for feeding back to the growth and material properties.

  9. Lipid hydroperoxide levels in plant tissues.

    Science.gov (United States)

    Griffiths, G; Leverentz, M; Silkowski, H; Gill, N; Sánchez-Serrano, J J

    2000-08-01

    Hydroperoxides are the primary oxygenated products of polyunsaturated fatty acids and are key intermediates in the octadecanoid signalling pathway in plants. Lipid hydroperoxides (LHPO) were determined spectrophotometrically based on their reaction with an excess of Fe(2+)at low pH in the presence of the dye xylenol orange. Triphenylphosphine-mediated hydroxide formation was used to authenticate the signal generated by the hydroperoxides. The method readily detected lipid peroxidation in Phaseolus: microsomes, senescing potato leaves and in a range of other plant tissues including Phaseolus hypocotyls (26+/-5 nmol g(-1) FW), Alstroemeria floral tissues (sepals 66+/-13 nmol g(-1) FW petals 49+/-6 nmol g(-1) FW), potato leaves (334+/-75 nmol g(-1) FW), broccoli florets (568+/-68 nmol g(-1) FW) and Chlamydomonas cells (602+/-40 nmol g(-1) FW). Relative to the total fatty acid content of the tissues, the % LHPO was within the range of 0.6-1.7% for all tissue types (photosynthetic and non-photosynthetic) and represents the basal oxidation level of membrane fatty acids in plant cells. In order to relate the levels of LHPO to specific signalling pathways, transgenic potato plant lines were used in which lipoxygenase (LOX) (responsible for hydroperoxide biosynthesis) and hydroperoxide lyase (a route of hydroperoxide degradation) activities were largely reduced by an antisense-mediated approach. While the LHPO levels were similar to wild type in the individual LOX antisensed plants, basal LHPO levels, by contrast, were elevated by 38% in transgenic potato leaves antisensed in hydroperoxide lyase, indicating a role for this enzyme in the maintenance of cellular levels of LHPOs.

  10. Tissue-Culture Method of Cloning Rubber Plants

    Science.gov (United States)

    Ball, E. A.

    1983-01-01

    Guayule plant, a high-yield rubber plant cloned by tissue-culture method to produce multiple new plants that mature quickly. By adjusting culture medium, excised shoot tip produces up to 50 identical guayule plants. Varying concentration of cytokinin, single excised tip produces either 1 or several (up to 50) new plants.

  11. [Issues of large scale tissue culture of medicinal plant].

    Science.gov (United States)

    Lv, Dong-Mei; Yuan, Yuan; Zhan, Zhi-Lai

    2014-09-01

    In order to increase the yield and quality of the medicinal plant and enhance the competitive power of industry of medicinal plant in our country, this paper analyzed the status, problem and countermeasure of the tissue culture of medicinal plant on large scale. Although the biotechnology is one of the most efficient and promising means in production of medicinal plant, it still has problems such as stability of the material, safety of the transgenic medicinal plant and optimization of cultured condition. Establishing perfect evaluation system according to the characteristic of the medicinal plant is the key measures to assure the sustainable development of the tissue culture of medicinal plant on large scale.

  12. [Tissue culture of medicinal plant and abscisic acid].

    Science.gov (United States)

    Fang, Hui-Yong; Zhu, Hong; Yao, Jian-Xun; Jia, Cai-Feng; Shan, Gao-Wei; Li, Min-Hui

    2013-01-01

    Abscisic acid (ABA) plays a key role in many physiological processes of plants, and it was also applied to fields of medicinal plant biotechnology. The article presents a review of some recent application of ABA in enhancing the production of secondary metabolites of medicinal plants, improving the in vitro conservation in medicinal plant tissue culture system.

  13. Repressor-mediated tissue-specific gene expression in plants

    Science.gov (United States)

    Meagher, Richard B.; Balish, Rebecca S.; Tehryung, Kim; McKinney, Elizabeth C.

    2009-02-17

    Plant tissue specific gene expression by way of repressor-operator complexes, has enabled outcomes including, without limitation, male sterility and engineered plants having root-specific gene expression of relevant proteins to clean environmental pollutants from soil and water. A mercury hyperaccumulation strategy requires that mercuric ion reductase coding sequence is strongly expressed. The actin promoter vector, A2pot, engineered to contain bacterial lac operator sequences, directed strong expression in all plant vegetative organs and tissues. In contrast, the expression from the A2pot construct was restricted primarily to root tissues when a modified bacterial repressor (LacIn) was coexpressed from the light-regulated rubisco small subunit promoter in above-ground tissues. Also provided are analogous repressor operator complexes for selective expression in other plant tissues, for example, to produce male sterile plants.

  14. RDX in Plant Tissue: Leading to Humification in Surface Soils

    Science.gov (United States)

    2013-01-01

    aromatics in plant tissue may control or alter plant-related transformations and photodegradation. Bio - available carbon from decaying plant tissue may be...TR-13-4 39 Agronomists have shown that high-organic-matter soils reduce the efficacy of the herbicide 2-chloro-4-ethylamino-6-isopropylamino-1,3,5...resulting in the formation of nitroamine and formaldehyde (Hawari et al. 2000). These intermediates can then be further bio -transformed to either

  15. Crossing kingdoms: Using decellularized plants as perfusable tissue engineering scaffolds.

    Science.gov (United States)

    Gershlak, Joshua R; Hernandez, Sarah; Fontana, Gianluca; Perreault, Luke R; Hansen, Katrina J; Larson, Sara A; Binder, Bernard Y K; Dolivo, David M; Yang, Tianhong; Dominko, Tanja; Rolle, Marsha W; Weathers, Pamela J; Medina-Bolivar, Fabricio; Cramer, Carole L; Murphy, William L; Gaudette, Glenn R

    2017-05-01

    Despite significant advances in the fabrication of bioengineered scaffolds for tissue engineering, delivery of nutrients in complex engineered human tissues remains a challenge. By taking advantage of the similarities in the vascular structure of plant and animal tissues, we developed decellularized plant tissue as a prevascularized scaffold for tissue engineering applications. Perfusion-based decellularization was modified for different plant species, providing different geometries of scaffolding. After decellularization, plant scaffolds remained patent and able to transport microparticles. Plant scaffolds were recellularized with human endothelial cells that colonized the inner surfaces of plant vasculature. Human mesenchymal stem cells and human pluripotent stem cell derived cardiomyocytes adhered to the outer surfaces of plant scaffolds. Cardiomyocytes demonstrated contractile function and calcium handling capabilities over the course of 21 days. These data demonstrate the potential of decellularized plants as scaffolds for tissue engineering, which could ultimately provide a cost-efficient, "green" technology for regenerating large volume vascularized tissue mass. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

  16. Methods to Quantify Nickel in Soils and Plant Tissues

    Directory of Open Access Journals (Sweden)

    Bruna Wurr Rodak

    2015-06-01

    Full Text Available In comparison with other micronutrients, the levels of nickel (Ni available in soils and plant tissues are very low, making quantification very difficult. The objective of this paper is to present optimized determination methods of Ni availability in soils by extractants and total content in plant tissues for routine commercial laboratory analyses. Samples of natural and agricultural soils were processed and analyzed by Mehlich-1 extraction and by DTPA. To quantify Ni in the plant tissues, samples were digested with nitric acid in a closed system in a microwave oven. The measurement was performed by inductively coupled plasma/optical emission spectrometry (ICP-OES. There was a positive and significant correlation between the levels of available Ni in the soils subjected to Mehlich-1 and DTPA extraction, while for plant tissue samples the Ni levels recovered were high and similar to the reference materials. The availability of Ni in some of the natural soil and plant tissue samples were lower than the limits of quantification. Concentrations of this micronutrient were higher in the soil samples in which Ni had been applied. Nickel concentration differed in the plant parts analyzed, with highest levels in the grains of soybean. The grain, in comparison with the shoot and leaf concentrations, were better correlated with the soil available levels for both extractants. The methods described in this article were efficient in quantifying Ni and can be used for routine laboratory analysis of soils and plant tissues.

  17. Oxygen radical microscopy in living plant tissues

    DEFF Research Database (Denmark)

    Kristiansen, Kim Anker; Møller, Ian Max; Schulz, Alexander

    Reactive oxygen species (ROS) play a crucial role in a wide variety of processes. Initiation of many different cellular pathways, crosstalk between cells, developmental signalling in planta, programmed cell death and hypersensitive response in connection with plant-pathogen interactions are among...... the different roles ROS play. On the other hand ROS also cause damage to cellular components at sub-lethal to lethal levels. In photosynthesizing plants the major production of ROS origin from the chloroplast. ROS is a by product from the Photosystem I/II handling of light energy. In nonphotosynthesizing plants...... the ROS production stems from the mitochondria and peroxisomes as is seen in animal cells. At the Bioimaging Center at KVL we employ different techniques to induce, detect and monitor ROS production, distribution and in and among living plant cells. Both confocal laser scanning microscopy and 2-photon...

  18. Using Tissue Culture To Investigate Plant Cell Differentiation and Dedifferentiation.

    Science.gov (United States)

    Bozzone, Donna M.

    1997-01-01

    Describes an experimental project that uses plant tissue culture techniques to examine cell differentiation in the carrot. Allows students to gain experience in some important techniques and to explore fundamental questions about cell differentiation. (DDR)

  19. Banana Musa tissue culture plants enhanced by endophytic fungi

    African Journals Online (AJOL)

    Mo

    In vivo screenhouse experiments using tissue culture plants revealed that the endophytic strain ... some of these strains against both C. sordidus eggs and vermiform R. similis stages ... A 5 µl drop of spore suspension or sterile PDB was added.

  20. Using Tissue Culture To Investigate Plant Cell Differentiation and Dedifferentiation.

    Science.gov (United States)

    Bozzone, Donna M.

    1997-01-01

    Describes an experimental project that uses plant tissue culture techniques to examine cell differentiation in the carrot. Allows students to gain experience in some important techniques and to explore fundamental questions about cell differentiation. (DDR)

  1. A Multiscale Mechanical Model for Plant Tissue Stiffness

    Directory of Open Access Journals (Sweden)

    Damiano Pasini

    2013-06-01

    Full Text Available Plant petioles and stems are hierarchical cellular structures, displaying structuralfeatures defined at multiple length scales. The current work focuses on the multi-scalemodelling of plant tissue, considering two orders of structural hierarchy, cell wall and tissue.The stiffness of plant tissue is largely governed by the geometry of the tissue cells, thecomposition of the cell wall and the structural properties of its constituents. The cell wallis analogous to a fiber reinforced composite, where the cellulose microfibril (CMF is theload bearing component. For multilayered cell wall, the microfibril angle (MFA in themiddle layer of the secondary cell wall (S2 layer largely affects the longitudinal stiffnessfor values up to 40o. The MFA in turn influences the overall wall stiffness. In this work,the effective stiffness of a model system based on collenchyma cell wall of a dicotyledonousplant, the Rheum rhabarbarum, is computed considering generic MFA and volume fractions.At the cellular level, a 2-D Finite Edge Centroidal Voronoi tessellation (FECVT has beendeveloped and implemented to generate the non-periodic microstructure of the plant tissue.The effective elastic properties of the cellular tissue are obtained through finite elementanalysis (FEA of the Voronoi model coupled with the cell wall properties. The stiffness ofthe hierarchically modeled tissue is critically important in determining the overall structuralproperties of plant petioles and stems.

  2. Development of germ-free plants and tissue culture

    Science.gov (United States)

    Venketeswaran, S.

    1973-01-01

    The botanical program is reported for experiments performed at the Lunar Receiving Laboratory. Papers prepared during this program are listed. The studies reported include: tissues cultured on various mediums, nutritional studies, preparation of plant cultures for Apollo 15, and pine tissue cultures.

  3. Quantitative permeability imaging of plant tissues

    NARCIS (Netherlands)

    Sibgatullin, T.; Vergeldt, F.J.; Gerkema, E.; As, van H.

    2010-01-01

    A method for mapping tissue permeability based on time-dependent diffusion measurements is presented. A pulsed field gradient sequence to measure the diffusion encoding time dependence of the diffusion coefficients based on the detection of stimulated spin echoes to enable long diffusion times is co

  4. DNA damage in plant herbarium tissue.

    NARCIS (Netherlands)

    Staats, M.; Cuenca, A.; Richardson, J.E.; Ginkel, R.V.; Petersen, G.; Seberg, O.; Bakker, F.T.

    2011-01-01

    Dried plant herbarium specimens are potentially a valuable source of DNA. Efforts to obtain genetic information from this source are often hindered by an inability to obtain amplifiable DNA as herbarium DNA is typically highly degraded. DNA post-mortem damage may not only reduce the number of amplif

  5. Water retention capacity of tissue cultured plants

    NARCIS (Netherlands)

    Klerk, de G.J.M.; Wijnhoven, F.

    2005-01-01

    Leaves rapidly close their stomata after detachment resulting in a strong reduction of water loss. It has been reported that detached leaves of in vitro produced plants show continuous water loss indicating that they are unable to close the stomata properly and/or that their cuticle is

  6. Water retention capacity of tissue cultured plants

    NARCIS (Netherlands)

    Klerk, de G.J.M.; Wijnhoven, F.

    2005-01-01

    Leaves rapidly close their stomata after detachment resulting in a strong reduction of water loss. It has been reported that detached leaves of in vitro produced plants show continuous water loss indicating that they are unable to close the stomata properly and/or that their cuticle is malfunctionin

  7. Commercializing plant tissue culture processes: economics, problems and prospects

    Energy Technology Data Exchange (ETDEWEB)

    Sahai, O.; Knuth, M.

    1985-03-01

    Novel tissue culture techniques and a range of process schemes may be considered for commercial production of plant derived drugs, chemicals, flavors and cosmetics. Plant cell immobilization, in conjunction with strain selection and product leakage, represents a major technological advancement, with significant economic implications. Conventional batch processes produce high value products at low production capacities, whereas continuous biocatalytic processes can potentially enable production of plant derived chemicals in the $20-$25/kg price range.

  8. The role of silicon in plant tissue culture.

    Science.gov (United States)

    Sivanesan, Iyyakkannu; Park, Se Won

    2014-01-01

    Growth and morphogenesis of in vitro cultures of plant cells, tissues, and organs are greatly influenced by the composition of the culture medium. Mineral nutrients are necessary for the growth and development of plants. Several morpho-physiological disorders such as hooked leaves, hyperhydricity, fasciation, and shoot tip necrosis are often associated with the concentration of inorganic nutrient in the tissue culture medium. Silicon (Si) is the most abundant mineral element in the soil. The application of Si has been demonstrated to be beneficial for growth, development and yield of various plants and to alleviate various stresses including nutrient imbalance. Addition of Si to the tissue culture medium improves organogenesis, embryogenesis, growth traits, morphological, anatomical, and physiological characteristics of leaves, enhances tolerance to low temperature and salinity, protects cells and against metal toxicity, prevents oxidative phenolic browning and reduces the incidence of hyperhydricity in various plants. Therefore, Si possesses considerable potential for application in a wide range of plant tissue culture studies such as cryopreservation, organogenesis, micropropagation, somatic embryogenesis and secondary metabolites production.

  9. The Role of Silicon in Plant Tissue Culture

    Directory of Open Access Journals (Sweden)

    Iyyakkannu eSivanesan

    2014-10-01

    Full Text Available Growth and morphogenesis of in vitro cultures of plant cells, tissues and organs are greatly influenced by the composition of the culture medium. Mineral nutrients are necessary for the growth and development of plants. Several morpho-physiological disorders such as hooked leaves, hyperhydricity, fasciation and shoot tip necrosis are often associated with the concentration of inorganic nutrient in the tissue culture medium. Silicon (Si is the most abundant mineral element in the soil. The application of Si has been demonstrated to be beneficial for growth, development and yield of various plants and to alleviate various stresses including nutrient imbalance. Addition of Si to the tissue culture medium improves organogenesis, embryogenesis, growth traits, morphological, anatomical and physiological characteristics of leaves, enhances tolerance to low temperature and salinity, protects cells and against metal toxicity, prevents oxidative phenolic browning and reduces the incidence of hyperhydricity in various plants. Therefore, Si possesses considerable potential for application in a wide range of plant tissue culture studies such as cryopreservation, organogenesis, micropropagation, somatic embryogenesis and secondary metabolites production.

  10. DNA damage in plant herbarium tissue.

    Directory of Open Access Journals (Sweden)

    Martijn Staats

    Full Text Available Dried plant herbarium specimens are potentially a valuable source of DNA. Efforts to obtain genetic information from this source are often hindered by an inability to obtain amplifiable DNA as herbarium DNA is typically highly degraded. DNA post-mortem damage may not only reduce the number of amplifiable template molecules, but may also lead to the generation of erroneous sequence information. A qualitative and quantitative assessment of DNA post-mortem damage is essential to determine the accuracy of molecular data from herbarium specimens. In this study we present an assessment of DNA damage as miscoding lesions in herbarium specimens using 454-sequencing of amplicons derived from plastid, mitochondrial, and nuclear DNA. In addition, we assess DNA degradation as a result of strand breaks and other types of polymerase non-bypassable damage by quantitative real-time PCR. Comparing four pairs of fresh and herbarium specimens of the same individuals we quantitatively assess post-mortem DNA damage, directly after specimen preparation, as well as after long-term herbarium storage. After specimen preparation we estimate the proportion of gene copy numbers of plastid, mitochondrial, and nuclear DNA to be 2.4-3.8% of fresh control DNA and 1.0-1.3% after long-term herbarium storage, indicating that nearly all DNA damage occurs on specimen preparation. In addition, there is no evidence of preferential degradation of organelle versus nuclear genomes. Increased levels of C→T/G→A transitions were observed in old herbarium plastid DNA, representing 21.8% of observed miscoding lesions. We interpret this type of post-mortem DNA damage-derived modification to have arisen from the hydrolytic deamination of cytosine during long-term herbarium storage. Our results suggest that reliable sequence data can be obtained from herbarium specimens.

  11. DNA Damage in Plant Herbarium Tissue

    Science.gov (United States)

    Staats, Martijn; Cuenca, Argelia; Richardson, James E.; Vrielink-van Ginkel, Ria; Petersen, Gitte; Seberg, Ole; Bakker, Freek T.

    2011-01-01

    Dried plant herbarium specimens are potentially a valuable source of DNA. Efforts to obtain genetic information from this source are often hindered by an inability to obtain amplifiable DNA as herbarium DNA is typically highly degraded. DNA post-mortem damage may not only reduce the number of amplifiable template molecules, but may also lead to the generation of erroneous sequence information. A qualitative and quantitative assessment of DNA post-mortem damage is essential to determine the accuracy of molecular data from herbarium specimens. In this study we present an assessment of DNA damage as miscoding lesions in herbarium specimens using 454-sequencing of amplicons derived from plastid, mitochondrial, and nuclear DNA. In addition, we assess DNA degradation as a result of strand breaks and other types of polymerase non-bypassable damage by quantitative real-time PCR. Comparing four pairs of fresh and herbarium specimens of the same individuals we quantitatively assess post-mortem DNA damage, directly after specimen preparation, as well as after long-term herbarium storage. After specimen preparation we estimate the proportion of gene copy numbers of plastid, mitochondrial, and nuclear DNA to be 2.4–3.8% of fresh control DNA and 1.0–1.3% after long-term herbarium storage, indicating that nearly all DNA damage occurs on specimen preparation. In addition, there is no evidence of preferential degradation of organelle versus nuclear genomes. Increased levels of C→T/G→A transitions were observed in old herbarium plastid DNA, representing 21.8% of observed miscoding lesions. We interpret this type of post-mortem DNA damage-derived modification to have arisen from the hydrolytic deamination of cytosine during long-term herbarium storage. Our results suggest that reliable sequence data can be obtained from herbarium specimens. PMID:22163018

  12. Methanol fixation of plant tissue for Scanning Electron Microscopy improves preservation of tissue morphology and dimensions.

    Science.gov (United States)

    Talbot, Mark J; White, Rosemary G

    2013-10-02

    It is well known that preparation of biological (plant and animal) tissues for Scanning Electron Microscopy (SEM) by chemical fixation and critical point drying results in shrinkage of tissues, often by up to 20-30%, depending on the tissue type and fixation protocol used. We sought to identify a protocol that would preserve tissue size and morphology better than standard chemical fixatives and dehydration regimes. We compared a range of processing techniques by quantifying changes in tissue size and recording details of surface morphology using leaf tissues from three commonly studied species; Arabidopsis thaliana, barley and cotton. All processing protocols altered tissue dimensions. Methanol fixation and dehydration, followed by a further short (1 h) dehydration step in ethanol and critical point drying (which was based on a previously published method), preserved tissue dimensions most consistently of all protocols tested, although it did cause 8% shrinkage in all three species. This protocol was also best for preservation of surface morphology in all three species. We outline a recommended protocol and advise that the method is best trialled for different tissues, especially thicker or larger samples. This study shows that simultaneous fixation and dehydration in methanol followed by ethanol results in better preservation of dimensions and morphology of critical point dried plant tissues than other fixation and dehydration procedures. It is a quick and simple method, and requires standard SEM preparation equipment.

  13. Effect of lunar materials on plant tissue culture.

    Science.gov (United States)

    Walkinshaw, C. H.; Venketeswaran, S.; Baur, P. S.; Croley, T. E.; Scholes, V. E.; Weete, J. D.; Halliwell, R. S.; Hall, R. H.

    1973-01-01

    Lunar material collected during the Apollo 11, 12, 14, and 15 missions has been used to treat 12 species of higher plant tissue cultures. Biochemical and morphological studies have been conducted on several of these species. Tobacco tissue cultures treated with 0.22 g of lunar material exhibited increased greening more complex chloroplasts, less cytoplasmic vacuolation and greater vesiculation. Pine tissue cultures reacted to treatment by an increased deposition of tannin-like materials. The percentage of dry weight and soluble protein was increased in cultures treated with either lunar or terrestrial rock materials.

  14. Cloning of medicinal plants through tissue culture--a review.

    Science.gov (United States)

    Chaturvedi, H C; Jain, Madhu; Kidwai, N R

    2007-11-01

    In order to have standardized formulations, the chemical constituents from plants and their parts are required to be uniform both qualitatively and quantitatively. Furthermore, an ever increasing demand of uniform medicinal plants based medicines warrants their mass cloning through plant tissue culture strategy. A good number of medicinal plants have been reported to regenerate in vitro from their various parts, but a critical evaluation of such reports reveals that only a few complete medicinal plants have been regenerated and still fewer have actually been grown in soil, while their micropropagation on a mass scale has rarely been achieved, particularly in those medicinal plants where conventional propagation is inadequate, like, the mass clonal propagation of Dioscorea floribunda leading to its successful field trials. Such facts make it imperative to document the factual position of micropropagation of medicinal plants bringing out the advancements made along with the short falls, in this important area. The present review deals with the futuristic view on the said subject restricted to higher plants.

  15. Application of plant cell and tissue culture for the production of phytochemicals in medicinal plants.

    Science.gov (United States)

    Pant, Bijaya

    2014-01-01

    Approximately 80% of the world inhabitants depend on the medicinal plants in the form of traditional formulations for their primary health care system well as in the treatment of a number of diseases since the ancient time. Many commercially used drugs have come from the information of indigenous knowledge of plants and their folk uses. Linking of the indigenous knowledge of medicinal plants to modern research activities provides a new reliable approach, for the discovery of novel drugs much more effectively than with random collection. Increase in population and increasing demand of plant products along with illegal trade are causing depletion of medicinal plants and many are threatened in natural habitat. Plant tissue culture technique has proved potential alternative for the production of desirable bioactive components from plants, to produce the enough amounts of plant material that is needed and for the conservation of threatened species. Different plant tissue culture systems have been extensively studied to improve and enhance the production of plant chemicals in various medicinal plants.

  16. Dynamics of Long-distance Signaling via Plant Vascular Tissues

    Directory of Open Access Journals (Sweden)

    Michitaka eNotaguchi

    2015-03-01

    Full Text Available Plant vascular systems are constructed by specific cell wall modifications through which cells are highly specialized to make conduits for water and nutrients. Xylem vessels are formed by thickened cell walls that remain after programmed cell death, and serve as water conduits from the root to the shoot. In contrast, phloem tissues consist of a complex of living cells, including sieve tube elements and their neighboring companion cells, and translocate photosynthetic assimilates from mature leaves to developing young tissues. Intensive studies on the content of vascular flow fluids have unveiled that plant vascular tissues transport various types of gene product, and the transport of some provides the molecular basis for the long-distance communications. Analysis of xylem sap has demonstrated the presence of proteins in the xylem transpiration stream. Recent studies have revealed that CLE and CEP peptides secreted in the roots are transported to above ground via the xylem in response to plant-microbe interaction and soil nitrogen starvation, respectively. Their leucine-rich repeat transmembrane receptors localized in the shoot phloem are required for relaying the signal from the shoot to the root. These findings well fit to the current scenario of root-to-shoot-to-root feedback signaling, where peptide transport achieves the root-to-shoot signaling, the first half of the signaling process. Meanwhile, it is now well evidenced that proteins and a range of RNAs are transported via the phloem translocation system, and some of those can exert their physiological functions at their destinations, including roots. Thus, plant vascular systems may serve not only as conduits for the translocation of essential substances but also as long-distance communication pathways that allow plants to adapt to changes in internal and external environments at the whole plant level.

  17. Virtual Plant Tissue: Building Blocks for Next-Generation Plant Growth Simulation.

    Science.gov (United States)

    De Vos, Dirk; Dzhurakhalov, Abdiravuf; Stijven, Sean; Klosiewicz, Przemyslaw; Beemster, Gerrit T S; Broeckhove, Jan

    2017-01-01

    Motivation: Computational modeling of plant developmental processes is becoming increasingly important. Cellular resolution plant tissue simulators have been developed, yet they are typically describing physiological processes in an isolated way, strongly delimited in space and time. Results: With plant systems biology moving toward an integrative perspective on development we have built the Virtual Plant Tissue (VPTissue) package to couple functional modules or models in the same framework and across different frameworks. Multiple levels of model integration and coordination enable combining existing and new models from different sources, with diverse options in terms of input/output. Besides the core simulator the toolset also comprises a tissue editor for manipulating tissue geometry and cell, wall, and node attributes in an interactive manner. A parameter exploration tool is available to study parameter dependence of simulation results by distributing calculations over multiple systems. Availability: Virtual Plant Tissue is available as open source (EUPL license) on Bitbucket (https://bitbucket.org/vptissue/vptissue). The project has a website https://vptissue.bitbucket.io.

  18. Dynamics of long-distance signaling via plant vascular tissues

    Science.gov (United States)

    Notaguchi, Michitaka; Okamoto, Satoru

    2015-01-01

    Plant vascular systems are constructed by specific cell wall modifications through which cells are highly specialized to make conduits for water and nutrients. Xylem vessels are formed by thickened cell walls that remain after programmed cell death, and serve as water conduits from the root to the shoot. In contrast, phloem tissues consist of a complex of living cells, including sieve tube elements and their neighboring companion cells, and translocate photosynthetic assimilates from mature leaves to developing young tissues. Intensive studies on the content of vascular flow fluids have unveiled that plant vascular tissues transport various types of gene product, and the transport of some provides the molecular basis for the long-distance communications. Analysis of xylem sap has demonstrated the presence of proteins in the xylem transpiration stream. Recent studies have revealed that CLE and CEP peptides secreted in the roots are transported to above ground via the xylem in response to plant–microbe interaction and soil nitrogen starvation, respectively. Their leucine-rich repeat transmembrane receptors localized in the shoot phloem are required for relaying the signal from the shoot to the root. These findings well-fit to the current scenario of root-to-shoot-to-root feedback signaling, where peptide transport achieves the root-to-shoot signaling, the first half of the signaling process. Meanwhile, it is now well-evidenced that proteins and a range of RNAs are transported via the phloem translocation system, and some of those can exert their physiological functions at their destinations, including roots. Thus, plant vascular systems may serve not only as conduits for the translocation of essential substances but also as long-distance communication pathways that allow plants to adapt to changes in internal and external environments at the whole plant level. PMID:25852714

  19. Anatomic composition of plant tissues of highly metamorphosed coals

    Energy Technology Data Exchange (ETDEWEB)

    Kizil' shtein, L.Ya.; Shpitsgluz, A.L.

    1985-09-01

    Method is described to improve microscopic study of highly metamorphosed coals (anthracite). Study of such coals with aid of reflected polarized light is enhanced by means of ionic etching of surface of slides that enables observation not only of structures of basic microcomponents but also of finest structural details of individual cells by reflected non-polarized light. Figures illustrate results of studying many samples by ionic etching (bombing a polished surface in a vacuum with ions and pulverizing material of microcomponents to reveal heterogeneity of crystal chemistry of surface) which reveals great variety of structures of plant tissues and their component cells. Pictures of 35 slides depict gelified coal-forming plants of Donbass and central Ural coal fields; fusainized coal-forming plants of Donetsk, Gorlovsk and Tungus basins; organs of Donbass plants; structure of cells and organs of plants of Donbass. Method of ionic etching opens new perspectives for studying anatomy and histology in area of classical paleobotany by making available a large number of samples of plant material and components of highly metamorphosed coals compared with the rare samples obtained by using the polarized light method. 14 references.

  20. Plant-Derived Human Collagen Scaffolds for Skin Tissue Engineering

    Science.gov (United States)

    Willard, James J.; Drexler, Jason W.; Das, Amitava; Roy, Sashwati; Shilo, Shani; Shoseyov, Oded

    2013-01-01

    Tissue engineering scaffolds are commonly formed using proteins extracted from animal tissues, such as bovine hide. Risks associated with the use of these materials include hypersensitivity and pathogenic contamination. Human-derived proteins lower the risk of hypersensitivity, but possess the risk of disease transmission. Methods engineering recombinant human proteins using plant material provide an alternate source of these materials without the risk of disease transmission or concerns regarding variability. To investigate the utility of plant-derived human collagen (PDHC) in the development of engineered skin (ES), PDHC and bovine hide collagen were formed into tissue engineering scaffolds using electrospinning or freeze-drying. Both raw materials were easily formed into two common scaffold types, electrospun nonwoven scaffolds and lyophilized sponges, with similar architectures. The processing time, however, was significantly lower with PDHC. PDHC scaffolds supported primary human cell attachment and proliferation at an equivalent or higher level than the bovine material. Interleukin-1 beta production was significantly lower when activated THP-1 macrophages where exposed to PDHC electrospun scaffolds compared to bovine collagen. Both materials promoted proper maturation and differentiation of ES. These data suggest that PDHC may provide a novel source of raw material for tissue engineering with low risk of allergic response or disease transmission. PMID:23298216

  1. Classification of explosives transformation products in plant tissue

    Energy Technology Data Exchange (ETDEWEB)

    Larson, S.L.; Jones, R.P. (Army Corps of Engineers, Vicksburg, MI (United States). Waterways Experiment Station); Escalon, L.; Parker, D. (AScI Corp., McLean, VA (United States))

    1999-06-01

    Explosives contamination in surface or groundwater used for the irrigation of food crops and phytoremediation of explosives-contaminated soil or water using plant-assisted biodegradation have brought about concerns as to the fate of explosives in plants. Liquid scintillation counting, high-performance liquid chromatography, and gel permeation chromatography were utilized to characterize explosives (hexahydro-1,3,5-trinitro-1,3,5-triazine and trinitrotoluene) and their metabolites in plant tissues obtained from three separate studies. Analyzing tissues of yellow nutsedge (Cyperus esculentus), corn (Zea mays), lettuce (Lacuta sativa), tomato (Lyopersicum esculentum), radish (Raphanus sativus), and parrot feather (Myriophyllum aquaticum) from three studies where exposure to explosives at nontoxic levels occurred showed that extensive transformation of the explosive contaminant occurred, variations were noted in uptake and transformation between terrestrial and aquatic plants, the products had significantly higher polarity and water solubility than the parent compounds, and the molecular sizes of the transformation products were significantly greater than those of the parent compounds.

  2. High-throughput determination of malondialdehyde in plant tissues.

    Science.gov (United States)

    Davey, M W; Stals, E; Panis, B; Keulemans, J; Swennen, R L

    2005-12-15

    Malondialdehyde (MDA) is a widely used marker of oxidative lipid injury whose concentration varies in response to biotic and abiotic stress. Commonly, MDA is quantified as a strong light-absorbing and fluorescing adduct following reaction with thiobarbituric acid (TBA). However, plant tissues in particular contain many compounds that potentially interfere with this reaction and whose concentrations also vary according to the tissue type and stress conditions. As part of our studies into the stress responses of plant tissues, we were interested in developing a rapid, accurate, and robust protocol for MDA analysis using reverse-phased HPLC to avoid these problems with reaction specificity. We demonstrate that a partitioning step into n-butanol during sample preparation is essential and that gradient HPLC analysis is necessary to prevent sample carryover between injections. Furthermore, the starting composition of the mobile phase must be sufficiently hydrophobic to allow direct injection of the n-butanol extracts without peak splitting, tailing, and other artifacts. To minimize analysis times, we used a short, so-called "Rocket" HPLC column and high flow rates. The optimized HPLC separation has a turnaround time of 2.5 min per sample. Butanolic extracts of MDA(TBA)(2) were stable for at least 48 h, and recoveries were linear between 0.38 and 7.5 pmol MDA added. Importantly, this procedure proved to be compatible with existing extraction procedures for l-ascorbate and glutathione analysis in different plant species, allowing multiple "stress metabolite" analyses to be carried out on a single tissue extract.

  3. New Insights into Fe Localization in Plant Tissues

    Directory of Open Access Journals (Sweden)

    Hannetz eRoschzttardtz

    2013-09-01

    Full Text Available Deciphering cellular iron (Fe homeostasis requires having access to both quantitative and qualitative information on the subcellular pools of Fe in tissues and their dynamics within the cells. We have taken advantage of the Perls/DAB Fe staining procedure to perform a systematic analysis of Fe distribution in roots, leaves and reproductive organs of the model plant Arabidopsis thaliana, using wild-type and mutant genotypes affected in iron transport and storage. Roots of soil-grown plants accumulate iron in the apoplast of the central cylinder, a pattern that is strongly intensified when the citrate effluxer FRD3 is not functional, thus stressing the importance of citrate in the apoplastic movement of Fe. In leaves, Fe level is low and only detected in and around vascular tissues. In contrast, Fe staining in leaves of iron-treated plants extends in the surrounding mesophyll cells where Fe deposits, likely corresponding to Fe-ferritin complexes, accumulate in the chloroplasts. The loss of ferritins in the fer1,3,4 triple mutant provoked a massive accumulation of Fe in the apoplastic space, suggesting that in the absence of iron buffering in the chloroplast, cells activate iron efflux and/or repress iron influx to limit the amount of iron in the cell. In flowers, Perls/DAB staining has revealed a major sink for Fe in the anthers. In particular, developing pollen grains accumulate detectable amounts of Fe in small-size intracellular bodies that aggregate around the vegetative nucleus at the binuclear stage and that were identified as amyloplasts. In conclusion, using the Perls/DAB procedure combined to selected mutant genotypes, this study has established a reliable atlas of Fe distribution in the main Arabidopsis organs, proving and refining long-assumed intracellular locations and uncovering new ones. This iron map of Arabidopsis will serve as a basis for future studies of possible actors of iron movement in plant tissues and cell compartments.

  4. Laser Capture Microdissection Protocol for Xylem Tissues of Woody Plants

    Science.gov (United States)

    Blokhina, Olga; Valerio, Concetta; Sokołowska, Katarzyna; Zhao, Lei; Kärkönen, Anna; Niittylä, Totte; Fagerstedt, Kurt

    2017-01-01

    Laser capture microdissection (LCM) enables precise dissection and collection of individual cell types from complex tissues. When applied to plant cells, and especially to woody tissues, LCM requires extensive optimization to overcome such factors as rigid cell walls, large central vacuoles, intercellular spaces, and technical issues with thickness and flatness of the sections. Here we present an optimized protocol for the laser-assisted microdissection of developing xylem from mature trees: a gymnosperm (Norway spruce, Picea abies) and an angiosperm (aspen, Populus tremula) tree. Different cell types of spruce and aspen wood (i.e., ray cells, tracheary elements, and fibers) were successfully microdissected from tangential, cross and radial cryosections of the current year’s growth ring. Two approaches were applied to achieve satisfactory flatness and anatomical integrity of the spruce and aspen specimens. The commonly used membrane slides were ineffective as a mounting surface for the wood cryosections. Instead, in the present protocol we use glass slides, and introduce a glass slide sandwich assembly for the preparation of aspen sections. To ascertain that not only the anatomical integrity of the plant tissue, but also the molecular features were not compromised during the whole LCM procedure, good quality total RNA could be extracted from the microdissected cells. This showed the efficiency of the protocol and established that our methodology can be integrated in transcriptome analyses to elucidate cell-specific molecular events regulating wood formation in trees. PMID:28101088

  5. The role of activated charcoal in plant tissue culture.

    Science.gov (United States)

    Thomas, T Dennis

    2008-01-01

    Activated charcoal has a very fine network of pores with large inner surface area on which many substances can be adsorbed. Activated charcoal is often used in tissue culture to improve cell growth and development. It plays a critical role in micropropagation, orchid seed germination, somatic embryogenesis, anther culture, synthetic seed production, protoplast culture, rooting, stem elongation, bulb formation etc. The promotary effects of AC on morphogenesis may be mainly due to its irreversible adsorption of inhibitory compounds in the culture medium and substancially decreasing the toxic metabolites, phenolic exudation and brown exudate accumulation. In addition to this activated charcoal is involved in a number of stimulatory and inhibitory activities including the release of substances naturally present in AC which promote growth, alteration and darkening of culture media, and adsorption of vitamins, metal ions and plant growth regulators, including abscisic acid and gaseous ethylene. The effect of AC on growth regulator uptake is still unclear but some workers believe that AC may gradually release certain adsorbed products, such as nutrients and growth regulators which become available to plants. This review focuses on the various roles of activated charcoal in plant tissue culture and the recent developments in this area.

  6. Response of plant nutrient stoichiometry to fertilization varied with plant tissues in a tropical forest.

    Science.gov (United States)

    Mo, Qifeng; Zou, Bi; Li, Yingwen; Chen, Yao; Zhang, Weixin; Mao, Rong; Ding, Yongzhen; Wang, Jun; Lu, Xiankai; Li, Xiaobo; Tang, Jianwu; Li, Zhian; Wang, Faming

    2015-09-29

    Plant N:P ratios are widely used as indices of nutrient limitation in terrestrial ecosystems, but the response of these metrics in different plant tissues to altered N and P availability and their interactions remains largely unclear. We evaluated changes in N and P concentrations, N:P ratios of new leaves (1 yr), stems and mixed fine roots of seven species after 3-years of an N and P addition experiment in a tropical forest. Nitrogen addition only increased fine root N concentrations. P addition increased P concentrations among all tissues. The N × P interaction reduced leaf and stem P concentrations, suggesting a negative effect of N addition on P concentrations under P addition. The reliability of using nutrient ratios as indices of soil nutrient availability varied with tissues: the stoichiometric metrics of stems and older leaves were more responsive indicators of changed soil nutrient availability than those of new leaves and fine roots. However, leaf N:P ratios can be a useful indicator of inter-specific variation in plant response to nutrients availability. This study suggests that older leaf is a better choice than other tissues in the assessment of soil nutrient status and predicting plant response to altered nutrients using nutrients ratios.

  7. In situ hybridization for the detection of rust fungi in paraffin embedded plant tissue sections

    Science.gov (United States)

    Rust fungi infect a wide range of plant species making them of particular interest to plant pathologists. In order to study the interactions between these important pathogenic fungi and their host plants it is useful to be able to differentiate fungal tissue from plant tissue. This can be accomplish...

  8. NADPH Oxidase-Dependent Superoxide Production in Plant Reproductive Tissues.

    Science.gov (United States)

    Jiménez-Quesada, María J; Traverso, José Á; Alché, Juan de Dios

    2016-01-01

    In the life cycle of a flowering plant, the male gametophyte (pollen grain) produced in the anther reaches the stigmatic surface and initiates the pollen-pistil interaction, an important step in plant reproduction, which ultimately leads to the delivery of two sperm cells to the female gametophyte (embryo sac) inside the ovule. The pollen tube undergoes a strictly apical expansion characterized by a high growth rate, whose targeting should be tightly regulated. A continuous exchange of signals therefore takes place between the haploid pollen and diploid tissue of the pistil until fertilization. In compatible interactions, theses processes result in double fertilization to form a zygote (2n) and the triploid endosperm. Among the large number of signaling mechanisms involved, the redox network appears to be particularly important. Respiratory burst oxidase homologs (Rbohs) are superoxide-producing enzymes involved in a broad range of processes in plant physiology. In this study, we review the latest findings on understanding Rboh activity in sexual plant reproduction, with a particular focus on the male gametophyte from the anther development stages to the crowning point of fertilization. Rboh isoforms have been identified in both the male and female gametophyte and have proven to be tightly regulated. Their role at crucial points such as proper growth of pollen tube, self-incompatibility response and eventual fertilization is discussed.

  9. Plant cell, tissue and organ culture: the most flexible foundations for plant metabolic engineering applications.

    Science.gov (United States)

    Ogita, Shinjiro

    2015-05-01

    Significant advances in plant cell, tissue and organ culture (PCTOC) have been made in the last five decades. PCTOC is now thought to be the underlying technique for understanding general or specific biological functions of the plant kingdom, and it is one of the most flexible foundations for morphological, physiological and molecular biological applications of plants. Furthermore, the recent advances in the field of information technology (IT) have enabled access to a large amount of information regarding all aspects of plant biology. For example, sequencing information is stored in mega repositories such as the National Center for Biotechnology Information (NCBI), which can be easily accessed by researchers worldwide. To date, the PCTOC and IT combination strategy for regulation of target plant metabolism and the utilization of bioactive plant metabolites for commercial purposes is essential. In this review, the advantages and the limitations of these methodologies, especially regarding the production of bioactive plant secondary metabolites and metabolic engineering in target plants are discussed mainly from the phenotypic view point.

  10. What makes plants different? Principles of extracellular matrix function in 'soft' plant tissues.

    Science.gov (United States)

    Peters, W S; Hagemann, W; Deri Tomos, A

    2000-02-01

    An overview of the biomechanic and morphogenetic function of the plant extracellular matrix (ECM) in its primary state is given. ECMs can play a pivotal role in cellular osmo- and volume-regulation, if they enclose the cell hermetically and constrain hydrostatic pressure evoked by osmotic gradients between the cell and its environment. From an engineering viewpoint, such cell walls turn cells into hydraulic machines, which establishes a crucial functional differences between cell walls and other cellular surface structures. Examples of such hydraulic machineries are discussed. The function of cell walls in the control of pressure, volume, and shape establishes constructional evolutionary constraints, which can explain aspects commonly considered typical of plants (sessility, autotrophy). In plants, 'cell division' by insertion of a new cell wall is a process of internal cytoplasmic differentiation. As such it differs fundamentally from cell separation during cytokinesis in animals, by leaving the coherence of the dividing protoplast basically intact. The resulting symplastic coherence appears more important for plant morphogenesis than histological structure; similar morphologies are realized on the basis of distinct tissue architectures in different plant taxa. The shape of a plant cell is determined by the shape its cell wall attains under multiaxial tensile stress. Consequently, the development of form in plants is achieved by a differential plastic deformation of the complex ECM in response to this multiaxial force (hydrostatic pressure). Current concepts of the regulation of these deformation processes are briefly evaluated.

  11. Extraction of DNA from plant and fungus tissues in situ

    Directory of Open Access Journals (Sweden)

    Abu Almakarem Amal S

    2012-06-01

    Full Text Available Abstract Background When samples are collected in the field and transported to the lab, degradation of the nucleic acids contained in the samples is frequently observed. Immediate extraction and precipitation of the nucleic acids reduces degradation to a minimum, thus preserving accurate sequence information. An extraction method to obtain high quality DNA in field studies is described. Findings DNA extracted immediately after sampling was compared to DNA extracted after allowing the sampled tissues to air dry at 21°C for 48 or 72 hours. While DNA extracted from fresh tissues exhibited little degradation, DNA extracted from all tissues exposed to 21°C air for 48 or 72 hours exhibited varying degrees of degradation. Yield was higher for extractions from fresh tissues in most cases. Four microcentrifuges were compared for DNA yield: one standard electric laboratory microcentrifuge (max rcf = 16,000×g, two battery-operated microcentrifuges (max rcf = 5,000 and 3,000 ×g, and one manually-operated microcentrifuge (max rcf = 120×g. Yields for all centrifuges were similar. DNA extracted under simulated field conditions was similar in yield and quality to DNA extracted in the laboratory using the same equipment. Conclusions This CTAB (cetyltrimethylammonium bromide DNA extraction method employs battery-operated and manually-operated equipment to isolate high quality DNA in the field. The method was tested on plant and fungus tissues, and may be adapted for other types of organisms. The method produced high quality DNA in laboratory tests and under simulated field conditions. The field extraction method should prove useful for working in remote sites, where ice, dry ice, and liquid nitrogen are unavailable; where degradation is likely to occur due to the long distances between the sample site and the laboratory; and in instances where other DNA preservation and transportation methods have been unsuccessful. It may be possible to adapt

  12. Cocowood fibrovascular tissue system – another wonder of plant evolution

    Directory of Open Access Journals (Sweden)

    Oswaldo Mauricio González

    2016-08-01

    Full Text Available The coconut palm (Cocos nucifera L. stem tissue (referred to as cocowood in this study is a complex fibrovascular system that is made up of fibrovascular bundles embedded into a parenchymatous ground tissue. The complex configuration of fibrovascular bundles along with the non-uniform distribution of the material properties likely allow senile coconut stems to optimize their biomechanical performance per unit mass (i.e. mechanical efficiency and grow into tall, slender and very flexible plants with minimum resources of biomass and water. For the first time, to the best of the authors’ knowledge, this paper examines, from the integral (i.e. stem structure and macroscopic (i.e. tissue structure levels of hierarchy, the characteristic triple helix formation depicted by the fibrovascular bundles within the monocotyledon cocowood. The natural course of the tangential orientation of the axial fibrovascular bundles is mapped for the whole cocowood structure by quantifying 264 cocowood discs, corresponding to 41 senile coconut palms estimated to be greater than 70 years old. The observed variations were modelled in this paper by simple equations that partially enabled characterization of the cocowood fibrovascular tissue system. Furthermore, 11 finite element analyses (FEA were performed over a three dimensional (3D finite element (FE model resembling a characteristic coconut palm stem of 25 m in height to analyze the biomaterial reactions produced by the progressive deviation of the tangential fibrovascular bundles on the cocowood mechanical response (i.e. on the material compressive strength and the bending stiffness. The analyses in this study were carried out for the critical wind speed of 23 m/s (i.e. Gale tornado according to the Fujita tornado scale. For each analysis, the characteristic average maxima degree of orientation of the cocowood fibrovascular bundles was varied from 0˚ to 51˚. The acquired results provided a deep understanding of

  13. Plant Generation of TM-1 via Tissue Culture

    Institute of Scientific and Technical Information of China (English)

    LIU Chuan-liang; LI Fu-guang; XU Leng-chun

    2008-01-01

    @@ Plant generation of TM-1 via tissue culture was established.The hypocotyledon sections as explants which were cultured in a series of improved MS media containing 0.05~0.10 mg · L-1 IAA, 0.1~0.15 mg · L-1 Kt,0.07~0.14 mg · L-1 2,4-D could produce a large number of calli which were easier to regeneration in this system.The calli,which were subcuhured in another MS media containing 0.03~0.05 mg · L-1 Kt for 3-4 times produced embryoid callus in a rate of 35%.Fifty-six somatic embryoid calli were subcultured in an improved MS medium containing 0.1 mg·L-1 BA and 0.1~0.15 mg · L-1 IAA for plant regenerating,and 47 cotton plantlets were regenerated from them.

  14. Measuring NO Production by Plant Tissues and Suspension Cultured Cells

    Institute of Scientific and Technical Information of China (English)

    Jan Vitecek; Vilem Reinohl; Russell L.Jones

    2008-01-01

    We describe an inexpensive and reliable detector for measuring NO emitted in the gas phase from plants.The method relies on the use of a strong oxidizer to convert NO to NO2 and subsequent capture of NO2 by a Griess reagent trap.The set-up approaches the sensitivity for NO comparable to that of instruments based on chemiluminescence and photoacoustic detectors.We demonstrate the utility of our set-up by measuring NO produced by a variety of well established plant sources.NO produced by nitrate reductase (NR) in tobacco leaves and barley aleurone was readily detected,as was the production of NO from nitrite by the incubation medium of barley aleurone.Arabidopsis mutants that overproduce NO or lack NO-synthase (AtNOS1) also displayed the expected NO synthesis phenotype when assayed by our set-up.We could also measure NO production from elicitor-treated suspension cultured cells using this set-up.Further,we have focused on the detection of NO by a widely used fluorescent probe 4-amino-5-methylamino-2',7'-difluorofluorescein (DAF-FM).Our work points to the pitfalls that must be avoided when using DAF-FM to detect the production of NO by plant tissues.In addition to the dramatic effects that pH can have on fluorescence from DAF-FM,the widely used NO scavengers 2-phenyl-4,4,5,5-tetramethylimidazoline-l-oxyl-3-oxide (PTIO) and 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (cPTIO) can produce anomalous and unexpected results.Perhaps the most serious drawback of DAF-FM is its ability to bind to dead cells and remain NO-sensitive.

  15. How-To-Do-It: Using Cauliflower to Demonstrate Plant Tissue Culture.

    Science.gov (United States)

    Haldeman, Janice H.; Ellis, Jane P.

    1988-01-01

    Presents techniques used for disinfestation of plant material, preparation of equipment and media, and laboratory procedures for tissue culture using cauliflower. Details methods for preparing solutions and plant propagation by cloning. (CW)

  16. Effect of vanadium on plant growth and its accumulation in plant tissues

    Directory of Open Access Journals (Sweden)

    Narumol Vachirapatama

    2011-06-01

    Full Text Available Hydroponic experiments were conducted to investigate vanadium uptake by Chinese green mustard and tomato plantsand its effect on their growth. Twenty-eight (Chinese green mustard and 79 days (tomato after germination, the plants wereexposed for a further seven days to a solution containing six different concentrations of ammonium metavanadate (0-80 mg/lNH4VO3. The vanadium accumulated in the plant tissues were determined by ion-interaction high performance liquid chromatography,with confirmation by magnetic sector ICP-MS.The results indicated that nutrient solution containing more than 40 mg/l NH4VO3 affected plant growth for bothChinese green mustard and tomato plant. Chinese green mustard grown in the solution containing NH4VO3 at the concentrationsof 40 and 80 mg/l had stem length, number of leaves, dry weight of leaf, stem and root significantly lower than those ofplants grown in the solution containing 0-20 mg/l NH4VO3. Tomato plants were observed to wilt after four days in contactwith the nutrient solutions containing 40 and 80 mg/l NH4VO3. As the vanadium concentrations increased, a resultantdecrease in the stem length, root fresh weight, and fruit fresh weight were noted. The accumulation of vanadium was higher inthe root compared with leaf, stem, or fruit. Measured levels of vanadium, from a nutrient solution containing 40 mg/l NH4VO3,were 328, 340, and 9.66x103 g/g in the leaf, stem and root for Chinese green mustard, and 4.04 and 4.01x103 g/g in the fruitand roots for tomato plants, respectively.

  17. Embryogenesis and plant regeneration of Medicago spp. in tissue culture.

    Science.gov (United States)

    Nagarajan, P; McKenzie, J S; Walton, P D

    1986-02-01

    Ten cultivars and breeding lines from two species of alfalfa (Medicago media and M. sativa) were screened for their ability to produce embryos and plantlets from the root and hypocotyl under three different tissue culture protocols. The three protocols differed in basal salt composition, vitamins, hormones and cytokinin additions. That protocol having a high 2-4,D low cytokinin induction step gave the highest percentage of embryogenic calli in some cultivars and lines. M. media cultivars and breeding lines had a high percentage of embryoid formation. M. sativa cultivars gave no embryoid formation. Two M. media breeding lines (Br1 and Le1), which were intermediate in the percentage of embryogenic calli formed from explants, had the highest number of regenerated plants established in soil. The creeping rooted M. media cultivar Heinrichs produced the highest percentage of embryogenic calli from explants but most of these embryoids were abnormal and failed to grow in soil or vermiculite. Accordingly, successful regeneration is directly related to the quality and quantity of the embryoids produced.

  18. Stachyose synthesis in source leaf tissues of the CAM plant Xerosicyos danguyi H. Humb

    Energy Technology Data Exchange (ETDEWEB)

    Madore, M.A.; Mitchell, D.E.; Boyd, C.M. (Univ. of California, Riverside (USA))

    1988-07-01

    Leaf tissues from Xerosicyos danguyi H. Humb., a succulent member of the Cucurbitaceae, were found to possess both galactinol synthase activity and the capacity for photosynthetic production of stachyose, the phloem transport oligosaccahride common to other nonsucculent cucurbits, the amounts of stachyose isolated from leaf tissues, and the extractable activity of galactinol synthase, were somewhat higher in leaf tissues obtained from plants operating in the Crassulacean acid metabolism (CAM) mode (well watered plants) compared to leaf tissues from plants operating in the CAM-idling mode (water-stressed plants). In contrast, in leaf discs, the photosynthetic incorporation of label into stachyose following pulse labeling with {sup 14}CO{sub 2} was similar for stressed and for nonstressed tissues. Stachyose could be extracted from, and was synthesized photosynthetically by, leaf discs which contained no vascular tissues, indicating that synthesis of stachyose can occur in photosynthetic mesophyll cells of Xerosicyos.

  19. Bacterial Communities Associated with Different Anthurium andraeanum L. Plant Tissues

    OpenAIRE

    Sarria-Guzmán, Yohanna; Chávez-Romero, Yosef; Gómez-Acata, Selene; Montes-Molina, Joaquín Adolfo; Morales-Salazar, Eleacin; Dendooven,Luc; Yendi E. Navarro-Noya

    2016-01-01

    Plant-associated microbes have specific beneficial functions and are considered key drivers for plant health. The bacterial community structure of healthy Anthurium andraeanum L. plants was studied by 16S rRNA gene pyrosequencing associated with different plant parts and the rhizosphere. A limited number of bacterial taxa, i.e., Sinorhizobium, Fimbriimonadales, and Gammaproteobacteria HTCC2089 were enriched in the A. andraeanum rhizosphere. Endophytes were more diverse in the roots than in th...

  20. Pattern matching and adaptive image segmentation applied to plant reproduction by tissue culture

    Science.gov (United States)

    Vazquez Rueda, Martin G.; Hahn, Federico

    1999-03-01

    This paper shows the results obtained in a system vision applied to plant reproduction by tissue culture using adaptive image segmentation and pattern matching algorithms, this analysis improves the number of tissue obtained and minimize errors, the image features of tissue are considered join to statistical analysis to determine the best match and results. Tests make on potato plants are used to present comparative results with original images processed with adaptive segmentation algorithm and non adaptive algorithms and pattern matching.

  1. Esau's Plant anatomy: meristems, cells, and tissues of the plant body : their structure, function, and development

    National Research Council Canada - National Science Library

    Evert, Ray Franklin; Esau, Katherine; Eichhorn, Susan E

    2006-01-01

    ... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xix Chapter 1 Structure and Development of the Plant Body- An Overview . . . . . . . . . . . . . . . . . . . . . . . . 1 Internal Organization of the Plant Body...

  2. Application of plant tissue cultures in phytoremediation research: incentives and limitations.

    Science.gov (United States)

    Doran, Pauline M

    2009-05-01

    The aim of this review is to critically assess the benefits and limitations associated with the use of in vitro plant cell and organ cultures as research tools in phytoremediation studies. Plant tissue cultures such as callus, cell suspensions, and hairy roots are applied frequently in phytoremediation research as model plant systems. In vitro cultures offer a range of experimental advantages in studies aimed at examining the intrinsic metabolic capabilities of plant cells and their capacity for toxicity tolerance. The ability to identify the contributions of plant cells to pollutant uptake and detoxification without interference from microorganisms is of particular significance in the search for fundamental knowledge about plants. However, if the ultimate goal of plant tissue culture experiments is the development of practical phytoremediation technology, the limitations inherent in the use of in vitro cultures as a representative of whole plants in the field must be recognized. The bioavailability of contaminants and the processes of pollutant uptake and metabolite distribution are likely to be substantially different in the two systems; this can lead to qualitative as well as quantitative differences in metabolic profiles and tolerance characteristics. Yet, many studies have demonstrated that plant tissue cultures are an extremely valuable tool in phytoremediation research. The results derived from tissue cultures can be used to predict the responses of plants to environmental contaminants, and to improve the design and thus reduce the cost of subsequent conventional whole plant experiments.

  3. Comparative study on Allium schoenoprasum cultivated plant and Allium schoenoprasum tissue culture organs antioxidant status.

    Science.gov (United States)

    Stajner, D; Popović, B M; Calić-Dragosavac, D; Malenčić, D; Zdravković-Korać, S

    2011-11-01

    This study was designed to examine Allium schoenoprasum tissue culture organs antioxidant and scavenging activity and to make a comparison between Allium schoenoprasum cultivated plant and Allium schoenoprasum tissue culture organs antioxidant activity. This study reports the results on the root, stalk and leaf antioxidant enzyme activities (superoxide dismutase, catalase, guaiacol peroxidase and glutathione peroxidase), reduced glutathione quantity, flavonoids and soluble protein contents and quantities of malonyldialdehyde and ·OH radical. In Allium schoenoprasum tissue culture organs the total antioxidant capacity was determined by the FRAP method and scavenger activity by the DPPH method. The present results indicated that the crude extract of Allium schoenoprasum tissue culture exhibited antioxidant and scavenging abilities in all investigated plant parts, especially in the roots. According to our results, the tissue culture plants exhibited the highest activities in the roots in contrast to the cultivated plants where highest activities were observed in the leaves.

  4. A high-resolution method for the localization of proanthocyanidins in plant tissues

    Directory of Open Access Journals (Sweden)

    Panter Stephen

    2011-05-01

    Full Text Available Abstract Background Histochemical staining of plant tissues with 4-dimethylaminocinnamaldehyde (DMACA or vanillin-HCl is widely used to characterize spatial patterns of proanthocyanidin accumulation in plant tissues. These methods are limited in their ability to allow high-resolution imaging of proanthocyanidin deposits. Results Tissue embedding techniques were used in combination with DMACA staining to analyze the accumulation of proanthocyanidins in Lotus corniculatus (L. and Trifolium repens (L. tissues. Embedding of plant tissues in LR White or paraffin matrices, with or without DMACA staining, preserved the physical integrity of the plant tissues, allowing high-resolution imaging that facilitated cell-specific localization of proanthocyanidins. A brown coloration was seen in proanthocyanidin-producing cells when plant tissues were embedded without DMACA staining and this was likely to have been due to non-enzymatic oxidation of proanthocyanidins and the formation of colored semiquinones and quinones. Conclusions This paper presents a simple, high-resolution method for analysis of proanthocyanidin accumulation in organs, tissues and cells of two plant species with different patterns of proanthocyanidin accumulation, namely Lotus corniculatus (birdsfoot trefoil and Trifolium repens (white clover. This technique was used to characterize cell type-specific patterns of proanthocyanidin accumulation in white clover flowers at different stages of development.

  5. Therapeutically important proteins from in vitro plant tissue culture systems.

    Science.gov (United States)

    Doran, Pauline M

    2013-01-01

    Plant cells cultured in liquid medium in bioreactors are now being used commercially to produce biopharmaceutical proteins. The emergence of in vitro plant cell culture as a production vehicle reflects the importance of key biosafety and biocontainment concerns affecting the competitiveness of alternative systems such as mammalian cell culture and agriculture. Food plant species are particularly attractive as hosts for in vitro protein production: the risk of transgene escape and food chain contamination is eliminated using containment facilities, while regulatory approval for oral delivery of drugs may be easier than if non-edible species were used. As in whole plants, proteolysis in cultured plant cells can lead to significant degradation of foreign proteins after synthesis; however, substantial progress has been made to counter the destructive effects of proteases in plant systems. Although protein secretion into the culture medium is advantageous for product recovery and purification, measures are often required to minimise extracellular protease activity and product losses due to irreversible surface adsorption. Disposable plastic bioreactors, which are being used increasingly in mammalian cell bioprocessing, are also being adopted for plant cell culture to allow rapid scale-up and generation of saleable product. This review examines a range of technical and regulatory issues affecting the choice of industrial production platform for foreign proteins, and assesses progress in the development of in vitro plant systems for biopharmaceutical production.

  6. Bacterial Communities Associated with Different Anthurium andraeanum L. Plant Tissues

    Science.gov (United States)

    Sarria-Guzmán, Yohanna; Chávez-Romero, Yosef; Gómez-Acata, Selene; Montes-Molina, Joaquín Adolfo; Morales-Salazar, Eleacin; Dendooven, Luc; Navarro-Noya, Yendi E.

    2016-01-01

    Plant-associated microbes have specific beneficial functions and are considered key drivers for plant health. The bacterial community structure of healthy Anthurium andraeanum L. plants was studied by 16S rRNA gene pyrosequencing associated with different plant parts and the rhizosphere. A limited number of bacterial taxa, i.e., Sinorhizobium, Fimbriimonadales, and Gammaproteobacteria HTCC2089 were enriched in the A. andraeanum rhizosphere. Endophytes were more diverse in the roots than in the shoots, whereas all shoot endophytes were found in the roots. Streptomyces, Flavobacterium succinicans, and Asteroleplasma were only found in the roots, Variovorax paradoxus only in the stem, and Fimbriimonas 97%-OTUs only in the spathe, i.e., considered specialists, while Brevibacillus, Lachnospiraceae, Pseudomonas, and Pseudomonas pseudoalcaligenes were generalist and colonized all plant parts. The anaerobic diazotrophic bacteria Lachnospiraceae, Clostridium sp., and Clostridium bifermentans colonized the shoot system. Phylotypes belonging to Pseudomonas were detected in the rhizosphere and in the substrate (an equiproportional mixture of soil, cow manure, and peat), and dominated the endosphere. Pseudomonas included nine 97%-OTUs with different patterns of distribution and phylogenetic affiliations with different species. P. pseudoalcaligenes and P. putida dominated the shoots, but were also found in the roots and rhizosphere. P. fluorescens was present in all plant parts, while P. resinovorans, P. denitrificans, P. aeruginosa, and P. stutzeri were only detected in the substrate and rhizosphere. The composition of plant-associated bacterial communities is generally considered to be suitable as an indicator of plant health. PMID:27524305

  7. An assay for secologanin in plant tissues based on enzymatic conversion into strictosidine

    DEFF Research Database (Denmark)

    Hallard, Didier; van der Heijden, Robert; Contin, Adriana

    1998-01-01

    The secoiridoid glucoside secologanin is the terpenoid building block in the biosynthesis of terpenoid indole alkaloids. A method for its determination in plant tissues and cell suspension cultures has been developed. This assay is based on the condensation of secologanin with tryptamine, yielding...... of STR for secologanin, in combination with a sensitive and selective HPLC system, allows a simple extraction of secologanin from plant tissue. The detection limit of this methos is 15 ng secologanin. Using this assay, secologanin contents were determined in tissues of various plant species; Lonicera...... xylosteum hairy roots were found to contain 1 % of secologanin on a dry weight basis....

  8. Vascular tissue differentiation and pattern formation in plants.

    Science.gov (United States)

    Ye, Zheng-Hua

    2002-01-01

    Vascular tissues, xylem and phloem, are differentiated from meristematic cells, procambium, and vascular cambium. Auxin and cytokinin have been considered essential for vascular tissue differentiation; this is supported by recent molecular and genetic analyses. Xylogenesis has long been used as a model for study of cell differentiation, and many genes involved in late stages of tracheary element formation have been characterized. A number of mutants affecting vascular differentiation and pattern formation have been isolated in Arabidopsis. Studies of some of these mutants have suggested that vascular tissue organization within the bundles and vascular pattern formation at the organ level are regulated by positional information.

  9. Plants regenerated from tissue culture contain stable epigenome changes in rice.

    Science.gov (United States)

    Stroud, Hume; Ding, Bo; Simon, Stacey A; Feng, Suhua; Bellizzi, Maria; Pellegrini, Matteo; Wang, Guo-Liang; Meyers, Blake C; Jacobsen, Steven E

    2013-03-19

    Most transgenic crops are produced through tissue culture. The impact of utilizing such methods on the plant epigenome is poorly understood. Here we generated whole-genome, single-nucleotide resolution maps of DNA methylation in several regenerated rice lines. We found that all tested regenerated plants had significant losses of methylation compared to non-regenerated plants. Loss of methylation was largely stable across generations, and certain sites in the genome were particularly susceptible to loss of methylation. Loss of methylation at promoters was associated with deregulated expression of protein-coding genes. Analyses of callus and untransformed plants regenerated from callus indicated that loss of methylation is stochastically induced at the tissue culture step. These changes in methylation may explain a component of somaclonal variation, a phenomenon in which plants derived from tissue culture manifest phenotypic variability. DOI:http://dx.doi.org/10.7554/eLife.00354.001.

  10. Yield improvement strategies for the production of secondary metabolites in plant tissue culture: silymarin from Silybum marianum tissue culture.

    Science.gov (United States)

    AbouZid, S

    2014-01-01

    Plant cell culture can be a potential source for the production of important secondary metabolites. This technology bears many advantages over conventional agricultural methods. The main problem to arrive at a cost-effective process is the low productivity. This is mainly due to lack of differentiation in the cultured cells. Many approaches have been used to maximise the yield of secondary metabolites produced by cultured plant cells. Among these approaches: choosing a plant with a high biosynthetic capacity, obtaining efficient cell line for growth and production of metabolite of interest, manipulating culture conditions, elicitation, metabolic engineering and organ culture. This article gives an overview of the various approaches used to maximise the production of pharmaceutically important secondary metabolites in plant cell cultures. Examples of using these different approaches are shown for the production of silymarin from Silybum marianum tissue culture.

  11. The effect of plant growth regulators on optimization of tissue culture ...

    African Journals Online (AJOL)

    USER

    2010-04-05

    Apr 5, 2010 ... tissue culture system in Malaysian upland rice. E. Shahsavari1*, A. A. .... The experiments were arranged in a completely randomized design with four replications. ..... Plant Cell Tissue Organ Cult. 37: 217-242. Ge XJ, Chu ZH, ...

  12. Monitoring programmed cell death of living plant tissues in microfluidics using electrochemical and optical techniques

    DEFF Research Database (Denmark)

    Mark, Christina; Zor, Kinga; Heiskanen, Arto

    This project focuses on developing and applying a tissue culture system with electrochemical and optical detection techniques for tissue culture of barley aleurone layer to increase understanding of the underlying mechanisms of programmed cell death (PCD) in plants. The major advantage of electro...

  13. Dental wax impressions of plant tissues for viewing with scanning electron microscopy (SEM).

    Science.gov (United States)

    Beermann, Anke; Hülskamp, Martin

    2010-09-01

    Scanning electron microscopy (SEM) is a valuable method for examining surface structures. Taking wax impressions of plant structures, such as leaves, is a nondestructive procedure that makes it possible to view changes in surface structures over time, such as during development. This protocol describes a method for making dental wax impressions of plant tissues.

  14. Adaptive image segmentation applied to plant reproduction by tissue culture

    Science.gov (United States)

    Vazquez Rueda, Martin G.; Hahn, Federico; Zapata, Jose L.

    1997-04-01

    This paper presents that experimental results obtained on indoor tissue culture using the adaptive image segmentation system. The performance of the adaptive technique is contrasted with different non-adaptive techniques commonly used in the computer vision field to demonstrate the improvement provided by the adaptive image segmentation system.

  15. Production of therapeutic proteins through plant tissue and cell culture

    Directory of Open Access Journals (Sweden)

    Reza S. Gharelo

    2016-04-01

    Full Text Available Nowadays, pharmaceutical recombinant protein is increasingly used in treatment of many diseases such as hepatitis, anemia, diabetes and cancer. Different protein expression systems have been used for the expression of recombinant proteins in which each of them face obstacles that make utilizing them as comprehensive expression system in order to express wide variety of proteins difficult. Plant cell as a eukaryotic expression system have many advantages compared to other hosts. They are very "safe" and significantly decrease concerns about the contamination of recombinant proteins with human pathogens. In addition to this, plants as eukaryotic expression system perform proper post-translational modification, in case of eukaryotic proteins, and appropriate folding resulting in right function in biological environments. Therefore, the production of pharmaceutical protein through plant cells can be absolutely promising approach. In this review, the production of pharmaceutical protein in plant cells, advantages and disadvantages, offered methods and techniques for developing recombinant protein yields, and affective factors on the whole process of pharmaceutical protein expression in the molecular level will be reviewed.

  16. A simple way to identify non-viable cells within living plant tissue using confocal microscopy

    Directory of Open Access Journals (Sweden)

    Truernit Elisabeth

    2008-06-01

    Full Text Available Abstract Background Plant cell death is a normal process during plant development. Mutant plants may exhibit misregulation of this process, which can lead to severe growth defects. Simple ways of visualising cell death in living plant tissues can aid the study of plant development and physiology. Results Spectral variants of the fluorescent SYTOX dyes were tested for their usefulness for the detection of non-viable cells within plant embryos and roots using confocal laser-scanning microscopy. The dyes were selective for non-viable cells and showed very little background staining in living cells. Simultaneous detection of SYTOX dye and fluorescent protein (e.g. GFP fluorescence was possible. Conclusion The fluorescent SYTOX dyes are useful for an easy and quick first assay of plant cell viability in living plant samples using fluorescence and confocal laser-scanning microscopy.

  17. Enhancing plant regeneration in tissue culture: a molecular approach through manipulation of cytokinin sensitivity.

    Science.gov (United States)

    Hill, Kristine; Schaller, G Eric

    2013-10-01

    Micropropagation is used for commercial purposes worldwide, but the capacity to undergo somatic organogenesis and plant regeneration varies greatly among species. The plant hormones auxin and cytokinin are critical for plant regeneration in tissue culture, with cytokinin playing an instrumental role in shoot organogenesis. Type-B response regulators govern the transcriptional output in response to cytokinin and are required for plant regeneration. In our paper published in Plant Physiology, we explored the functional redundancy among the 11 type-B Arabidopsis response regulators (ARRs). Interestingly, we discovered that the enhanced expression of one family member, ARR10, induced hypersensitivity to cytokinin in multiple assays, including callus greening and shoot induction of explants. Here we 1) discuss the hormone dependence for in vitro plant regeneration, 2) how manipulation of the cytokinin response has been used to enhance plant regeneration, and 3) the potential of the ARR10 transgene as a tool to increase the regeneration capacity of agriculturally important crop plants. The efficacy of ARR10 for enhancing plant regeneration likely arises from its ability to transcriptionally regulate key cytokinin responsive genes combined with an enhanced protein stability of ARR10 compared with other type-B ARRs. By increasing the capacity of key tissues and cell types to respond to cytokinin, ARR10, or other type-B response regulators with similar properties, could be used as a tool to combat the recalcitrance of some crop species to tissue culture techniques.

  18. Mapping element distributions in plant tissues using synchrotron X-ray fluorescence techniques.

    Science.gov (United States)

    Donner, Erica; de Jonge, Martin D; Kopittke, Peter M; Lombi, Enzo

    2013-01-01

    Synchrotron-based X-ray fluorescence (XRF) is allowing substantial advances in several disciplines of plant science by allowing the in situ examination of elements within plant tissues. Continual improvements in detector speed, sensitivity, and resolution are increasing the diversity of questions that can be addressed using this technique, including the in situ analysis of elements (such as nutrients or toxicants) within fresh and hydrated tissues. Here, we describe the general principles for designing and conducting experiments for the examination of elemental distributions in plant material using micro-XRF.

  19. Cloning higher plants from aseptically cultured tissues and cells

    Science.gov (United States)

    Krikorian, A. D.

    1982-01-01

    A review of aseptic culture methods for higher plants is presented, which focuses on the existing problems that limit or prevent the full realization of cloning plants from free cells. It is shown that substantial progress in clonal multiplication has been made with explanted stem tips or lateral buds which can be stimulated to produce numerous precocious axillary branches. These branches can then be separated or subdivided and induced to root in order to yield populations of genetically and phenotypically uniorm plantlets. Similarly, undifferentiated calluses can sometimes be induced to form shoots and/or roots adventitiously. Although the cell culture techniques required to produce somatic embryos are presently rudimentary, steady advances are being made in learning how to stimulate formation of somatic or adventive embryos from totipotent cells grown in suspension cultures. It is concluded that many problems exist in the producing and growing of totipotent or morphogenetically competent cell suspensions, but the potential benefits are great.

  20. Surviving freezing in plant tissues by oomycetous snow molds.

    Science.gov (United States)

    Murakami, Ryo; Yajima, Yuka; Kida, Ken-ichi; Tokura, Katsuyuki; Tojo, Motoaki; Hoshino, Tamotsu

    2015-04-01

    Oomyceteous snow molds, Pythium species, were reported to be less tolerant to chilling and freezing temperatures than other snow mold taxa. However, Pythium species are often found to be pathogenic on mosses in Polar Regions. We investigated the frost resistance of Pythium species from Temperate (Hokkaido, Japan) and Subantarctic Regions. Free mycelia and hyphal swellings, structures for survival, of Pythium iwayamai and Pythium paddicum lost viability within freeze-thaw 3 cycles; however, mycelia in host plants survived the treatment. It was reported that fungi in permafrost are characterized both by the presence of natural cryoprotectants in these ecotopes and by the ability to utilize their inherent mechanisms of protection. It is conceivable that plant substrates or derivatives thereof are natural cryoprotectants, enabling them to provide advantageous conditions to microorganisms under freezing conditions. Our results are the first to experimentally support this hypothesis.

  1. Improved method for HPLC analysis of polyamines, agmatine and aromatic monoamines in plant tissue

    Science.gov (United States)

    Slocum, R. D.; Flores, H. E.; Galston, A. W.; Weinstein, L. H.

    1989-01-01

    The high performance liquid chromatographic (HPLC) method of Flores and Galston (1982 Plant Physiol 69: 701) for the separation and quantitation of benzoylated polyamines in plant tissues has been widely adopted by other workers. However, due to previously unrecognized problems associated with the derivatization of agmatine, this important intermediate in plant polyamine metabolism cannot be quantitated using this method. Also, two polyamines, putrescine and diaminopropane, also are not well resolved using this method. A simple modification of the original HPLC procedure greatly improves the separation and quantitation of these amines, and further allows the simulation analysis of phenethylamine and tyramine, which are major monoamine constituents of tobacco and other plant tissues. We have used this modified HPLC method to characterize amine titers in suspension cultured carrot (Daucas carota L.) cells and tobacco (Nicotiana tabacum L.) leaf tissues.

  2. Improved method for HPLC analysis of polyamines, agmatine and aromatic monoamines in plant tissue

    Science.gov (United States)

    Slocum, R. D.; Flores, H. E.; Galston, A. W.; Weinstein, L. H.

    1989-01-01

    The high performance liquid chromatographic (HPLC) method of Flores and Galston (1982 Plant Physiol 69: 701) for the separation and quantitation of benzoylated polyamines in plant tissues has been widely adopted by other workers. However, due to previously unrecognized problems associated with the derivatization of agmatine, this important intermediate in plant polyamine metabolism cannot be quantitated using this method. Also, two polyamines, putrescine and diaminopropane, also are not well resolved using this method. A simple modification of the original HPLC procedure greatly improves the separation and quantitation of these amines, and further allows the simulation analysis of phenethylamine and tyramine, which are major monoamine constituents of tobacco and other plant tissues. We have used this modified HPLC method to characterize amine titers in suspension cultured carrot (Daucas carota L.) cells and tobacco (Nicotiana tabacum L.) leaf tissues.

  3. REGULATION OF CHLOROPHY LL DEGRADATION IN PLANT TISSUES

    Directory of Open Access Journals (Sweden)

    Syvash O. O.

    2017-06-01

    Full Text Available The purpose of the review was to analyze the basic biochemical processes leading to the chlorophyll degradation and ways to control this process in plant product storage. First of all, this is a complex of enzymatic reactions starting with the hydrolysis of chlorophyll with the formation of acyclic diterpene phytol and water-soluble chlorophyllide. An alternative primary reaction is the removal of magnesium from the chlorophyll tetrapyrrole ring to form pheophytin with the participation of Mg2+-dechelatase and/or low-molecular Mg2+-dechelating substances. The chlorophyll breakdown can also be caused by free radicals formed in the peroxidase-catalyzed reaction of Н2О2 with phenolic compounds or fatty acids. The unstable product of chlorophyll peroxidation, C132 –hydroxychlorophyll a decomposes to colorless low-molecular compounds. Expression of the genes of chlorophyll catabolism enzymes is controlled by phytohormones. Methods for controlling the pigment decomposition during storage of plant products are associated with the use of activators and inhibitors of chlorophyll decomposition. The best known inductor of the synthesis of catabolic enzymes is ethylene, widely used to accelerate fruit ripening. Gibberellins, cytokinins and nitric oxide, on the contrary, slow down the loss of chlorophyll.

  4. Demonstration of the economic feasibility of plant tissue culture for jojoba (Simmondsia chinensis) and Euphorbia spp

    Energy Technology Data Exchange (ETDEWEB)

    Sluis, C.

    1980-09-01

    The economic feasibility of plant tissue culture was demonstrated as applied to two plants: jojoba (Simmondsia chinensis) and Euphorbia spp. The gopher weed (Euphorbia lathyris) was selected as the species of Euphorbia to research due to the interest in this plant as a potential source of hydrocarbon-like compounds. High yield female selections of jojoba were chosen from native stands and were researched to determine the economic feasibility of mass producing these plants via a tissue culture micropropagation program. The female jojoba selection was successfully mass produced through tissue culture. Modifications in initiation techniques, as well as in multiplication media and rooting parameters, were necessary to apply the tissue culture system, which had been developed for juvenile seedling tissue, to mature jojobas. Since prior attempts at transfer of tissue cultured plantlets were unsuccessful, transfer research was a major part of the project and has resulted in a system for transfer of rooted jojoba plantlets to soil. Euphorbia lathyris was successfully cultured using shoot tip cultures. Media and procedures were established for culture initiation, multiplication of shoots, callus induction and growth, and root initiation. Well-developed root systems were not attained and root initiation percentages should be increased if the system is to become commercially feasible.

  5. Embryoidogenesis and plant regeneration from leaf tissue of Gloriosa superba.

    Science.gov (United States)

    Sivakumar, G; Krishnamurthy, K V; Rajendran, T D

    2003-05-01

    The induction, maturation and germination of embryoids from leaf tissue of Gloriosa superba L. were developed by exploiting solid and liquid culture. Nodular calli were obtained from SH medium supplemented with 2,4-D and 2iP. In solid culture, the nodular calli when transferred to 2,4-D along with glycerol gave the best response (68.4 %) in embryoid induction after 20 days. After two subcultures at 7-day intervals in a medium with thiamine instead of glycerol, the embryoids matured. When mature embryoids were transferred to BAP and IBA medium, they gave rise to plantlets with single shoots and roots. In liquid culture, the medium supplemented with NAA and L-glutamine with continuous agitation, the embryoidogenic calli produced embryoids (85 %) after 21 days. The mature embryoids began to turn green and produced shoots and elongated "radicles" after 35 days.

  6. Illuminating a plant's tissue-specific metabolic diversity using computational metabolomics and information theory.

    Science.gov (United States)

    Li, Dapeng; Heiling, Sven; Baldwin, Ian T; Gaquerel, Emmanuel

    2016-11-22

    Secondary metabolite diversity is considered an important fitness determinant for plants' biotic and abiotic interactions in nature. This diversity can be examined in two dimensions. The first one considers metabolite diversity across plant species. A second way of looking at this diversity is by considering the tissue-specific localization of pathways underlying secondary metabolism within a plant. Although these cross-tissue metabolite variations are increasingly regarded as important readouts of tissue-level gene function and regulatory processes, they have rarely been comprehensively explored by nontargeted metabolomics. As such, important questions have remained superficially addressed. For instance, which tissues exhibit prevalent signatures of metabolic specialization? Reciprocally, which metabolites contribute most to this tissue specialization in contrast to those metabolites exhibiting housekeeping characteristics? Here, we explore tissue-level metabolic specialization in Nicotiana attenuata, an ecological model with rich secondary metabolism, by combining tissue-wide nontargeted mass spectral data acquisition, information theory analysis, and tandem MS (MS/MS) molecular networks. This analysis was conducted for two different methanolic extracts of 14 tissues and deconvoluted 895 nonredundant MS/MS spectra. Using information theory analysis, anthers were found to harbor the most specialized metabolome, and most unique metabolites of anthers and other tissues were annotated through MS/MS molecular networks. Tissue-metabolite association maps were used to predict tissue-specific gene functions. Predictions for the function of two UDP-glycosyltransferases in flavonoid metabolism were confirmed by virus-induced gene silencing. The present workflow allows biologists to amortize the vast amount of data produced by modern MS instrumentation in their quest to understand gene function.

  7. A Protocol for Rapid, Measurable Plant Tissue Culture Using Stem Disc Meristem Micropropagation of Garlic ("Allium Sativum L.")

    Science.gov (United States)

    Peat, Gerry; Jones, Meriel

    2012-01-01

    Plant tissue culture is becoming an important technique for the mass propagation of plants. Problems with existing techniques, such as slow growth and contamination, have restricted the practical work in plant tissue culture carried out in schools. The new protocol using garlic meristematic stem discs explained in this article addresses many of…

  8. A Protocol for Rapid, Measurable Plant Tissue Culture Using Stem Disc Meristem Micropropagation of Garlic ("Allium Sativum L.")

    Science.gov (United States)

    Peat, Gerry; Jones, Meriel

    2012-01-01

    Plant tissue culture is becoming an important technique for the mass propagation of plants. Problems with existing techniques, such as slow growth and contamination, have restricted the practical work in plant tissue culture carried out in schools. The new protocol using garlic meristematic stem discs explained in this article addresses many of…

  9. Enhanced electroporation in plant tissues via low frequency pulsed electric fields: influence of cytoplasmic streaming.

    Science.gov (United States)

    Asavasanti, Suvaluk; Stroeve, Pieter; Barrett, Diane M; Jernstedt, Judith A; Ristenpart, William D

    2012-01-01

    Pulsed electric fields (PEF) are known to be effective at permeabilizing plant tissues. Prior research has demonstrated that lower pulse frequencies induce higher rates of permeabilization, but the underlying reason for this response is unclear. Intriguingly, recent microscopic observations with onion tissues have also revealed a correlation between PEF frequency and the subsequent speed of intracellular convective motion, i.e., cytoplasmic streaming. In this paper, we investigate the effect of cytoplasmic streaming on the efficacy of plant tissue permeabilization via PEF. Onion tissue samples were treated with Cytochalasin B, a known inhibitor of cytoplasmic streaming, and changes in cellular integrity and viability were measured over a wide range of frequencies and field strengths. We find that at low frequencies (f streaming results in a 19% decrease in the conductivity disintegration index compared with control samples. Qualitatively, similar results were observed using a microscopic cell viability assay. The results suggest that at low frequencies convection plays a statistically significant role in distributing more conductive fluid throughout the tissue, making subsequent pulses more efficacious. The key practical implication is that PEF pretreatment at low frequency can increase the rate of tissue permeabilization in dehydration or extraction processes, and that the treatment will be most effective when cytoplasmic streaming is most active, i.e., with freshly prepared plant tissues.

  10. Glow in the dark: fluorescent proteins as cell and tissue-specific markers in plants.

    Science.gov (United States)

    Ckurshumova, Wenzislava; Caragea, Adriana E; Goldstein, Rochelle S; Berleth, Thomas

    2011-09-01

    Since the hallmark discovery of Aequorea victoria's Green Fluorescent Protein (GFP) and its adaptation for efficient use in plants, fluorescent protein tags marking expression profiles or genuine proteins of interest have been used to recognize plant tissues and cell types, to monitor dynamic cell fate selection processes, and to obtain cell type-specific transcriptomes. Fluorescent tagging enabled visualization in living tissues and the precise recordings of dynamic expression pattern changes. The resulting accurate recording of cell fate acquisition kinetics in space and time has strongly stimulated mathematical modeling of self-organizing feedback mechanisms. In developmental studies, the use of fluorescent proteins has become critical, where morphological markers of tissues, cell types, or differentiation stages are either not known or not easily recognizable. In this review, we focus on the use of fluorescent markers to identify and illuminate otherwise invisible cell states in plant development.

  11. Glow in the Dark: Fluorescent Proteins as Cell and Tissue-Specific Markers in Plants

    Institute of Scientific and Technical Information of China (English)

    Wenzislava Ckurshumova; Adriana E. Caragea; Rochelle S. Goldstein; Thomas Berleth

    2011-01-01

    Since the hallmark discovery of Aequorea victoria's Green Fluorescent Protein (GFP) and its adaptation for efficient use in plants,fluorescent protein tags marking expression profiles or genuine proteins of interest have been used to recognize plant tissues and cell types,to monitor dynamic cell fate selection processes,and to obtain cell type-specific transcriptomes.Fluorescent tagging enabled visualization in living tissues and the precise recordings of dynamic expression pattern changes.The resulting accurate recording of cell fate acquisition kinetics in space and time has strongly stimulated mathematical modeling of self-organizing feedback mechanisms.In developmental studies,the use of fluorescent proteins has become critical,where morphological markers of tissues,cell types,or differentiation stages are either not known or not easily recognizable.In this review,we focus on the use of fluorescent markers to identify and illuminate otherwise invisible cell states in plant development.

  12. A particle based model to simulate microscale morphological changes of plant tissues during drying.

    Science.gov (United States)

    Karunasena, H C P; Senadeera, W; Brown, R J; Gu, Y T

    2014-08-07

    Fundamental understanding on microscopic physical changes of plant materials is vital to optimize product quality and processing techniques, particularly in food engineering. Although grid-based numerical modelling can assist in this regard, it becomes quite challenging to overcome the inherited complexities of these biological materials especially when such materials undergo critical processing conditions such as drying, where the cellular structure undergoes extreme deformations. In this context, a meshfree particle based model was developed which is fundamentally capable of handling extreme deformations of plant tissues during drying. The model is built by coupling a particle based meshfree technique: Smoothed Particle Hydrodynamics (SPH) and a Discrete Element Method (DEM). Plant cells were initiated as hexagons and aggregated to form a tissue which also accounts for the characteristics of the middle lamella. In each cell, SPH was used to model cell protoplasm and DEM was used to model the cell wall. Drying was incorporated by varying the moisture content, the turgor pressure, and cell wall contraction effects. Compared to the state of the art grid-based microscale plant tissue drying models, the proposed model can be used to simulate tissues under excessive moisture content reductions incorporating cell wall wrinkling. Also, compared to the state of the art SPH-DEM tissue models, the proposed model better replicates real tissues and the cell-cell interactions used ensure efficient computations. Model predictions showed good agreement both qualitatively and quantitatively with experimental findings on dried plant tissues. The proposed modelling approach is fundamentally flexible to study different cellular structures for their microscale morphological changes at dehydration.

  13. Single-step protocol for preparation of plant tissue for analysis by PCR.

    Science.gov (United States)

    Thomson, D; Henry, R

    1995-09-01

    PCR has many applications in the isolation and analysis of plant DNA. The influence of salt and EDTA concentration, pH, incubation time and temperature on the preparation of plant material for PCR was evaluated. A general single-step method was developed in which a small amount of plant tissue was heated in a simple solution. The DNA in the supernatant was found to be suitable for most PCR applications including arbitrarily primed PCR (random-amplified polymorphic DNA) and PCR with specific primers for both single- and multiple-copy genes. The technique is much simpler than those generally used for plant DNA preparation and was successful with tissues from a wide range of species.

  14. Imaging Nuclear Morphology and Organization in Cleared Plant Tissues Treated with Cell Cycle Inhibitors.

    Science.gov (United States)

    de Souza Junior, José Dijair Antonino; de Sa, Maria Fatima Grossi; Engler, Gilbert; Engler, Janice de Almeida

    2016-01-01

    Synchronization of root cells through chemical treatment can generate a large number of cells blocked in specific cell cycle phases. In plants, this approach can be employed for cell suspension cultures and plant seedlings. To identify plant cells in the course of the cell cycle, especially during mitosis in meristematic tissues, chemical inhibitors can be used to block cell cycle progression. Herein, we present a simplified and easy-to-apply protocol to visualize mitotic figures, nuclei morphology, and organization in whole Arabidopsis root apexes. The procedure is based on tissue clearing, and fluorescent staining of nuclear DNA with DAPI. The protocol allows carrying out bulk analysis of nuclei and cell cycle phases in root cells and will be valuable to investigate mutants like overexpressing lines of genes disturbing the plant cell cycle.

  15. Estimation of total Terpenoids concentration in plant tissues using a monoterpene, Linalool as standard reagent.

    OpenAIRE

    sprotocols

    2015-01-01

    Authors: Narayan Ghorai, Sondipon Chakraborty, Shamik Gucchait, Samir Kumar Saha & Suman Biswas ### Abstract Terpenes and terpenoids are primary constituents of essential oils of different type of plants and flowers. Some qualitative estimation methods of terpenoids in plant tissue have been previously described but there is no protocol of estimating the same quantitatively till date. In the present study a protocol has been attempted to estimate the total terpenoids concentrati...

  16. Increase of homologous recombination frequency in vascular tissue of Arabidopsis plants exposed to salt stress.

    Science.gov (United States)

    Boyko, Alex; Hudson, Darryl; Bhomkar, Prasanna; Kathiria, Palak; Kovalchuk, Igor

    2006-06-01

    Here we analyzed the influence of salt stress on plant genome stability. Homologous recombination events were detected in transgenic Arabidopsis plants that carried in their genome a beta-glucuronidase recombination marker. Recombination events were scored as blue sectors using a stereo microscope. Exposure to 50 mM salt resulted in a 3.0-fold increase in recombination frequency. To analyze the organ and tissue specificity of recombination events, we examined cross-sections of leaves, stems and roots. We found that nearly 30% of recombination events in plants grown under normal conditions and nearly 50% of events in plants grown on salt were undetected by the conventional method. Most of the recombination events represented a cluster/group of cells (12 on average), although events with single cells were also detected. Recombination events were very frequent in leaf mesophyll cells. On average, individual recombination events located on leaves contained more cells than events located on roots or stems. Analysis of recombination events in cross-sectioned tissue of salt-treated plants revealed a shift in the distribution of recombination events towards the vascular tissue. We discuss the significance of the finding for plant stress physiology.

  17. Plant tissue culture of fast-growing trees for phytoremediation research.

    Science.gov (United States)

    Couselo, José Luis; Corredoira, Elena; Vieitez, Ana M; Ballester, Antonio

    2012-01-01

    The ability of plants to remove pollutants from the environment is currently used in a simple and low-cost cleaning technology known as phytoremediation. Unfortunately, little is known about the metabolic pathways involved in the transformation of xenobiotic compounds and the ability of certain plants to tolerate, detoxify, and store high concentrations of heavy metals. Plant cell and tissue culture is considered an important tool for fundamental studies that provide information about the plant-contaminant relationships, help to predict plant responses to environmental contaminants, and improve the design of plants with enhanced characteristics for phytoremediation. Callus, cell suspensions, hairy roots, and shoot multiplication cultures are used to study the interactions between plants and pollutants under aseptic conditions. Many plant species have an inherent ability to accumulate/metabolize a variety of pollutants, but they normally produce little biomass. However, fast-growing trees are excellent candidates for phytoremediation because of their rapid growth, extensive root system, and high water uptake. This chapter outlines the in vitro plant production of both somaclonal variants and transgenic plants of Populus spp. that exhibit high tolerance to heavy metals.

  18. Carbon Fluxes between Primary Metabolism and Phenolic Pathway in Plant Tissues under Stress

    Directory of Open Access Journals (Sweden)

    Sofia Caretto

    2015-11-01

    Full Text Available Higher plants synthesize an amazing diversity of phenolic secondary metabolites. Phenolics are defined secondary metabolites or natural products because, originally, they were considered not essential for plant growth and development. Plant phenolics, like other natural compounds, provide the plant with specific adaptations to changing environmental conditions and, therefore, they are essential for plant defense mechanisms. Plant defensive traits are costly for plants due to the energy drain from growth toward defensive metabolite production. Being limited with environmental resources, plants have to decide how allocate these resources to various competing functions. This decision brings about trade-offs, i.e., promoting some functions by neglecting others as an inverse relationship. Many studies have been carried out in order to link an evaluation of plant performance (in terms of growth rate with levels of defense-related metabolites. Available results suggest that environmental stresses and stress-induced phenolics could be linked by a transduction pathway that involves: (i the proline redox cycle; (ii the stimulated oxidative pentose phosphate pathway; and, in turn, (iii the reduced growth of plant tissues.

  19. Tissue culture-induced DNA methylation polymorphisms in repetitive DNA of tomato calli and regenerated plants.

    Science.gov (United States)

    Smulders, M J; Rus-Kortekaas, W; Vosman, B

    1995-12-01

    The propagation of plants through tissue culture can induce a variety of genetic and epigenetic changes. Variation in DNA methylation has been proposed as a mechanism that may explain at least a part of these changes. In the present study, the methylation of tomato callus DNA was compared with that of leaf DNA, from control or regenerated plants, at MspI/HpaII sites around five middle-repetitive sequences. Although the methylation of the internal cytosine in the recognition sequence CCGG varied from zero to nearly full methylation, depending on the probe used, no differences were found between callus and leaf DNA. For the external cytosine, small differences were revealed between leaf and callus DNA with two probes, but no polymorphisms were detected among DNA samples of calli or DNA samples of leaves of regenerated plants. When callus DNA cut with HindIII was studied with one of the probes, H9D9, most of the signal was found in high-molecular-weight DNA, as opposed to control leaf DNA where almost all the signal was in a fragment of 530 bp. Also, an extra fragment of 630 bp was found in the callus DNA that was not present in control leaf DNA. Among leaves of plants regenerated from tissue culture, the 630-bp fragment was found in 10 of 68 regenerated plants. This 630-bp fragment was present among progeny of only 4 of these 10 plants after selfing, i.e. it was partly inherited. In these cases, the fragment was not found in all progeny plants, indicating heterozygosity of the regenerated plants. The data are interpreted as indicating that a HindIII site becomes methylated in callus tissue, and that some of this methylation persists in regenerated plants and is partly transmitted to their progeny.

  20. Identification of dioxin and dioxin-like polychlorbiphenyls in plant tissues and contaminated soils.

    Science.gov (United States)

    Jou, Jin-Juh; Chung, Jen-Chir; Weng, Ying-Ming; Liaw, Shu-Liang; Wang, Ming Kuang

    2007-10-01

    The environmental analysis laboratory (EAL) of the Taiwan environmental protection administration (TEPA) has been monitoring certain sites polluted in southern Taiwan by pentachlorophenol manufacture. The analytical results revealed peculiarities in the concentration distributions in plant tissues. There are no available data on dioxin and dioxin-like polychlorbiphenyls (DL-PCBs), which can be taken up from contaminated soils by plant tissues. Thus, the aims of this study were to identify, understand, and to validate these dioxin and DL-PCBs concentrations in plant tissues of the contaminated soils. This research analyzed ten species of plant tissues, including tappa (Boussonetia papyrifera) and common jasmin orange (Murraya paniculata) from sites in southern Taiwan, with different levels of contamination. Dioxin concentrations in these plant tissues ranged from 12.7 to 2919 ng WHO-TEQ(DF)/kg dry weight (d.w.), with average of 463 ng WHO-TEQ(DF)/kg d.w. (n=16). The DL-PCBs concentrations ranged from 0.236 to 1.75 ng WHO-TEQp/kg d.w., with an average of 0.605 ng WHO-TEQp/kg d.w. (n=8). Tappa is one of the most common and fastest growing plants in Taiwan. It also shows the highest tolerance to environmental contaminants and accumulates dioxin and DL-PCBs. This is one of the best species to take up dioxins and DL-PCBs effectively. It can be recommended as a candidate for dioxin and DL-PCB phyto-remediation. These data are useful to evaluate bioaccumulation of dioxin and DL-PCBs, and to study the capability of phyto-remediation in contaminated soils.

  1. Permeabilization of plant tissues by monopolar pulsed electric fields: effect of frequency.

    Science.gov (United States)

    Asavasanti, Suvaluk; Ristenpart, William; Stroeve, Pieter; Barrett, Diane M

    2011-01-01

    Pulsed electric fields (PEF) nonthermally induce cell membrane permeabilization and thereby improve dehydration and extraction efficiencies in food plant materials. Effects of electrical field strength and number of pulses on plant tissue integrity have been studied extensively. Two previous studies on the effect of pulse frequency, however, did not provide a clear view: one study suggested no effect of frequency, while the other found a greater impact on tissue integrity at lower frequency. This study establishes the effect of pulse frequency on integrity of onion tissues. Changes in electrical characteristics, ion leakage, texture parameters, and percent weight loss were quantified for a wide range of pulse frequencies under conditions of fixed field strength and pulse number. Optical microscopy and viable-cell staining provided direct visualization of effects on individual cells. The key finding is that lower frequencies (f plant tissue. We hypothesize that cytoplasmic streaming plays a significant role in moving conductive ionic species from permeabilized cells to the intercellular space between plant cells, making subsequent pulses more efficacious at sufficiently low frequencies. The results suggest that decreasing the pulse frequency in PEF may minimize the number of pulses needed to achieve a desired amount of permeabilization, thus lowering the total energy consumption. Practical Application: PEF cause pores to be formed in plant cell membranes, thereby improve moisture removal and potential extraction of desirable components. This study used in situ microscopic evaluation of onion cells, as they were pulsed with electric fields at different frequencies, to determine whether frequency was an important parameter. We illustrate that membranes were more effectively broken at lower frequencies as compared to higher frequencies. Application of this information will allow for improved design of PEF systems for more energy efficient dehydration or extraction of

  2. Visualizing metabolite distribution and enzymatic conversion in plant tissues by desorption electrospray ionization mass spectrometry imaging

    DEFF Research Database (Denmark)

    Li, Bin; Baden, Camilla Knudsen; Hansen, Natascha Kristine Krahl

    2013-01-01

    In comparison to the technology platforms developed to localize transcripts and proteins, imaging tools for visualization of metabolite distributions in plant tissues are less well developed and lack versatility. This hampers our understanding of plant metabolism and dynamics. In this study we......) tubers. The hydroxynitrile glucoside levels were highest in the outer cell layers, as verified by LC-MS analyses. The unexpected discovery of a hydroxynitrile derived di-glycoside shows the potential of DESI-MSI to discover and guide investigations into new metabolic routes. © 2013 The Authors. The Plant...

  3. Target detect system in 3D using vision apply on plant reproduction by tissue culture

    Science.gov (United States)

    Vazquez Rueda, Martin G.; Hahn, Federico

    2001-03-01

    This paper presents the preliminary results for a system in tree dimension that use a system vision to manipulate plants in a tissue culture process. The system is able to estimate the position of the plant in the work area, first calculate the position and send information to the mechanical system, and recalculate the position again, and if it is necessary, repositioning the mechanical system, using an neural system to improve the location of the plant. The system use only the system vision to sense the position and control loop using a neural system to detect the target and positioning the mechanical system, the results are compared with an open loop system.

  4. PLANT TISSUE CULTURE IN BIOTECHNOLOGY: RECENT ADVANCES IN TRANSFORMATION THROUGH SOMATIC EMBRYOGENESIS

    Directory of Open Access Journals (Sweden)

    V. A. Sidorov

    2012-08-01

    Full Text Available Plant genetic transformation has become an important biotechnology tool for the improvement of many crops. A solid foundation for the fast development and implementation of biotechnology in agriculture was provided by achievements in plant tissue culture. On the 30th anniversary of plant transformation, I report the advancements, recent challenges and shifts in methodology of transformation. The main focus of this paper will be on conventional and novel approaches for genetic improvements of soybean, cotton and corn. I will also highlight results on the transformation of these crops that have considerably been improved by modern biotechnology.

  5. Prolific plant regeneration from protoplast-derived tissues of Lotus corniculatus L. (birdsfoot trefoil).

    Science.gov (United States)

    Ahuja, P S; Hadiuzzaman, S; Davey, M R; Cocking, E C

    1983-04-01

    Protoplasts isolated enzymatically from seedling roots, hypocotyls and cotyledons of Lotus corniculatus L. produced callus which underwent prolific shoot regeneration. The rapidity and ease of recovering plants from protoplast-derived tissues makes this forage legume an attractive experimental system for genetic manipulation.

  6. Tissue culture-induced DNA methylation polymorphisms in repetitive DNA of tomato calli and regenerated plants

    NARCIS (Netherlands)

    Smulders, M.J.M.; Rus-Kortekaas, W.; Vosman, B.

    1995-01-01

    The propagation of plants through tissue culture can induce a variety of genetic and epigenetic changes. Variation in DNA methylation has been proposed as a mechanism that may explain at least a part of these changes. In the present study, the methylation of tomato callus DNA was compared with that

  7. Lactococcus lactis metabolism and gene expression during growth on plant tissues.

    Science.gov (United States)

    Golomb, Benjamin L; Marco, Maria L

    2015-01-01

    Lactic acid bacteria have been isolated from living, harvested, and fermented plant materials; however, the adaptations these bacteria possess for growth on plant tissues are largely unknown. In this study, we investigated plant habitat-specific traits of Lactococcus lactis during growth in an Arabidopsis thaliana leaf tissue lysate (ATL). L. lactis KF147, a strain originally isolated from plants, exhibited a higher growth rate and reached 7.9-fold-greater cell densities during growth in ATL than the dairy-associated strain L. lactis IL1403. Transcriptome profiling (RNA-seq) of KF147 identified 853 induced and 264 repressed genes during growth in ATL compared to that in GM17 laboratory culture medium. Genes induced in ATL included those involved in the arginine deiminase pathway and a total of 140 carbohydrate transport and metabolism genes, many of which are involved in xylose, arabinose, cellobiose, and hemicellulose metabolism. The induction of those genes corresponded with L. lactis KF147 nutrient consumption and production of metabolic end products in ATL as measured by gas chromatography-time of flight mass spectrometry (GC-TOF/MS) untargeted metabolomic profiling. To assess the importance of specific plant-inducible genes for L. lactis growth in ATL, xylose metabolism was targeted for gene knockout mutagenesis. Wild-type L. lactis strain KF147 but not an xylA deletion mutant was able to grow using xylose as the sole carbon source. However, both strains grew to similarly high levels in ATL, indicating redundancy in L. lactis carbohydrate metabolism on plant tissues. These findings show that certain strains of L. lactis are well adapted for growth on plants and possess specific traits relevant for plant-based food, fuel, and feed fermentations.

  8. RNA extraction from plant tissues: the use of calcium to precipitate contaminating pectic sugars.

    Science.gov (United States)

    Dal Cin, Valeriano; Danesin, Marcello; Rizzini, Fabio Massimo; Ramina, Angelo

    2005-10-01

    Several protocols and commercial kits are used for the extraction of nucleic acids from different plant tissues. Although there are several procedures available to remove sugars, which hinder the extraction of clean genomic DNA, there are few to assist with extraction of RNA. Those presently used include precipitations with ethylene glycol monobutyl ether or lithium chloride (LiCl), or centrifugation in cesium chloride (CsCl) gradients, but these generally either do not allow high recovery of RNA, are time consuming, rely on hazardous chemicals or need special equipment. Here we present the use of the simple cation, Ca2+, which has been tested and shown to be very efficient for the precipitation of high molecular weight pectic sugars during RNA extraction. Results are presented for different plant tissues, especially tissues of peach and apple fruits at varying ripening stages.

  9. A novel approach for studying programmed cell death in living plant tissues

    DEFF Research Database (Denmark)

    Mark, Christina

    using the inhibitor DPI. The new incubation system for immobilised aleurone layers enabled simple, user friendly handling of plant tissue incubations and facilitated transient expression studies in plant tissues by particle bombardment as well as time course studies on the same population of cells......Programmed cell death (PCD) is a highly regulated process in which cells are killed as part of developmental programmes or as defence mechanisms against pathogens, but the process is less well understood in plant cells compared to animal cells. Reactive oxygen species (ROS) are involved in PCD...... and quality of crops and thus contribute to solving the increasing food demands of the planet. Examples of this could be the development of cultivars with enhanced and/or faster response to pathogen attacks, or cultivars with increased grain filling and hence increased starch content through delayed cell...

  10. In vitro propagation of plant virus using different forms of plant tissue culture and modes of culture operation.

    Science.gov (United States)

    Shih, Sharon M-H; Doran, Pauline M

    2009-09-10

    Plant virus accumulation was investigated in vitro using three different forms of plant tissue culture. Suspended cells, hairy roots and shooty teratomas of Nicotiana benthamiana were infected with tobacco mosaic virus (TMV) using the same initial virus:biomass ratio. Viral infection did not affect tissue growth or morphology in any of the three culture systems. Average maximum virus concentrations in hairy roots and shooty teratomas were similar and about an order of magnitude higher than in suspended cells. Hairy roots were considered the preferred host because of their morphological stability in liquid medium and relative ease of culture. The average maximum virus concentration in the hairy roots was 0.82+/-0.14 mg g(-1) dry weight; viral coat protein represented a maximum of approximately 6% of total soluble protein in the biomass. Virus accumulation in hairy roots was investigated further using different modes of semi-continuous culture operation aimed at prolonging the root growth phase and providing nutrient supplementation; however, virus concentrations in the roots were not enhanced compared with simple batch culture. The relative infectivity of virus in the biomass declined by 80-90% during all the cultures tested, irrespective of the form of plant tissue used or mode of culture operation. Hairy root cultures inoculated with a transgenic TMV-based vector in batch culture accumulated green fluorescent protein (GFP); however, maximum GFP concentrations in the biomass were relatively low at 39 microg g(-1) dry weight, probably due to genetic instability of the vector. This work highlights the advantages of using hairy roots for in vitro propagation of TMV compared with shooty teratomas and suspended plant cells, and demonstrates that batch root culture is more effective than semi-continuous operations for accumulation of high virus concentrations in the biomass.

  11. Plant Tissue Culture Development and Biotechnology, Chapter 10: Molecular Tools for Studying Plant Genetic Diversity

    Science.gov (United States)

    The ubiquitous nature of DNA is a central theme for all biology. The nucleus of each cell that makes up an organism contains genomic DNA, which is the blueprint for life. The differential expression of genes within each cell gives rise to different tissues, organs and, ultimately, different organism...

  12. Plant species differ in early seedling growth and tissue nutrient responses to arbuscular and ectomycorrhizal fungi.

    Science.gov (United States)

    Holste, Ellen K; Kobe, Richard K; Gehring, Catherine A

    2017-04-01

    Experiments with plant species that can host both arbuscular mycorrhizal fungi (AMF) and ectomycorrhizal fungi (EMF) are important to separating the roles of fungal type and plant species and understanding the influence of the types of symbioses on plant growth and nutrient acquisition. We examined the effects of mycorrhizal fungal type on the growth and tissue nutrient content of two tree species (Eucalyptus grandis and Quercus costaricensis) grown under four nutrient treatments (combinations of low versus high nitrogen (N) and phosphorus (P) with different N:P ratios) in the greenhouse. Trees were inoculated with unidentified field mixtures of AMF or EMF species cultivated on root fragments of AMF- or EMF-specific bait plants. In E. grandis, inoculation with both AMF and EMF positively affected belowground plant dry weight and negatively affected aboveground dry weight, while only inoculation with AMF increased tissue nutrient content. Conversely, Q. costaricensis dry weight and nutrient content did not differ significantly among inoculation treatments, potentially due to its dependence on cotyledon reserves for growth. Mineral nutrition of both tree species differed with the ratio of N to P applied while growth did not. Our results demonstrate that both tree species' characteristics and the soil nutrient environment can affect how AMF and EMF interact with their host plants. This research highlights the importance of mycorrhizal fungal-tree-soil interactions during early seedling growth and suggests that differences between AMF and EMF associations may be crucial to understanding forest ecosystem functioning.

  13. Plant tissue culture--an opportunity for the production of nutraceuticals.

    Science.gov (United States)

    Lucchesini, Mariella; Mensuali-Sodi, Anna

    2010-01-01

    This chapter provides a short discussion about the opportunity to cultivate in vitro plant tissue of species which synthesize secondary metabolites of nutraceutical interest. The introduction of species of particular interest in cultivation and domestication, can be an alternative to the harvest of wild species. In vitro culture techniques are a useful tool to improve production and marketing nutraceutical species which allows to make a rapid clonal propagation of plants selected for their active principles. The techniques of tissue culture are described in detail. In particular, it is underlined the necessity to clone selected plants and produce true-type plants when standardized plant products are the main goal. This can be reached by conventional micropropagation protocols culturing plants in vitro through the five culture phases. Another approach consists in applying unconventional systems in the last phase of in vitro culture which permit to develop autotrophy of the explants. Autotrophic growth improves the quality of the multiplied shoots and facilitates the acclimatization of the plantlets.

  14. Wounding in the plant tissue: the defense of a dangerous passage

    Directory of Open Access Journals (Sweden)

    Daniel Valentin Savatin

    2014-09-01

    Full Text Available Plants are continuously exposed to agents such as herbivores and environmental mechanical stresses that cause wounding and open the way to the invasion by microbial pathogens. Wounding provides nutrients to pathogens and facilitates their entry into the tissue and subsequent infection. Plants have evolved constitutive and induced defense mechanisms to properly respond to wounding and prevent infection. The constitutive defenses are represented by physical barriers, i.e. the presence of cuticle or lignin, or by metabolites that act as toxins or deterrents for herbivores. Plants are also able to sense the injured tissue as an altered self and induce responses similar to those activated by pathogen infection. Endogenous molecules released from wounded tissue may act as Damage-Associated Molecular Patterns (DAMPs that activate the plant innate immunity. Wound-induced responses are both rapid, such as the oxidative burst and the expression of defense-related genes, and late, such as the callose deposition, the accumulation of proteinase inhibitors and of hydrolytic enzymes (i.e. chitinases and gluganases. Typical examples of DAMPs involved in the response to wounding are the peptide systemin and the oligogalacturonides, which are oligosaccharides released from the pectic component of the cell wall. Responses to wounding take place both at the site of damage (local response and systemically (systemic response and are mediated by hormones such as jasmonic acid, ethylene, salicylic acid and abscisic acid.

  15. Molecular farming of pharmaceutical proteins using plant suspension cell and tissue cultures.

    Science.gov (United States)

    Schillberg, Stefan; Raven, Nicole; Fischer, Rainer; Twyman, Richard M; Schiermeyer, Andreas

    2013-01-01

    Plants have been used for more than 20 years to produce recombinant proteins but only recently has the focus shifted away from proof-of-principle studies (i.e. is my protein expressed and is it functional?) to a serious consideration of the requirements for sustainable productivity and the regulatory approval of pharmaceutical products (i.e. is my protein safe, is it efficacious, and does the product and process comply with regulatory guidelines?). In this context, plant tissue and cell suspension cultures are ideal production platforms whose potential has been demonstrated using diverse pharmaceutical proteins. Typically, cell/tissue cultures are grown in containment under defined conditions, allowing process controls to regulate growth and product formation, thus ensuring regulatory compliance. Recombinant proteins can also be secreted to the culture medium, facilitating recovery and subsequent purification because cells contain most of the contaminating proteins and can be removed from the culture broth. Downstream processing costs are therefore lower compared to whole plant systems, balancing the higher costs of the fermentation equipment. In this article, we compare different approaches for the production of valuable proteins in plant cell suspension and tissue cultures, describing the advantages and disadvantages as well as challenges that must be overcome to make this platform commercially viable. We also present novel strategies for system and process optimization, helping to increase yields and scalability.

  16. Visualizing metabolite distribution and enzymatic conversion in plant tissues by desorption electrospray ionization mass spectrometry imaging.

    Science.gov (United States)

    Li, Bin; Knudsen, Camilla; Hansen, Natascha Krahl; Jørgensen, Kirsten; Kannangara, Rubini; Bak, Søren; Takos, Adam; Rook, Fred; Hansen, Steen H; Møller, Birger Lindberg; Janfelt, Christian; Bjarnholt, Nanna

    2013-06-01

    In comparison with the technology platforms developed to localize transcripts and proteins, imaging tools for visualization of metabolite distributions in plant tissues are less well developed and lack versatility. This hampers our understanding of plant metabolism and dynamics. In this study, we demonstrate that desorption electrospray ionization mass spectrometry imaging (DESI-MSI) of tissue imprints on porous Teflon may be used to accurately image the distribution of even labile plant metabolites such as hydroxynitrile glucosides, which normally undergo enzymatic hydrolysis by specific β-glucosidases upon cell disruption. This fast and simple sample preparation resulted in no substantial differences in the distribution and ratios of all hydroxynitrile glucosides between leaves from wild-type Lotus japonicus and a β-glucosidase mutant plant that lacks the ability to hydrolyze certain hydroxynitrile glucosides. In wild-type, the enzymatic conversion of hydroxynitrile glucosides and the concomitant release of glucose were easily visualized when a restricted area of the leaf tissue was damaged prior to sample preparation. The gene encoding the first enzyme in hydroxynitrile glucoside biosynthesis in L. japonicus leaves, CYP79D3, was found to be highly expressed during the early stages of leaf development, and the hydroxynitrile glucoside distribution in mature leaves reflected this early expression pattern. The utility of direct DESI-MSI of plant tissue was demonstrated using cryo-sections of cassava (Manihot esculenta) tubers. The hydroxynitrile glucoside levels were highest in the outer cell layers, as verified by LC-MS analyses. The unexpected discovery of a hydroxynitrile-derived di-glycoside shows the potential of DESI-MSI to discover and guide investigations into new metabolic routes.

  17. Pathogen and biological contamination management in plant tissue culture: phytopathogens, vitro pathogens, and vitro pests.

    Science.gov (United States)

    Cassells, Alan C

    2012-01-01

    The ability to establish and grow plant cell, organ, and tissue cultures has been widely exploited for basic and applied research, and for the commercial production of plants (micro-propagation). Regardless of whether the application is for research or commerce, it is essential that the cultures be established in vitro free of biological contamination and be maintained as aseptic cultures during manipulation, growth, and storage. The risks from microbial contamination are spurious experimental results due to the effects of latent contaminants or losses of valuable experimental or commercial cultures. Much of the emphasis in culture contamination management historically focussed on the elimination of phytopathogens and the maintenance of cultures free from laboratory contamination by environmental bacteria, fungi (collectively referred to as "vitro pathogens", i.e. pathogens or environmental micro-organisms which cause culture losses), and micro-arthropods ("vitro pests"). Microbial contamination of plant tissue cultures is due to the high nutrient availability in the almost universally used Murashige and Skoog (Physiol Plant 15:473-497, 1962) basal medium or variants of it. In recent years, it has been shown that many plants, especially perennials, are at least locally endophytically colonized intercellularly by bacteria. The latter, and intracellular pathogenic bacteria and viruses/viroids, may pass latently into culture and be spread horizontally and vertically in cultures. Growth of some potentially cultivable endophytes may be suppressed by the high salt and sugar content of the Murashige and Skoog basal medium and suboptimal temperatures for their growth in plant tissue growth rooms. The management of contamination in tissue culture involves three stages: disease screening (syn. disease indexing) of the stock plants with disease and endophyte elimination where detected; establishment and pathogen and contaminant screening of established initial cultures

  18. Ectopic KNOX Expression Affects Plant Development by Altering Tissue Cell Polarity and Identity.

    Science.gov (United States)

    Richardson, Annis Elizabeth; Rebocho, Alexandra B; Coen, Enrico S

    2016-08-23

    Plant development involves two polarity types: tissue cell (asymmetries within cells are coordinated across tissues) and regional (identities vary spatially across tissues) polarity. Both appear altered in the barley (Hordeum vulgare) Hooded mutant, in which ectopic expression of the KNOTTED1-like Homeobox (KNOX) gene, BKn3, causes inverted polarity of differentiated hairs and ectopic flowers, in addition to wing-shaped outgrowths. These lemma-specific effects allow the spatiotemporal analysis of events following ectopic BKn3 expression, determining the relationship between KNOXs, polarity, and shape. We show that tissue cell polarity, based on localization of the auxin transporter SISTER OF PINFORMED1 (SoPIN1), dynamically reorients as ectopic BKn3 expression increases. Concurrently, ectopic expression of the auxin importer LIKE AUX1 and boundary gene NO APICAL MERISTEM is activated. The polarity of hairs reflects SoPIN1 patterns, suggesting that tissue cell polarity underpins oriented cell differentiation. Wing cell files reveal an anisotropic growth pattern, and computational modeling shows how polarity guiding growth can account for this pattern and wing emergence. The inverted ectopic flower orientation does not correlate with SoPIN1, suggesting that this form of regional polarity is not controlled by tissue cell polarity. Overall, the results suggest that KNOXs trigger different morphogenetic effects through interplay between tissue cell polarity, identity, and growth.

  19. Ectopic KNOX Expression Affects Plant Development by Altering Tissue Cell Polarity and Identity[OPEN

    Science.gov (United States)

    Rebocho, Alexandra B.

    2016-01-01

    Plant development involves two polarity types: tissue cell (asymmetries within cells are coordinated across tissues) and regional (identities vary spatially across tissues) polarity. Both appear altered in the barley (Hordeum vulgare) Hooded mutant, in which ectopic expression of the KNOTTED1-like Homeobox (KNOX) gene, BKn3, causes inverted polarity of differentiated hairs and ectopic flowers, in addition to wing-shaped outgrowths. These lemma-specific effects allow the spatiotemporal analysis of events following ectopic BKn3 expression, determining the relationship between KNOXs, polarity, and shape. We show that tissue cell polarity, based on localization of the auxin transporter SISTER OF PINFORMED1 (SoPIN1), dynamically reorients as ectopic BKn3 expression increases. Concurrently, ectopic expression of the auxin importer LIKE AUX1 and boundary gene NO APICAL MERISTEM is activated. The polarity of hairs reflects SoPIN1 patterns, suggesting that tissue cell polarity underpins oriented cell differentiation. Wing cell files reveal an anisotropic growth pattern, and computational modeling shows how polarity guiding growth can account for this pattern and wing emergence. The inverted ectopic flower orientation does not correlate with SoPIN1, suggesting that this form of regional polarity is not controlled by tissue cell polarity. Overall, the results suggest that KNOXs trigger different morphogenetic effects through interplay between tissue cell polarity, identity, and growth. PMID:27553356

  20. Turmeric powder (Curcuma longa Linn. as an antifungal agent in plant tissue culture studies

    Directory of Open Access Journals (Sweden)

    R.S. Upendra

    2011-11-01

    Full Text Available Culturing the individual plant cells, tissues (explants and organs in laboratory or in vitro on synthetic media (MS media under aseptic conditions is a usual process in plant tissue culture studies. The medium is rich in nutrients, also supports the growth of variety of microorganisms especially bacteria and fungi, which causecontamination of the medium, though the media is sterilized by autoclaving. During the process of cooling and transferring the media, the chances of fungal contamination remain high. This is avoided to the maximum extent following the good laboratory practices. A novel means could be incorporating turmeric, a well -known antifungal agent, into the media. In the present study, attempts were made to avoid fungal contamination using the media with various concentration of turmeric powder. Results of the investigation revealed that turmeric powder used at the concentrations of 0.8 g/L and 1.0 g/L in the media resulted in appreciable control of fungal contamination.

  1. Temperature responses of substrate carbon conversion efficiencies and growth rates of plant tissues.

    Science.gov (United States)

    Hansen, Lee D; Thomas, Nathan R; Arnholdt-Schmitt, Birgit

    2009-12-01

    Growth rates of plant tissues depend on both the respiration rate and the efficiency with which carbon is incorporated into new structural biomass. Calorespirometric measurement of respiratory heat and CO2 rates, from which both efficiency and growth rate can be calculated, is a well established method for determining the effects of rapid temperature changes on the respiratory and growth properties of plant tissues. The effect of the alternative oxidase/cytochrome oxidase activity ratio on efficiency is calculated from first principles. Data on the temperature dependence of the substrate carbon conversion efficiency are tabulated. These data show that epsilon is maximum and approximately constant through the optimum growth temperature range and decreases rapidly as temperatures approach temperature limits to growth. The width of the maximum and the slopes of decreasing epsilon at high and low temperatures vary greatly with species, cultivars and accessions.

  2. Evaluation of Protein Extraction Methods for Proteomic Analysis of Non-Model Recalcitrant Plant Tissues

    OpenAIRE

    2012-01-01

    Plant tissues contain relatively low amounts of proteins whose extraction is often difficult due to the presence of interfering compounds such as rigid cellulosic cell wall, storage polysaccharides, lipids and other contaminants that can cause protein degradation or modification. Therefore it is important to optimize protein extraction and to establish a robust protocol for two-dimensional gel electrophoresis (2-DE) and downstream processing. In this study, acetone, trichloroacetic acid/aceto...

  3. The origin of herbivory on land: Initial patterns of plant tissue consumption by arthropods

    Institute of Scientific and Technical Information of China (English)

    CONRAD LABANDEIRA

    2007-01-01

    The early fossil record of terrestrial arthropod herbivory consists of two pulses.The first pulse was concentrated during the latest Silurian to Early Devonian (417 to 403 Ma),and consists of the earliest evidence for consumption of sporangia and stems (and limited fungivore borings). Herbivorization of most of these tissues was rapid, representing 0 to 20 million-year (m.y.) lags from the earliest occurrences of these organs in the fossil record to their initial consumption (Phase 1). For approximately the next 75 m.y., there was a second,more histologically varied origination and expansion of roots, leaves, wood and seeds,whose earliest evidence for herbivorization occurred from the Middle-Late Mississippian boundary to the Middle Pennsylvanian (327 to 309 Ma). The appearance of this second herbivory pulse during the later Paleozoic (Phase 2) is accompanied by major lags of 98 to 54 m.y. between times of appearance of each of the four organ and tissue types and their subsequent herbivory. Both pulses provide a context for three emerging questions. First is an explanation for the contrast between the near instantaneous consumption of plant tissues during Phase 1, versus the exceptionally long lags between the earliest occurrences of plant tissues and their subsequent herbivorization during Phase 2. Second is the identity of arthropod herbivores for both phases. Third is the cause behind the overwhelming targeting of seed-fern plant hosts during Phase 2. Regardless of the answers to these questions, the trace fossil record of plant-arthropod associations provides primary ecological data that remain unaddressed by the body-fossil record alone.

  4. Simple preparation of plant epidermal tissue for laser microdissection and downstream quantitative proteome and carbohydrate analysis.

    Science.gov (United States)

    Falter, Christian; Ellinger, Dorothea; von Hülsen, Behrend; Heim, René; Voigt, Christian A

    2015-01-01

    The outwardly directed cell wall and associated plasma membrane of epidermal cells represent the first layers of plant defense against intruding pathogens. Cell wall modifications and the formation of defense structures at sites of attempted pathogen penetration are decisive for plant defense. A precise isolation of these stress-induced structures would allow a specific analysis of regulatory mechanism and cell wall adaption. However, methods for large-scale epidermal tissue preparation from the model plant Arabidopsis thaliana, which would allow proteome and cell wall analysis of complete, laser-microdissected epidermal defense structures, have not been provided. We developed the adhesive tape - liquid cover glass technique (ACT) for simple leaf epidermis preparation from A. thaliana, which is also applicable on grass leaves. This method is compatible with subsequent staining techniques to visualize stress-related cell wall structures, which were precisely isolated from the epidermal tissue layer by laser microdissection (LM) coupled to laser pressure catapulting. We successfully demonstrated that these specific epidermal tissue samples could be used for quantitative downstream proteome and cell wall analysis. The development of the ACT for simple leaf epidermis preparation and the compatibility to LM and downstream quantitative analysis opens new possibilities in the precise examination of stress- and pathogen-related cell wall structures in epidermal cells. Because the developed tissue processing is also applicable on A. thaliana, well-established, model pathosystems that include the interaction with powdery mildews can be studied to determine principal regulatory mechanisms in plant-microbe interaction with their potential outreach into crop breeding.

  5. Simple preparation of plant epidermal tissue for laser microdissection and downstream quantitative proteome and carbohydrate analysis

    Directory of Open Access Journals (Sweden)

    Christian eFalter

    2015-03-01

    Full Text Available The outwardly directed cell wall and associated plasma membrane of epidermal cells represent the first layers of plant defense against intruding pathogens. Cell wall modifications and the formation of defense structures at sites of attempted pathogen penetration are decisive for plant defense. A precise isolation of these stress-induced structures would allow a specific analysis of regulatory mechanism and cell wall adaption. However, methods for large-scale epidermal tissue preparation from the model plant Arabidopsis thaliana, which would allow proteome and cell wall analysis of complete, laser-microdissected epidermal defense structures, have not been provided. We developed the adhesive tape – liquid cover glass technique for simple leaf epidermis preparation from A. thaliana, which is also applicable on grass leaves. This method is compatible with subsequent staining techniques to visualize stress-related cell wall structures, which were precisely isolated from the epidermal tissue layer by laser microdissection coupled to laser pressure catapulting. We successfully demonstrated that these specific epidermal tissue samples could be used for quantitative downstream proteome and cell wall analysis. The development of the adhesive tape – liquid cover glass technique for simple leaf epidermis preparation and the compatibility to laser microdissection and downstream quantitative analysis opens new possibilities in the precise examination of stress- and pathogen-related cell wall structures in epidermal cells. Because the developed tissue processing is also applicable on A. thaliana, well-established, model pathosystems that include the interaction with powdery mildews can be studied to determine principal regulatory mechanisms in plant-microbe interaction with their potential outreach into crop breeding.

  6. Toxicity of molybdenum and its trace analysis in animal tissues and plants.

    Science.gov (United States)

    Abbasi, S A

    1981-01-01

    A sensitive, selective, rapid and reproducible method is presented for the analysis of submicrogram levels of molybdenum in animal tissues (Liver) and plants. The method is based on solvent extraction of Molybdenum (VI) using isoamyl alcohol solution of N-o-tolyl-o-methoxy-benzohydroxamic acid at pH 1.5-2.5, and subsequent spectrophotometric determination of the yellow extract at 350 nm.

  7. Tissue culture system using a PANDA ring resonator and wavelength router for hydroponic plant.

    Science.gov (United States)

    Kamoldilok, Surachart; Suwanpayak, Nathaporn; Suttirak, Saisudawan; Yupapin, Preecha P

    2012-06-01

    A novel system of nanofluidics trapping and delivery, which is known as a tissue culture system is proposed. By using the intense optical pulse(i.e., a soliton pulse) and a system constructed by a liquid core waveguide, the optical vortices (gradient optical fields/wells) can be generated, where the trapping tools in the same way as the optical tweezers in the PANDA ring resonator can be formed. By controlling the suitable parameters, the intense optical vortices can be generated within the PANDA ring resonator, in which the nanofluidics can be trapped and moved (transported) dynamically within the Tissue culture system(a wavelength router), which can be used for tissue culture and delivery in the hydroponic plant system.

  8. Spatial organization and correlation properties quantify structural changes on mesoscale of parenchymatous plant tissue

    Science.gov (United States)

    Valous, N. A.; Delgado, A.; Drakakis, K.; Sun, D.-W.

    2014-02-01

    The study of plant tissue parenchyma's intercellular air spaces contributes to the understanding of anatomy and physiology. This is challenging due to difficulty in making direct measurements of the pore space and the complex mosaic of parenchymatous tissue. The architectural complexity of pore space has shown that single geometrical measurements are not sufficient for characterization. The inhomogeneity of distribution depends not only on the percentage content of phase, but also on how the phase fills the space. The lacunarity morphometric, as multiscale measure, provides information about the distribution of gaps that correspond to degree of spatial organization in parenchyma. Additionally, modern theories have suggested strategies, where the focus has shifted from the study of averages and histograms to the study of patterns in data fluctuations. Detrended fluctuation analysis provides information on the correlation properties of the parenchyma at different spatial scales. The aim is to quantify (with the aid of the aforementioned metrics), the mesostructural changes—that occur from one cycle of freezing and thawing—in the void phase of pome fruit parenchymatous tissue, acquired with X-ray microcomputed tomography. Complex systems methods provide numerical indices and detailed insights regarding the freezing-induced modifications upon the arrangement of cells and voids. These structural changes have the potential to lead to physiological disorders. The work can further stimulate interest for the analysis of internal plant tissue structures coupled with other physico-chemical processes or phenomena.

  9. Spatial organization and correlation properties quantify structural changes on mesoscale of parenchymatous plant tissue

    Energy Technology Data Exchange (ETDEWEB)

    Valous, N. A.; Delgado, A.; Sun, D.-W., E-mail: dawen.sun@ucd.ie [School of Biosystems Engineering, University College Dublin, National University of Ireland, Belfield, Dublin 4, Dublin (Ireland); Drakakis, K. [Complex and Adaptive Systems Laboratory, University College Dublin, National University of Ireland, Belfield, Dublin 4, Dublin (Ireland)

    2014-02-14

    The study of plant tissue parenchyma's intercellular air spaces contributes to the understanding of anatomy and physiology. This is challenging due to difficulty in making direct measurements of the pore space and the complex mosaic of parenchymatous tissue. The architectural complexity of pore space has shown that single geometrical measurements are not sufficient for characterization. The inhomogeneity of distribution depends not only on the percentage content of phase, but also on how the phase fills the space. The lacunarity morphometric, as multiscale measure, provides information about the distribution of gaps that correspond to degree of spatial organization in parenchyma. Additionally, modern theories have suggested strategies, where the focus has shifted from the study of averages and histograms to the study of patterns in data fluctuations. Detrended fluctuation analysis provides information on the correlation properties of the parenchyma at different spatial scales. The aim is to quantify (with the aid of the aforementioned metrics), the mesostructural changes—that occur from one cycle of freezing and thawing—in the void phase of pome fruit parenchymatous tissue, acquired with X-ray microcomputed tomography. Complex systems methods provide numerical indices and detailed insights regarding the freezing-induced modifications upon the arrangement of cells and voids. These structural changes have the potential to lead to physiological disorders. The work can further stimulate interest for the analysis of internal plant tissue structures coupled with other physico-chemical processes or phenomena.

  10. Plant regeneration from petiole segments of some species in tissue culture

    Directory of Open Access Journals (Sweden)

    Krystyna Klimaszewska

    2013-12-01

    Full Text Available The regeneration ability of 21 plant species belonging to 14 families was tested. The method of tissue culture in vitro was applied, on basic MS medium with an addition of growth regulators from the auxin and cytokinin groups. From among the investigated plant groups Peperomia scandens and Caladium × hortulanum were capable of plant regeneration, Passiilora coerulea regenerated shoots, Hedera helix, Begonia glabra, Coleus blumei, Fuchsia hybrida, Passiflora suberosa and Peperomia eburnea formed callus and roots, Kalanchoe blossfeldiana, Pelargonium grandiflorum, P. peltatum, P. radula, Coleus shirensis and Magnolia soulangeana produced callus, Philodendron scandens, Rhododendron smirnovii, Hibiscus rosa-sinensis, Coprosma baueri, Cestrum purpureum and Solanum rantonnetii did not exhibit any regeneration reactions.

  11. Amelioration of iron mine soils with biosolids: Effects on plant tissue metal content and earthworms.

    Science.gov (United States)

    Cele, Emmanuel Nkosinathi; Maboeta, Mark

    2016-11-01

    The achievement of environmentally sound and economically feasible disposal strategies for biosolids is a major issue in the wastewater treatment industry around the world, including Swaziland. Currently, an iron ore mine site, which is located within a wildlife sanctuary, is being considered as a suitable place where controlled disposal of biosolids may be practiced. Therefore, this study was conducted to investigate the effects of urban biosolids on iron mine soils with regard to plant metal content and ecotoxicological effects on earthworms. This was done through chemical analysis of plants grown in biosolid-amended mine soil. Earthworm behaviour, reproduction and bioaccumulation tests were also conducted on biosolid-amended mine soil. According to the results obtained, the use of biosolids led to creation of soil conditions that were generally favourable to earthworms. However, plants were found to have accumulated Zn up to 346 mg kg(-1) (in shoots) and 462 mg kg(-1) (in roots). This was more than double the normal Zn content of plants. It was concluded that while biosolids can be beneficial to mine soils and earthworms, they can also lead to elevated metal content in plant tissues, which might be a concern to plant-dependant wildlife species. Nonetheless, it was not possible to satisfactorily estimate risks to forage quality since animal feeding tests with hyperaccumulator plants have not been reported. Quite possibly, there may be no cause for alarm since the uptake of metals from soil is greater in plants grown in pots in the greenhouse than from the same soil in the field since pot studies fail to mimic field conditions where the soil is heterogeneous and where the root system possesses a complex topology. It was thought that further field trials might assist in arriving at more satisfactory conclusions.

  12. Application of GC-MS for the detection of lipophilic compounds in diverse plant tissues

    Directory of Open Access Journals (Sweden)

    Hellmann Hanjo

    2009-04-01

    Full Text Available Abstract Background The concept of metabolite profiling has been around for decades and technical innovations are now enabling it to be carried out on a large scale with respect to the number of both metabolites measured and experiments carried out. However, studies are generally confined to polar compounds alone. Here we describe a simple method for lipophilic compounds analysis in various plant tissues. Results We choose the same preparative and instrumental platform for lipophilic profiling as that we routinely use for polar metabolites measurements. The method was validated in terms of linearity, carryover, reproducibility and recovery rates, as well as using various plant tissues. As a first case study we present metabolic profiling of Arabidopsis root and shoot tissue of wild type (C24 and mutant (rsr4-1 plants deficient on vitamin B6. We found significant alterations in lipid constituent contents, especially in the roots, which were characterised by dramatic increases in several fatty acids, thus providing further hint for the role of pyridoxine in oxidative stress and lipid peroxidation. The second example is the lipophilic profiling of red and green tomato fruit cuticles of wild type (Alisa Craig and the DFD (delayed fruit deterioration mutant, which we compared and contrasted with the more focused wax analysis of these plants reported before. Conclusion We can rapidly and reliably detect and quantify over 40 lipophilic metabolites including fatty acids, fatty alcohols, alkanes, sterols and tocopherols. The method presented here affords a simple and rapid, yet robust complement to previously validated methods of polar metabolite profiling by gas-chromatography mass-spectrometry.

  13. Application of supercritical CO2 for extraction of polyisoprenoid alcohols and their esters from plant tissues.

    Science.gov (United States)

    Jozwiak, Adam; Brzozowski, Robert; Bujnowski, Zygmunt; Chojnacki, Tadeusz; Swiezewska, Ewa

    2013-07-01

    In this study, a method of supercritical fluid extraction (SFE) with carbon dioxide of polyisoprenoids from plant photosynthetic tissues is described. SFE was an effective extraction method for short- and medium-chain compounds with even higher yield than that observed for the "classical extraction" method with organic solvents. Moreover, SFE-derived extracts contained lower amounts of impurities (e.g., chlorophylls) than those obtained by extraction of the same tissue with organic solvents. Elevated temperature and extended extraction time of SFE resulted in a higher rate of extraction of long-chain polyisoprenoids. Ethanol cofeeding did not increase the extraction efficiency of polyisoprenoids; instead, it increased the content of impurities in the lipid extract. Optimization of SFE time and temperature gives the opportunity of prefractionation of complex polyisoprenoid mixtures accumulated in plant tissues. Extracts obtained with application of SFE are very stable and free from organic solvents and can further be used directly in experimental diet supplementation or as starting material for preparation of semisynthetic polyisoprenoid derivatives, e.g., polyisoprenoid phosphates.

  14. Application of the chloramphenicol acetyltransferase (CAT) diffusion assay to transgenic plant tissues.

    Science.gov (United States)

    Peach, C; Velten, J

    1992-02-01

    Chloramphenicol acetyltransferase (CAT) activity was quantified in crude extracts from tobacco callus tissues using a modification of a previously reported diffusion assay. We describe here the alterations necessary in applying this rapid and simple assay procedure to plant materials. Due to the high concentration of nonspecific oxidases present in most plant tissues, some type of protective agent is required to maintain enzyme activity. We have tested beta-mercaptoethanol, cysteine, dithiothreitol, ascorbic acid and polyvinyl pyrrolidone as protective agents within the initial extraction buffer. We also investigated the effect of heat (60 degrees C, 10 min) and 5 mM EDTA on CAT activity. The highest CAT activity was obtained using 5 mM cysteine plus 5 mM EDTA in 40 mM Tris-HCl (pH 7.8) as the initial extraction buffer followed by a heat treatment. Using this buffer, CAT activity was stable on ice for more than two hours. In our hands, total acetyl-coenzyme A concentration within the assay mixture was found to be saturating at 250 microM and the Km determined to be 100 microM. Assays performed using the same crude plant extract indicate that 1) duplicate assays show less than 1.5% variation in activities and 2) CAT activity increases linearly with respect to volume of extract used.

  15. Detection, isolation, and preliminary characterization of bacteria contaminating plant tissue cultures

    Directory of Open Access Journals (Sweden)

    Monika Kałużna

    2014-01-01

    Full Text Available In order to limit the contamination problem in plant tissue cultures experiments on selection of media suitable for detection and isolation of bacteria contaminating plant tissue explants, and preliminary characterization of isolates were made. In the first experiment aiming at detection of bacteria in plant explants four strains representing genera most often occurring at our survey of plant tissue cultures, and earlier isolated and identified (Bacillus, Methylobacterium, Pseudomonas and Xanthomonas were streaked on five bacteriological media (NA, King B, K, R2A and 523 and on the medium used for plant culture initiation – ½ MS with milk albumin (IM. All strains grew on all media but on K and IM at the slowest rate and on 523 medium at the fastest. The IM medium proved to be useful for immediate bacteria detection at the initial stage of culture. In the second experiment, aiming at characterization of isolates on the basis of colony growth and morphology 14 strains (Agrobacterium, Bacillus, Curtobacterium, Flavobacterium, Lactobacillus, Methylobacterium – 2 strains Mycobacterium, Paenibacillus, Plantibacterium, Pseudomonas, Stenotrophomonas, Xanthomonas, and species Serratia marcescens were streaked on five microbiological media: KB, NBY, YDC, YNA and YPGA. All strains grew on all those media but at different rates. The only exception was the strain of Lactobacillus spp., which did not grow on King B medium. This medium allowed the detection of such characteristic traits as fluorescence (Pseudomonas and secretion of inclusions (Stenotrophomonas. The third experiment was focussed on assessment of the sensitivity of detection of specific bacteria in pure cultures and in plant tis- sue cultures using standard PCR and BIO-PCR techniques with genus specific primers and 2 methods of DNA isolation. Results showed that the use of Genomic Mini kit enabled an increase of the sensitivity by 100 times as compared to extraction of DNA by boiling

  16. Histology, histochemistry and SEM are useful tools to study regeneration processes in plant tissue culture

    Directory of Open Access Journals (Sweden)

    Piotr Żabicki

    2013-04-01

    Full Text Available Tissue cultures in vitroare used for the multiplication of plants via direct and indirect (via callus regeneration. This approach is commonly applied in the protection of endangered species by the introduction of regenerated in vitro plantlets to botanical gardens and to the nature (so called ex situ plant conservation. In vitroconditions, especially the supplementation of tissue culture media with plant growth regulators, cause a somaclonal variation, resulting in genetic differences among regenerated plants. To analyze callus structure, including cell shapes and sizes, cell differentiation (e.g. the presence of xylem vessels and regeneration processes (organogenesis, somatic embryogenesis, the histological, histochemical and SEM techniques are applied. In this study, to obtain regeneration of plants in culture conditions, we have used three Viola species (V. epipsilaLedeb., V. stagnina Kit. and V. uliginosaBesser, indicated to be critically endangered according to Polish Red Book of Plants (Kazmierczakowa & Zarzycki 2001 and two genotypes of a model plant Arabidopsis thaliana(L. Heynh. (Columbia-0 and an insertional cdkg ;2mutant line. An Arabidopsis homozygous cdkg ;2 knock-out originated from a T 3 generation of T-DNA insertional line SALK_090262 (Alonso et al. 2003 and has been selected from a subsequent T 4 generation based on PCR analysis using primers complementary to flanking positions of full-length cDNA of CDKG;2gene product (a clone isolated by Seki et al. 2002. The aims of the study were: 1 to select the most convenient method to obtain regenerated Violaplants with maternal genotype i.e., via direct organogenesis or somatic embryogenesis; 2 to determine the effect of mutation in CDKG;2 gene on the explant response to in vitroconditions, including callus proliferation and regeneration. In three Viola species organogenesis was induced on MS (Murashige and Skoog basal medium supplied with thidiazuron (TDZ in concentrations 0.5 mg

  17. A non-phenol-chloroform extraction of double-stranded RNA from plant and fungal tissues.

    Science.gov (United States)

    Balijja, Alitukiriza; Kvarnheden, Anders; Turchetti, Tullio

    2008-09-01

    Double-stranded RNA (dsRNA) molecules of viruses are found in nature at a very high frequency. Their detection in plants and fungi has been carried out with difficulty due to the complicated dsRNA extraction techniques used commonly which includes phenol-chloroform extractions. In this study, an extraction method for isolation of dsRNA is described that is free of phenol and chloroform. A lysis buffer, containing beta-mercaptoethanol and polyvinylpolypyrrolidone (PVPP-40), was added to homogenised tissues and the subsequent supernatant was filtered through a cellulose CF-11 mini-column. DsRNA molecules were separated based on the differing affinity of nucleic acids for the cellulose CF-11 resin in 20% ethanol buffer. This easy, rapid and cheap technique has been successfully tested on fungi and plants containing different dsRNA virus molecules, indicating the possibility of a wide use of the method.

  18. Simplified heavy metal staining techniques demonstrated with Fast Plant leaf tissue

    Institute of Scientific and Technical Information of China (English)

    HARRISJOSEPHB; THOMASG.GUILLIAMS; 等

    1992-01-01

    Fast Plant (Brassica rapa,Cruciferae)leaf tissue fixed in glutaradehyde-acrolein and post-fixed in osmium,was examined for response to several easilyprepared heavy metal stains.Lead and uranium,separately and in combination,gave typical results across the spectrum of cell orgeanelles.As s single stain following osmium,bismuth produced images seemingly equivalent to lead and uranium.Phosphotungstic acid produced very good membrane delineation but produced a washed-out background image similar to that from lead staining .Carbohydrate compounds were especially responsive to ruthenium;the cytoplasm and the matrix of all organelles were also stained very well.The procedures were no more demanding than traditional staining methods and may be easily used in research and teaching .Fast Plant materials are a reliable,quick nand easy source of living material.

  19. Application of 3D printing to prototype and develop novel plant tissue culture systems.

    Science.gov (United States)

    Shukla, Mukund R; Singh, Amritpal S; Piunno, Kevin; Saxena, Praveen K; Jones, A Maxwell P

    2017-01-01

    Due to the complex process of designing and manufacturing new plant tissue culture vessels through conventional means there have been limited efforts to innovate improved designs. Further, development and availability of low cost, energy efficient LEDs of various spectra has made it a promising light source for plant growth in controlled environments. However, direct replacement of conventional lighting sources with LEDs does not address problems with uniformity, spectral control, or the challenges in conducting statistically valid experiments to assess the effects of light. Prototyping using 3D printing and LED based light sources could help overcome these limitations and lead to improved culture systems. A modular culture vessel design in which the fluence rate and spectrum of light are independently controlled was designed, prototyped using 3D printing, and evaluated for plant growth. This design is compatible with semi-solid and liquid based culture systems. Observations on morphology, chlorophyll content, and chlorophyll fluorescence based stress parameters from in vitro plants cultured under different light spectra with similar overall fluence rate indicated different responses in Nicotiana tabacum and Artemisia annua plantlets. This experiment validates the utility of 3D printing to design and test functional vessels and demonstrated that optimal light spectra for in vitro plant growth is species-specific. 3D printing was successfully used to prototype novel culture vessels with independently controlled variable fluence rate/spectra LED lighting. This system addresses several limitations associated with current lighting systems, providing more uniform lighting and allowing proper replication/randomization for experimental plant biology while increasing energy efficiency. A complete procedure including the design and prototyping of a culture vessel using 3D printing, commercial scale injection molding of the prototype, and conducting a properly replicated

  20. Ecosystem engineering and manipulation of host plant tissues by the insect borer Oncideres albomarginata chamela.

    Science.gov (United States)

    Calderón-Cortés, Nancy; Uribe-Mú, Claudia A; Martínez-Méndez, A Karen; Escalera-Vázquez, Luis H; Cristobal-Pérez, E Jacob; García-Oliva, Felipe; Quesada, Mauricio

    2016-01-01

    Ecosystem engineering by insect herbivores occurs as the result of structural modification of plants manipulated by insects. However, only few studies have evaluated the effect of these modifications on the plant responses induced by stem-borers that act as ecosystem engineers. In this study, we evaluated the responses induced by the herbivory of the twig-girdler beetle Oncideres albomarginata chamela (Cerambycidae: Lamiinae) on its host plant Spondias purpurea (Anacardiaceae), and its relationship with the ecosystem engineering process carried out by this stem-borer. Our results demonstrated that O. albomarginata chamela branch removal induced the development of lateral branches increasing the resources needed for the development of future insect generations, of its own offspring and of many other insect species. Detached branches represent habitats with high content of nitrogen and phosphorous, which eventually can be incorporated into the ecosystem, increasing nutrient cycling efficiency. Consequently, branch removal and the subsequent plant tissue regeneration induced by O. albomarginata chamela represent key mechanisms underlying the ecosystem engineering process carried out by this stem-borer, which enhances arthropod diversity in the ecosystem.

  1. Plant tissues in 3D via X-ray tomography: simple contrasting methods allow high resolution imaging.

    Directory of Open Access Journals (Sweden)

    Yannick M Staedler

    Full Text Available Computed tomography remains strongly underused in plant sciences despite its high potential in delivering detailed 3D phenotypical information because of the low X-ray absorption of most plant tissues. Existing protocols to study soft tissues display poor performance, especially when compared to those used on animals. More efficient protocols to study plant material are therefore needed. Flowers of Arabidopsis thaliana and Marcgravia caudata were immersed in a selection of contrasting agents used to treat samples for transmission electron microscopy. Grayscale values for floral tissues and background were measured as a function of time. Contrast was quantified via a contrast index. The thick buds of Marcgravia were scanned to determine which contrasting agents best penetrate thick tissues. The highest contrast increase with cytoplasm-rich tissues was obtained with phosphotungstate, whereas osmium tetroxide and bismuth tatrate displayed the highest contrast increase with vacuolated tissues. Phosphotungstate also displayed the best sample penetration. Furthermore, infiltration with phosphotungstate allowed imaging of all plants parts at a high resolution of 3 µm, which approaches the maximum resolution of our equipment: 1.5 µm. The high affinity of phosphotungstate for vasculature, cytoplasm-rich tissue, and pollen causes these tissues to absorb more X-rays than the surrounding tissues, which, in turn, makes these tissues appear brighter on the scan data. Tissues with different brightness can then be virtually dissected from each other by selecting the bracket of grayscale to be visualized. Promising directions for the future include in silico phenotyping and developmental studies of plant inner parts (e.g., ovules, vasculature, pollen, and cell nuclei via virtual dissection as well as correlations of quantitative phenotypes with omics datasets. Therefore, this work represents a crucial improvement of previous methods, allowing new directions of

  2. Plant tissues in 3D via X-ray tomography: simple contrasting methods allow high resolution imaging.

    Science.gov (United States)

    Staedler, Yannick M; Masson, David; Schönenberger, Jürg

    2013-01-01

    Computed tomography remains strongly underused in plant sciences despite its high potential in delivering detailed 3D phenotypical information because of the low X-ray absorption of most plant tissues. Existing protocols to study soft tissues display poor performance, especially when compared to those used on animals. More efficient protocols to study plant material are therefore needed. Flowers of Arabidopsis thaliana and Marcgravia caudata were immersed in a selection of contrasting agents used to treat samples for transmission electron microscopy. Grayscale values for floral tissues and background were measured as a function of time. Contrast was quantified via a contrast index. The thick buds of Marcgravia were scanned to determine which contrasting agents best penetrate thick tissues. The highest contrast increase with cytoplasm-rich tissues was obtained with phosphotungstate, whereas osmium tetroxide and bismuth tatrate displayed the highest contrast increase with vacuolated tissues. Phosphotungstate also displayed the best sample penetration. Furthermore, infiltration with phosphotungstate allowed imaging of all plants parts at a high resolution of 3 µm, which approaches the maximum resolution of our equipment: 1.5 µm. The high affinity of phosphotungstate for vasculature, cytoplasm-rich tissue, and pollen causes these tissues to absorb more X-rays than the surrounding tissues, which, in turn, makes these tissues appear brighter on the scan data. Tissues with different brightness can then be virtually dissected from each other by selecting the bracket of grayscale to be visualized. Promising directions for the future include in silico phenotyping and developmental studies of plant inner parts (e.g., ovules, vasculature, pollen, and cell nuclei) via virtual dissection as well as correlations of quantitative phenotypes with omics datasets. Therefore, this work represents a crucial improvement of previous methods, allowing new directions of research to be

  3. 3D Reconstruction of Frozen Plant Tissue: a unique histological analysis to image post-freeze responses

    Science.gov (United States)

    Winter hardiness in plants is the result of a complex interaction between genes, the tissue where those genes are expressed and the environment. The light microscope is a valuable tool to understand this complexity which will ultimately help researchers improve the tolerance of plants to freezing st...

  4. Steroid-inducible BABY BOOM system for development of fertile Arabidopsis thaliana plants after prolonged tissue culture.

    Science.gov (United States)

    Lutz, Kerry A; Martin, Carla; Khairzada, Sahar; Maliga, Pal

    2015-10-01

    We describe a steroid-inducible BABY BOOM system that improves plant regeneration in Arabidopsis leaf cultures and yields fertile plants. Regeneration of Arabidopsis thaliana plants for extended periods of time in tissue culture may result in sterile plants. We report here a novel approach for A. thaliana regeneration using a regulated system to induce embryogenic cultures from leaf tissue. The system is based on BABY BOOM (BBM), a transcription factor that turns on genes involved in embryogenesis. We transformed the nucleus of A. thaliana plants with BBM:GR, a gene in which the BBM coding region is fused with the glucocorticoid receptor (GR) steroid-binding domain. In the absence of the synthetic steroid dexamethasone (DEX), the BBM:GR fusion protein is localized in the cytoplasm. Only when DEX is included in the culture medium does the BBM transcription factor enter the nucleus and turn on genes involved in embryogenesis. BBM:GR plant lines show prolific shoot regeneration from leaf pieces on media containing DEX. Removal of DEX from the culture media allowed for flowering and seed formation. Therefore, use of BBM:GR leaf tissue for regeneration of plants for extended periods of time in tissue culture will facilitate the recovery of fertile plants.

  5. Early diagenesis of vascular plant tissues: Lignin and cutin decomposition and biogeochemical implications

    Science.gov (United States)

    Opsahl, Stephen; Benner, Ronald

    1995-12-01

    Long-term subaqueous decomposition patterns of five different vascular plant tissues including mangrove leaves and wood ( Avicennia germinans), cypress needles and wood ( Taxodium distichum) and smooth cordgrass ( Spartina alternifora) were followed for a period of 4.0 years, representing the longest litter bag decomposition study to date. All tissues decomposed under identical conditions and final mass losses were 97, 68, 86, 39, and 93%, respectively. Analysis of the lignin component of herbaceous tissues using alkaline CuO oxidation was complicated by the presence of a substantial ester-bound phenol component composed primarily of cinnamyl phenols. To overcome this problem, we introduce a new parameter to represent lignin, Λ6. Λ6 is comprised only of the six syringyl and vanillyl phenols and was found to be much less sensitive to diagenetic variation than the commonly used parameter Λ, which includes the cinnamyl phenols. Patterns of change in lignin content were strongly dependent on tissue type, ranging from 77% enrichment in smooth cordgrass to 6% depletion in cypress needles. In contrast, depletion of cutin was extensive (65-99%) in all herbaceous tissues. Despite these differences in the overall reactivity of lignin and cutin, both macromolecules were extensively degraded during the decomposition period. The long-term decomposition series also provided very useful information about the compositional parameters which are derived from the specific oxidation products of both lignin and cutin. The relative lability of ester-bound cinnamyl phenols compromised their use in parameters to distinguish woody from herbaceous plant debris. The dimer to monomer ratios of lignin-derived phenols indicated that most intermonomeric linkages in lignin degraded at similar rates. Acid to aldehyde ratios of vanillyl and syringyl phenols became elevated, particularly during the latter stages of decomposition supporting the use of these parameters as indicators of diagenetic

  6. [Diversity and tissue distribution of fungal endophytes in Alpinia officinarum: an important south-China medicinal plant].

    Science.gov (United States)

    Zhou, Ren-Chao; Huang, Juan; Li, Ze-En; Li, Shu-Bin

    2014-08-01

    In the present study, terminal-restriction fragment length polymorphism (T-RFLP) technique was applied to assess the diversity and tissue distribution of the fungal endophyte communities of Alpinia officinarum collected from Longtang town in Xuwen county, Guangdong province, China, at which the pharmacological effect of the medicine plant is traditional considered to be the significantly higher than that in any other growth areas in China. A total of 28 distinct Terminal-Restriction Fragment (T-RFs) were detected with HhaI Mono-digestion targeted amplified fungal nuclear ribosomal internal transcribed spacer region sequences (rDNA ITS) from the root, rhizome, stem, and leaf internal tissues of A. officinarum plant, indicating that at least 28 distinct fungal species were able to colonize the internal tissue of the host plant. The rDNA ITS-T-RFLP profiles obtained from different tissues of the host plant were obvious distinct. And the numbers of total T-RFs, and the dominant T-RFs detected from various tissues were significantly different. Based on the obtained T-RFLP profiles, Shannon's diversity index and the Shannon's evenness index were calculated, which were significantly different among tissues (P endophyte communities of the host plant (P = 0, Pearson correlation coefficient ≤ -0.962), and significant positive correlations between both of the tested active components contents and 325 bp dominant T-RF linkage to Pestalotiopsis (P = 0, Pearson correlation coefficient ≥ 0.975). In conclusion, A. officinarum is colonized by diverse fungal endophytes communities. The diversity of the fungal endophytes was found in the A. officinarum varied with differences of the tissue types of the host plants and was closely correlated with the accumulation of main active components, total volatile oils and galangin contents in the host plant tissue.

  7. A Case Study of Native Plants at Industrially Contaminated Area in Bhopal and Their Tissue Culture Approach for Phytoremediation

    OpenAIRE

    Ashwini A Waoo; Sujata Ganguly; Swati Khare

    2014-01-01

    In phytoremediation techniques,experiments were done with normal soil-grown or hydroponically grown plants. In recent days, more and more effort is directed toward research to understand and improve the performance of plants in remediation technologies, the number of results obtained with the help of in vitro plant cell and tissue cultures is rapidly increasing.This review focuses on the report of various phytoremediation technologies, paying special attention to removal of pollutants from so...

  8. Comparison Of Cd And Zn Accumulation In Tissues Of Different Vascular Plants: A Radiometric Study

    Directory of Open Access Journals (Sweden)

    Dürešová Zuzana

    2015-12-01

    Full Text Available The aim of the present work was to compare the accumulation and translocation of Cd and Zn in plants of tobacco (Nicotiana tabacum L., celery (Apium graveolens L., maize (Zea mays L., giant reed (Arundo donax L., and alpine pennycress (Noccaea caerulescens L. under conditions of short-term hydroponic experiments using nutrient solutions spiked with radionuclides 109Cd or 65Zn, and direct gamma-spectrometry. It was found that the time-course of metals accumulation in studied plants was not different in terms of target metal, but it was significantly different on the level of plant species. The highest values of Cd accumulation showed plants of giant reed, whereby the accumulation decreased in the order: giant reed > tobacco > alpine pennycress >> maize and celery. On the basis of concentration ratios (CR [Me]shoot / [Me]root calculation for both metals, it was found that Cd and Zn were in prevailing part accumulated in the root tissues and only partially accumulated in the shoots, where the amount of accumulated Cd and Zn increased from the oldest developed leaves to the youngest developed leaves. The CR values corresponding to these facts were calculated in the range 0.06 – 0.27 for Cd and for Zn 0.06 – 0.48. In terms of plant species, the CR values obtained for Cd decreased in the order: maize > celery > tobacco and giant reed > alpine pennycress. The similarity between studied objects – individual plant species on the basis of the obtained variables defining Cd or Zn accumulation at different conditions of the experiments as well as the relationships between obtained variables and conditions of the experiments were subjected to multivariate analysis method – cluster analysis (CA. According to the findings and this analysis, it can be expected that plants of tobacco and giant reed will dispose with similar characteristics as plants of alpine pennycress, which are classified as Zn/Cd hyperaccumulators, in terms of Cd or Zn accumulation

  9. A facile means for the identification of indolic compounds from plant tissues.

    Science.gov (United States)

    Yu, Peng; Hegeman, Adrian D; Cohen, Jerry D

    2014-09-01

    The bulk of indole-3-acetic acid (IAA) in plants is found in the form of conjugated molecules, yet past research on identifying these compounds has largely relied on methods that were both laborious and inefficient. Using recent advances in analytical instrumentation, we have developed a simple yet powerful liquid chromatography-mass spectrometry (LC-MS)-based method for the facile characterization of the small IAA conjugate profile of plants. The method uses the well-known quinolinium ion (m/z 130.0651) generated in MS processes as a signature with high mass accuracy that can be used to screen plant extracts for indolic compounds, including IAA conjugates. We reinvestigated Glycine max (soybean) for its indoles and found indole-3-acetyl-trytophan (IA-Trp) in addition to the already known indole-3-acetyl-aspartic acid (IA-Asp) and indole-3-acetyl-glutamic acid (IA-Glu) conjugates. Surprisingly, several organic acid conjugates of tryptophan were also discovered, many of which have not been reported in planta before. These compounds may have important physiological roles in tryptophan metabolism, which in turn can affect human nutrition. We also demonstrated the general applicability of this method by identifying indolic compounds in different plant tissues of diverse phylogenetic origins. It involves minimal sample preparation but can work in conjunction with sample enrichment techniques. This method enables quick screening of IAA conjugates in both previously characterized as well as uncharacterized species, and facilitates the identification of indolic compounds in general. © 2014 The Authors The Plant Journal © 2014 John Wiley & Sons Ltd.

  10. Development of Rapid Isothermal Amplification Assays for Detection of Phytophthora spp. in Plant Tissue.

    Science.gov (United States)

    Miles, Timothy D; Martin, Frank N; Coffey, Michael D

    2015-02-01

    Several isothermal amplification techniques recently have been developed that are tolerant of inhibitors present in many plant extracts, which can reduce the need for obtaining purified DNA for running diagnostic assays. One such commercially available technique that has similarities with real-time polymerase chain reaction (PCR) for designing primers and a labeled probe is recombinase polymerase amplification (RPA). This technology was used to develop two simple and rapid approaches for detection of Phytophthora spp.: one genus-specific assay multiplexed with a plant internal control and the other species-specific assays for Phytophthora ramorum and P. kernoviae. All assays were tested for sensitivity (ranging from 3 ng to 1 fg of DNA) and specificity using DNA extracted from more than 136 Phytophthora taxa, 21 Pythium spp., 1 Phytopythium sp., and a wide range of plant species. The lower limit of linear detection using purified DNA was 200 to 300 fg of DNA in all pathogen RPA assays. Six different extraction buffers were tested for use during plant tissue maceration and the assays were validated in the field by collecting 222 symptomatic plant samples from over 50 different hosts. Only 56 samples were culture positive for Phytophthora spp. whereas 91 were positive using the Phytophthora genus-specific RPA test and a TaqMan real-time PCR assay. A technique for the generation of sequencing templates from positive RPA amplifications to confirm species identification was also developed. These RPA assays have added benefits over traditional technologies because they are rapid (results can be obtained in as little as 15 min), do not require DNA extraction or extensive training to complete, use less expensive portable equipment than PCR-based assays, and are significantly more specific than current immunologically based methods. This should provide a rapid, field-deployable capability for pathogen detection that will facilitate point-of-sample collection processing

  11. An electrochemical approach to monitor pH change in agar media during plant tissue culture.

    Science.gov (United States)

    Wang, Min; Ha, Yang

    2007-05-15

    In this work, metal oxide microelectrodes were developed to monitor pH change in agar media during plant tissue culture. An antimony wire was produced by a new approach "capillary melt method". The surface of the obtained antimony wire was oxidized in a potassium nitrate melt to fabricate an antimony oxide film for pH sensing. Characterization results show that the oxide layer grown on the wire surface consists of Sb(2)O(3) crystal phase. The sensing response, open-circuit potential, of the electrode has a good linear relationship (R(2)=1.00) with pH value of the test solution. Adding organic compounds into the test media would not affect the linear relationship, although the slope of the lines varied with different ingredients added. The antimony oxide electrodes were employed to continuously monitor pH change of agar culture media during a 2-week plant tissue culture of Dendrobium candidum. The antimony oxide electrode fabricated this way has the advantages of low cost, easy fabrication, fast response, and almost no contamination introduced into the system. It would be suitable for in situ and continuous pH measurement in many bio applications.

  12. Nitrate dynamics in natural plants: Insights based on the concentration and natural isotope abundances of tissue nitrate

    Directory of Open Access Journals (Sweden)

    Xue Yan Liu

    2014-07-01

    Full Text Available The dynamics of nitrate (NO3-, a major nitrogen (N source for natural plants, has been studied mostly through experimental N addition, enzymatic assay, isotope labeling, and genetic expression. However, artificial N supply may not reasonably reflect the N strategies in natural plants because NO3- uptake and reduction may vary with external N availability. Abrupt application and short operation times, field N addition, and isotopic labeling hinder the elucidation of in situ NO3--use mechanisms. The concentration and natural isotopes of tissue NO3- can offer insights into the plant NO3- sources and dynamics in a natural context. Furthermore, they facilitate the exploration of plant NO3- utilization and its interaction with N pollution and ecosystem N cycles without disturbing the N pools. The present study was conducted to review the application of the denitrifier method for concentration and isotope analyses of NO3- in plants. Moreover, this study highlights the utility and benefits of these parameters in interpreting NO3- sources and dynamics in natural plants. We summarize the major sources and reduction processes of NO3- in plants, and discuss the implications of NO3- concentration in plant tissues based on existing data. Particular emphasis was laid on the regulation of soil NO3 - and plant ecophysiological functions in interspecific and intra-plant NO3- variations. We introduce N and O isotope systematics of NO3- in plants and discusse the principles and feasibilities of using isotopic enrichment and fractionation factors; the correlation between concentration and isotopes (N and O isotopes: δ18O and ∆17O; and isotope mass-balance calculations to constrain sources and reduction of NO3- in possible scenarios for natural plants are deliberated. Finally, we construct a preliminary framework of intraplant δ18O-NO3- variation, and summarize the uncertainties in using tissue NO3- parameters to interpret plant NO3- utilization.

  13. How to store plant tissues in the absence of liquid nitrogen? Ethanol preserves the RNA integrity of Cannabis sativa stem tissues

    Directory of Open Access Journals (Sweden)

    Lauralie Mangeot-Peter

    2016-09-01

    Full Text Available The preservation of intact RNA is a limiting step when gene expression profiling is performed using field-collected plant material. The use of liquid nitrogen ensures the optimal preservation of RNA, however it is not always practical, especially if the plant material has to be sampled in remote locations. Ethanol is known to preserve DNA in plant tissues even after a long storage period and here its suitability to preserve the RNA of textile hemp cortical tissues was tested. Hemp (Cannabis sativa L. is an economically important fibre crop because it supplies cellulosic bast fibres used in different industrial sectors. In this study we demonstrate the suitability of ethanol for RNA preservation by analyzing tissues stored at 4 °C for 1, 2, 4 and 8 days. We show that in all the cases the extracted RNA is intact. We finally analyze hemp stem tissues stored in ethanol for 1 month and demonstrate the preservation of the tissue structure, particularly of bast fibres.

  14. Development of a fluorescent in situ hybridization (FISH) technique for visualizing CGMMV in plant tissues.

    Science.gov (United States)

    Shargil, D; Zemach, H; Belausov, E; Lachman, O; Kamenetsky, R; Dombrovsky, A

    2015-10-01

    Cucumber green mottle mosaic virus (CGMMV), which belongs to the genus Tobamovirus, is a major pathogen of cucurbit crops grown indoors and in open fields. Currently, immunology (e.g., ELISA) and molecular amplification techniques (e.g., RT-PCR) are employed extensively for virus detection in plant tissues and commercial seed lots diagnostics. In this study, a fluorescent in situ hybridization (FISH) technique, using oligonucleotides whose 5'-terminals were labeled with red cyanine 3 (Cy3) or green fluorescein isothiocyanate (FITC), was developed for the visualization of the pathogen in situ. This simple and reliable method allows detection and localization of CGMMV in the vegetative and reproductive tissues of cucumber and melon. When this technique was applied in male flowers, anther tissues were found to be infected; whereas the pollen grains were found to be virus-free. These results have meaningful epidemiological implications for the management of CGMMV, particularly with regard to virus transfer via seed and the role of insects as CGMMV vectors.

  15. An RNA isolation system for plant tissues rich in secondary metabolites

    Directory of Open Access Journals (Sweden)

    Bhardwaj Pardeep K

    2011-03-01

    Full Text Available Abstract Background Secondary metabolites are reported to interfere with the isolation of RNA particularly with the recipes that use guanidinium-based salt. Such interference was observed in isolation of RNA with medicinal plants rheum (Rheum australe and arnebia (Arnebia euchroma. A rapid and less cumbersome system for isolation of RNA was essential to facilitate any study related to gene expression. Findings An RNA isolation system free of guanidinium salt was developed that successfully isolated RNA from rheum and arnebia. The method took about 45 min and was successfully evaluated on twenty one tissues with varied secondary metabolites. The A260/280 ratio ranged between 1.8 - 2.0 with distinct 28 S and 18 S rRNA bands visible on a formaldehyde-agarose gel. Conclusions The present manuscript describes a rapid protocol for isolation of RNA, which works well with all the tissues examined so far. The remarkable feature was the success in isolation of RNA with those tissues, wherein the most commonly used methods failed. Isolated RNA was amenable to downstream applications such as reverse transcription-polymerase chain reaction (RT-PCR, differential display (DD, suppression subtractive hybridization (SSH library construction, and northern hybridization.

  16. Volatile Profiling of Aromatic Traditional Medicinal Plant, Polygonum minus in Different Tissues and Its Biological Activities

    Directory of Open Access Journals (Sweden)

    Rafidah Ahmad

    2014-11-01

    Full Text Available The aim of this research was to identify the volatile metabolites produced in different organs (leaves, stem and roots of Polygonum minus, an important essential oil producing crop in Malaysia. Two methods of extraction have been applied: Solid Phase Microextraction (SPME and hydrodistillation coupled with Gas Chromatography-Mass Spectrometry (GC-MS. Approximately, 77 metabolites have been identified and aliphatic compounds contribute significantly towards the aroma and flavour of this plant. Two main aliphatic compounds: decanal and dodecanal were found to be the major contributor. Terpenoid metabolites were identified abundantly in leaves but not in the stem and root of this plant. Further studies on antioxidant, total phenolic content, anticholinesterase and antimicrobial activities were determined in the essential oil and five different extracts. The plant showed the highest DPPH radical scavenging activity in polar (ethanol extract for all the tissues tested. For anti-acetylcholinesterase activity, leaf in aqueous extract and methanol extract showed the best acetylcholinesterase inhibitory activities. However, in microbial activity, the non-polar extracts (n-hexane showed high antimicrobial activity against Methicillin-resistant Staphylococcus aureus (MRSA compared to polar extracts. This study could provide the first step in the phytochemical profiles of volatile compounds and explore the additional value of pharmacology properties of this essential oil producing crop Polygonum minus.

  17. Volatile profiling of aromatic traditional medicinal plant, Polygonum minus in different tissues and its biological activities.

    Science.gov (United States)

    Ahmad, Rafidah; Baharum, Syarul Nataqain; Bunawan, Hamidun; Lee, Minki; Mohd Noor, Normah; Rohani, Emelda Roseleena; Ilias, Norashikin; Zin, Noraziah Mohamad

    2014-11-20

    The aim of this research was to identify the volatile metabolites produced in different organs (leaves, stem and roots) of Polygonum minus, an important essential oil producing crop in Malaysia. Two methods of extraction have been applied: Solid Phase Microextraction (SPME) and hydrodistillation coupled with Gas Chromatography-Mass Spectrometry (GC-MS). Approximately, 77 metabolites have been identified and aliphatic compounds contribute significantly towards the aroma and flavour of this plant. Two main aliphatic compounds: decanal and dodecanal were found to be the major contributor. Terpenoid metabolites were identified abundantly in leaves but not in the stem and root of this plant. Further studies on antioxidant, total phenolic content, anticholinesterase and antimicrobial activities were determined in the essential oil and five different extracts. The plant showed the highest DPPH radical scavenging activity in polar (ethanol) extract for all the tissues tested. For anti-acetylcholinesterase activity, leaf in aqueous extract and methanol extract showed the best acetylcholinesterase inhibitory activities. However, in microbial activity, the non-polar extracts (n-hexane) showed high antimicrobial activity against Methicillin-resistant Staphylococcus aureus (MRSA) compared to polar extracts. This study could provide the first step in the phytochemical profiles of volatile compounds and explore the additional value of pharmacology properties of this essential oil producing crop Polygonum minus.

  18. Tissue culture and regeneration of an antimalarial plant, Artemisia sieberi Besser

    Directory of Open Access Journals (Sweden)

    A. Sharafi

    2014-10-01

    Full Text Available WHO recommends artemisinin-based combination therapies (ACTs as the most effective choice to treat malaria. For developing transgenic plants with high accumulation of artemisinin (by introducing genes encoding enzymes which regulate the biosynthetic pathway of artemisinin, an efficient protocol for tissue culture and plant regeneration is necessary. In the present study, leaf explants of Artemisia sieberi were cultivated in Murashige & Skoog based medium supplemented by combination of different plant growth regulators including 6-benzyl-aminopurine (BA, α-naphthalene-acetic acid (NAA, indole-3-acetic acid (IAA, picloram (Pic and 2,4-dichlorophenoxyacetic acid (2,4-D. The highest frequency of shoot induction was obtained on MS medium supplemented with 2 mg/L BA plus 0.05 mg/L NAA (95% regeneration and MS medium supplemented with 2 mg/L BA plus 0.5 mg/L IAA (85% regeneration. Rooting was obtained on MS medium supplemented with 0.05 mg/L NAA. The present study has revealed a simple, reliable, rapid and high efficient regeneration system for A. sieberi Besser as a source of artemisinin in short period via adventitious shoot induction procedure.

  19. TOR pathway activation in Zea mays L. tissues: conserved function between animal and plant kingdoms.

    Science.gov (United States)

    Garrocho-Villegas, Verónica; de Jiménez, Estela Sánchez

    2012-06-01

    In most non-photosynthetic eukaryotes it has been demonstrated a conserved signal transduction pathway, namely TOR-S6K, that coordinates growth and cell proliferation. This pathway targets the translational apparatus to induce selective translation of ribosomal mRNAs as well as stimulate the cell cycle transition through the G1/S phase. Thus, by activation of this pathway through environmental signals, nutrients, stress, or specific growth factors, such as insulin or insulin-like growth factors (IGF), this pathway allows organisms to regulate growth and cell division. In plants, evidence has shown that TOR protein has been highly conserved through evolution, being involved in growth and cell proliferation control as well. Particularly in maize, a peptide named ZmIGF has been found in actively growing tissues. It targets the maize TOR pathway at the same extent as insulin and, by doing so it induces growth, as well as ribosomal proteins and DNA synthesis. Thus, higher metazoans and plants seem to conserve similar biochemical paths to regulate cell growth through equivalent targets that conduce to activation of the TOR-S6K pathway. Recent research shows evidence that supports this proposal by uncovering the ZmIGF receptor in maize, providing further means for analyzing the role of the conserved TOR signaling pathway in this plant.

  20. Effect of Thermal and Nonthermal Processing on Textural Quality of Plant Tissues.

    Science.gov (United States)

    Ranganathan, Kumar; Subramanian, Vijayalakshmi; Shanmugam, Nadanasabapathi

    2016-12-09

    In the current fast revolving world, the consumption of processed food is increasing drastically. The population who depend on these processed foods are also cautious about the quality and safety of what they consume. This being the case, in order to satisfy the consumer it is the responsibility of the researcher and the manufacturer to check what happens to food on processing. Plant-derived foods such as fruits and vegetables are sensitive producers which are to be handled cautiously through each steps involved in processing, starting from harvest to storage, processing to package, transportation to distribution, till it reaches the consumer. During processing, the plant materials, which are made up of complex structural components such as lignin, cellulose, pectin, etc. undergo changes which has its effect on the quality attributes of the final product. Texture is an important quality parameter of all the sensory properties. The relation between the structure of the plant tissue and the texture of the final product is reviewed in this paper comprehensively.

  1. Effect of radiation-degraded chitosan on growth promotion of flower plant in tissue culture

    Energy Technology Data Exchange (ETDEWEB)

    Luan, Le Quang; Ha, Vo Thi Thu; Hai, Le; Hien, Nguyen Quoc [Vietnam Atomic Energy Commission, Nuclear Research Institute, Dalat (Viet Nam); Nagasawa, Naotsugu; Yoshii, Fumio; Kume, Tamikazu [Japan Atomic Energy Research Inst., Takasaki, Gunma (Japan). Takasaki Radiation Chemistry Research Establishment

    2002-03-01

    Radiation is a useful tool for degradation of polysaccharides, such as starch, carrageenan, alginate and chitin/chitosan. The viscosity molecular weight (Mw) of chitosan with 80% degree of deacetylation was reduced to 1.5 x 10{sup 5} by irradiation of 50kGy in solid phase. The solution of 10% of chitosan with Mw ca. 15 x 10{sup 5} was then irradiated at doses ranging 10-250kGy for further degradation and the products were supplemented into cultural media for testing of plant growth promotion effect. The results indicated that irradiated chitosan showed a strong growth-promotion effect on the increase of the length of shoot, the length of root and fresh biomass for flower plants namely Limonium latifolium, Eustoma grandiflorum and Chrysanthemum morifolium in tissue culture. The growth-promotion effect was obtained by the treatments with 50ppm of chitosan irradiated at the doses of 75-100kGy in 10% solution. The suitable concentrations of chitosan irradiated at 100kGy are ca. 100ppm for C. morifolium, 30ppm for E. grandiflorum and 40ppm for L. latifolium. In addition, our study also indicated that the survival ratio of transferred flower plantlets treated with irradiated chitosan was improved after acclimatizing for 30 days in the greenhouse. Accordingly, it is concluded that degraded chitosan obtained by radiation degradation technique is effective as a plant growth promoter as well as irradiated alginate. (author)

  2. Two New Complete Genome Sequences Offer Insight into Host and Tissue Specificity of Plant Pathogenic Xanthomonas spp.▿†

    Science.gov (United States)

    Bogdanove, Adam J.; Koebnik, Ralf; Lu, Hong; Furutani, Ayako; Angiuoli, Samuel V.; Patil, Prabhu B.; Van Sluys, Marie-Anne; Ryan, Robert P.; Meyer, Damien F.; Han, Sang-Wook; Aparna, Gudlur; Rajaram, Misha; Delcher, Arthur L.; Phillippy, Adam M.; Puiu, Daniela; Schatz, Michael C.; Shumway, Martin; Sommer, Daniel D.; Trapnell, Cole; Benahmed, Faiza; Dimitrov, George; Madupu, Ramana; Radune, Diana; Sullivan, Steven; Jha, Gopaljee; Ishihara, Hiromichi; Lee, Sang-Won; Pandey, Alok; Sharma, Vikas; Sriariyanun, Malinee; Szurek, Boris; Vera-Cruz, Casiana M.; Dorman, Karin S.; Ronald, Pamela C.; Verdier, Valérie; Dow, J. Maxwell; Sonti, Ramesh V.; Tsuge, Seiji; Brendel, Volker P.; Rabinowicz, Pablo D.; Leach, Jan E.; White, Frank F.; Salzberg, Steven L.

    2011-01-01

    Xanthomonas is a large genus of bacteria that collectively cause disease on more than 300 plant species. The broad host range of the genus contrasts with stringent host and tissue specificity for individual species and pathovars. Whole-genome sequences of Xanthomonas campestris pv. raphani strain 756C and X. oryzae pv. oryzicola strain BLS256, pathogens that infect the mesophyll tissue of the leading models for plant biology, Arabidopsis thaliana and rice, respectively, were determined and provided insight into the genetic determinants of host and tissue specificity. Comparisons were made with genomes of closely related strains that infect the vascular tissue of the same hosts and across a larger collection of complete Xanthomonas genomes. The results suggest a model in which complex sets of adaptations at the level of gene content account for host specificity and subtler adaptations at the level of amino acid or noncoding regulatory nucleotide sequence determine tissue specificity. PMID:21784931

  3. Two new complete genome sequences offer insight into host and tissue specificity of plant pathogenic Xanthomonas spp.

    Science.gov (United States)

    Bogdanove, Adam J; Koebnik, Ralf; Lu, Hong; Furutani, Ayako; Angiuoli, Samuel V; Patil, Prabhu B; Van Sluys, Marie-Anne; Ryan, Robert P; Meyer, Damien F; Han, Sang-Wook; Aparna, Gudlur; Rajaram, Misha; Delcher, Arthur L; Phillippy, Adam M; Puiu, Daniela; Schatz, Michael C; Shumway, Martin; Sommer, Daniel D; Trapnell, Cole; Benahmed, Faiza; Dimitrov, George; Madupu, Ramana; Radune, Diana; Sullivan, Steven; Jha, Gopaljee; Ishihara, Hiromichi; Lee, Sang-Won; Pandey, Alok; Sharma, Vikas; Sriariyanun, Malinee; Szurek, Boris; Vera-Cruz, Casiana M; Dorman, Karin S; Ronald, Pamela C; Verdier, Valérie; Dow, J Maxwell; Sonti, Ramesh V; Tsuge, Seiji; Brendel, Volker P; Rabinowicz, Pablo D; Leach, Jan E; White, Frank F; Salzberg, Steven L

    2011-10-01

    Xanthomonas is a large genus of bacteria that collectively cause disease on more than 300 plant species. The broad host range of the genus contrasts with stringent host and tissue specificity for individual species and pathovars. Whole-genome sequences of Xanthomonas campestris pv. raphani strain 756C and X. oryzae pv. oryzicola strain BLS256, pathogens that infect the mesophyll tissue of the leading models for plant biology, Arabidopsis thaliana and rice, respectively, were determined and provided insight into the genetic determinants of host and tissue specificity. Comparisons were made with genomes of closely related strains that infect the vascular tissue of the same hosts and across a larger collection of complete Xanthomonas genomes. The results suggest a model in which complex sets of adaptations at the level of gene content account for host specificity and subtler adaptations at the level of amino acid or noncoding regulatory nucleotide sequence determine tissue specificity.

  4. Parameters affecting the efficient delivery of mesoporous silica nanoparticle materials and gold nanorods into plant tissues by the biolistic method.

    Science.gov (United States)

    Martin-Ortigosa, Susana; Valenstein, Justin S; Sun, Wei; Moeller, Lorena; Fang, Ning; Trewyn, Brian G; Lin, Victor S-Y; Wang, Kan

    2012-02-06

    Applying nanotechnology to plant science requires efficient systems for the delivery of nanoparticles (NPs) to plant cells and tissues. The presence of a cell wall in plant cells makes it challenging to extend the NP delivery methods available for animal research. In this work, research is presented which establishes an efficient NP delivery system for plant tissues using the biolistic method. It is shown that the biolistic delivery of mesoporous silica nanoparticle (MSN) materials can be improved by increasing the density of MSNs through gold plating. Additionally, a DNA-coating protocol is used based on calcium chloride and spermidine for MSN and gold nanorods to enhance the NP-mediated DNA delivery. Furthermore, the drastic improvement of NP delivery is demonstrated when the particles are combined with 0.6 μm gold particles during bombardment. The methodology described provides a system for the efficient delivery of NPs into plant cells using the biolistic method.

  5. Advancements in mass spectrometry for biological samples: Protein chemical cross-linking and metabolite analysis of plant tissues

    Energy Technology Data Exchange (ETDEWEB)

    Klein, Adam [Iowa State Univ., Ames, IA (United States)

    2015-01-01

    This thesis presents work on advancements and applications of methodology for the analysis of biological samples using mass spectrometry. Included in this work are improvements to chemical cross-linking mass spectrometry (CXMS) for the study of protein structures and mass spectrometry imaging and quantitative analysis to study plant metabolites. Applications include using matrix-assisted laser desorption/ionization-mass spectrometry imaging (MALDI-MSI) to further explore metabolic heterogeneity in plant tissues and chemical interactions at the interface between plants and pests. Additional work was focused on developing liquid chromatography-mass spectrometry (LC-MS) methods to investigate metabolites associated with plant-pest interactions.

  6. Plant maturity and nitrogen fertilization affected fructan metabolism in harvestable tissues of timothy (Phleum pratense L.).

    Science.gov (United States)

    Ould-Ahmed, Marouf; Decau, Marie-Laure; Morvan-Bertrand, Annette; Prud'homme, Marie-Pascale; Lafrenière, Carole; Drouin, Pascal

    2014-10-15

    Timothy (Phleum pratense L.) is an important grass forage used for pasture, hay, and silage in regions with cool and humid growth seasons. One of the factors affecting the nutritive value of this grass is the concentration of non-structural carbohydrates (NSC), mainly represented by fructans. NSC concentration depends on multiple factors, making it hardly predictable. To provide a better understanding of NSC metabolism in timothy, the effects of maturity stage and nitrogen (N) fertilization level on biomass, NSC and N-compound concentrations were investigated in the tissues used for forage (leaf blades and stems surrounded by leaf sheaths) of hydroponically grown plants. Moreover, activities and relative expression level of enzymes involved in fructan metabolism were measured in the same tissues. Forage biomass was not altered by the fertilization level but was strongly modified by the stage of development. It increased from vegetative to heading stages while leaf-to-stem biomass ratio decreased. Total NSC concentration, which was not altered by N fertilization level, increased between heading and anthesis due to an accumulation of fructans in leaf blades. Fructan metabolizing enzyme activities (fructosyltransferase-FT and fructan exohydrolase-FEH) were not or only slightly altered by both maturity stage and N fertilization level. Conversely, the relative transcript levels of genes coding for enzymes involved in fructan metabolism were modified by N supply (PpFT1 and Pp6-FEH1) or maturity stage (PpFT2). The relative transcript level of PpFT1 was the highest in low N plants while that of Pp6-FEH1 was the highest in high N plants. Morevoer, transcript level of PpFT1 was negatively correlated with nitrate concentration while that of PpFT2 was positively correlated with sucrose concentration. This distinct regulation of the two genes coding for 6-sucrose:fructan fructosyltransferase (6-SFT) may allow a fine adequation of C allocation towards fructan synthesis in

  7. Isolation of functional RNA from plant tissues rich in phenolic compounds.

    Science.gov (United States)

    Schneiderbauer, A; Sandermann, H; Ernst, D

    1991-08-15

    A method for the isolation of RNA from different tissues of trees (seedlings, saplings, and adult trees) is described. Using this procedure it is possible to remove large amounts of disturbing polyphenolic compounds from nucleic acids. The method involves an acetone treatment of the freeze-dried and powdered plant material, the use of high salt concentrations in the extraction buffer and an aqueous two-phase system. These steps were combined with the conventional phenol/chloroform extraction and CsCl centrifugation. The method has been successfully applied to the isolation and purification of RNA from pine (Pinus sylvestris L. and Pinus mugo Turr.), Norway spruce (Picea abies L.), and beech (Fagus sylvatica L.). The functional quality of RNA extracted by this procedure has been characterized by its uv spectrum, by agarose gel electrophoresis with ethidium bromide staining, Northern blot hybridization, and in vitro translation.

  8. Application of SEM and EDX in studying biomineralization in plant tissues.

    Science.gov (United States)

    He, Honghua; Kirilak, Yaowanuj

    2014-01-01

    This chapter describes protocols using formalin-acetic acid-alcohol (FAA) to fix plant tissues for studying biomineralization by means of scanning electron microscopy (SEM) and qualitative energy-dispersive X-ray microanalysis (EDX). Specimen preparation protocols for SEM and EDX mainly include fixation, dehydration, critical point drying (CPD), mounting, and coating. Gold-coated specimens are used for SEM imaging, while gold- and carbon-coated specimens are prepared for qualitative X-ray microanalyses separately to obtain complementary information on the elemental compositions of biominerals. During the specimen preparation procedure for SEM, some biominerals may be dislodged or scattered, making it difficult to determine their accurate locations, and light microscopy is used to complement SEM studies. Specimen preparation protocols for light microscopy generally include fixation, dehydration, infiltration and embedding with resin, microtome sectioning, and staining. In addition, microwave processing methods are adopted here to speed up the specimen preparation process for both SEM and light microscopy.

  9. The safety assessment of food ingredients derived from plant cell, tissue and organ cultures: a review.

    Science.gov (United States)

    Murthy, Hosakatte Niranjana; Georgiev, Milen I; Park, So-Young; Dandin, Vijayalaxmi S; Paek, Kee-Yoeup

    2015-06-01

    Plant cell, tissue and organ cultures (PCTOC) have become an increasingly attractive alternative for the production of various high molecular weight molecules which are used as flavourings, fragrances, colouring agents and food additives. Although PCTOC products are cultivated in vitro in a contamination free environment, the raw material produced from PCTOC may contain many components apart from the target compound. In some cases, PCTOC raw materials may also carry toxins, which may be naturally occurring or accumulated during the culture process. Assessment of the safety of PCTOC products is, therefore, a priority of the biotech industries involved in their production. The safety assessment involves the evaluation of starting material, production process and the end product. Before commercialisation, PCTOC products should be evaluated for their chemical and biological properties, as well as for their toxicity. In this review, measures and general criteria for biosafety evaluation of PCTOC products are addressed and thoroughly discussed.

  10. Potential application of urea-derived herbicides as cytokinins in plant tissue culture.

    Science.gov (United States)

    Srinivasan, Malathi; Nachiappan, Vasanthi; Rajasekharan, Ram

    2006-12-01

    Various urea-derived herbicides and different cytokinin analogues were used to determine their effects on callusing response and shoot regenerating capacity of alfalfa (Medicago sativa L.) and Coleus (Coleus forskohlii Briq.). The herbicides monuron and diuron evoked profuse callusing response from Coleus leaf segments and alfalfa petiole explants on Murashige and Skoog medium. Shoot regeneration by monuron (2.0 mg/l) showed a maximum of 3 multiple shoots both in alfalfa and Coleus with a frequency of 92% and 75%, respectively. Whereas diuron (0.5 mg/l) showed a high frequency of shoot regeneration (89%)with a mean number of 5 shoots in alfalfa, in C.forskohlii, the frequency of regeneration was 90%with a mean number of 6 shoots. Diuron with two chloride groups in the phenyl ring showed significantly higher cytokinin-like activity than single chloride substitution monuron. This study demonstrates the potential use of monuron and diuron as cytokinins in plant tissue culture.

  11. 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.

  12. Potential application of urea-derived herbicides as cytokinins in plant tissue culture

    Indian Academy of Sciences (India)

    Malathi Srinivasan; Vasanthi Nachiappan; Ram Rajasekharan

    2006-12-01

    Various urea-derived herbicides and different cytokinin analogues were used to determine their effects on callusing response and shoot regenerating capacity of alfalfa (Medicago sativa L.) and Coleus (Coleus forskohlii Briq.). The herbicides monuron and diuron evoked profuse callusing response from Coleus leaf segments and alfalfa petiole explants on Murashige and Skoog medium. Shoot regeneration by monuron (2.0 mg/l) showed a maximum of 3 multiple shoots both in alfalfa and Coleus with a frequency of 92% and 75%, respectively. Whereas diuron (0.5 mg/l) showed a high frequency of shoot regeneration (89%) with a mean number of 5 shoots in alfalfa, in C. forskohlii, the frequency of regeneration was 90% with a mean number of 6 shoots. Diuron with two chloride groups in the phenyl ring showed significantly higher cytokinin-like activity than single chloride substitution monuron. This study demonstrates the potential use of monuron and diuron as cytokinins in plant tissue culture.

  13. In vivo monitoring of seeds and plant-tissue water absorption using optical coherence tomography and optical coherence microscopy

    Science.gov (United States)

    Sapozhnikova, Veronika V.; Kutis, Irina S.; Kutis, Sergey D.; Kuranov, Roman V.; Gelikonov, Grigory V.; Shabanov, Dmitry V.; Kamensky, Vladislav A.

    2004-07-01

    First experimental results on OCT imaging of internal structure of plant tissues and in situ OCT monitoring of plant tissue regeneration at different water supply are reported. Experiments for evaluating OCT capabilities were performed on Tradescantia. The investigation of seeds swelling was performed on wheat seeds (Triticum L.), barley seeds (Hordeum L.), long-fibred flax seeds (Linum usitatissimum L.) and cucumber seeds (Cucumis sativus L.). These OCT images correlate with standard microscopy data from the same tissue regions. Seeds were exposed to a low-intensity physical factor-the pulsed gradient magnetic field (GMF) with pulse duration 0.1 s and maximum amplitude 5 mT (4 successive pulses during 0.4 s). OCT and OCM enable effective monitoring of fast reactions in plants and seeds at different water supply.

  14. Screenhouse and field persistence of nonpathogenic endophytic Fusarium oxysporum in Musa tissue culture plants.

    Science.gov (United States)

    Paparu, Pamela; Dubois, Thomas; Gold, Clifford S; Niere, Björn; Adipala, Ekwamu; Coyne, Daniel

    2008-04-01

    Two major biotic constraints to highland cooking banana (Musa spp., genome group AAA-EA) production in Uganda are the banana weevil Cosmopolites sordidus and the burrowing nematode Radopholus similis. Endophytic Fusarium oxysporum strains inoculated into tissue culture banana plantlets have shown control of the banana weevil and the nematode. We conducted screenhouse and field experiments to investigate persistence in the roots and rhizome of two endophytic Fusarium oxysporum strains, V2w2 and III4w1, inoculated into tissue-culture banana plantlets of highland cooking banana cultivars Kibuzi and Nabusa. Re-isolation of F. oxysporum showed that endophyte colonization decreased faster from the rhizomes than from the roots of inoculated plants, both in the screenhouse and in the field. Whereas rhizome colonization by F. oxysporum decreased in the screenhouse (4-16 weeks after inoculation), root colonization did not. However, in the field (17-33 weeks after inoculation), a decrease was observed in both rhizome and root colonization. The results show a better persistence in the roots than rhizomes of endophytic F. oxysporum strains V2w2 and III4w1.

  15. An evolutionary view of plant tissue culture: somaclonal variation and selection.

    Science.gov (United States)

    Wang, Qin-Mei; Wang, Li

    2012-09-01

    Plants regenerated from in vitro cultures possess an array of genetic and epigenetic changes. This phenomenon is known as 'somaclonal variation' and the frequency of somaclonal variation (SV) is usually elevated far beyond that expected in nature. Initially, the relationship between time in culture and detected SV was found to support the widespread belief that SV accumulates with culture age. However, a few studies indicated that older cultures yielded regenerants with less SV. What leads to this seemed contradiction? In this article, we have proposed a novel in vitro callus selection hypothesis, differentiation bottleneck (D-bottleneck) and dedifferentiation bottleneck (Dd-bottleneck), which consider natural selection theory to be fit for cell population in vitro. The results of multiplication races between the cells with the true-to-type phenotype and the deleterious cells determine the increase/decrease of SV frequencies in calli or regenerants as in vitro culture time goes on. The possibility of interpreting the complex situation of time-related SV by the evolutionary theory is discussed in this paper. In addition, the SV threshold, space-determined hypothesis and D-bottleneck are proposed to interpret the loss of the regenerability through a long period of plant tissue culture (PTC).

  16. SOIL EXCHANGEABLE ALUMINUM INFLUENCING THE GROWTH AND LEAF TISSUE MACRONUTRIENTS CONTENT OF CASTOR PLANTS

    Directory of Open Access Journals (Sweden)

    ROSIANE DE LOURDES SILVA DE LIMA

    2014-01-01

    Full Text Available Three castor ( Ricinus communis genotypes were studied regarding tolerance to high exchange factorial distribution of five doses of exchangeable aluminum added to the soil (0, 0.15, 0.30, 0.60, and 1.20 cmol c dm - 3 and three castor genotypes (BRS Nordestina, BRS Paraguaçu, and Lyra. The plants were raised in pots in a greenhouse. At 53 days after emergence, data were taken on plant height, leaf area, dry mass of shoot and root, and leaf tissue content of macronutrients. The most sensitive genotype was the cv. BRS Nordestina, in which the shoot and root dry weight in the highest aluminum content were reduced to 12.9% and 16.2% of the control treatment, respectively. The most tolerant genotype was the hybrid Lyra, in which the shoot and root dry weight in the maximum content of aluminum were reduced to 43.5% and 42.7% of the control treatment, respectively.The increased exchangeable aluminum affected the leaf nutrient content, and the intensity of the response was different among cultivars. The aluminum toxicity increased N, Ca, and Mg contents and reduced on P, K, and S contents. The cv. BRS Nordestina had a drastic shoot dry weight reduction associated with an intense increment in the N leaf content. Thus, the N increment was caused by a concentration effect caused by the limited growth.

  17. Multiscale characterization of pyritized plant tissues in blueschist facies metamorphic rocks

    Science.gov (United States)

    Bernard, Sylvain; Benzerara, Karim; Beyssac, Olivier; Brown, Gordon E., Jr.

    2010-09-01

    Pyritized plant tissues with well-preserved morphology were studied in rocks from Vanoise (western Alps, France) that experienced high-pressure, low-temperature metamorphic conditions in the blueschist facies during the Alpine orogeny. Organic and inorganic phases composing these fossils were characterized down to the nanometer scale by Raman microspectroscopy, scanning transmission X-ray microscopy and transmission electron microscopy. The graphitic but disordered organic matter composing these fossils is chemically and structurally homogeneous and mostly contains aromatic functional groups. Its original chemistry remains undefined likely because it was significantly transformed by diagenetic processes and/or thermal degradation during metamorphism. Various mineral phases are closely associated with this organic matter, including sulphides such as pyrite and pyrrhotite, carbonates such as ankerite and calcite, and iron oxides. A tentative time sequence of formation of these diverse mineral phases relative to organic matter decay is proposed. The absence of traces of organic matter sulphurization, the pervasive pyritization of the vascular tissues and the presence of ankerite suggest that the depositional/diagenetic environment of these metasediments was likely rich in reactive iron. Fe-sulphides and ankerite likely precipitated early and might have promoted the preservation of the fossilized biological soft tissues by providing mechanical resistance to compaction during diagenesis and subsequent metamorphism. In contrast, iron oxides which form rims of 100-nm in thickness at the interface between organic matter and Fe-sulphides may result from metamorphic processes. This study illustrates that it may be possible in some instances to deconvolve metamorphic from diagenetic imprints and opens new avenues to better constrain processes that may allow the preservation of organic fossils during diagenesis and metamorphism.

  18. Simultaneous extraction and derivatization of carbohydrates from green plant tissues for analysis by gas-liquid chromatography.

    Science.gov (United States)

    Streeter, J G; Strimbu, C E

    1998-06-01

    Simultaneous extraction and derivatization of carbohydrates was performed by mixing dry ground plant tissue with derivatization reagents in pyridine; trimethylsilyl derivatives were analyzed by gas-liquid chromatography. This "direct analysis" was compared to analysis of samples prepared by exhaustive ethanol extraction of the same ground plant tissues. Comparisons included leaf blades from apple, grape, corn, and tomato and leaf blade, petiole, stem, and pod tissues from soybean plants. Direct analysis gave superior quantification of sucrose, glucose, and fructose because of sucrose hydrolysis during ethanol extraction. Sucrose hydrolysis was highly variable among plant species and use of hot ethanol at the first extraction step reduced sucrose hydrolysis but did not always abolish it. Sucrose hydrolysis was probably due to the activity of hydrolytic enzymes in 75% ethanol at room temperature. Direct analysis was inferior for the quantification of cyclitols in the fibrous tissues of soybean but provided acceptable results for cyclitol analysis in leaf blade tissue. When the time for extraction/reaction was extended from 40 to 60 min, some improvement in recovery of cyclitols was observed, but recovery remained 10 to 20% below that obtained with exhaustive ethanol extraction. Mannitol was vacuum infiltrated into the five types of leaf tissue and recovery averaged 100% by the direct method relative to ETOH extraction for apple, grape, corn, and soybean leaves but was only 76% for tomato leaves. Direct analysis provides very large time savings and is clearly the method of choice when the analysis of large numbers of samples of plant tissues for carbohydrate composition is required.

  19. Qualitative and quantitative analysis of anthraquinone derivatives in rhizomes of tissue culture-raised Rheum emodi Wall. plants.

    Science.gov (United States)

    Malik, Sonia; Sharma, Nandini; Sharma, Upendra K; Singh, Narendra P; Bhushan, Shashi; Sharma, Madhu; Sinha, Arun K; Ahuja, Paramvir S

    2010-06-15

    This paper presents quantification of five anthraquinone derivatives (emodin glycoside, chrysophanol glycoside, emodin, chrysophanol and physcion) in rhizomes of hardened micro-propagated Rheum emodi plants using high-performance liquid chromatography (HPLC). Aseptic shoot cultures were raised using rhizome buds. Shoot multiplication occurred in both agar gelled and liquid Murashige and Skoog (MS) medium supplemented with 10.0 microM 6-benzylaminopurine (BAP) and 5.0 microM indole-3-butyric acid (IBA). Rooted plantlets obtained on plant growth regulator (PGR)-free medium were transferred to soil with 92% survival. HPLC analysis revealed the presence of five anthraquinone derivatives: emodin glycoside, chrysophanol glycoside, emodin, chrysophanol and physcion in rhizomes of tissue culture-raised plants. Only emodin glycoside (1) and chrysophanol glycoside (2) were present in 6-month-old hardened tissue cultured plants. In addition, the other three derivatives (emodin (3), chrysophanol (4) and physcion (5)) were also detected after 9 months.

  20. GENETIC VARIABILITY OF CULTURED PLANT TISSUES UNDER NORMAL CONDITIONS AND UNDER STRESS

    Directory of Open Access Journals (Sweden)

    Dolgikh Yu.I.

    2012-08-01

    Full Text Available The genetic variability induced by in vitro conditions known as somaclonal variation is of practical interest due to its potential uses in plant breeding but, on the other hand, if clonal propagation or transformation is main goal, it becomes an unwelcome phenomenon. Thus, it is important to know frequency, the genomic distribution, the mechanisms and factors influencing somaclonal variation. We studied variability of PCR-based DNA markers of cultured tissues and regenerated plants of maize and bread wheat. The original A188 line of maize and the somaclones obtained were tested using 38 RAPD and 10 ISSR primers. None of the A188 plants showed variation in the RAPD and ISSR spectra for any of the primers used. However, the PCR spectra obtained from the somaclones demonstrated some variations, i.e., 22 RAPD primers and 6 ISSR primers differentiated at least one somaclonal variant from the progenitor line. Six SCAR markers were developed based on several RAPD and ISSR fragments. The inheritance of these SCAR markers was verified in the selfing progeny of each somaclone in the R1–R4 generations and in the hybrids, with A188 as the parental line in the F1 and F2 generations. These markers were sequenced and bioinformatic searches were performed to understand the molecular events that may underlie the variability observed in the somaclones. All changes were found in noncoding sequences and were induced by different molecular events, such as the insertion of long terminal repeat transposon, precise miniature inverted repeat transposable element (MITE excision, microdeletion, recombination, and a change in the pool of mitochondrial DNA. In two groups of independently produced somaclones, the same features (morphological, molecular were variable, which confirms the theory of ‘hot spots’ occurring in the genome. The presence of the same molecular markers in the somaclones and in different non-somaclonal maize variants suggests that in some cases

  1. Tissue Culture as a Source of Replicates in Nonmodel Plants: Variation in Cold Response in Arabidopsis lyrata ssp. petraea.

    Science.gov (United States)

    Kenta, Tanaka; Edwards, Jessica E M; Butlin, Roger K; Burke, Terry; Quick, W Paul; Urwin, Peter; Davey, Matthew P

    2016-12-07

    While genotype-environment interaction is increasingly receiving attention by ecologists and evolutionary biologists, such studies need genetically homogeneous replicates-a challenging hurdle in outcrossing plants. This could be potentially overcome by using tissue culture techniques. However, plants regenerated from tissue culture may show aberrant phenotypes and "somaclonal" variation. Here, we examined somaclonal variation due to tissue culturing using the response to cold treatment of photosynthetic efficiency (chlorophyll fluorescence measurements for Fv/Fm, Fv'/Fm', and ΦPSII, representing maximum efficiency of photosynthesis for dark- and light-adapted leaves, and the actual electron transport operating efficiency, respectively, which are reliable indicators of photoinhibition and damage to the photosynthetic electron transport system). We compared this to variation among half-sibling seedlings from three different families of Arabidopsis lyrata ssp. petraea Somaclonal variation was limited, and we could detect within-family variation in change in chlorophyll fluorescence due to cold shock successfully with the help of tissue-culture derived replicates. Icelandic and Norwegian families exhibited higher chlorophyll fluorescence, suggesting higher performance after cold shock, than a Swedish family. Although the main effect of tissue culture on Fv/Fm, Fv'/Fm', and ΦPSII was small, there were significant interactions between tissue culture and family, suggesting that the effect of tissue culture is genotype-specific. Tissue-cultured plantlets were less affected by cold treatment than seedlings, but to a different extent in each family. These interactive effects, however, were comparable to, or much smaller than the single effect of family. These results suggest that tissue culture is a useful method for obtaining genetically homogenous replicates for studying genotype-environment interaction related to adaptively-relevant phenotypes, such as cold response, in

  2. Tissue Culture as a Source of Replicates in Nonmodel Plants: Variation in Cold Response in Arabidopsis lyrata ssp. petraea

    Directory of Open Access Journals (Sweden)

    Tanaka Kenta

    2016-12-01

    Full Text Available While genotype–environment interaction is increasingly receiving attention by ecologists and evolutionary biologists, such studies need genetically homogeneous replicates—a challenging hurdle in outcrossing plants. This could be potentially overcome by using tissue culture techniques. However, plants regenerated from tissue culture may show aberrant phenotypes and “somaclonal” variation. Here, we examined somaclonal variation due to tissue culturing using the response to cold treatment of photosynthetic efficiency (chlorophyll fluorescence measurements for Fv/Fm, Fv′/Fm′, and ΦPSII, representing maximum efficiency of photosynthesis for dark- and light-adapted leaves, and the actual electron transport operating efficiency, respectively, which are reliable indicators of photoinhibition and damage to the photosynthetic electron transport system. We compared this to variation among half-sibling seedlings from three different families of Arabidopsis lyrata ssp. petraea. Somaclonal variation was limited, and we could detect within-family variation in change in chlorophyll fluorescence due to cold shock successfully with the help of tissue-culture derived replicates. Icelandic and Norwegian families exhibited higher chlorophyll fluorescence, suggesting higher performance after cold shock, than a Swedish family. Although the main effect of tissue culture on Fv/Fm, Fv′/Fm′, and ΦPSII was small, there were significant interactions between tissue culture and family, suggesting that the effect of tissue culture is genotype-specific. Tissue-cultured plantlets were less affected by cold treatment than seedlings, but to a different extent in each family. These interactive effects, however, were comparable to, or much smaller than the single effect of family. These results suggest that tissue culture is a useful method for obtaining genetically homogenous replicates for studying genotype–environment interaction related to adaptively

  3. Differences on Pb accumulation among plant tissues of 25 varieties of maize (Zea mays)

    Institute of Scientific and Technical Information of China (English)

    DAI Quanlin; YUAN Jiangang; FANG Wei; YANG Zhongyi

    2007-01-01

    Pollution of agricultural land by heavy metals has imposed an increasingly serious risk to environmental and human health in recent years.Heavy metal pollutants may enter the human food chain through agricultural products and groundwater from the polluted soils.Progress has been made in the past decade on phytoremediation,a safe and inexpensive approach to remove contaminants from soil and water using plants.However,in most cases,agricultural land in China cannot afford to grow phytoremediator plants instead of growing crops due to food supply for the great population.Therefore,new and effective methods to decrease the risk of heavy metal pollution in crops and to clean the contaminated soils are urgently needed.If we can find crop germplasms (including species and varieties) that accumulate heavy metals in their edible parts,such as the leaves of vegetables or grains of cereals,at a level low enough for safe consumption,then we can grow these selected species or varieties in the lands contaminated or potentially contaminated by heavy metals.If we can find crop germplasms that take in low concentrations of heavy metals in their edible parts and high content of the metals in their inedible parts,then we can use these selected species or varieties for soil remediation.In this study,the feasibility of the method is assessed by analyzing Pb concentrations in edible and inedible parts of 25 varieties of maize (Zea mays) grown in Pb-contaminated soils.The soil concentrations of Pb were 595.55 mg/kg in the high Pb exposed treatment and 195.55 mg/kg in the control.The results showed that the Pb concentrations in different tissues were in the order of root > shoot ≌ leaf> grain.Compared with the control,the Pb concentrations in root,shoot and leaf were greatly increased under the high Pb exposed condition,while the increments of Pb concentration in grain were relatively lower.Under the high Pb exposure,the grain Pb concentrations of 12 varieties exceeded the maximal

  4. Fungal endophytes in aboveground tissues of desert plants: infrequent in culture, but highly diverse and distinctive symbionts.

    Science.gov (United States)

    Massimo, Nicholas C; Nandi Devan, M M; Arendt, Kayla R; Wilch, Margaret H; Riddle, Jakob M; Furr, Susan H; Steen, Cole; U'Ren, Jana M; Sandberg, Dustin C; Arnold, A Elizabeth

    2015-07-01

    In hot deserts, plants cope with aridity, high temperatures, and nutrient-poor soils with morphological and biochemical adaptations that encompass intimate microbial symbioses. Whereas the root microbiomes of arid-land plants have received increasing attention, factors influencing assemblages of symbionts in aboveground tissues have not been evaluated for many woody plants that flourish in desert environments. We evaluated the diversity, host affiliations, and distributions of endophytic fungi associated with photosynthetic tissues of desert trees and shrubs, focusing on nonsucculent woody plants in the species-rich Sonoran Desert. To inform our strength of inference, we evaluated the effects of two different nutrient media, incubation temperatures, and collection seasons on the apparent structure of endophyte assemblages. Analysis of >22,000 tissue segments revealed that endophytes were isolated four times more frequently from photosynthetic stems than leaves. Isolation frequency was lower than expected given the latitude of the study region and varied among species a function of sampling site and abiotic factors. However, endophytes were very species-rich and phylogenetically diverse, consistent with less arid sites of a similar latitudinal position. Community composition differed among host species, but not as a function of tissue type, sampling site, sampling month, or exposure. Estimates of abundance, diversity, and composition were not influenced by isolation medium or incubation temperature. Phylogenetic analyses of the most commonly isolated genus (Preussia) revealed multiple evolutionary origins of desert-plant endophytism and little phylogenetic structure with regard to seasonality, tissue preference, or optimal temperatures and nutrients for growth in vitro. Together, these results provide insight into endophytic symbioses in desert-plant communities and can be used to optimize strategies for capturing endophyte biodiversity at regional scales.

  5. Routine sample preparation and HPLC analysis for ascorbic acid (vitamin C) determination in wheat plants and Arabidopsis leaf tissues.

    Science.gov (United States)

    Szalai, Gabriella; Janda, T; Pál, Magda

    2014-06-01

    Plants have developed various mechanisms to protect themselves against oxidative stress. One of the most important non-enzymatic antioxidants is ascorbic acid. There is thus a need for a rapid, sensitive method for the analysis of the reduced and oxidised forms of ascorbic acid in crop plants. In this paper a simple, economic, selective, precise and stable HPLC method is presented for the detection of ascorbate in plant tissue. The sensitivity, the short retention time and the simple isocratic elution mean that the method is suitable for the routine quantification of ascorbate in a high daily sample number. The method has been found to be better than previously reported methods, because of the use of an economical, readily available mobile phase, UV detection and the lack of complicated extraction procedures. The method has been tested on Arabidopsis plants with different ascorbate levels and on wheat plants during Cd stress.

  6. Herbivores alter plant-wind interactions by acting as a point mass on leaves and by removing leaf tissue.

    Science.gov (United States)

    Kothari, Adit R; Burnett, Nicholas P

    2017-09-01

    In nature, plants regularly interact with herbivores and with wind. Herbivores can wound and alter the structure of plants, whereas wind can exert aerodynamic forces that cause the plants to flutter or sway. While herbivory has many negative consequences for plants, fluttering in wind can be beneficial for plants by facilitating gas exchange and loss of excess heat. Little is known about how herbivores affect plant motion in wind. We tested how the mass of an herbivore resting on a broad leaf of the tulip tree Liriodendron tulipifera, and the damage caused by herbivores, affected the motion of the leaf in wind. For this, we placed mimics of herbivores on the leaves, varying each herbivore's mass or position, and used high-speed video to measure how the herbivore mimics affected leaf movement and reconfiguration at two wind speeds inside a laboratory wind tunnel. In a similar setup, we tested how naturally occurring herbivore damage on the leaves affected leaf movement and reconfiguration. We found that the mass of an herbivore resting on a leaf can change that leaf's orientation relative to the wind and interfere with the ability of the leaf to reconfigure into a smaller, more streamlined shape. A large herbivore load slowed the leaf's fluttering frequency, while naturally occurring damage from herbivores increased the leaf's fluttering frequency. We conclude that herbivores can alter the physical interactions between wind and plants by two methods: (1) acting as a point mass on the plant while it is feeding and (2) removing tissue from the plant. Altering a plant's interaction with wind can have physical and physiological consequences for the plant. Thus, future studies of plants in nature should consider the effect of herbivory on plant-wind interactions, and vice versa.

  7. VirtualLeaf: an open source framework for cell-based modeling of plant tissue growth and development

    NARCIS (Netherlands)

    R.M.H. Merks (Roeland); M.A. Guravage (Michael); D. Inze; G.T.S. Beemster

    2011-01-01

    htmlabstractPlant organs, including leaves and roots, develop by means of a multi-level crosstalk between gene regulation, patterned cell division and cell expansion, and tissue mechanics. The multi-level regulatory mechanisms complicate classic molecular genetics or functional genomics approaches t

  8. VirtualLeaf: an open source framework for cell-based modeling of plant tissue growth and development

    NARCIS (Netherlands)

    Merks, R.M.H.; Guravage, M.A.; Inze, D.; Beemster, G.T.S.

    2011-01-01

    Plant organs, including leaves and roots, develop by means of a multi-level crosstalk between gene regulation, patterned cell division and cell expansion, and tissue mechanics. The multi-level regulatory mechanisms complicate classic molecular genetics or functional genomics approaches to biological

  9. Salt stress enhances xylem development and expression of S-adenosyl-L-methionine synthase in lignifying tissues of tomato plants.

    Science.gov (United States)

    Sánchez-Aguayo, Inmaculada; Rodríguez-Galán, José Manuel; García, Remedios; Torreblanca, José; Pardo, José Manuel

    2004-12-01

    S-Adenosyl-L-methionine synthase (SAM; ATP: L-methionine adenosyltransferase, EC 2.5.1.6) catalyzes the biosynthesis of S-adenosyl-L-methionine (AdoMet), a universal methyl-group donor. This enzyme is induced by salinity stress in tomato (Lycopersicon esculentum Mill.). To elucidate the role of SAM and AdoMet in the adaptation of plants to a saline environment, the expression pattern and histological distribution of SAM was investigated in control and salt-stressed tomato plants. Immunohistochemical analysis showed that SAM proteins were expressed in all cell types and plant organs, albeit with preferential accumulation in lignified tissues. Lignin deposition was estimated by histochemical tests and the extent of tissue lignification in response to salinity was quantified by image analysis. The average number of lignified cells in vascular bundles was significantly greater in plants under salt stress, with a maximal expansion of the lignified area found in the root vasculature. Accordingly, the greatest abundance of SAM gene transcripts and proteins occurred in roots. These results indicate that increased SAM activity correlated with a greater deposition of lignin in the vascular tissues of plants under salinity stress. A model is proposed in which an increased number of lignified tracheary elements in tomato roots under salt stress may enhance the cell-to-cell pathway for water transport, which would impart greater selectivity and reduced ion uptake, and compensate for diminished bulk flow of water and solutes along the apoplastic pathway.

  10. Water exchange in plant tissue studied by proton NMR in the presence of paramagnetic centers.

    Science.gov (United States)

    Bacić, G; Ratković, S

    1984-04-01

    The proton NMR relaxation of water in maize roots in the presence of paramagnetic centers, Mn2+, Mn- EDTA2 -, and dextran-magnetite was measured. It was shown that the NMR method of Conlon and Outhred (1972, Biochem. Biophys. Acta. 288:354-361) can be applied to a heterogenous multicellular system, and the water exchange time between cortical cells and the extracellular space can be calculated. The water exchange is presumably controlled by the intracellular unstirred layers. The Mn- EDTA2 - complex is a suitable paramagnetic compound for complex tissue, while the application of dextran-magnetite is probably restricted to studies of water exchange in cell suspensions. The water free space of the root and viscosity of the cells cytoplasm was estimated with the use of Mn- EDTA2 -. The convenience of proton NMR for studying the multiphase uptake of paramagnetic ions by plant root as well as their transport to leaves is demonstrated. A simple and rapid NMR technique (spin-echo recovery) for continuous measurement of the uptake process is presented.

  11. Tissue culture study of the medicinal plant leek (allium ampeloprasum L).

    Science.gov (United States)

    Monemi, Mohammad Bagher; Kazemitabar, S Kamal; Bakhshee Khaniki, Gholamreza; Yasari, Esmaeil; Sohrevardi, Firouzeh; Pourbagher, Roghayeh

    2014-01-01

    Persian shallot, also called leek (Allium ampeloprasum), is a monocotyledon plant of the lily family (Liliaceae). It belongs to the genus Allium, has a characteristic taste and morphological features, making it to be considered as one of the popular herbal medicine. This research was conducted with the purpose of obtaining optimal conditions for tissue culture of Persian shallot and comparing its active ingredient production in vitro versus in vivo. In this study, the auxin 2, 4-D and benzyl aminopurine- 6 (BAP) hormones, each at two concentrations (0.5 and 0.1 mg/ L) and Kin at 0.5 mg/ L were used in the format of a randomized complete block design in three replications. Results showed that the best culture media for callus formation for leaf and seed explants were the MS cultures with the hormonal compositions (0.5 mg/ L of 2, 4- D, 0.1 mg/ L of BAP) and (0.5 mg/ L of Kin and 0.1 mg/ L of 2, 4- D). Identification of the chemical composition of the essential oils, extracted either from leek callus or leaf was carried out using GC mass analysis. Twenty one compounds were detected in the GC mass spectra, seven of which constitutv about 51.5% of the total amount of compounds present in the essential oils were identified. Our data demonstrate that the leek essential oil constituents as well as callus formation can be affected by culture medium condition.

  12. Species and tissue type regulate long-term decomposition of brackish marsh plants grown under elevated CO2 conditions

    Science.gov (United States)

    Jones, Joshua A.; Cherry, Julia A.; McKee, Karen L.

    2016-02-01

    Organic matter accumulation, the net effect of plant production and decomposition, contributes to vertical soil accretion in coastal wetlands, thereby playing a key role in whether they keep pace with sea-level rise. Any factor that affects decomposition may affect wetland accretion, including atmospheric CO2 concentrations. Higher CO2 can influence decomposition rates by altering plant tissue chemistry or by causing shifts in plant species composition or biomass partitioning. A combined greenhouse-field experiment examined how elevated CO2 affected plant tissue chemistry and subsequent decomposition of above- and belowground tissues of two common brackish marsh species, Schoenoplectus americanus (C3) and Spartina patens (C4). Both species were grown in monoculture and in mixture under ambient (350-385 μL L-1) or elevated (ambient + 300 μL L-1) atmospheric CO2 conditions, with all other growth conditions held constant, for one growing season. Above- and belowground tissues produced under these treatments were decomposed under ambient field conditions in a brackish marsh in the Mississippi River Delta, USA. Elevated CO2 significantly reduced nitrogen content of S. americanus, but not sufficiently to affect subsequent decomposition. Instead, long-term decomposition (percent mass remaining after 280 d) was controlled by species composition and tissue type. Shoots of S. patens had more mass remaining (41 ± 2%) than those of S. americanus (12 ± 2%). Belowground material decomposed more slowly than that placed aboveground (62 ± 1% vs. 23 ± 3% mass remaining), but rates belowground did not differ between species. Increases in atmospheric CO2 concentration will likely have a greater effect on overall decomposition in this brackish marsh community through shifts in species dominance or biomass allocation than through effects on tissue chemistry. Consequent changes in organic matter accumulation may alter marsh capacity to accommodate sea-level rise through vertical

  13. An UPLC-MS/MS method for highly sensitive high-throughput analysis of phytohormones in plant tissues

    Directory of Open Access Journals (Sweden)

    Balcke Gerd Ulrich

    2012-11-01

    Full Text Available Abstract Background Phytohormones are the key metabolites participating in the regulation of multiple functions of plant organism. Among them, jasmonates, as well as abscisic and salicylic acids are responsible for triggering and modulating plant reactions targeted against pathogens and herbivores, as well as resistance to abiotic stress (drought, UV-irradiation and mechanical wounding. These factors induce dramatic changes in phytohormone biosynthesis and transport leading to rapid local and systemic stress responses. Understanding of underlying mechanisms is of principle interest for scientists working in various areas of plant biology. However, highly sensitive, precise and high-throughput methods for quantification of these phytohormones in small samples of plant tissues are still missing. Results Here we present an LC-MS/MS method for fast and highly sensitive determination of jasmonates, abscisic and salicylic acids. A single-step sample preparation procedure based on mixed-mode solid phase extraction was efficiently combined with essential improvements in mobile phase composition yielding higher efficiency of chromatographic separation and MS-sensitivity. This strategy resulted in dramatic increase in overall sensitivity, allowing successful determination of phytohormones in small (less than 50 mg of fresh weight tissue samples. The method was completely validated in terms of analyte recovery, sensitivity, linearity and precision. Additionally, it was cross-validated with a well-established GC-MS-based procedure and its applicability to a variety of plant species and organs was verified. Conclusion The method can be applied for the analyses of target phytohormones in small tissue samples obtained from any plant species and/or plant part relying on any commercially available (even less sensitive tandem mass spectrometry instrumentation.

  14. Plant DNA Detection from Grasshopper Guts: A Step-by-Step Protocol, from Tissue Preparation to Obtaining Plant DNA Sequences

    Directory of Open Access Journals (Sweden)

    Alina Avanesyan

    2014-02-01

    Full Text Available Premise of the study: A PCR-based method of identifying ingested plant DNA in gut contents of Melanoplus grasshoppers was developed. Although previous investigations have focused on a variety of insects, there are no protocols available for plant DNA detection developed for grasshoppers, agricultural pests that significantly influence plant community composition. Methods and Results: The developed protocol successfully used the noncoding region of the chloroplast trnL (UAA gene and was tested in several feeding experiments. Plant DNA was obtained at seven time points post-ingestion from whole guts and separate gut sections, and was detectable up to 12 h post-ingestion in nymphs and 22 h post-ingestion in adult grasshoppers. Conclusions: The proposed protocol is an effective, relatively quick, and low-cost method of detecting plant DNA from the grasshopper gut and its different sections. This has important applications, from exploring plant “movement” during food consumption, to detecting plant–insect interactions.

  15. Recent progress in the understanding of tissue culture-induced genome level changes in plants and potential applications.

    Science.gov (United States)

    Neelakandan, Anjanasree K; Wang, Kan

    2012-04-01

    In vitro cell and tissue-based systems have tremendous potential in fundamental research and for commercial applications such as clonal propagation, genetic engineering and production of valuable metabolites. Since the invention of plant cell and tissue culture techniques more than half a century ago, scientists have been trying to understand the morphological, physiological, biochemical and molecular changes associated with tissue culture responses. Establishment of de novo developmental cell fate in vitro is governed by factors such as genetic make-up, stress and plant growth regulators. In vitro culture is believed to destabilize the genetic and epigenetic program of intact plant tissue and can lead to chromosomal and DNA sequence variations, methylation changes, transposon activation, and generation of somaclonal variants. In this review, we discuss the current status of understanding the genomic and epigenomic changes that take place under in vitro conditions. It is hoped that a precise and comprehensive knowledge of the molecular basis of these variations and acquisition of developmental cell fate would help to devise strategies to improve the totipotency and embryogenic capability in recalcitrant species and genotypes, and to address bottlenecks associated with clonal propagation.

  16. Phytoplasma detection in tissue culture of Gladiolus plants grown under various conditions

    Directory of Open Access Journals (Sweden)

    Maria Kamińska

    2014-01-01

    Full Text Available To test whether phytoplasmas are sensitive to temperature, phytoplasma affected micropropagated gladiolus plants were grown under varying conditions of media content and temperature, in the presence or absence of light. PCR analysis indicated that phytoplasma detection was more successful in plants grown at low temperatures. Plants kept from one to three months at reduced temperature tended to have higher titre of phytoplasma than the plants maintained in stable 20oC high temperature. The best detection was in plants grown on medium containing kinetin+NAA and in the presence of light. In those plants phytoplasmas were detected in direct PCR after one month of culture.

  17. Why are Nitrogen Concentrations in Plant Tissues Lower under Elevated CO2? A Critical Examination of the Hypotheses

    Institute of Scientific and Technical Information of China (English)

    Daniel R. Taub; Xianzhong Wang

    2008-01-01

    Plants grown under elevated atmospheric [CO2] typically have decreased tissue concentrations of N compared with plants grown under current ambient [CO2]. The physiological mechanisms responsible for this phenomenon have not been definitely established, although a considerable number of hypotheses have been advanced to account for it. In this review we discuss and critically evaluate these hypotheses. One contributing factor to the decreases in tissue N concentrations clearly is dilution of N by increased photosynthetic assimilation of C. In addition, studies on intact plants show strong evidence for a general decrease in the specific uptake rates (uptake per unit mass or length of root) of N by roots under elevated CO2. This decreased root uptake appears likely to be the result both of decreased N demand by shoots and of decreased ability of the soil-root system to supply N. The best-supported mechanism for decreased N supply is a decrease in transpiration-driven mass flow of N in soils due to decreased stomatal conductance at elevated CO2, although some evidence suggests that altered root system architecture may also play a role. There is also limited evidence suggesting that under elevated CO2, plants may exhibit increased rates of N loss through volatilization and/or root exudation, further contributing to lowering tissue N concentrations.

  18. Growth of plant tissue cultures in simulated lunar soil: Implications for a lunar base CELSS (Controlled Ecological Life Support System)

    Science.gov (United States)

    Venketeswaran, S.

    1988-01-01

    Experiments were carried out on plant tissue cultures, seed germination, seedling development and plants grown on Simulated Lunar Soil to evaluate the potential of future development of lunar based agriculture. The studies done to determine the effect of the placement of SLS on tissue cultures showed no adverse effect of SLS on tissue cultures. Although statistically insignificant, SLS in suspension showed a comparatively higher growth rate. Observations indicate the SLS, itself cannot support calli growth but was able to show a positive effect on growth rate of calli when supplemented with MS salts. This positive effect related to nutritive value of the SLS was found to have improved at high pH levels, than at the recommended low pH levels for standard media. Results from seed germination indicated that there is neither inhibitory, toxicity nor stimulatory effect of SLS, even though SLS contains high amounts of aluminum compounds compared to earth soil. Analysis of seeding development and growth data showed significant reduction in growth rate indicating that, SLS was a poor growth medium for plant life. This was confirmed by the studies done with embryos and direct plant growth on SLS. Further observations attributed this poor quality of SLS is due to it's lack of essential mineral elements needed for plant growth. By changing the pH of the soil, to more basic conditions, the quality of SLS for plant growth could be improved up to a significant level. Also it was found that the quality of SLS could be improved by almost twice, by external supply of major mineral elements, directly to SLS.

  19. Seasonal variation in nitrogen pools and 15N/13C natural abundances in different tissues of grassland plants

    Directory of Open Access Journals (Sweden)

    J. K. Schjoerring

    2011-12-01

    Full Text Available Seasonal changes in nitrogen (N pools, carbon (C content and natural abundance of 13C and 15N in different tissues of ryegrass plants were investigated in two intensively managed grassland fields in order to address their ammonia (NH3 exchange potential. Green leaves generally had the largest total N concentration followed by stems and inflorescences. Senescent leaves had the lowest N concentration, indicating N re-allocation. The seasonal pattern of the Γ value, i.e. the ratio between NH4+ and H+ concentrations, was similar for the various tissues of the ryegrass plants but the magnitude of Γ differed considerably among the different tissues. Green leaves and stems generally had substantially lower Γ values than senescent leaves and litter. Substantial peaks in Γ were observed during spring and summer in response to fertilization and grazing. These peaks were associated with high NH4+ rather than with low H+ concentrations. Peaks in Γ also appeared during the winter, coinciding with increasing δ15N values, indicating absorption of N derived from mineralization of soil organic matter. At the same time, δ13C values were declining, suggesting reduced photosynthesis and capacity for N assimilation. δ15N and δ13C values were more influenced by mean monthly temperature than by the accumulated monthly precipitation. In conclusion, ryegrass plants showed a clear seasonal pattern in N pools. Green leaves and stems of ryegrass plants generally seem to constitute a sink for NH3, while senescent leaves have a large potential for NH3 emission. However, management events such as fertilisation and grazing may create a high NH3 emission potential even in green plant parts. The obtained results provide input for future modelling of plant-atmosphere NH3 exchange.

  20. Ricinosomes: an organelle for developmentally regulated programmed cell death in senescing plant tissues

    Science.gov (United States)

    Gietl, C.; Schmid, M.

    2001-02-01

    This review describes aspects of programmed cell death (PCD). Present research maps the enzymes involved and explores the signal transduction pathways involved in their synthesis. A special organelle (the ricinosome) has been discovered in the senescing endosperm of germinating castor beans (Ricinus communis) that develops at the beginning of PCD and delivers large amounts of a papain-type cysteine endopeptidase (CysEP) in the final stages of cellular disintegration. Castor beans store oil and proteins in a living endosperm surrounding the cotyledons. These stores are mobilized during germination and transferred into the cotyledons. PCD is initiated after this transfer is complete. The CysEP is synthesized in the lumen of the endoplasmic reticulum (ER) where it is retained by its C-terminal KDEL peptide as a rather inactive pro-enzyme. Large number of ricinosomes bud from the ER at the same time as the nuclear DNA is characteristically fragmented during PCD. The mitochondria, glyoxysomes and ribosomes are degraded in autophagic vacuoles, while the endopeptidase is activated by removal of the propeptide and the KDEL tail and enters the cytosol. The endosperm dries and detaches from the cotyledons. A homologous KDEL-tailed cysteine endopeptidase has been found in several senescing tissues; it has been localized in ricinosomes of withering day-lily petals and dying seed coats. Three genes for a KDEL-tailed cysteine endopeptidase have been identified in Arabidopsis. One is expressed in senescing ovules, the second in the vascular vessels and the third in maturing siliques. These genes open the way to exploring PCD in plants.

  1. Ultrastructural changes in aster yellows phytoplasma affected Limonium sinuatum Mill. plants.I Pathology of conducting tissues

    Directory of Open Access Journals (Sweden)

    Anna Rudzińska-Langwald

    2014-01-01

    Full Text Available Changes in anatomy and cytology of conducting tissues of Limonium sinuatum Mill. plants affected by aster yellows phytoplasma were investigated. In the phloem tissues of affected plants stem necrosis takes place. In necrotic regions no sieve tubes were observed only necrotic cells and parenchyma cells. The sieve tubes present on the border of necrosis showed collapsed walls and were rich in vesicles. Phytoplasma cells were observed in sieve tubes present in nonnecrotic regions of the phloem. Various structural changes in sieve elements were investigated. The endoplasmic reticulum cistemae were often localised in the lumen of the sieve element without contact with the walls. Such localisation of endoplasmic reticulum was never observed in healthy plants. Vesicles of different size, fuzzy material and clumping of p-proteins were characteristic for sieve elements from nonnecrotic part of phloem. No correlation with the sieve tube structure and the appearance of phytoplasma in a single sieve element was found. In control plants of L. sinuatum phloem observed were phloem parenchyma cells with spiny vesicles (SV. In infected plants there were a remarkable increase in cells with SV. Also the SV itself had not only a vesicular but also a tubular or extended cistern shape.

  2. Analysis of Explosives in Plant Tissues: Modifications to Method 8330 for Soil.

    Science.gov (United States)

    1998-05-01

    containing tomato plant 30 Figure 13. Chromatograms of RDX containing radish root 31 Figure 14. Chromatograms of RDX containing lettuce 32...grown hydroponically in TNT solutions have shown to both retain TNT and reductively transform TNT (Palazzo and Leggett 1986a,b). Con- siderable...quantities of bound monoaminodinitrotoluenes were recovered from hydroponically grown plants using acid hydrolysis of plant material following benzene

  3. Preparation and electrochemical application of a new biosensor based on plant tissue/polypyrrole for determination of acetaminophen

    Indian Academy of Sciences (India)

    Gholamhossein Rounaghi; Roya Mohammadzadeh Kakhki

    2012-10-01

    Banana tissue containing polyphenol oxidase was incorporated into polypyrrole matrix to make a biosensor for the analysis of acetaminophen (ACT). The electrocatalytic behaviour of oxidized acetaminophen was studied at the surface of the biosensor, using various electrochemical methods. The advantages of this biosensor for the determination of acetaminophen are excellent catalytic activity, good detection limit and high exchange current density. The electrochemical and structural properties of the electrode were assessed using cyclic voltammetry, differential voltammetry, chronoamperometric techniques. The analytical properties (sensitivity, p) of this biosensor increased with plant tissue loading. Also this new biosensor was successfully applied for determination of acetaminophen in biologic samples.

  4. Minimizing Nitrate Reduction during Kjeldahl Digestion of Plant Tissue Extracts and Stem Exudates : APPLICATION TO N STUDIES.

    Science.gov (United States)

    Pace, G M; Mackown, C T; Volk, R J

    1982-01-01

    From 10 to 60% of the nitrate present in plant tissue extracts and stem exudates of corn (Zea mays L.) was found to be reduced during Kjeldahl digestion, even in the absence of added reducing agents. This reduction is of particular concern in [(15)N]nitrate assimilation studies, because it results in an overestimate of nitrate reduction. To overcome this problem, a method was developed for removing nitrate prior to Kjeldahl digestion, thereby preventing nitrate reduction. The procedure utilizes hydrogen peroxide for partial oxidation of organic matter in order to minimize the nitration of organic compounds. The free nitrates are then volatilized as nitric acid from concentrated sulfuric acid at 95 degrees C. When the proposed method was used as a pretreatment to Kjeldahl digestion, less than 0.5% of the applied nitrate was recovered in the reduced nitrogen fraction of plant tissue extracts and stem exudates.

  5. Phenolic compounds in cultures of tissues of tea plants and the effect of light on their synthesis

    Energy Technology Data Exchange (ETDEWEB)

    Koretskaya, T.F.; Zaprometov, M.N.

    1975-01-01

    Stem and leaf calluses of tea plants (Camellia sinensis) retain the capacity for synthesis of phenolic compounds. The content of phenolic compounds comprises 2 to 5 percent of dry weight, the main share (80 to 95 percent) belonging to catechins and leucoanthocyans, including their polymeric forms. The following compounds were detected in callus tissue: (--)-epicatechin, (+)-catechin, two leucoanthocyans, and several unidentified phenolic compounds that fluoresce in UV. (--)-Epicatechin is predominant. In contrast to tissues of an intact plant, the callus does not contain gallocatechins or free gallic acid under the given cultivation conditions. The content of phenolic compounds changes in proportion to callus growth, their greatest amount being formed during the phase of intensive growth. Light stimulates synthesis of phenolic compounds, including the most reduced group of flavonoids, viz., leucoanthocyans and catechins.

  6. Expression of a plant-associated human cancer antigen in normal,premalignant and malignant esophageal tissues

    Institute of Scientific and Technical Information of China (English)

    Jun Fu; Ping Qu; Mo Li; Hai-Mei Tian; Zhen-Hai Zheng; Xin-Wen Zheng; Wei Zhang

    2003-01-01

    AIM: To study the relationship between the expression profiles of a plant-associated human cancer antigen and carcinogenesis of esophagus and its significance. METHODS: We analyzed expression of a plant-associated human cancer antigen in biopsy specimens of normal (n=29),mildly hyperplastic (n=29), mildly (n=30), moderately (n=27)and severely dysplastic (n=29) and malignant esophageal (n=30) tissues by immunohistochemistry. RESULTS: The plant-associated human cancer antigen was mainly confined to the cytoplasm and showed diffuse type of staining. Positive staining was absent or weak in normal (0/30) and mildly hyperplastic tissue samples (2/29), while strong staining was observed in severe dysplasia (23/29) and carcinoma in situ (24/30). There was significant difference of its expression between normal mucosa and severely dysplastic tissues (P<0.001) or carcinoma in situ (P<0.001). Significant difference was also observed between mild dysplasia and severe dysplasia (P<0.001) or carcinomain situ (P<0.001). An overall trend toward increased staining intensity with increasing grade of dysplasia was found. There was a linear correlation between grade of lesions and staining intensity (r=0.794,P<0.001). Samples from esophageal cancer showed no higher levels of expression than those in severely dysplastic lesions (P>0.05). CONCLUSION: The abnormal expression of this plantassociated human cancer antigen in esophageal lesions is a frequent and early finding in the normal-dysplasiacarcinoma sequence in esophageal carcinogenesis. It might contribute to the carcinogenesis of esophageal cancer. The abnormal expression of this plant-associated human cancer antigen in esophageal lesion tissues may serve as a potential new biomarker for early identification of esophageal cancer.

  7. Manipulation of host plant cells and tissues by gall-inducing insects and adaptive strategies used by different feeding guilds.

    Science.gov (United States)

    Oliveira, D C; Isaias, R M S; Fernandes, G W; Ferreira, B G; Carneiro, R G S; Fuzaro, L

    2016-01-01

    Biologists who study insect-induced plant galls are faced with the overwhelming diversity of plant forms and insect species. A challenge is to find common themes amidst this diversity. We discuss common themes that have emerged from our cytological and histochemical studies of diverse neotropical insect-induced galls. Gall initiation begins with recognition of reactive plant tissues by gall inducers, with subsequent feeding and/or oviposition triggering a cascade of events. Besides, to induce the gall structure insects have to synchronize their life cycle with plant host phenology. We predict that reactive oxygen species (ROS) play a role in gall induction, development and histochemical gradient formation. Controlled levels of ROS mediate the accumulation of (poly)phenols, and phytohormones (such as auxin) at gall sites, which contributes to the new cell developmental pathways and biochemical alterations that lead to gall formation. The classical idea of an insect-induced gall is a chamber lined with a nutritive tissue that is occupied by an insect that directly harvests nutrients from nutritive cells via its mouthparts, which function mechanically and/or as a delivery system for salivary secretions. By studying diverse gall-inducing insects we have discovered that insects with needle-like sucking mouthparts may also induce a nutritive tissue, whose nutrients are indirectly harvested as the gall-inducing insects feeds on adjacent vascular tissues. Activity of carbohydrate-related enzymes across diverse galls corroborates this hypothesis. Our research points to the importance of cytological and histochemical studies for elucidating mechanisms of induced susceptibility and induced resistance. Copyright © 2015 Elsevier Ltd. All rights reserved.

  8. Rhizospheric bacterial strain Brevibacterium casei MH8a colonizes plant tissues and enhances Cd, Zn, Cu phytoextraction by white mustard

    Directory of Open Access Journals (Sweden)

    Tomasz ePłociniczak

    2016-02-01

    Full Text Available Environmental pollution by heavy metals has become a serious problem in the world. Phytoextraction, which is one of the plant-based technologies, has attracted the most attention for the bioremediation of soils polluted with these contaminants.The aim of this study was to determine whether the multiple-tolerant bacterium, Brevibacterium casei MH8a isolated from the heavy metal-contaminated rhizosphere soil of Sinapis alba L., is able to promote plant growth and enhance Cd, Zn and Cu uptake by white mustard under laboratory conditions. Additionally, the ability of the rifampicin-resistant spontaneous mutant of MH8a to colonize plant tissues and its mechanisms of plant growth promotion were also examined. In order to assess the ecological consequences of bioaugmentation on autochthonous bacteria, the phospholipid fatty acid (PLFA analysis was used. The MH8a strain exhibited the ability to produce ammonia, 1-amino-cyclopropane-1-carboxylic acid deaminase, indole 3-acetic acid and HCN but was not able to solubilize inorganic phosphate and produce siderophores. Introduction of MH8a into soil significantly increased S. alba biomass and the accumulation of Cd (208%, Zn (86% and Cu (39% in plant shoots in comparison with those grown in non-inoculated soil. Introduced into the soil, MH8a was able to enter the plant and was found in the roots and leaves of inoculated plants thus indicating its endophytic features. PLFA analysis revealed that the MH8a that was introduced into soil had a temporary influence on the structure of the autochthonous bacterial communities. The plant growth-promoting features of the MH8a strain and its ability to enhance the metal uptake by white mustard and its long-term survival in soil as well as its temporary impact on autochthonous microorganisms make the strain a suitable candidate for the promotion of plant growth and the efficiency of phytoextraction.

  9. Down-regulation of tissue N:P ratios in terrestrial plants by elevated CO2.

    Science.gov (United States)

    Deng, Qi; Hui, Dafeng; Luo, Yiqi; Elser, James; Wang, Ying-ping; Loladze, Irakli; Zhang, Quanfa; Dennis, Sam

    2015-12-01

    Increasing atmospheric CO2 concentrations generally alter element stoichiometry in plants. However, a comprehensive evaluation of the elevated CO2 impact on plant nitrogen: phosphorus (N:P) ratios and the underlying mechanism has not been conducted. We synthesized the results from 112 previously published studies using meta-analysis to evaluate the effects of elevated CO2 on the N:P ratio of terrestrial plants and to explore the underlying mechanism based on plant growth and soil P dynamics. Our results show that terrestrial plants grown under elevated CO2 had lower N:P ratios in both above- and belowground biomass across different ecosystem types. The response ratio for plant N:P was negatively correlated with the response ratio for plant growth in croplands and grasslands, and showed a stronger relationship for P than for N. In addition, the CO2-induced down-regulation of plant N:P was accompanied by 19.3% and 4.2% increases in soil phosphatase activity and labile P, respectively, and a 10.1% decrease in total soil P. Our results show that down-regulation of plant N:P under elevated CO2 corresponds with accelerated soil P cycling. These findings should be useful for better understanding of terrestrial plant stoichiometry in response to elevated CO2 and of the underlying mechanisms affecting nutrient dynamics under climate change.

  10. Acquisition and Evolution of Plant Pathogenesis–Associated Gene Clusters and Candidate Determinants of Tissue-Specificity in Xanthomonas

    Science.gov (United States)

    Van Sluys, Marie-Anne; White, Frank F.; Ryan, Robert P.; Dow, J. Maxwell; Rabinowicz, Pablo; Salzberg, Steven L.; Leach, Jan E.; Sonti, Ramesh; Brendel, Volker; Bogdanove, Adam J.

    2008-01-01

    Background Xanthomonas is a large genus of plant-associated and plant-pathogenic bacteria. Collectively, members cause diseases on over 392 plant species. Individually, they exhibit marked host- and tissue-specificity. The determinants of this specificity are unknown. Methodology/Principal Findings To assess potential contributions to host- and tissue-specificity, pathogenesis-associated gene clusters were compared across genomes of eight Xanthomonas strains representing vascular or non-vascular pathogens of rice, brassicas, pepper and tomato, and citrus. The gum cluster for extracellular polysaccharide is conserved except for gumN and sequences downstream. The xcs and xps clusters for type II secretion are conserved, except in the rice pathogens, in which xcs is missing. In the otherwise conserved hrp cluster, sequences flanking the core genes for type III secretion vary with respect to insertion sequence element and putative effector gene content. Variation at the rpf (regulation of pathogenicity factors) cluster is more pronounced, though genes with established functional relevance are conserved. A cluster for synthesis of lipopolysaccharide varies highly, suggesting multiple horizontal gene transfers and reassortments, but this variation does not correlate with host- or tissue-specificity. Phylogenetic trees based on amino acid alignments of gum, xps, xcs, hrp, and rpf cluster products generally reflect strain phylogeny. However, amino acid residues at four positions correlate with tissue specificity, revealing hpaA and xpsD as candidate determinants. Examination of genome sequences of xanthomonads Xylella fastidiosa and Stenotrophomonas maltophilia revealed that the hrp, gum, and xcs clusters are recent acquisitions in the Xanthomonas lineage. Conclusions/Significance Our results provide insight into the ancestral Xanthomonas genome and indicate that differentiation with respect to host- and tissue-specificity involved not major modifications or wholesale

  11. Acquisition and evolution of plant pathogenesis-associated gene clusters and candidate determinants of tissue-specificity in xanthomonas.

    Directory of Open Access Journals (Sweden)

    Hong Lu

    Full Text Available BACKGROUND: Xanthomonas is a large genus of plant-associated and plant-pathogenic bacteria. Collectively, members cause diseases on over 392 plant species. Individually, they exhibit marked host- and tissue-specificity. The determinants of this specificity are unknown. METHODOLOGY/PRINCIPAL FINDINGS: To assess potential contributions to host- and tissue-specificity, pathogenesis-associated gene clusters were compared across genomes of eight Xanthomonas strains representing vascular or non-vascular pathogens of rice, brassicas, pepper and tomato, and citrus. The gum cluster for extracellular polysaccharide is conserved except for gumN and sequences downstream. The xcs and xps clusters for type II secretion are conserved, except in the rice pathogens, in which xcs is missing. In the otherwise conserved hrp cluster, sequences flanking the core genes for type III secretion vary with respect to insertion sequence element and putative effector gene content. Variation at the rpf (regulation of pathogenicity factors cluster is more pronounced, though genes with established functional relevance are conserved. A cluster for synthesis of lipopolysaccharide varies highly, suggesting multiple horizontal gene transfers and reassortments, but this variation does not correlate with host- or tissue-specificity. Phylogenetic trees based on amino acid alignments of gum, xps, xcs, hrp, and rpf cluster products generally reflect strain phylogeny. However, amino acid residues at four positions correlate with tissue specificity, revealing hpaA and xpsD as candidate determinants. Examination of genome sequences of xanthomonads Xylella fastidiosa and Stenotrophomonas maltophilia revealed that the hrp, gum, and xcs clusters are recent acquisitions in the Xanthomonas lineage. CONCLUSIONS/SIGNIFICANCE: Our results provide insight into the ancestral Xanthomonas genome and indicate that differentiation with respect to host- and tissue-specificity involved not major

  12. Inhibition of phenylpropanoid biosynthesis in Artemisia annua L.: a novel approach to reduce oxidative browning in plant tissue culture.

    Science.gov (United States)

    Jones, Andrew Maxwell Phineas; Saxena, Praveen Kumar

    2013-01-01

    Oxidative browning is a common and often severe problem in plant tissue culture systems caused by the accumulation and oxidation of phenolic compounds. The current study was conducted to investigate a novel preventative approach to address this problem by inhibiting the activity of the phenylalanine ammonia lyase enzyme (PAL), thereby reducing the biosynthesis of phenolic compounds. This was accomplished by incorporating 2-aminoindane-2-phosphonic acid (AIP), a competitive PAL inhibitor, into culture media of Artemisia annua as a model system. Addition of AIP into culture media resulted in significant reductions in visual tissue browning, a reduction in total phenol content, as well as absorbance and autoflourescence of tissue extracts. Reduced tissue browning was accompanied with a significant increase in growth on cytokinin based medium. Microscopic observations demonstrated that phenolic compounds accumulated in discrete cells and that these cells were more prevalent in brown tissue. These cells were highly plasmolyzed and often ruptured during examination, demonstrating a mechanism in which phenolics are released into media in this system. These data indicate that inhibiting phenylpropanoid biosynthesis with AIP is an effective approach to reduce tissue browning in A. annua. Additional experiments with Ulmus americana and Acer saccharum indicate this approach is effective in many species and it could have a wide application in systems where oxidative browning restricts the development of biotechnologies.

  13. Diversity of endophytic fungi associated with the foliar tissue of a hemi-parasitic plant Macrosolen cochinchinensis.

    Science.gov (United States)

    Zhou, Sheng-Liang; Yan, Shu-Zhen; Liu, Qi-Sha; Chen, Shuang-Lin

    2015-01-01

    Foliar fungal endophytes are an important plant-associated fungal group. However, little is known about these fungi in hemi-parasitic plants, a unique plant group which derive nutrients from living plants of its hosts by haustoria while are photosynthetic to some degree. In this paper, the endophytic fungi in the leaves of a species of hemi-parasitic plant, Macrosolen cochinchinensis, were studied by both culture-dependent and culture-independent methods. By culture-dependent method, a total of 511 isolates were recovered from 452 of 600 leaf fragments (colonization rate = 75.3 %) and were identified to be 51 taxa. Valsa sp. was the most abundant (relative abundance = 38.4 %), followed by Cladosporium sp. 1 (13.5 %), Ulocladium sp. (4.3 %), Phomopsis sp. 2 (3.7 %), Hendersonia sp. (3.5 %), and Diaporthe sp. 4 (3.5 %). The Shannon index (H') of the isolated endophytic fungi was 2.628, indicating a moderate diversity. By culture-independent method, Aspergillus spp., Cladosporium sp., Mycosphaerella sp., Acremonium strictum, and Tremella sp. were detected. To our knowledge, the Tremella species have never been detected as endophytes so far. In addition, a cloned sequence was not similar with any current sequence in the Genbank, which may represent a novel species. Altogether, this study documented endophytic fungal assemble in the leaves of M. cochinchinensis which was worthy of our attention, and may expand our knowledge about endophytic fungi within the photosynthetic tissues of plants.

  14. Prediction of atmospheric δ13CO2 using fossil plant tissues

    Energy Technology Data Exchange (ETDEWEB)

    Jahren, A. Hope [Johns Hopkins Univ., Baltimore, MD (United States); Arens, Nan Crystal [Hobart and William Smith Colleges, Geneva, NY (United States); Harbeson, Stephanie A. [Johns Hopkins Univ., Baltimore, MD (United States); Univ. of Virginia, Charlottesville, VA (United States)

    2008-06-30

    To summarize the content: we presented the results of laboratory experiments designed to quantify the relationship between plant tissue δ13C and δ13CO2 values under varying environmental conditions, including differential pCO2 ranging from 1 to 3 times today’s levels. As predicted, plants grown under elevated pCO2 showed increased average biomass compared to controls grown at the same temperature. Across a very large range in δ13Ca (≈ 24 ‰) and pCO2 (≈ 740 ppmv) we observed a consistent correlation between δ13Ca and δ13Cp (p<0.001). We show an average isotopic depletion of -25.4 ‰ for above-ground tissue and -23.2 ‰ for below-ground tissue of Raphanus sativus L. relative to the composition of the atmosphere under which it formed. For both above- and below-ground tissue, grown at both ~23 °C and ~29 °C, correlation was strong and significant (r2 ≥ 0.98, p<0.001); variation in pCO2 level had little or no effect on this relationship.

  15. Growth response and tissue accumulation trends of herbaceous wetland plant species exposed to elevated aqueous mercury levels.

    Science.gov (United States)

    Willis, Jonathan M; Gambrell, Robert P; Hester, Mark W

    2010-08-01

    The impacts of elevated aqueous mercury levels (0, 2, and 4 ppm) on the growth status and mercury tissue concentrations of Eleocharis parvula, Saururus cernuus, Juncus effuses, Typha latifolia, and Panicum hemitomon were determined. Both short-term (net CO2 assimilation) and long-term (biomass) indicators of plant growth status suggest that Eleocharis parvula, Saururus cernuus, and Juncus effuses were relatively unimpacted by elevated mercury levels, whereas Typha latifolia and Panicum hemitomon were somewhat impacted at elevated mercury levels. Eleocharis parvula, Panicum hemitomon, and Typha latifolia generally had the greatest overall belowground tissue concentrations of mercury (2 ppm treatment: 7.21, 7.32, and 9.64 ppm respectively; 4 ppm treatment: 16.23, 18.23, and 13.98 ppm, respectively) and aboveground tissue concentrations of mercury (2 ppm treatment: 0.01, 0.04, 0.02; 4 ppm treatment: 0.26; 0.11; 0.17 ppm, respectively). However, the species investigated in this study demonstrated lower levels of mercury accumulation into tissues when compared with similar investigations of other aquatic plants, suggesting that the above species are not optimal for phytoremediation efforts.

  16. Dianthin 30 and 32 from Dianthus caryophyllus: two inhibitors of plant protein synthesis and their tissue distribution.

    Science.gov (United States)

    Reisbig, R R; Bruland, O

    1983-07-15

    The ability of dianthin 30 and 32 to inhibit translation in reticulocyte lysates and wheat germ extracts has been studied. The dianthins, like the A chains of the toxins abrin and ricin, inhibited protein synthesis in reticulocyte lysates by inactivating the 60S ribosomal subunit. They also inhibited, at concentrations of 10 ng/ml, a protein-synthesizing system from wheat germ and inactivated isolated wheat germ ribosomes. The concentration of the dianthins in different tissues of the plant was determined by rocket immunoelectrophoresis and by the dianthin's ability to inhibit protein synthesis. Dianthin 32 was found only in the leaves and in growing shoots, while dianthin 30 was present throughout the plant. In the older parts of the plant, the dianthins constituted between 1 and 3% of the total extractable protein whereas much less was found in the younger parts.

  17. An immunocytochemical procedure for protein localization in various nematode life stages combined with plant tissues using methylacrylate-embedded specimens.

    Science.gov (United States)

    Vieira, Paulo; Banora, Mohamed Youssef; Castagnone-Sereno, Philippe; Rosso, Marie-Noëlle; Engler, Gilbert; de Almeida Engler, Janice

    2012-10-01

    Plant-parasitic nematodes possess a large number of proteins that are secreted in planta, allowing them to be successful parasites of plants. The majority of these proteins are synthesized mainly in the nematode subventral and dorsal glands as well as in other organs. To improve the immunovisualization of these proteins, we adapted a methacrylate embedding method for the localization of proteins inside nematode tissues, and extracellularly when secreted in planta or within plant cells. An important advantage is that the method is applicable for all nematode stages: preparasitic as well as parasitic stages, including large mature females. Herein, the method has been successfully applied for the localization of four nematode secreted proteins, such as Mi-MAP-1, Mi-CBM2-bearing proteins, Mi-PEL3, and Mi-6D4. In addition, we could also localize 14-3-3 proteins, as well as two cytoskeletal proteins, by double-immunolabeling on preparasitic juveniles. Superior preservation of nematode and plant morphology, allowed more accurate protein localization as compared with other methods. Besides excellent epitope preservation, dissolution of methacrylate from tissue sections unmasks target proteins and thereby drastically increases antibody access.

  18. Species and tissue type regulate long-term decomposition of brackish marsh plants grown under elevated CO2 conditions

    Science.gov (United States)

    Jones, Joshua A; Cherry, Julia A; Mckee, Karen L.

    2016-01-01

    Organic matter accumulation, the net effect of plant production and decomposition, contributes to vertical soil accretion in coastal wetlands, thereby playing a key role in whether they keep pace with sea-level rise. Any factor that affects decomposition may affect wetland accretion, including atmospheric CO2 concentrations. Higher CO2 can influence decomposition rates by altering plant tissue chemistry or by causing shifts in plant species composition or biomass partitioning. A combined greenhouse-field experiment examined how elevated CO2 affected plant tissue chemistry and subsequent decomposition of above- and belowground tissues of two common brackish marsh species, Schoenoplectus americanus (C3) and Spartina patens (C4). Both species were grown in monoculture and in mixture under ambient (350-385 μL L-1) or elevated (ambient + 300 μL L-1) atmospheric CO2 conditions, with all other growth conditions held constant, for one growing season. Above- and belowground tissues produced under these treatments were decomposed under ambient field conditions in a brackish marsh in the Mississippi River Delta, USA. Elevated CO2 significantly reduced nitrogen content of S. americanus, but not sufficiently to affect subsequent decomposition. Instead, long-term decomposition (percent mass remaining after 280 d) was controlled by species composition and tissue type. Shoots of S. patens had more mass remaining (41 ± 2%) than those of S. americanus (12 ± 2 %). Belowground material decomposed more slowly than that placed aboveground (62 ± 1% vs. 23 ± 3% mass remaining), but rates belowground did not differ between species. Increases in atmospheric CO2concentration will likely have a greater effect on overall decomposition in this brackish marsh community through shifts in species dominance or biomass allocation than through effects on tissue chemistry. Consequent changes in organic matter accumulation may alter marsh capacity to accommodate sea-level rise

  19. Three-dimensional reconstruction of frozen and thawed plant tissues from microscopic images

    Science.gov (United States)

    Histological analysis of frozen and thawed plants has been conducted for many years but the observation of individual sections only provides a 2 dimensional representation of a 3 dimensional phenomenon. Most techniques for viewing internal plant structure in 3 dimensions is either low in resolution...

  20. ANTIOXIDANT ACTIVITY OF TISSUE CULTURE-RAISED BALLOTA NIGRA L. PLANTS GROWN EX VITRO.

    Science.gov (United States)

    Makowczyńska, Joanna; Grzegorczyk-KAROLAK, Izabela; Wysokińska, Halina

    2015-01-01

    Antioxidant properties and total phenolic and flavonoid contents were evaluated in methanolic extracts of shoots from Ballota nigra plants initiated in vitro (from nodal explants) and in vivo (from seeds). The plants were grown in greenhouse and in the field, and were analyzed at the vegetative and flowering stages. The shoot extract of wild-grown plants of B. nigra was also investigated. The results indicate that antioxidant potential of the B. nigra extracts seems to be due to their scavenging of free radicals (DPPH assay) and metal reducing (FRAP test), while they were less effective at the prevention of linoleic acid peroxidation (LPO test). The extracts from shoots of in vitro derived plants were found to exhibit the greatest antioxidant properties. The extracts were also characterized by the highest content of phenolic compounds and their level was affected by plant developmental stage. The extracts of shoots collected at the flowering period exhibited higher amounts of phenolics and flavonoids than in the extracts of immature plants. A close correlation between the total phenolic content and flavonoid content and antioxidant activity using the DPPH and FRAP assays was obtained. The results of the present study suggest the use in vitro-derived plants of B. nigra instead of using wild plants for pharmaceutical purposes.

  1. Microfluidic monitoring of programmed cell death in living plant seed tissue

    DEFF Research Database (Denmark)

    Mark, Christina; Heiskanen, Arto; Zor, Kinga

    Programmed cell death (PCD) is a highly regulated process in which cells are dismantled. Reactive oxygen species (ROS) are involved in PCD in plants, but the relationship between and mechanisms behind ROS and PCD are only poorly understood in plant cells compared to in animal cells (Gechev, Tsanko...

  2. Microfluidic monitoring of programmed cell death in living plant seed tissue

    DEFF Research Database (Denmark)

    Mark, Christina; Heiskanen, Arto; Zor, Kinga

    Programmed cell death (PCD) is a highly regulated process in which cells are dismantled. Reactive oxygen species (ROS) are involved in PCD in plants, but the relationship between and mechanisms behind ROS and PCD are only poorly understood in plant cells compared to in animal cells (Gechev, Tsank...

  3. Short Communication An efficient method for simultaneous extraction of high-quality RNA and DNA from various plant tissues.

    Science.gov (United States)

    Oliveira, R R; Viana, A J C; Reátegui, A C E; Vincentz, M G A

    2015-12-29

    Determination of gene expression is an important tool to study biological processes and relies on the quality of the extracted RNA. Changes in gene expression profiles may be directly related to mutations in regulatory DNA sequences or alterations in DNA cytosine methylation, which is an epigenetic mark. Correlation of gene expression with DNA sequence or epigenetic mark polymorphism is often desirable; for this, a robust protocol to isolate high-quality RNA and DNA simultaneously from the same sample is required. Although commercial kits and protocols are available, they are mainly optimized for animal tissues and, in general, restricted to RNA or DNA extraction, not both. In the present study, we describe an efficient and accessible method to extract both RNA and DNA simultaneously from the same sample of various plant tissues, using small amounts of starting material. The protocol was efficient in the extraction of high-quality nucleic acids from several Arabidopsis thaliana tissues (e.g., leaf, inflorescence stem, flower, fruit, cotyledon, seedlings, root, and embryo) and from other tissues of non-model plants, such as Avicennia schaueriana (Acanthaceae), Theobroma cacao (Malvaceae), Paspalum notatum (Poaceae), and Sorghum bicolor (Poaceae). The obtained nucleic acids were used as templates for downstream analyses, such as mRNA sequencing, quantitative real time-polymerase chain reaction, bisulfite treatment, and others; the results were comparable to those obtained with commercial kits. We believe that this protocol could be applied to a broad range of plant species, help avoid technical and sampling biases, and facilitate several RNA- and DNA-dependent analyses.

  4. A Review of Mid-Infrared and Near-Infrared Imaging: Principles, Concepts and Applications in Plant Tissue Analysis

    Directory of Open Access Journals (Sweden)

    Sevgi Türker-Kaya

    2017-01-01

    Full Text Available Plant cells, tissues and organs are composed of various biomolecules arranged as structurally diverse units, which represent heterogeneity at microscopic levels. Molecular knowledge about those constituents with their localization in such complexity is very crucial for both basic and applied plant sciences. In this context, infrared imaging techniques have advantages over conventional methods to investigate heterogeneous plant structures in providing quantitative and qualitative analyses with spatial distribution of the components. Thus, particularly, with the use of proper analytical approaches and sampling methods, these technologies offer significant information for the studies on plant classification, physiology, ecology, genetics, pathology and other related disciplines. This review aims to present a general perspective about near-infrared and mid-infrared imaging/microspectroscopy in plant research. It is addressed to compare potentialities of these methodologies with their advantages and limitations. With regard to the organization of the document, the first section will introduce the respective underlying principles followed by instrumentation, sampling techniques, sample preparations, measurement, and an overview of spectral pre-processing and multivariate analysis. The last section will review selected applications in the literature.

  5. A Review of Mid-Infrared and Near-Infrared Imaging: Principles, Concepts and Applications in Plant Tissue Analysis.

    Science.gov (United States)

    Türker-Kaya, Sevgi; Huck, Christian W

    2017-01-20

    Plant cells, tissues and organs are composed of various biomolecules arranged as structurally diverse units, which represent heterogeneity at microscopic levels. Molecular knowledge about those constituents with their localization in such complexity is very crucial for both basic and applied plant sciences. In this context, infrared imaging techniques have advantages over conventional methods to investigate heterogeneous plant structures in providing quantitative and qualitative analyses with spatial distribution of the components. Thus, particularly, with the use of proper analytical approaches and sampling methods, these technologies offer significant information for the studies on plant classification, physiology, ecology, genetics, pathology and other related disciplines. This review aims to present a general perspective about near-infrared and mid-infrared imaging/microspectroscopy in plant research. It is addressed to compare potentialities of these methodologies with their advantages and limitations. With regard to the organization of the document, the first section will introduce the respective underlying principles followed by instrumentation, sampling techniques, sample preparations, measurement, and an overview of spectral pre-processing and multivariate analysis. The last section will review selected applications in the literature.

  6. Regeneration of plants from callus tissues of hairy roots induced by Agrobacterium rhizogenes on Alhagi pseudoalhagi

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    The legume forage Alhagi pseudoalhagi was transformed by the Agrobacterium rhizogenes strain A4 using cotyledon and hypocotyl segments as infection materials. Regenerated plants were achieved from sterile calli derived from hairy roots, which occurred at or near the infection sites. The regenerated plants from hairy root were characterized by normal leaf morphology and stem growth but a shallow and more extensive root system than normal plants. Opine synthesis, PCR and Southern blot confirmed that TDNA had been integrated into the A. pseudoalhagi genome. Acetosyringone (AS) was found to be vital for successful transformation of A. pseudoalhagi.

  7. Using Bulk Magnetic Susceptibility to Resolve Internal and External Signals in the NMR Spectra of Plant Tissues

    Science.gov (United States)

    Shachar-Hill, Yair; Befroy, Douglas E.; Pfeffer, Philip E.; Ratcliffe, R. George

    1997-07-01

    Internal and external NMR signals from a variety of plant cells and plant tissues can be resolved by changing the bulk magnetic susceptibility (BMS) of the perfusing medium with [Gd (EDTA)]-or Dy(DTPA-BMA). This separation is observed in samples consisting of cylindrical cells oriented along theB0field, and is consistent with established theoretical predictions about BMS effects. Evidence is presented that the shifted signals represent material outside the tissue as well as some contribution from intercellular spaces and cell walls, while intracellular signals are unshifted. The paramagnetic complexes used to separate the signals are shown to be nontoxic and to have no effect on a number of transport processes. The method has been applied to roots, shoots, and giant algal cells, facilitating the interpretation of thein vivospectra from a range of biologically important magnetic isotopes. The potential of the method for studies of transport is illustrated with experiments showing: (i)14N/15N isotopic exchange of nitrate in roots; (ii) the influx of HDO into root and shoot segments; and (iii) the use of saturation transfer to follow water movement into and out of plant cells.

  8. Tissue culture and generation of autotetraploid plants of Sophora flavescens Aiton

    Directory of Open Access Journals (Sweden)

    Wei Kun-Hua

    2010-01-01

    Full Text Available Background: Sophora flavescens Aiton is an important medicinal plant in China. Early in vitro researches of S. flavescens were focused on callus induction and cell suspension culture, only a few were concerned with in vitro multiplication. Objective: To establish and optimize the rapid propagation technology of S. flavescens and to generate and characterize polyploid plants of S. flavescens. Materials and Methods: The different concentrations of 6-benzylaminopurine (BAP, indole-3-acetic acid (IAA and kinetin (KT were used to establish and screen the optimal rapid propagation technology of S. flavescens by orthogonal test; 0.2% colchicine solution was used to induce polyploid plants and the induced buds were identified by root-tip chromosome determination and stomatal apparatus observation. Results: A large number of buds could be induced directly from epicotyl and hypocotyl explants on the Murashige and Skoog medium (MS; 1962 supplemented with 1.4-1.6 mg/l 6-benzylaminopurine (BAP and 0.3 mg/l indole-3-acetic acid (IAA. More than 50 lines of autotetraploid plants were obtained. The chromosome number of the autotetraploid plantlet was 2n = 4x = 36. All tetraploid plants showed typical polyploid characteristics. Conclusion: Obtained autotetraploid lines will be of important genetic and breeding value and can be used for further selection and plant breeding.

  9. Multi-instrumental Analysis of Tissues of Sunflower Plants Treated with Silver(I) Ions – Plants as Bioindicators of Environmental Pollution

    Science.gov (United States)

    Krizkova, Sona; Ryant, Pavel; Krystofova, Olga; Adam, Vojtech; Galiova, Michaela; Beklova, Miroslava; Babula, Petr; Kaiser, Jozef; Novotny, Karel; Novotny, Jan; Liska, Miroslav; Malina, Radomir; Zehnalek, Josef; Hubalek, Jaromir; Havel, Ladislav; Kizek, Rene

    2008-01-01

    The aim of this work is to investigate sunflower plants response on stress induced by silver(I) ions. The sunflower plants were exposed to silver(I) ions (0, 0.1, 0.5, and 1 mM) for 96 h. Primarily we aimed our attention to observation of basic physiological parameters. We found that the treated plants embodied growth depression, coloured changes and lack root hairs. Using of autofluorescence of anatomical structures, such as lignified cell walls, it was possible to determine the changes of important shoot and root structures, mainly vascular bungles and development of secondary thickening. The differences in vascular bundles organisation, parenchymatic pith development in the root centre and the reduction of phloem part of vascular bundles were well observable. Moreover with increasing silver(I) ions concentration the vitality of rhizodermal cells declined; rhizodermal cells early necrosed and were replaced by the cells of exodermis. Further we employed laser induced breakdown spectroscopy for determination of spatial distribution of silver(I) ions in tissues of the treated plants. The Ag is accumulated mainly in near-root part of the sample. Moreover basic biochemical indicators of environmental stress were investigated. The total content of proteins expressively decreased with increasing silver(I) ions dose and the time of the treatment. As we compare the results obtained by protein analysis – the total protein contents in shoot as well as root parts – we can assume on the transport of the proteins from the roots to shoots. This phenomenon can be related with the cascade of processes connecting with photosynthesis. The second biochemical parameter, which we investigated, was urease activity. If we compared the activity in treated plants with control, we found out that presence of silver(I) ions markedly enhanced the activity of urease at all applied doses of this toxic metal. Finally we studied the effect of silver(I) ions on activity of urease in in vitro

  10. Multi-instrumental Analysis of Tissues of Sunflower Plants Treated with Silver(I Ions – Plants as Bioindicators of Environmental Pollution

    Directory of Open Access Journals (Sweden)

    Rene Kizek

    2007-01-01

    Full Text Available The aim of this work is to investigate sunflower plants response on stressinduced by silver(I ions. The sunflower plants were exposed to silver(I ions (0, 0.1, 0.5,and 1 mM for 96 h. Primarily we aimed our attention to observation of basic physiologicalparameters. We found that the treated plants embodied growth depression, coloured changes and lack root hairs. Using of autofluorescence of anatomical structures, such aslignified cell walls, it was possible to determine the changes of important shoot and rootstructures, mainly vascular bungles and development of secondary thickening. Thedifferences in vascular bundles organisation, parenchymatic pith development in the rootcentre and the reduction of phloem part of vascular bundles were well observable.Moreover with increasing silver(I ions concentration the vitality of rhizodermal cellsdeclined; rhizodermal cells early necrosed and were replaced by the cells of exodermis.Further we employed laser induced breakdown spectroscopy for determination of spatialdistribution of silver(I ions in tissues of the treated plants. The Ag is accumulated mainlyin near-root part of the sample. Moreover basic biochemical indicators of environmentalstress were investigated. The total content of proteins expressively decreased withincreasing silver(I ions dose and the time of the treatment. As we compare the resultsobtained by protein analysis – the total protein contents in shoot as well as root parts – wecan assume on the transport of the proteins from the roots to shoots. This phenomenon canbe related with the cascade of processes connecting with photosynthesis. The secondbiochemical parameter, which we investigated, was urease activity. If we compared theactivity in treated plants with control, we found out that presence of silver(I ions markedlyenhanced the activity of urease at all applied doses of this toxic metal. Finally we studiedthe effect of silver(I ions on activity of urease

  11. Levels of organochlorine pesticides in soils and rye plant tissues in a field study.

    Science.gov (United States)

    Waliszewski, Stefan M; Carvajal, Octavio; Infanzon, Rosa M; Trujillo, Patricia; Aguirre, Angel A; Maxwell, Mary

    2004-11-17

    The organochlorine pesticides are lipophilic and persistent and tend to accumulate in soils and growing plants. The contamination of growing plants occurs by adhesion of volatile substances from the air to the plant surface and by the migration of contaminants through xylem in inner ascendant transport. Persistent organochlorine pesticides (HCB, alpha,gamma-HCH, pp'DDE, op'DDT, pp'DDT) levels were determined in soils and rye plants. The aims of the study were the monitoring of organochlorine pesticide concentrations and the comparison of these levels among soil, rye straw, and rye grains. Fifty soil samples and 50 rye plant (50 straw and 50 grains) samples were taken. The GLC-ECD chromatographic results indicated the following contamination levels distributed among soil, straw, and grains: HCB (0.7-1.2-0.7 microg.kg(-1)), alpha-HCH (0.6-3.4-1.2 microg.kg(-1)), gamma-HCH (1.8-27.3-4.4 microg.kg(-1)), Sigma-HCH (2.5-30.7-5.6 microg.kg(-1)), pp'DDE (1.0-7.8-5.5 microg.kg(-1)), op'DDT (16.1-20.4-17.0 microg.kg(-1)), pp'DDT (38.0-41.7-49.6 microg.kg(-1)), and Sigma-DDT (54.2-63.2-72.1 microg.kg(-1)). The study verified the presence of organochlorine pesticides in the Mexican agricultural environment and their migration from soil to the growing rye plants. However, DDT has been banned since 1999 for sanitary reasons, and Lindane is applied only in some cases as a seed dresser. The determined organochlorine pesticide levels in rye plants are low, at residual levels that are below Codex Alimentarius Commission maximum residue limits.

  12. Neuroprotection and enhanced neurogenesis by extract from the tropical plant Knema laurina after inflammatory damage in living brain tissue.

    Science.gov (United States)

    Häke, Ines; Schönenberger, Silvia; Neumann, Jens; Franke, Katrin; Paulsen-Merker, Katrin; Reymann, Klaus; Ismail, Ghazally; Bin Din, Laily; Said, Ikram M; Latiff, A; Wessjohann, Ludger; Zipp, Frauke; Ullrich, Oliver

    2009-01-03

    Inflammatory reactions in the CNS, resulting from a loss of control and involving a network of non-neuronal and neuronal cells, are major contributors to the onset and progress of several major neurodegenerative diseases. Therapeutic strategies should therefore keep or restore the well-controlled and finely-tuned balance of immune reactions, and protect neurons from inflammatory damage. In our study, we selected plants of the Malaysian rain forest by an ethnobotanic survey, and investigated them in cell-based-assay-systems and in living brain tissue cultures in order to identify anti-inflammatory and neuroprotective effects. We found that alcoholic extracts from the tropical plant Knema laurina (Black wild nutmeg) exhibited highly anti-inflammatory and neuroprotective effects in cell culture experiments, reduced NO- and IL-6-release from activated microglia cells dose-dependently, and protected living brain tissue from microglia-mediated inflammatory damage at a concentration of 30 microg/ml. On the intracellular level, the extract inhibited ERK-1/2-phosphorylation, IkB-phosphorylation and subsequently NF-kB-translocation in microglia cells. K. laurina belongs to the family of Myristicaceae, which have been used for centuries for treatment of digestive and inflammatory diseases and is also a major food plant of the Giant Hornbill. Moreover, extract from K. laurina promotes also neurogenesis in living brain tissue after oxygen-glucose deprivation. In conclusion, extract from K. laurina not only controls and limits inflammatory reaction after primary neuronal damage, it promotes moreover neurogenesis if given hours until days after stroke-like injury.

  13. The signature of seeds in resurrection plants: a molecular and physiological comparison of desiccation tolerance in seeds and vegetative tissues.

    Science.gov (United States)

    Illing, Nicola; Denby, Katherine J; Collett, Helen; Shen, Arthur; Farrant, Jill M

    2005-11-01

    Desiccation-tolerance in vegetative tissues of angiosperms has a polyphyletic origin and could be due to 1) appropriation of the seed-specific program of gene expression that protects orthodox seeds against desiccation, and/or 2) a sustainable version of the abiotic stress response. We tested these hypotheses by comparing molecular and physiological data from the development of orthodox seeds, the response of desiccation-sensitive plants to abiotic stress, and the response of desiccation-tolerant plants to extreme water loss. Analysis of publicly-available gene expression data of 35 LEA proteins and 68 anti-oxidant enzymes in the desiccation-sensitive Arabidopsis thaliana identified 13 LEAs and 4 anti-oxidants exclusively expressed in seeds. Two (a LEA6 and 1-cys-peroxiredoxin) are not expressed in vegetative tissues in A. thaliana, but have orthologues that are specifically activated in desiccating leaves of Xerophyta humilis. A comparison of antioxidant enzyme activity in two desiccation-sensitive species of Eragrostis with the desiccation-tolerant E. nindensis showed equivalent responses upon initial dehydration, but activity was retained at low water content in E. nindensis only. We propose that these antioxidants are housekeeping enzymes and that they are protected from damage in the desiccation-tolerant species. Sucrose is considered an important protectant against desiccation in orthodox seeds, and we show that sucrose accumulates in drying leaves of E. nindensis, but not in the desiccation-sensitive Eragrostis species. The activation of "seed-specific" desiccation protection mechanisms (sucrose accumulation and expression of LEA6 and 1-cys-peroxiredoxin genes) in the vegetative tissues of desiccation-tolerant plants points towards acquisition of desiccation tolerance from seeds.

  14. Aplicações da cultura de tecidos em plantas medicinais Applications of tissue culture in medicinal plants

    Directory of Open Access Journals (Sweden)

    T.P. Morais

    2012-01-01

    Full Text Available Esta revisão tem por objetivo levantar dados de literatura sobre o histórico e a situação atual das técnicas de cultura de tecidos em plantas medicinais. Para tanto, foi realizada uma revisão de publicações do período de 1976 a 2009. A cultura de tecidos é muito utilizada em pesquisas envolvendo plantas medicinais, com destaque para a técnica de micropropagação. A aplicação das técnicas de cultura de tecidos em plantas medicinais tem como perspectivas a obtenção de germoplasma competitivo e adaptado a diversos métodos de cultivo, escolha de novas espécies que servirão como fonte de compostos biologicamente ativos e aprimoramento da produção de fitofármacos, a fim de assegurar exploração sustentável destas espécies.The aim of this literature review is to conduct a survey concerning the history and current situation of tissue culture techniques in medicinal plants. Therefore, a review was done considering the period from 1976 to 2009. Tissue culture is widely applied in medicinal plants researches, especially micropropagation. The perspectives of tissue culture techniques in medicinal plants are related to the development of competitive germoplasm adapted to diverse methods of cultivation, the election of new species that will serve as source of biological active composts, and the improvement of phytochemicals production, in order to assure sustainable exploration of these species.

  15. Agrobacterium tumefaciens-mediated transformation of embryogenic tissue and transgenic plant regeneration in Chamaecyparis obtusa Sieb. et Zucc.

    Science.gov (United States)

    Taniguchi, T; Kurita, M; Ohmiya, Y; Kondo, T

    2005-03-01

    A genetic transformation procedure for Chamaecyparis obtusa was developed after co-cultivation of embryogenic tissues with disarmed Agrobacterium tumefaciens strain C58/pMP90, which harbours the sgfp (synthetic green fluorescent protein) visual reporter and nptII (neomycin phoshotransferase II) selectable marker genes. The highest transformation frequency was 22.5 independent transformed lines per dish (250 mg embryogenic tissue) following selection on kanamycin medium. Transgenic plantlets were regenerated through the maturation and germination of somatic embryos. The intensity of GFP fluorescence, observed under a fluorescence microscope, varied from very faint to relatively strong, depending on the transgenic line or part of the transgenic plant. The integration of the genes into the genome of regenerated plantlets was confirmed by Southern blot analysis.

  16. Tissue-specific expression and post-translational modifications of plant- and bacterial-type phosphoenolpyruvate carboxylase isozymes of the castor oil plant, Ricinus communis L.

    Science.gov (United States)

    O'Leary, Brendan; Fedosejevs, Eric T; Hill, Allyson T; Bettridge, James; Park, Joonho; Rao, Srinath K; Leach, Craig A; Plaxton, William C

    2011-11-01

    This study employs transcript profiling together with immunoblotting and co-immunopurification to assess the tissue-specific expression, protein:protein interactions, and post-translational modifications (PTMs) of plant- and bacterial-type phosphoenolpyruvate carboxylase (PEPC) isozymes (PTPC and BTPC, respectively) in the castor plant, Ricinus communis. Previous studies established that the Class-1 PEPC (PTPC homotetramer) of castor oil seeds (COS) is activated by phosphorylation at Ser-11 and inhibited by monoubiquitination at Lys-628 during endosperm development and germination, respectively. Elimination of photosynthate supply to developing COS by depodding caused the PTPC of the endosperm and cotyledon to be dephosphorylated, and then subsequently monoubiquitinated in vivo. PTPC monoubiquitination rather than phosphorylation is widespread throughout the castor plant and appears to be the predominant PTM of Class-1 PEPC that occurs in planta. The distinctive developmental patterns of PTPC phosphorylation versus monoubiquitination indicates that these two PTMs are mutually exclusive. By contrast, the BTPC: (i) is abundant in the inner integument, cotyledon, and endosperm of developing COS, but occurs at low levels in roots and cotyledons of germinated COS, (ii) shows a unique developmental pattern in leaves such that it is present in leaf buds and young expanding leaves, but undetectable in fully expanded leaves, and (iii) tightly interacts with co-expressed PTPC to form the novel and allosterically-desensitized Class-2 PEPC heteromeric complex. BTPC and thus Class-2 PEPC up-regulation appears to be a distinctive feature of rapidly growing and/or biosynthetically active tissues that require a large anaplerotic flux from phosphoenolpyruvate to replenish tricarboxylic acid cycle C-skeletons being withdrawn for anabolism.

  17. Growth of plant tissue cultures in simulated lunar soil: Implications for a lunar base Controlled Ecological Life Support System (CELSS)

    Science.gov (United States)

    Venketeswaran, S.

    1987-01-01

    Experiments to determine whether plant tissue cultures can be grown in the presence of simulated lunar soil (SLS) and the effect of simulated lunar soil on the growth and morphogenesis of such cultures, as well as the effect upon the germination of seeds and the development of seedlings were carried out . Preliminary results on seed germination and seedling growth of rice and calli growth of winged bean and soybean indicate that there is no toxicity or inhibition caused by SLS. SLS can be used as a support medium with supplements of certain major and micro elements.

  18. Effect of Nitrogen and Phosphorus on Tissue Nutrition and Biomass of Freshwater Wetland Plant in Sanjiang Plain, Northeast China

    Institute of Scientific and Technical Information of China (English)

    XU Zhiguo; YAN Baixing; HE Yan; ZHAI Jinliang; SONG Changchun

    2006-01-01

    Nitrogen (4, 10, 20 and 40g/m2) and phosphorus (1.2, 4.8 and 9.6g/m2) were applied to tanks to evaluating the effects of N and P additions on plant tissue nutrition and the biomass of two freshwater wetland plants in the Sanjiang Plain of Northeast China, namely Carex lasiocarpa and Carex meyeriana. For C. lasiocarpa, the total N concentration (TN) of plant tissues under the treatment of 10g/m2 was lower compared with the other N treatments. Initially, C.lasiocarpa exhibited a significant increase of biomass as compared with the control value, reaching the maximum of 31.20±4.01 g/tank under the treatment of 10g/m2, and then dropped to 18.02±1.53g/tank under the treatment of 40g/m2.For C. meyeriana, TN generally increased with increasing amount of N applied. High N applied produced more aboveground biomass than low N applied. C. meyeriana, as the accompanying species, can adapt itself to the wetland enriched by N, and it may replace C. lasiocarpa as the dominant species of wetland. The total P concentration (TP) in tissues of C. lasiocarpa increased with P addition. The aboveground biomass of C. lasiocarpa increased with P addition,and it changed from 18.77±3.29g/tank to 46.03±3.95g/tank. However, TP of tissue may accelerate the C. meyeriana development under the treatment of 1.2g/m2. P accumulation contributes to the dominance of C. lasiocarpa but limits the production of C. meyeriana, and the latter may disappear gradually from the wetland enriched by P. Increased input of N and P might have an influence on wetland plant community composition and structure, so the effects of nutrient inputs and accumulation should be considered to protect the freshwater wetland.

  19. Phylogenic diversity and tissue specificity of fungal endophytes associated with the pharmaceutical plant, Stellera chamaejasme L. revealed by a cultivation-independent approach.

    Science.gov (United States)

    Jin, Hui; Yang, Xiaoyan; Lu, Dengxue; Li, Chunjie; Yan, Zhiqiang; Li, Xiuzhuang; Zeng, Liming; Qin, Bo

    2015-10-01

    The fungal endophytes associated with medicinal plants have been demonstrated as a reservoir with novel natural products useful in medicine and agriculture. It is desirable to explore the species composition, diversity and tissue specificity of endophytic fungi that inhabit in different tissues of medicinal plants. In this study, a culture-independent survey of fungal diversity in the rhizosphere, leaves, stems and roots of a toxic medicinal plant, Stellera chamaejasme L., was conducted by sequence analysis of clone libraries of the partial internal transcribed spacer region. Altogether, 145 fungal OTUs (operational taxonomic units), represented by 464 sequences, were found in four samples, of these 109 OTUs (75.2 %) belonging to Ascomycota, 20 (13.8 %) to Basidiomycota, 14 (9.7 %) to Zygomycota, 1 (0.7 %) to Chytridiomycota, and 1 (0.7 %) to Glomeromycota. The richness and diversity of fungal communities were strongly influenced by plant tissue environments, and the roots are associated with a surprisingly rich endophyte community. The endophyte assemblages associated with S. chamaejasme were strongly shaped by plant tissue environments, and exhibited a certain degree of tissue specificity. Our results suggested that a wide variety of fungal assemblages inhabit in S. chamaejasme, and plant tissue environments conspicuously influence endophyte community structure.

  20. An Effective System to Produce Smoke Solutions from Dried Plant Tissue for Seed Germination Studies

    OpenAIRE

    Janice Coons; Nancy Coutant; Barbara Lawrence; Daniel Finn; Stephanie Finn

    2014-01-01

    • Premise of the study: An efficient and inexpensive system was developed to produce smoke solutions from plant material to research the influence of water-soluble compounds from smoke on seed germination. • Methods and Results: Smoke solutions (300 mL per batch) were produced by burning small quantities (100–200 g) of dried plant material from a range of species in a bee smoker attached by a heater hose to a side-arm flask. The flask was attached to a vacuum water aspirator, to pull the smok...

  1. Phenotype analysis of Russian dandelion root tissues from the national plant germplasm system collection

    Science.gov (United States)

    Russian dandelion (Taraxacum kok-saghyz) (TKS) produces high quality natural rubber (NR), cis-1,4 polyisoprene, by biosynthesis, and has been used historically as a source of NR during times of short supply or high prices for Hevea NR. The rubber is primarily located in root tissues along with appre...

  2. Mercury Concentrations in Plant Tissues as Affected by FGDG Application to Soil

    Science.gov (United States)

    Flue Gas Desulfurization Gypsum (FGDG) is produced by reducing sulfur dioxide emissions from themo-electric coal-fired power plants. The most common practice of FGDG production may trap some of the Mercury (Hg) present in the coal that normally would escape as vapor in the stack gases. Concern for t...

  3. Botrytis cinerea endopolygalacturonase genes are differentially expressed in various plant tissues

    NARCIS (Netherlands)

    Have, ten A.; Oude Breuil, W.; Wubben, J.P.; Visser, J.; Kan, van J.A.L.

    2001-01-01

    Botrytis cinerea, the causal agent of blight, rot, and gray mold on many plant species, secretes various endopolygalacturonases during all stages of infection. The expression pattern of the encoding genes (Bcpg 1-6) was studied on four hosts: tomato, broad bean, apple, and courgette (also known as z

  4. Development of rapid isothermal amplification assays for Phytophthora species from plant tissue

    Science.gov (United States)

    Several isothermal amplification techniques recently have been developed that are tolerant of inhibitors present in many plant extracts, which can reduce the need for obtaining purified DNA for running diagnostic assays. One such commercially available technique that has similarities with real time ...

  5. Botrytis cinerea endopolygalacturonase genes are differentially expressed in various plant tissues

    NARCIS (Netherlands)

    Have, ten A.; Oude Breuil, W.; Wubben, J.P.; Visser, J.; Kan, van J.A.L.

    2001-01-01

    Botrytis cinerea, the causal agent of blight, rot, and gray mold on many plant species, secretes various endopolygalacturonases during all stages of infection. The expression pattern of the encoding genes (Bcpg 1-6) was studied on four hosts: tomato, broad bean, apple, and courgette (also known as z

  6. Monitoring programmed cell death of living plant tissues in microfluidics using electrochemical and optical techniques

    DEFF Research Database (Denmark)

    Mark, Christina; Heiskanen, Arto; Svensson, Birte

    Programmed cell death (PCD) in plants can influence the outcome of yield and quality of crops through its important role in seed germination and the defence process against pathogens. The main scope of the project is to apply microfluidic cell culture for the measurement of electrochemically or o...

  7. Extraction and analysis of inositols and other carbohydrates from soybean plant tissues

    Science.gov (United States)

    An outstanding characteristic of soybean plants is their ability to produce large amounts of the carbohydrate pinitol. Pinitol and the closely related inositols are currently undergoing widespread investigation for their biological and nutritional value. These and all the carbohydrates are typical...

  8. Proton-dependent coniferin transport, a common major transport event in differentiating xylem tissue of woody plants.

    Science.gov (United States)

    Tsuyama, Taku; Kawai, Ryo; Shitan, Nobukazu; Matoh, Toru; Sugiyama, Junji; Yoshinaga, Arata; Takabe, Keiji; Fujita, Minoru; Yazaki, Kazufumi

    2013-06-01

    Lignin biosynthesis is an essential physiological activity of vascular plants if they are to survive under various environmental stresses on land. The biosynthesis of lignin proceeds in the cell wall by polymerization of precursors; the initial step of lignin polymerization is the transportation of lignin monomers from the cytosol to the cell wall, which is critical for lignin formation. There has been much debate on the transported form of the lignin precursor, either as free monolignols or their glucosides. In this study, we performed biochemical analyses to characterize the membrane transport mechanism of lignin precursors using angiosperms, hybrid poplar (Populus sieboldii × Populus grandidentata) and poplar (Populus sieboldii), as well gymnosperms, Japanese cypress (Chamaecyparis obtusa) and pine (Pinus densiflora). Membrane vesicles prepared from differentiating xylem tissues showed clear ATP-dependent transport activity of coniferin, whereas less than 4% of the coniferin transport activity was seen for coniferyl alcohol. Bafilomycin A1 and proton gradient erasers markedly inhibited coniferin transport in hybrid poplar membrane vesicles; in contrast, vanadate had no effect. Cis-inhibition experiments suggested that this transport activity was specific for coniferin. Membrane fractionation of hybrid poplar microsomes demonstrated that transport activity was localized to the tonoplast- and endomembrane-rich fraction. Differentiating xylem of Japanese cypress exhibited almost identical transport properties, suggesting the involvement of a common endomembrane-associated proton/coniferin antiport mechanism in the lignifying tissues of woody plants, both angiosperms and gymnosperms.

  9. Protein Synthesis Inhibition Activity by Strawberry Tissue Protein Extracts during Plant Life Cycle and under Biotic and Abiotic Stresses

    Directory of Open Access Journals (Sweden)

    Walther Faedi

    2013-07-01

    Full Text Available Ribosome-inactivating proteins (RIPs, enzymes that are widely distributed in the plant kingdom, inhibit protein synthesis by depurinating rRNA and many other polynucleotidic substrates. Although RIPs show antiviral, antifungal, and insecticidal activities, their biological and physiological roles are not completely understood. Additionally, it has been described that RIP expression is augmented under stressful conditions. In this study, we evaluated protein synthesis inhibition activity in partially purified basic proteins (hereafter referred to as RIP activity from tissue extracts of Fragaria × ananassa (strawberry cultivars with low (Dora and high (Record tolerance to root pathogens and fructification stress. Association between the presence of RIP activity and the crop management (organic or integrated soil, growth stage (quiescence, flowering, and fructification, and exogenous stress (drought were investigated. RIP activity was found in every tissue tested (roots, rhizomes, leaves, buds, flowers, and fruits and under each tested condition. However, significant differences in RIP distribution were observed depending on the soil and growth stage, and an increase in RIP activity was found in the leaves of drought-stressed plants. These results suggest that RIP expression and activity could represent a response mechanism against biotic and abiotic stresses and could be a useful tool in selecting stress-resistant strawberry genotypes.

  10. Production of 2-hydroxy-4-methoxybenzaldehyde in roots of tissue culture raised and acclimatized plants of Decalepis hamiltonii Wight & Arn., an endangered shrub endemic to Southern India and evaluation of its performance vis-a-vis plants from natural habitat.

    Science.gov (United States)

    Giridhar, P; Rajasekaran, T; Nagarajan, S; Ravishankar, G A

    2004-01-01

    Axillary buds obtained from field grown plants of D. hamiltonii were used to initiate multiple shoots on Murashige and Skoog's medium (MS) supplemented with 2 mg L(-1) 6-benzyl aminopurine (BA) and 0.5 mg L(-1) indole-3-acetic acid (IAA). Profuse rooting was achieved when the actively growing shoots were cultured on MS medium supplemented with 1.0 mg l(-1) indole-3-butyric acid (IBA). Regenerated plants were grown successfully in the plains, in contrast to wild growth in high altitudes and rocky crevices of hilly regions. Roots of different sizes from one-year-old tissue culture raised field grown plants had the same profile of 2-hydroxy-4-methoxybenzaldehyde as that of wild plants. A maximum of 0.14% and 0.12% 2-hydroxy-4-methoxybenzaldehyde was produced in roots of one year old tissue culture derived plants and greenhouse grown plants respectively.

  11. Electron tomography of cryo-immobilized plant tissue: a novel approach to studying 3D macromolecular architecture of mature plant cell walls in situ.

    Science.gov (United States)

    Sarkar, Purbasha; Bosneaga, Elena; Yap, Edgar G; Das, Jyotirmoy; Tsai, Wen-Ting; Cabal, Angelo; Neuhaus, Erica; Maji, Dolonchampa; Kumar, Shailabh; Joo, Michael; Yakovlev, Sergey; Csencsits, Roseann; Yu, Zeyun; Bajaj, Chandrajit; Downing, Kenneth H; Auer, Manfred

    2014-01-01

    Cost-effective production of lignocellulosic biofuel requires efficient breakdown of cell walls present in plant biomass to retrieve the wall polysaccharides for fermentation. In-depth knowledge of plant cell wall composition is therefore essential for improving the fuel production process. The precise spatial three-dimensional (3D) organization of cellulose, hemicellulose, pectin and lignin within plant cell walls remains unclear to date since the microscopy techniques used so far have been limited to two-dimensional, topographic or low-resolution imaging, or required isolation or chemical extraction of the cell walls. In this paper we demonstrate that by cryo-immobilizing fresh tissue, then either cryo-sectioning or freeze-substituting and resin embedding, followed by cryo- or room temperature (RT) electron tomography, respectively, we can visualize previously unseen details of plant cell wall architecture in 3D, at macromolecular resolution (∼ 2 nm), and in near-native state. Qualitative and quantitative analyses showed that wall organization of cryo-immobilized samples were preserved remarkably better than conventionally prepared samples that suffer substantial extraction. Lignin-less primary cell walls were well preserved in both self-pressurized rapidly frozen (SPRF), cryo-sectioned samples as well as high-pressure frozen, freeze-substituted and resin embedded (HPF-FS-resin) samples. Lignin-rich secondary cell walls appeared featureless in HPF-FS-resin sections presumably due to poor stain penetration, but their macromolecular features could be visualized in unprecedented details in our cryo-sections. While cryo-tomography of vitreous tissue sections is currently proving to be instrumental in developing 3D models of lignin-rich secondary cell walls, here we confirm that the technically easier method of RT-tomography of HPF-FS-resin sections could be used immediately for routine study of low-lignin cell walls. As a proof of principle, we characterized the

  12. Electron tomography of cryo-immobilized plant tissue: a novel approach to studying 3D macromolecular architecture of mature plant cell walls in situ.

    Directory of Open Access Journals (Sweden)

    Purbasha Sarkar

    Full Text Available Cost-effective production of lignocellulosic biofuel requires efficient breakdown of cell walls present in plant biomass to retrieve the wall polysaccharides for fermentation. In-depth knowledge of plant cell wall composition is therefore essential for improving the fuel production process. The precise spatial three-dimensional (3D organization of cellulose, hemicellulose, pectin and lignin within plant cell walls remains unclear to date since the microscopy techniques used so far have been limited to two-dimensional, topographic or low-resolution imaging, or required isolation or chemical extraction of the cell walls. In this paper we demonstrate that by cryo-immobilizing fresh tissue, then either cryo-sectioning or freeze-substituting and resin embedding, followed by cryo- or room temperature (RT electron tomography, respectively, we can visualize previously unseen details of plant cell wall architecture in 3D, at macromolecular resolution (∼ 2 nm, and in near-native state. Qualitative and quantitative analyses showed that wall organization of cryo-immobilized samples were preserved remarkably better than conventionally prepared samples that suffer substantial extraction. Lignin-less primary cell walls were well preserved in both self-pressurized rapidly frozen (SPRF, cryo-sectioned samples as well as high-pressure frozen, freeze-substituted and resin embedded (HPF-FS-resin samples. Lignin-rich secondary cell walls appeared featureless in HPF-FS-resin sections presumably due to poor stain penetration, but their macromolecular features could be visualized in unprecedented details in our cryo-sections. While cryo-tomography of vitreous tissue sections is currently proving to be instrumental in developing 3D models of lignin-rich secondary cell walls, here we confirm that the technically easier method of RT-tomography of HPF-FS-resin sections could be used immediately for routine study of low-lignin cell walls. As a proof of principle, we

  13. Inducibility of chemical defences by two chewing insect herbivores in pine trees is specific to targeted plant tissue, particular herbivore and defensive trait.

    Science.gov (United States)

    Moreira, Xoaquín; Lundborg, Lina; Zas, Rafael; Carrillo-Gavilán, Amparo; Borg-Karlson, Anna-Karin; Sampedro, Luis

    2013-10-01

    There is increasing evidence that plants can react to biotic aggressions with highly specific responses. However, few studies have attempted to jointly investigate whether the induction of plant defences is specific to a targeted plant tissue, plant species, herbivore identity, and defensive trait. Here we studied those factors contributing to the specificity of induced defensive responses in two economically important pine species against two chewing insect pest herbivores. Juvenile trees of Pinus pinaster and P. radiata were exposed to herbivory by two major pest threats, the large pine weevil Hylobius abietis (a bark-feeder) and the pine processionary caterpillar Thaumetopoea pityocampa (a folivore). We quantified in two tissues (stem and needles) the constitutive (control plants) and herbivore-induced concentrations of total polyphenolics, volatile and non-volatile resin, as well as the profile of mono- and sesquiterpenes. Stem chewing by the pine weevil increased concentrations of non-volatile resin, volatile monoterpenes, and (marginally) polyphenolics in stem tissues. Weevil feeding also increased the concentration of non-volatile resin and decreased polyphenolics in the needle tissues. Folivory by the caterpillar had no major effects on needle defensive chemistry, but a strong increase in the concentration of polyphenolics in the stem. Interestingly, we found similar patterns for all these above-reported effects in both pine species. These results offer convincing evidence that induced defences are highly specific and may vary depending on the targeted plant tissue, the insect herbivore causing the damage and the considered defensive compound.

  14. On the micro-indentation of plant cells in a tissue context

    Science.gov (United States)

    Mosca, Gabriella; Sapala, Aleksandra; Strauss, Soeren; Routier-Kierzkowska, Anne-Lise; Smith, Richard S.

    2017-02-01

    The effect of geometry on cell stiffness measured with micro-indentation techniques has been explored in single cells, however it is unclear if results on single cells can be readily transferred to indentation experiments performed on a tissue in vivo. Here we explored this question by using simulation models of osmotic treatments and micro-indentation experiments on 3D multicellular tissues with the finite element method. We found that the cellular context does affect measured cell stiffness, and that several cells of context in each direction are required for optimal results. We applied the model to micro-indentation data obtained with cellular force microscopy on the sepal of A. thaliana, and found that differences in measured stiffness could be explained by cellular geometry, and do not necessarily indicate differences in cell wall material properties or turgor pressure.

  15. An Effective System to Produce Smoke Solutions from Dried Plant Tissue for Seed Germination Studies

    Directory of Open Access Journals (Sweden)

    Janice Coons

    2014-02-01

    Full Text Available Premise of the study: An efficient and inexpensive system was developed to produce smoke solutions from plant material to research the influence of water-soluble compounds from smoke on seed germination. Methods and Results: Smoke solutions (300 mL per batch were produced by burning small quantities (100–200 g of dried plant material from a range of species in a bee smoker attached by a heater hose to a side-arm flask. The flask was attached to a vacuum water aspirator, to pull the smoke through the water. The entire apparatus was operated in a laboratory fume hood. Conclusions: Compared with other smoke solution preparation systems, the system described is easy to assemble and operate, inexpensive to build, and effective at producing smoke solutions from desired species in a small indoor space. Quantitative measurements can be made when using this system, allowing for replication of the process.

  16. Basic procedures for epigenetic analysis in plant cell and tissue culture.

    Science.gov (United States)

    Rodríguez, José L; Pascual, Jesús; Viejo, Marcos; Valledor, Luis; Meijón, Mónica; Hasbún, Rodrigo; Yrei, Norma Yague; Santamaría, María E; Pérez, Marta; Fernández Fraga, Mario; Berdasco, María; Rodríguez Fernández, Roberto; Cañal, María J

    2012-01-01

    In vitro culture is one of the most studied techniques, and it is used to study many developmental processes, especially in forestry species, because of growth timing and easy manipulation. Epigenetics has been shown as an important influence on many research analyses such as cancer in mammals and developmental processes in plants such as flowering, but regarding in vitro culture, techniques to study DNA methylation or chromatin modifications were mainly limited to identify somaclonal variation of the micropropagated material. Because in vitro culture is not only a way to generate plant material but also a bunch of differentially induced developmental processes, an approach of techniques and some research carried out to study the different changes regarding DNA methylation and chromatin and translational modifications that take place during these processes is reviewed.

  17. Analysis of laser-induced fluorescence spectra of in vitro plant tissue cultures

    Science.gov (United States)

    Muñoz-Muñoz, Ana Celia; Gutiérrez-Pulido, Humberto; Rodríguez-Domínguez, José Manuel; Gutiérrez-Mora, Antonia; Rodríguez-Garay, Benjamín; Cervantes-Martínez, Jesús

    2007-04-01

    We demonstrate the effectiveness of laser-induced fluorescence (LIF) for monitoring the development and stress detection of in vitro tissue cultures in a nondestructive and noninvasive way. The changes in LIF spectra caused by the induction of organogenesis, the increase of the F690/F740 ratio as a result of the stress originated in the organogenic explants due to shoot emergence, and the relationship between fluorescence spectra and shoot development were detected by LIF through closed containers of Saintpaulia ionantha.

  18. Extraction and labeling methods for microarrays using small amounts of plant tissue.

    Science.gov (United States)

    Stimpson, Alexander J; Pereira, Rhea S; Kiss, John Z; Correll, Melanie J

    2009-03-01

    Procedures were developed to maximize the yield of high-quality RNA from small amounts of plant biomass for microarrays. Two disruption techniques (bead milling and pestle and mortar) were compared for the yield and the quality of RNA extracted from 1-week-old Arabidopsis thaliana seedlings (approximately 0.5-30 mg total biomass). The pestle and mortar method of extraction showed enhanced RNA quality at the smaller biomass samples compared with the bead milling technique, although the quality in the bead milling could be improved with additional cooling steps. The RNA extracted from the pestle and mortar technique was further tested to determine if the small quantity of RNA (500 ng-7 microg) was appropriate for microarray analyses. A new method of low-quantity RNA labeling for microarrays (NuGEN Technologies, Inc.) was used on five 7-day-old seedlings (approximately 2.5 mg fresh weight total) of Arabidopsis that were grown in the dark and exposed to 1 h of red light or continued dark. Microarray analyses were performed on a small plant sample (five seedlings; approximately 2.5 mg) using these methods and compared with extractions performed with larger biomass samples (approximately 500 roots). Many well-known light-regulated genes between the small plant samples and the larger biomass samples overlapped in expression changes, and the relative expression levels of selected genes were confirmed with quantitative real-time polymerase chain reaction, suggesting that these methods can be used for plant experiments where the biomass is extremely limited (i.e. spaceflight studies).

  19. Physiological and Anatomical Response of Plant Leaf Tissue to Designated Air Pollutants.

    Science.gov (United States)

    1982-01-07

    of the spongy mesophyll . Alternatively, cells associated with intercellular air chambers immediately beneath stomata may collapse or be disrupted. This...crenalate and plasmolyzed. Cells of the spongy mesophyll 75 were also plasmolyzed. Membrane damage was prevalent in both cell types and the plasmalemma was...and more extensively than palisade or spongy parenchyma cells. Because there are no detailed cytological ultrastructural studies of HCl-fumigated plant

  20. Unexpected behavior of some nitric oxide modulators under cadmium excess in plant tissue.

    Directory of Open Access Journals (Sweden)

    Jozef Kováčik

    Full Text Available Various nitric oxide modulators (NO donors--SNP, GSNO, DEA NONOate and scavengers--PTIO, cPTIO were tested to highlight the role of NO under Cd excess in various ontogenetic stages of chamomile (Matricaria chamomilla. Surprisingly, compared to Cd alone, SNP and PTIO elevated Cd uptake (confirmed also by PhenGreen staining but depleted glutathione (partially ascorbic acid and phytochelatins PC2 and PC3 in both older plants (cultured hydroponically and seedlings (cultured in deionised water. Despite these anomalous impacts, fluorescence staining of NO and ROS confirmed predictable assumptions and revealed reciprocal changes (decrease in NO but increase in ROS after PTIO addition and the opposite after SNP application. Subsequent tests using alternative modulators and seedlings confirmed changes to NO and ROS after application of GSNO and DEA NONOate as mentioned above for SNP while cPTIO altered only NO level (depletion. On the contrary to SNP and PTIO, GSNO, DEA NONOate and cPTIO did not elevate Cd content and phytochelatins (PC2, PC3 were rather elevated. These data provide evidence that various NO modulators are useful in terms of NO and ROS manipulation but interactions with intact plants affect metal uptake and must therefore be used with caution. In this view, cPTIO and DEA NONOate revealed the less pronounced side impacts and are recommended as suitable NO scavenger/donor in plant physiological studies under Cd excess.

  1. Fluorescence in situ hybridizations (FISH) for the localization of viruses and endosymbiotic bacteria in plant and insect tissues.

    Science.gov (United States)

    Kliot, Adi; Kontsedalov, Svetlana; Lebedev, Galina; Brumin, Marina; Cathrin, Pakkianathan Britto; Marubayashi, Julio Massaharu; Skaljac, Marisa; Belausov, Eduard; Czosnek, Henryk; Ghanim, Murad

    2014-02-24

    Fluorescence in situ hybridization (FISH) is a name given to a variety of techniques commonly used for visualizing gene transcripts in eukaryotic cells and can be further modified to visualize other components in the cell such as infection with viruses and bacteria. Spatial localization and visualization of viruses and bacteria during the infection process is an essential step that complements expression profiling experiments such as microarrays and RNAseq in response to different stimuli. Understanding the spatiotemporal infections with these agents complements biological experiments aimed at understanding their interaction with cellular components. Several techniques for visualizing viruses and bacteria such as reporter gene systems or immunohistochemical methods are time-consuming, and some are limited to work with model organisms and involve complex methodologies. FISH that targets RNA or DNA species in the cell is a relatively easy and fast method for studying spatiotemporal localization of genes and for diagnostic purposes. This method can be robust and relatively easy to implement when the protocols employ short hybridizing, commercially-purchased probes, which are not expensive. This is particularly robust when sample preparation, fixation, hybridization, and microscopic visualization do not involve complex steps. Here we describe a protocol for localization of bacteria and viruses in insect and plant tissues. The method is based on simple preparation, fixation, and hybridization of insect whole mounts and dissected organs or hand-made plant sections, with 20 base pairs short DNA probes conjugated to fluorescent dyes on their 5' or 3' ends. This protocol has been successfully applied to a number of insect and plant tissues, and can be used to analyze expression of mRNAs or other RNA or DNA species in the cell.

  2. A simple, rapid and reliable protocol to localize hydrogen peroxide in large plant organs by DAB-mediated tissue printing

    Directory of Open Access Journals (Sweden)

    Yonghua eLiu

    2014-12-01

    Full Text Available Hydrogen peroxide (H2O2 is a major reactive oxygen species (ROS and plays diverse roles in plant development and stress responses. However, its localization in large and thick plant organs (e.g. stem, roots and fruits, other than leaves, has proven to be challenging due to the difficulties for the commonly used H2O2-specific chemicals, such as 3, 3’-diaminobenzidine (DAB, cerium chloride (CeCl3 and 2’, 7’-dichlorofluorescin diacetate (H2DCF-DA, to penetrate those organs. Theoretically, the reaction of endogenous H2O2 with these chemicals could be facilitated by using thin organ sections. However, the rapid production of wound-induced H2O2 associated with this procedure inevitably disturbs the original distribution of H2O2 in vivo. Here, by employing tomato seedling stems and fruits as testing materials, we report a novel, simple and rapid protocol to localize H2O2 in those organs using DAB-mediated tissue printing. The rapidity of the protocol (within 15 s completely avoided the interference of wound-induced H2O2 during experimentation. Moreover, the H2O2 signal on the printing was stable for at least 1 h with no or little background produced. We conclude that DAB-mediated tissue printing developed here provide a new feasible and reliable method to localize H2O2 in large plant organs, hence should have broad applications in studying ROS biology.

  3. Micro-scaled high-throughput digestion of plant tissue samples for multi-elemental analysis

    Directory of Open Access Journals (Sweden)

    Husted Søren

    2009-09-01

    Full Text Available Abstract Background Quantitative multi-elemental analysis by inductively coupled plasma (ICP spectrometry depends on a complete digestion of solid samples. However, fast and thorough sample digestion is a challenging analytical task which constitutes a bottleneck in modern multi-elemental analysis. Additional obstacles may be that sample quantities are limited and elemental concentrations low. In such cases, digestion in small volumes with minimum dilution and contamination is required in order to obtain high accuracy data. Results We have developed a micro-scaled microwave digestion procedure and optimized it for accurate elemental profiling of plant materials (1-20 mg dry weight. A commercially available 64-position rotor with 5 ml disposable glass vials, originally designed for microwave-based parallel organic synthesis, was used as a platform for the digestion. The novel micro-scaled method was successfully validated by the use of various certified reference materials (CRM with matrices rich in starch, lipid or protein. When the micro-scaled digestion procedure was applied on single rice grains or small batches of Arabidopsis seeds (1 mg, corresponding to approximately 50 seeds, the obtained elemental profiles closely matched those obtained by conventional analysis using digestion in large volume vessels. Accumulated elemental contents derived from separate analyses of rice grain fractions (aleurone, embryo and endosperm closely matched the total content obtained by analysis of the whole rice grain. Conclusion A high-throughput micro-scaled method has been developed which enables digestion of small quantities of plant samples for subsequent elemental profiling by ICP-spectrometry. The method constitutes a valuable tool for screening of mutants and transformants. In addition, the method facilitates studies of the distribution of essential trace elements between and within plant organs which is relevant for, e.g., breeding programmes aiming at

  4. Application of a Novel and Automated Branched DNA in Situ Hybridization Method for the Rapid and Sensitive Localization of mRNA Molecules in Plant Tissues

    Directory of Open Access Journals (Sweden)

    Andrew J. Bowling

    2014-04-01

    Full Text Available Premise of the study: A novel branched DNA detection technology, RNAscope in situ hybridization (ISH, originally developed for use on human clinical and animal tissues, was adapted for use in plant tissue in an attempt to overcome some of the limitations associated with traditional ISH assays. Methods and Results: Zea mays leaf tissue was formaldehyde fixed and paraffin embedded (FFPE and then probed with the RNAscope ISH assay for two endogenous genes, phosphoenolpyruvate carboxylase (PEPC and phosphoenolpyruvate carboxykinase (PEPCK. Results from both manual and automated methods showed tissue- and cell-specific mRNA localization patterns expected from these well-studied genes. Conclusions: RNAscope ISH is a sensitive method that generates high-quality, easily interpretable results from FFPE plant tissues. Automation of the RNAscope method on the Ventana Discovery Ultra platform allows significant advantages for repeatability, reduction in variability, and flexibility of workflow processes.

  5. Elastic cavitation, tube hollowing, and differential growth in plants and biological tissues

    KAUST Repository

    Goriely, A.

    2010-07-01

    Elastic cavitation is a well-known physical process by which elastic materials under stress can open cavities. Usually, cavitation is induced by applied loads on the elastic body. However, growing materials may generate stresses in the absence of applied loads and could induce cavity opening. Here, we demonstrate the possibility of spontaneous growth-induced cavitation in elastic materials and consider the implications of this phenomenon to biological tissues and in particular to the problem of schizogenous aerenchyma formation. Copyright © EPLA, 2010.

  6. Polyamines inhibit biosynthesis of ethylene in higher plant tissue and fruit protoplasts.

    Science.gov (United States)

    Apelbaum, A; Burgoon, A C; Anderson, J D; Lieberman, M

    1981-08-01

    Ethylene production in apple fruit and protoplasts and in leaf tissue was inhibited by spermidine or spermine. These polyamines, as well as putrescine, inhibited auxin-induced ethylene production and the conversion of methionine and 1-aminocyclopropane-1-carboxylic acid to ethylene. Polyamines were more effective as inhibitors of ethylene synthesis at the early, rather than at the late, stages of fruit ripening. Ca(2+) in the incubation medium reduced the inhibitory effect caused by the amines. A possible mode of action by which polyamines inhibit ethylene production is discussed.

  7. Plasticity of Sorghum Stem Biomass Accumulation in Response to Water Deficit: A Multiscale Analysis from Internode Tissue to Plant Level.

    Science.gov (United States)

    Perrier, Lisa; Rouan, Lauriane; Jaffuel, Sylvie; Clément-Vidal, Anne; Roques, Sandrine; Soutiras, Armelle; Baptiste, Christelle; Bastianelli, Denis; Fabre, Denis; Dubois, Cécile; Pot, David; Luquet, Delphine

    2017-01-01

    Sorghum is increasingly used as a biomass crop worldwide. Its genetic diversity provides a large range of stem biochemical composition suitable for various end-uses as bioenergy or forage. Its drought tolerance enables it to reasonably sustain biomass production under water limited conditions. However, drought effect on the accumulation of sorghum stem biomass remains poorly understood which limits progress in crop improvement and management. This study aimed at identifying the morphological, biochemical and histological traits underlying biomass accumulation in the sorghum stem and its plasticity in response to water deficit. Two hybrids (G1, G4) different in stem biochemical composition (G4, more lignified, less sweet) were evaluated during 2 years in the field in Southern France, under two water treatments differentiated during stem elongation (irrigated; 1 month dry-down until an average soil water deficit of -8.85 bars). Plant phenology was observed weekly. At the end of the water treatment and at final harvest, plant height, stem and leaf dry-weight and the size, biochemical composition and tissue histology of internodes at 2-4 positions along the stem were measured. Stem biomass accumulation was significantly reduced by drought (in average 42% at the end of the dry-down). This was due to the reduction of the length, but not diameter, of the internodes expanded during water deficit. These internodes had more soluble sugar but lower lignin and cellulose contents. This was associated with a decrease of the areal proportion of lignified cell wall in internode outer zone whereas the areal proportion of this zone was not affected. All internodes for a given genotype and environment followed a common histochemical dynamics. Hemicellulose content and the areal proportion of inner vs. outer internode tissues were set up early during internode growth and were not drought responsive. G4 exhibited a higher drought sensitivity than G1 for plant height only. At final

  8. Improving knowledge of plant tissue culture and media formulation by neurofuzzy logic: a practical case of data mining using apricot databases.

    Science.gov (United States)

    Gago, Jorge; Pérez-Tornero, Olaya; Landín, Mariana; Burgos, Lorenzo; Gallego, Pedro P

    2011-10-15

    Plant tissue growth can be regulated and controlled via culture media composition. A number of different laborious and time-consuming approaches have been used to attempt development of optimized media for a wide range of species and applications. However, plant tissue culture is a very complex task, and the identification of the influences of process factors such as mineral nutrients or plant growth regulators on a wide spectrum of growth responses cannot always well comprehended. This study employs a new approach, data mining, to uncover and integrate knowledge hidden in multiple data from plant tissue culture media formulations using apricot micropropagation databases as an example. Neurofuzzy logic technology made it possible to identify relationships among several factors (cultivars, mineral nutrients and plant growth regulators) and growth parameters (shoots number, shoots length and productivity), extracting biologically useful information from each database and combining them to create a model. The IF-THEN rule sets generated by neurofuzzy logic were completely in agreement with previous findings based on statistical analysis, but advantageously generated understandable and reusable knowledge that can be applied in future plant tissue culture media optimization.

  9. Monitoring programmed cell death of living plant tissues in microfluidics using electrochemical and optical techniques

    DEFF Research Database (Denmark)

    Mark, Christina; Heiskanen, Arto; Svensson, Birte

    for online, real-time, parallel analysis of important parameters such as redox activity (NADPH:NADP ratio), H2O2 concentration, oxygen consumption, extracellular pH, cell viability and release of target enzymes (α-amylase and limit dextrinase). Probing the intracellular redox activity is of major importance......Programmed cell death (PCD) in plants can influence the outcome of yield and quality of crops through its important role in seed germination and the defence process against pathogens. The main scope of the project is to apply microfluidic cell culture for the measurement of electrochemically......, that the H2O2 concentration changes depending on phytohormone activation or inactivation of aleurone layer metabolism and subsequent PCD3. Currently, we are working on the optimization of an intracellular whole-cell redox activity (NADP:NADPH ratio) assay5 to be able to detect possible changes...

  10. Safer DNA extraction from plant tissues using sucrose buffer and glass fiber filter.

    Science.gov (United States)

    Takakura, Koh-Ichi; Nishio, Takayuki

    2012-11-01

    For some plant species, DNA extraction and downstream experiments are inhibited by various chemicals such as polysaccharides and polyphenols. This short communication proposed an organic-solvent free (except for ethanol) extraction method. This method consists of an initial washing step with STE buffer (0.25 M sucrose, 0.03 M Tris, 0.05 M EDTA), followed by DNA extraction using a piece of glass fiber filter. The advantages of this method are its safety and low cost. The purity of the DNA solution obtained using this method is not necessarily as high as that obtained using the STE/CTAB method, but it is sufficient for PCR experiments. These points were demonstrated empirically with two species, Japanese speedwell and common dandelion, for which DNA has proven difficult to amplify via PCR in past studies.

  11. Improved Plant-based Production of E1 endoglucanase Using Potato: Expression Optimization and Tissue Targeting

    Energy Technology Data Exchange (ETDEWEB)

    Dai, Ziyu [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Hooker, Brian S. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Anderson, Daniel B. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Thomas, Steven R. [National Renewable Energy Lab. (NREL), Golden, CO (United States)

    2000-06-01

    Optimization of Acidothermus cellulolyticus endoglucanase (E1) gene expression in transgenic potato (Solanum tuberosum L.) was examined in this study, where the E1 coding sequence was transcribed under control of a leaf specific promoter (tomato RbcS-3C) or the Mac promoter (a hybrid promoter of mannopine synthase promoter and cauliflower mosaic virus 35S promoter enhancer region). Average E1 activity in leaf extracts of potato transformants, in which E1 protein was targeted by a chloroplast signal peptide and an apoplast signal peptide were much higher than those by an E1 native signal peptide and a vacuole signal peptide. E1 protein accumulated up to 2.6% of total leaf soluble protein, where E1 gene was under control of the RbcS-3C promoter, alfalfa mosaic virus 5-untranslated leader, and RbcS-2A signal peptide. E1 protein production, based on average E1 activity and E1 protein accumulation in leaf extracts, is higher in potato than those measured previously in transgenic tobacco bearing the same transgene constructs. Comparisons of E1 activity, protein accumulation, and relative mRNA levels showed that E1 expression under control of tomato RbcS-3C promoter was specifically localized in leaf tissues, while E1 gene was expressed in both leaf and tuber tissues under control of Mac promoter. This suggests dual-crop applications in which potato vines serve as enzyme production `bioreactors' while tubers are preserved for culinary applications.

  12. Fluorescent in situ hybridization for the localization of viruses, bacteria and other microorganisms in insect and plant tissues.

    Science.gov (United States)

    Kliot, Adi; Ghanim, Murad

    2016-04-01

    Methods for the localization of cellular components such as nucleic acids, proteins, cellular vesicles and more, and the localization of microorganisms including viruses, bacteria and fungi have become an important part of any research program in biological sciences that enable the visualization of these components in fixed and live tissues without the need for complex processing steps. The rapid development of microscopy tools and technologies as well as related fluorescent markers and fluorophores for many cellular components, and the ability to design DNA and RNA sequence-based molecular probes and antibodies which can be visualized fluorescently, have rapidly advanced this field. This review will focus on some of the localizations methods which have been used in plants and insect pests in agriculture, and other microorganisms, which are rapidly advancing the research in agriculture-related fields.

  13. Optimization of cDNA amplification of Apricot Latent Virus (ApLV) from various plant tissues sources.

    Science.gov (United States)

    Gumus, M; Sipahioğlu, H M; Paylan, I C; Erkan, S

    2007-03-15

    Although the reverse transcriptase polymerase chain reaction (RT-PCR) procedure is basically simple operation, often it is not possible to achieve optimum results without optimizing the protocols. An RT-PCR method targeting a 200 bp sequence of the CP gene of Apricot Latent Virus (ApLV) was used as a model to improve the detection limit and to compare the behavior of three different plant tissues in a RT-PCR assay. A number of factors should be considered when selecting the optimal system for RT-PCR. Important considerations include the optimal concentrations of MgCl2, dNTP, Taq DNA polymerase enzyme, specific primer and the amount of cDNA for the downstream applications. This study therefore discusses a series of critical PCR parameters and feasible strategies for optimization of RT-PCR detection of ApLV.

  14. Tissue tests

    NARCIS (Netherlands)

    Sonneveld, C.; Voogt, W.

    2009-01-01

    Tissue tests are widely used in horticulture practice and have in comparison with soil or substrate testing advantages as well disadvantages in comparison with soil testing. One of the main advantages of tissue tests is the certainty that analysed nutrients in plant tissues are really present in the

  15. Tissue tests

    NARCIS (Netherlands)

    Sonneveld, C.; Voogt, W.

    2009-01-01

    Tissue tests are widely used in horticulture practice and have in comparison with soil or substrate testing advantages as well disadvantages in comparison with soil testing. One of the main advantages of tissue tests is the certainty that analysed nutrients in plant tissues are really present in the

  16. Determination of o,oEDDHA - a xenobiotic chelating agent used in Fe fertilizers - in plant tissues by liquid chromatography/electrospray mass spectrometry: overcoming matrix effects.

    Science.gov (United States)

    Orera, Irene; Abadía, Anunciación; Abadía, Javier; Alvarez-Fernández, Ana

    2009-06-01

    The Fe(III)-chelate of ethylenediamine-N,N'-bis(o-hydroxyphenylacetic) acid (o,oEDDHA) is generally considered as the most efficient and widespread Fe fertilizer for fruit crops and intensive horticulture. The determination of the xenobiotic chelating agent o,oEDDHA inside the plant is a key issue in the study of this fertilizer. Both the low concentrations of o,oEDDHA expected and the complexity of plant matrices have been important drawbacks in the development of analytical methods for the determination of o,oEDDHA in plant tissues. The determination of o,oEDDHA in plant materials has been tackled in this study by liquid chromatography coupled to mass spectrometry using several plant species and tissues. Two types of internal standards have been tested: Iron stable isotope labeled compounds and a structural analogue compound, the Fe(III) chelate of ethylenediamine-N,N'-bis(2-hydroxy-4-methylphenylacetic) acid (o,oEDDHMA). Iron stable isotope labeled internal standards did not appear to be suitable because of the occurrence of isobaric endogenous compounds and/or isotope exchange reactions between plant native Fe pools and the Fe stable isotope of the internal standard. However, the structural analogue Fe(III)-o,oEDDHMA is an adequate internal standard for the determination of both isomers of o,oEDDHA (racemic and meso) in plant tissues. The method was highly sensitive, with limits of detection and quantification in the range of 3-49 and 11-162 pmol g(-1) fresh weight, respectively, and analyte recoveries were in the range of 74-116%. Using this methodology, both o,oEDDHA isomers were found in all tissues of sugar beet and tomato plants treated with 90 microM Fe(III)-o,oEDDHA for 24 h, including leaves, roots and xylem sap. This methodology constitutes a useful tool for studies on o,oEDDHA plant uptake, transport and allocation.

  17. Direct observation of cell wall structure in living plant tissues by solid-state C NMR spectroscopy.

    Science.gov (United States)

    Jarvis, M C; Apperley, D C

    1990-01-01

    Solid-state (13)C nuclear magnetic resonance (NMR) spectra of the following intact plant tissues were recorded by the crosspolarization magic-angle spinning technique: celery (Apium graveolens L.) collenchyma; carob bean (Ceratonia siliqua L.), fenugreek (Trigonella foenum-graecum L.), and nasturtium (Tropaeolum majus L.) endosperm; and lupin (Lupinus polyphyllus Lindl.) seed cotyledons. All these tissues had thickened cell walls which allowed them to withstand the centrifugal forces of magic angle spinning and which, except in the case of lupin seeds, dominated the NMR spectra. The celery collenchyma cell walls gave spectra typical of dicot primary cell walls. The carob bean and fenugreek seed spectra were dominated by resonances from galactomannans, which showed little sign of crystalline order. Resonances from beta(1,4')-d galactan were visible in the lupin seed spectrum, but there was much interference from protein. The nasturtium seed spectrum was largely derived from a xyloglucan, in which the conformation of the glucan core chain appeared to be intermediate between the solution form and solid forms of cellulose.

  18. A new cadmium reduction device for the microplate determination of nitrate in water, soil, plant tissue, and physiological fluids.

    Science.gov (United States)

    Crutchfield, James D; Grove, John H

    2011-01-01

    A reusable catalytic reductor consisting of 96 copperized-cadmium pins attached to a microplate lid was developed to simultaneously reduce nitrate (NO3-) to nitrite (NO2-) in all wells of a standard microplate. The resulting NO2- is analyzed colorimetrically by the Griess reaction using a microplate reader. Nitrate data from groundwater samples analyzed using the new device correlated well with data obtained by ion chromatography (r2 = 0.9959). Soil and plant tissue samples previously analyzed for NO3- in an interlaboratory validation study sponsored by the Soil Science Society of America were also analyzed using the new technique. For the soil sample set, the data are shown to correlate well with the other methods used (r2 = 0.9976). Plant data correlated less well, especially for samples containing low concentrations of NO3-. Reasons for these discrepancies are discussed, and new techniques to increase the accuracy of the analysis are explored. In addition, a method is presented for analyzing NO3- in physiological fluids (blood serum and urine) after matrix modification with Somogyi's reagent. A protocol for statistical validation of data when analyzing samples with complex matrixes is also established. The simplicity, adaptability, and low cost of the device indicate its potential for widespread application.

  19. [Effects of alcohol extracts from three kinds of biomass energy plant tissues on biological activity of Bemisia tabaci].

    Science.gov (United States)

    Zhou, Fu-cai; Zhou, Gui-sheng; Li, Chuan-ming; Yang, Yi-zhong; Qin, Pei

    2009-03-01

    To test the feasibility of using raw extracts from the tissues of biomass energy plants Ricinus communi and Kosteletzkya virginica as plant protection agents, the alcohol extracts from R. communi seed and leaf and from K. virginica leaf were used to treat adult Bemisia tabaci by spraying. The glutathione S-transferase and carboxylesterase activities in B. tabaci body were measured after treated for 4 h, 24 h, 48 h, 72 h, and 96 h, and the olfaction responses of B. tabaci to the alcohol extracts were detected with a Y-tube olfactomet. All the three alcohol extracts obviously inhibited the glutathione S-transferase and carboxylesterase activities in a concentration-dependent manner. The inhibitory effect of the 250-times diluted alcohol extracts on the two enzyme activities was equivalent to that of 3000 times-diluted 1.8% avermectins. In addition, the 250-times diluted alcohol extracts had obvious repellent effect on B. tabaci, with the repellent coefficient of the alcohol extracts from R. communi seed and leaf and from K, virginica leaf being 100.0%, 96.7%, and 79.4%, respectively. All of these suggested that the test three alcohol extracts had repellent and other biological effects on B. tabaci.

  20. Endophytic bacteria in plant tissue culture: differences between easy- and difficult-to-propagate Prunus avium genotypes.

    Science.gov (United States)

    Quambusch, Mona; Pirttilä, Anna Maria; Tejesvi, Mysore V; Winkelmann, Traud; Bartsch, Melanie

    2014-05-01

    The endophytic bacterial communities of six Prunus avium L. genotypes differing in their growth patterns during in vitro propagation were identified by culture-dependent and culture-independent methods. Five morphologically distinct isolates from tissue culture material were identified by 16S rDNA sequence analysis. To detect and analyze the uncultivable fraction of endophytic bacteria, a clone library was established from the amplified 16S rDNA of total plant extract. Bacterial diversity within the clone libraries was analyzed by amplified ribosomal rDNA restriction analysis and by sequencing a clone for each identified operational taxonomic unit. The most abundant bacterial group was Mycobacterium sp., which was identified in the clone libraries of all analyzed Prunus genotypes. Other dominant bacterial genera identified in the easy-to-propagate genotypes were Rhodopseudomonas sp. and Microbacterium sp. Thus, the community structures in the easy- and difficult-to-propagate cherry genotypes differed significantly. The bacterial genera, which were previously reported to have plant growth-promoting effects, were detected only in genotypes with high propagation success, indicating a possible positive impact of these bacteria on in vitro propagation of P. avium, which was proven in an inoculation experiment.

  1. Membrane-bound ATPase of intact vacuoles and tonoplasts isolated from mature plant tissue

    Energy Technology Data Exchange (ETDEWEB)

    Lin, W.; Wagner, G.J.; Siegelman, H.W.; Hind, G.

    1977-01-01

    Intact vacuoles were isolated from petals of Hippeastrum and Tulipa (Wagner, G. J. and Siegelman, H. W. (1975) Science 190, 1298 to 1299). The ATPase activity of fresh vacuole suspensions was found to be 2 to 3 times that of protoplasts from the same tissue. 70 to 80% of the ATPase activity of intact vacuoles was recovered in tonoplast preparations. The antibiotic Dio-9 at 6 ..mu..g/10/sup 6/ vacuoles or protoplasts causes 40% inhibition. However, only the protoplast ATPase is sensitive to oligomycin. N,N'-dicyclohexylcarbondiimide (DCCD) slightly stimulates ATPase activity in both vacuole and protoplast suspensions, whereas ethyl-3-(3-dimethylaminopropyl carbodiimide) (EDAC) strongly inhibits. Spectrophotometric studies show that in the petal the vacuolar contents have a pH of 4.0 for Tulipa and 4.3 for Hippeastrum, whereas the intact isolated vacuole has an internal pH of 7.0 (in pH 8.0 buffer) for Tulipa and about 7.3 for Hippeastrum. Internal ion concentrations of 150, 46, 30, 30 and 6 mM were found for K/sup +/, Na/sup +/, Mg/sup 2 +/, Cl/sup -/, and Ca/sup 2 +/ respectively, which are about the same as those in protoplasts.

  2. Monitoring plant tissue nitrogen isotopes to assess nearshore inputs of nitrogen to Lake Crescent, Olympic National Park, Washington

    Science.gov (United States)

    Cox, Stephen E.; Moran, Patrick W.; Huffman, Raegan L.; Fradkin, Steven C.

    2016-05-31

    Mats of filamentous-periphytic algae present in some nearshore areas of Lake Crescent, Olympic National Park, Washington, may indicate early stages of eutrophication from nutrient enrichment of an otherwise highly oligotrophic lake. Natural abundance ratios of stable isotopes of nitrogen (δ15N) measured in plant tissue growing in nearshore areas of the lake indicate that the major source of nitrogen used by these primary producing plants is derived mainly from atmospherically fixed nitrogen in an undeveloped forested ecosystem. Exceptions to this pattern occurred in the Barnes Point area where elevated δ15N ratios indicate that effluent from septic systems also contribute nitrogen to filamentous-periphytic algae growing in the littoral zone of that area. Near the Lyre River outlet of Lake Crescent, the δ15N of filamentous-periphytic algae growing in close proximity to the spawning areas of a unique species of trout show little evidence of elevated δ15N indicating that nitrogen from on-site septic systems is not a substantial source of nitrogen for these plants. The δ15N data corroborate estimates that nitrogen input to Lake Crescent from septic sources is comparatively small relative to input from motor vehicle exhaust and vegetative sources in undeveloped forests, including litterfall, pollen, and symbiotic nitrogen fixation. The seasonal timing of blooms of filamentous-periphytic algal near the lake shoreline is also consistent with nitrogen exported from stands of red alder trees (Alnus rubra). Isotope biomonitoring of filamentous-periphytic algae may be an effective approach to monitoring the littoral zone for nutrient input to Lake Crescent from septic sources.

  3. Effect of fluazifop-p-butyl treatment on pigments and polyamines level within tissues of non-target maize plants.

    Science.gov (United States)

    Horbowicz, Marcin; Sempruch, Cezary; Kosson, Ryszard; Koczkodaj, Danuta; Walas, Dajana

    2013-09-01

    Fluazifop-p-butyl (FL) is one of the most popular graminicides from arylophenoxypropionate group. These herbicides act as inhibitors of acetyl-CoA carboxylase (ACCase) that catalyzes the formation of malonyl-CoA during metabolism of lipids and/or of some secondary compounds. On the other hand arylopropionates and cyclohexanediones cause phytotoxic effects by stimulating free-radicals generation and causing oxidative stress in susceptible plants. However, the importance of disturbances in plant pigments and polyamines accumulation for this effect is not clear. The aim of this work is to quantify the phytotoxicity of FL to non target maize plant and to explain how photosynthetic pigments, anthocyanins (ANC) and polyamines participate in this interaction. Obtained results showed reduction of chlorophyll a and b, but only in case of the highest herbicide dose. Lower FL concentrations caused increase of the photosynthetic pigments, or were not effective. A similar effect was stated for putrescine, while spermidine was reduced within epicotyl of leaf tissues. In case of 2-phenylethylamine (PEA), there was observed a lack of significant changes within leaves and an increase in epicotyl under the middle and the highest dose of the herbicide. Moreover, FL induced ANC accumulation in epicotyls of maize seedlings. The activity of such key enzymes of polyamine biosynthesis as: ornithine decarboxylase (ODC) and lysine decarboxylase (LDC), increased in leaves treated with herbicide at the lowest concentration and decreased under the highest. However, in case of epicotyls the decreasing tendency was observed with the exception of ODC under the highest FL dose. The activity of tyrosine decarboxylase (TyDC) was importantly elevated only within epicotyls under the lower FL concentrations. It was concluded that FL inhibits maize growth, and the intensity of the effect is positively correlated with the herbicide concentration. The phenomenon was related to changes in content of

  4. Human health risk assessment of pharmaceuticals and personal care products in plant tissue due to biosolids and manure amendments, and wastewater irrigation.

    Science.gov (United States)

    Prosser, R S; Sibley, P K

    2015-02-01

    Amending soil with biosolids or livestock manure provides essential nutrients in agriculture. Irrigation with wastewater allows for agriculture in regions where water resources are limited. However, biosolids, manure and wastewater have all been shown to contain pharmaceuticals and personal care products (PPCPs). Studies have shown that PPCPs can accumulate in the tissues of plants but the risk that accumulated residues may pose to humans via consumption of edible portions is not well documented. This study reviewed the literature for studies that reported residues of PPCPs in the edible tissue of plants grown in biosolids- or manure-amended soils or irrigated with wastewater. These residues were used to determine the estimated daily intake of PPCPs for an adult and toddler. Estimated daily intake values were compared to acceptable daily intakes to determine whether PPCPs in plant tissue pose a hazard to human health. For all three amendment practices, the majority of reported residues resulted in hazard quotients <0.1. Amendment with biosolids or manure resulted in hazard quotients ≥0.1 for carbamazepine, diphenhydramine, salbutamol, triclosan, and sulfamethazine. Irrigation with wastewater resulted in hazard quotients of ≥0.1 for ambrettolid, carbamazepine, diclofenac, flunixin, lamotrigine, metoprolol, naproxen, sildenafil and tonalide. [corrected]. Many of the residues that resulted in hazard quotients ≥0.1 were due to exposing plants to concentrations of PPCPs that would not be considered relevant based on concentrations reported in biosolids and manure or unrealistic methods of exposure, which lead to artificially elevated plant residues. Our assessment indicates that the majority of individual PPCPs in the edible tissue of plants due to biosolids or manure amendment or wastewater irrigation represent a de minimis risk to human health. Assuming additivity, the mixture of PPCPs could potentially present a hazard. Further work needs to be done to assess

  5. Binding affinity and capacities for ytterbium(3+) and hafinum(4+) by chemical entities of plant tissue fragments.

    Science.gov (United States)

    Worley, R; Clearfield, A; Ellis, W C

    2002-12-01

    The binding affinity of ytterbium (Yb3+) and hafinum (Hf4+) to ligands of chemical entities of fragments of bermudagrass tissues and their resistance to exchanging Yb with other ligands and to displacement by protons were investigated. Chemical entities of acid resistant NDF (ARNDF), 0.1 N acid detergent fiber (0.1 N ADF), and permanganate cellulose (CELL) were prepared from fragments of bermudagrass hay (Cynodon dactylon [L.] Pers.) obtained by grinding to pass a 2-mm sieve. 175Ytterbium and Yb, as YbCl3, were initially bound to each preparation by soaking for 12 h in pH 5.5 borate buffer to obtain Yb bound onto ligands having affinity constants for Yb equal to or greater than that for the weakly stable borate ligand, Yb > or = borate. The fraction of Yb > or = borate was measured and fragments then sequentially exposed to acetate, citrate, nitrotriacetate (NTA), and EDTA ions to allow exchange of Yb from Yb > or = borate with ligands having affinity constants for Yb equal to or greater than acetate (Yb > or = acetate), citrate (Yb > or = citrate), NTA (Yb > or = NTA), and EDTA (Yb > or = EDTA) ions. Binding of Yb > or = borate indicated the existence of two species of ligands: strong ligands binding essentially 100% of added Yb at levels of 1 to 1,300 ppm (0.1 N ADF) and at 1 to 7,000 ppm (ARNDF); and weaker ligands binding 4 and 8% of the Yb, respectively, at levels of added Yb greater than 1,300 ppm and 7,000 ppm. Ytterbium > or = acetate of ARNDF, but not 0.1 N ADF, was as resistant to exchange as Yb > or = citrate. Ytterbium > or = borate was exchanged extensively (85% or greater) with soluble ligands having affinity constants > or = NTA. Ytterbium resistance to proton displacement at pH of 1.5 increased with Yb > or = EDTA > Yb > or = NTA > Yb > or = citrate > Yb > or = acetate. Very efficient binding of Yb to CELL suggested that such chemical preparations are not representative of native cellulose. Hafnium (4+) was strongly bound to plant tissues rendering

  6. Application of New Technologies in Tissue Culture of Ornamental Plants%新技术在观赏植物组织培养中的应用

    Institute of Scientific and Technical Information of China (English)

    肖远志; 黄国林; 张平

    2013-01-01

    Plant tissue culture is a new technology which will promote vegetative propagation. This paper summarized the application of some new tissue culture technologies in ornamental plants, including sugar-free tissue culture technology, open tissue culture technology, application of new light sources and new hormones. Plant tissue culture can shorten the production period and the reduce production cost, thereby promoting the development of ornamental plant industry.%植物组织培养是一项加速无性繁殖的新技术。综述了无糖组培技术、开放组培技术、新型光源和新型激素的应用等组培新技术在观赏植物上的应用。植物组织培养能缩短观赏植物的生产周期,降低生产成本,有力推动了观赏植物产业的发展。

  7. RNA extraction from various recalcitrant plant tissues with a cethyltrimethylammonium bromide-containing buffer followed by an acid guanidium thiocyanate-phenol-chloroform treatment.

    Science.gov (United States)

    Suzuki, Yuji; Mae, Tadahiko; Makino, Amane

    2008-07-01

    High-quality total RNA was extracted using a cethyltrimethylammonium bromide-containing buffer followed by an acid guanidium thiocyanate-phenol-chloroform treatment from recalcitrant plant tissues such as tree leaves (pine, Norway spruce, ginkgo, Japanese cedar, rose), flowers (rose, Lotus japonicus) and storage tissues (seeds of Lotus japonicus and rice, sweet potato tuber, banana fruit). This protocol greatly reduced the time required for RNA extraction.

  8. Influence of atmospheric [CO2] on growth, carbon allocation and cost of plant tissues on leaf nitrogen concentration maintenance in nodulated Medicago sativa

    Science.gov (United States)

    Pereyra, Gabriela; Hartmann, Henrik; Ziegler, Waldemar; Michalzik, Beate; Gonzalez-Meler, Miquel; Trumbore, Susan

    2015-04-01

    Plant carbon (C) allocation and plant metabolic processes (i.e. photosynthesis and respiration) can be affected by changes in C availability, for example from changing atmospheric [CO2]. In nodulated plants, C availability may also influence nitrogen (N) fixation by bacteriods. But C allocation and N fixation are often studied independently and hence do not allow elucidating interactive effects. We investigated how different atmospheric [CO2] (Pleistocene: 170 ppm, ambient: 400 ppm and projected future: 700 ppm) influence plant growth, allocation to nodules, and the ratio of photosynthesis-to-respiration (R:A) as an indicator of C cost in Medicago sativa inoculated with Ensifer meliloti. M. sativa grew c. 38% more nodules at 400 ppm and 700 ppm than at 170 ppm. However, ratios of above- and belowground plant biomass to nodule biomass were constant over time and independent of atmospheric [CO2]. Total non-structural carbohydrate concentrations were not significantly different between plants grown at 400 and 700 ppm, but were four to five-fold higher than in 170 ppm plants. Leaf level N concentration was similar across treatments, but N-based photosynthetic rates were 82% and 93% higher in leaves of plants grown at 400 and 700 ppm, respectively, than plants grown at 170 ppm. In addition, leaf R:A was greater (48% or 55%) in plants grown at 170 ppm than plants grown at 400 and 700 ppm. Similarly, the greatest proportion of assimilated CO2 released by root respiration occurred in rhizobial plants growing at 170 ppm. Our results suggest that C limitation in nodulated Medicago sativa plants did not influence C allocation to nodule biomass but caused a proportionally greater allocation of C to belowground respiration, most likely to bacteriods. This suggests that N tissue concentration was maintained at low [CO2] by revving up bacteriod metabolism and at the expense of non-structural carbohydrate reserves.

  9. Tissue-specific, development-dependent phenolic compounds accumulation profile and gene expression pattern in tea plant [Camellia sinensis].

    Directory of Open Access Journals (Sweden)

    Xiaolan Jiang

    Full Text Available Phenolic compounds in tea plant [Camellia sinensis (L.] play a crucial role in dominating tea flavor and possess a number of key pharmacological benefits on human health. The present research aimed to study the profile of tissue-specific, development-dependent accumulation pattern of phenolic compounds in tea plant. A total of 50 phenolic compounds were identified qualitatively using liquid chromatography in tandem mass spectrometry technology. Of which 29 phenolic compounds were quantified based on their fragmentation behaviors. Most of the phenolic compounds were higher in the younger leaves than that in the stem and root, whereas the total amount of proanthocyanidins were unexpectedly higher in the root. The expression patterns of 63 structural and regulator genes involved in the shikimic acid, phenylpropanoid, and flavonoid pathways were analyzed by quantitative real-time polymerase chain reaction and cluster analysis. Based on the similarity of their expression patterns, the genes were classified into two main groups: C1 and C2; and the genes in group C1 had high relative expression level in the root or low in the bud and leaves. The expression patterns of genes in C2-2-1 and C2-2-2-1 groups were probably responsible for the development-dependent accumulation of phenolic compounds in the leaves. Enzymatic analysis suggested that the accumulation of catechins was influenced simultaneously by catabolism and anabolism. Further research is recommended to know the expression patterns of various genes and the reason for the variation in contents of different compounds in different growth stages and also in different organs.

  10. Tissue-specific, development-dependent phenolic compounds accumulation profile and gene expression pattern in tea plant [Camellia sinensis].

    Science.gov (United States)

    Jiang, Xiaolan; Liu, Yajun; Li, Weiwei; Zhao, Lei; Meng, Fei; Wang, Yunsheng; Tan, Huarong; Yang, Hua; Wei, Chaoling; Wan, Xiaochun; Gao, Liping; Xia, Tao

    2013-01-01

    Phenolic compounds in tea plant [Camellia sinensis (L.)] play a crucial role in dominating tea flavor and possess a number of key pharmacological benefits on human health. The present research aimed to study the profile of tissue-specific, development-dependent accumulation pattern of phenolic compounds in tea plant. A total of 50 phenolic compounds were identified qualitatively using liquid chromatography in tandem mass spectrometry technology. Of which 29 phenolic compounds were quantified based on their fragmentation behaviors. Most of the phenolic compounds were higher in the younger leaves than that in the stem and root, whereas the total amount of proanthocyanidins were unexpectedly higher in the root. The expression patterns of 63 structural and regulator genes involved in the shikimic acid, phenylpropanoid, and flavonoid pathways were analyzed by quantitative real-time polymerase chain reaction and cluster analysis. Based on the similarity of their expression patterns, the genes were classified into two main groups: C1 and C2; and the genes in group C1 had high relative expression level in the root or low in the bud and leaves. The expression patterns of genes in C2-2-1 and C2-2-2-1 groups were probably responsible for the development-dependent accumulation of phenolic compounds in the leaves. Enzymatic analysis suggested that the accumulation of catechins was influenced simultaneously by catabolism and anabolism. Further research is recommended to know the expression patterns of various genes and the reason for the variation in contents of different compounds in different growth stages and also in different organs.

  11. Differential tissue accumulation of 2,3,7,8-Tetrachlorinated dibenzo-p-dioxin in Arabidopsis thaliana affects plant chronology, lipid metabolism and seed yield.

    Science.gov (United States)

    Hanano, Abdulsamie; Almousally, Ibrahem; Shaban, Mouhnad; Moursel, Nour; Shahadeh, AbdAlbaset; Alhajji, Eskander

    2015-08-11

    Dioxins are one of the most toxic groups of persistent organic pollutants. Their biotransmission through the food chain constitutes a potential risk for human health. Plants as principal actors in the food chain can play a determinant role in removing dioxins from the environment. Due to the lack of data on dioxin/plant research, this study sets out to determine few responsive reactions adopted by Arabidopsis plant towards 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), the most toxic congener of dioxins. Using a high resolution gas chromatography/mass spectrometry, we demonstrated that Arabidopsis plant uptakes TCDD by the roots and accumulates it in the vegetative parts in a tissue-specific manner. TCDD mainly accumulated in rosette leaves and mature seeds and less in stem, flowers and immature siliques. Moreover, we observed that plants exposed to high doses of TCDD exhibited a delay in flowering and yielded fewer seeds of a reduced oil content with a low vitality. A particular focus on the plant fatty acid metabolism showed that TCDD caused a significant reduction in C18-unsaturated fatty acid level in plant tissues. Simultaneously, TCDD induced the expression of 9-LOX and 13-LOX genes and the formation of their corresponding hydroperoxides, 9- and 13-HPOD as well as 9- or 13-HPOT, derived from linoleic and linolenic acids, respectively. The current work highlights a side of toxicological effects resulting in the administration of 2,3,7,8-TCDD on the Arabidopsis plant. Similarly to animals, it seems that plants may accumulate TCDD in their lipids by involving few of the FA-metabolizing enzymes for sculpting a specific oxylipins "signature" typified to plant TCDD-tolerance. Together, our results uncover novel responses of Arabidopsis to dioxin, possibly emerging to overcome its toxicity.

  12. Draft Genome Sequence of Bacillus pumilus Strain GM3FR, an Endophyte Isolated from Aerial Plant Tissues of Festuca rubra L.

    Science.gov (United States)

    Hollensteiner, Jacqueline; Daniel, Rolf; Liesegang, Heiko; Vidal, Stefan

    2017-01-01

    ABSTRACT Here, we report the draft genome sequence of Bacillus pumilus GM3FR, an endophytic bacterium isolated from aerial plant tissues of Festuca rubra L. The draft genome consists of 3.5 Mb and harbors 3,551 predicted protein-encoding genes. The genome provides insights into the biocontrol potential of B. pumilus GM3FR. PMID:28360161

  13. Epiphyte-cover on seagrass (Zostera marina L. leaves impedes plant performance and radial O2 loss from the below-ground tissue

    Directory of Open Access Journals (Sweden)

    Kasper Elgetti Brodersen

    2015-08-01

    Full Text Available The O2 budget of seagrasses is a complex interaction between several sources and sinks, which is strongly regulated by light availability and mass transfer over the diffusive boundary layer (DBL surrounding the plant. Epiphyte growth on leaves may thus strongly affect the O2 availability of the seagrass plant and its capability to aerate its rhizosphere as a defence against plant toxins.We used electrochemical and fiber-optic microsensors to quantify the O2 flux, DBL and light microclimate around leaves with and without filamentous algal epiphytes. We also quantified the below-ground radial O2 loss from roots (~1 mm from the root-apex to elucidate how this below-ground oxic microzone was affected by the presence of epiphytes.Epiphyte-cover on seagrass leaves (~21% areal cover resulted in reduced light quality and quantity for photosynthesis, thus leading to reduced plant fitness. A ~4 times thicker diffusive boundary layer around leaves with epiphyte-cover impeded gas (and nutrient exchange with the surrounding water-column and thus the amount of O2 passively diffusing into the leaves in darkness. During light exposure of the leaves, radial oxygen loss from the below-ground tissue was ~2 times higher from plants without epiphyte-cover. In contrast, no O2 was detectable at the surface of the root-cap tissue of plants with epiphyte-cover during darkness, leaving the plants more susceptible to sulphide intrusion.Epiphyte growth on seagrass leaves thus negatively affects the light climate and O2 uptake in darkness, hampering the plants performance and thereby reducing the oxidation capability of its below-ground tissue.

  14. Efficient extraction of proteins from recalcitrant plant tissue for subsequent analysis by two-dimensional gel electrophoresis.

    Science.gov (United States)

    Parkhey, Suruchi; Chandrakar, Vibhuti; Naithani, S C; Keshavkant, S

    2015-10-01

    Protein extraction for two-dimensional electrophoresis from tissues of recalcitrant species is quite problematic and challenging due to the low protein content and high abundance of contaminants. Proteomics in Shorea robusta is scarcely conducted due to the lack of a suitable protein preparation procedure. To establish an effective protein extraction protocol suitable for two-dimensional electrophoresis in Shorea robusta, four procedures (borate buffer/trichloroacetic acid extraction, organic solvent/trichloroacetic acid precipitation, sucrose/Tris/phenol, and organic solvent/phenol/sodium dodecyl sulfate) were evaluated. Following these, proteins were isolated from mature leaves and were analyzed for proteomics, and also for potential contaminants, widely reported to hinder proteomics. The borate buffer/trichloroacetic acid extraction had the lowest protein yield and did not result in any banding even in one-dimensional electrophoresis. In contrast, organic solvent/phenol/sodium dodecyl sulfate extraction allowed the highest protein yield. Moreover, during proteomics, organic solvent/phenol/sodium dodecyl sulfate extracted protein resolved the maximum number (144) of spots. Further, when proteins were evaluated for contaminants, significant (77-95%) reductions in the nucleic acids, phenol, and sugars were discernible with refinement in extraction procedure. Accumulated data suggested that the organic solvent/phenol/sodium dodecyl sulfate extraction was the most effective protocol for protein isolation for proteomics of Shorea robusta and can be used for plants that have a similar set of contaminants.

  15. Variations in alkaloidal constituents of plant tissue cultures. [Nicotiana tabacum, Nicotiana rustica, Datura stramonium and Hyoscyamus niger

    Energy Technology Data Exchange (ETDEWEB)

    Essa, A.K.

    1987-01-01

    The accumulation of tobacco and Solanaceae-tropane alkaloids in calluses and suspensions of Nicotiana tabacum, Nicotiana rustica, Datura stramonium and Hyoscyamus niger was the main concern of this work. Nicotine, anabasine and anatabine had regularly been found in tobacco callus tissues grown for several passages. For the first time, N. tabacum suspensions were shown able to accumulate anabasine, anatabine, anatalline, myosmine and nicotelline whereas N. rustica calluses, N-methyl-, N,N-dimethyl- and N-methyl-N-nitroso anilines. The aim of these experiments was an attempt to affect the yields and types of alkaloids produced. The interrelationship between nicotine and protein contents in N. tabacum and N. rustica calluses was investigated. The possible role of urea and sodium propionate as precursors of nicotine in tobacco suspensions was checked by feeding the latter with carbon-14 radioactive substrates. The scope and quantum of the principle alkaloidal components present in the source N. tabacum plants and D. stramonium and H. niger seeds were investigated to stand as references against in vitro production. Identification of the products found was made by using gas chromatography-mass spectrometry (GC-MS) and/or MS.

  16. Effects of plant tissue factors of the acceptance of four greenhouse vegetable host plants by the greenhouse whitefly: an electrical penetration graph (EPG) study

    NARCIS (Netherlands)

    Lei, H.; Lenteren, van J.C.; Xu, R.M.

    2001-01-01

    A combination of biological control and host-plant resistance is needed to control greenhouse whitefly, Trialeurodes vaporariorum (Westwood). The high level of susceptibility of several host plants to whitefly, based on their performance on these plants, is well documented. These studies only

  17. In-Field, In Situ, and In Vivo 3-Dimensional Elemental Mapping for Plant Tissue and Soil Analysis Using Laser-Induced Breakdown Spectroscopy

    Directory of Open Access Journals (Sweden)

    Chunjiang Zhao

    2016-10-01

    Full Text Available Sensing and mapping element distributions in plant tissues and its growth environment has great significance for understanding the uptake, transport, and accumulation of nutrients and harmful elements in plants, as well as for understanding interactions between plants and the environment. In this study, we developed a 3-dimensional elemental mapping system based on laser-induced breakdown spectroscopy that can be deployed in- field to directly measure the distribution of multiple elements in living plants as well as in the soil. Mapping is performed by a fast scanning laser, which ablates a micro volume of a sample to form a plasma. The presence and concentration of specific elements are calculated using the atomic, ionic, and molecular spectral characteristics of the plasma emission spectra. Furthermore, we mapped the pesticide residues in maize leaves after spraying to demonstrate the capacity of this method for trace elemental mapping. We also used the system to quantitatively detect the element concentrations in soil, which can be used to further understand the element transport between plants and soil. We demonstrate that this method has great potential for elemental mapping in plant tissues and soil with the advantages of 3-dimensional and multi-elemental mapping, in situ and in vivo measurement, flexible use, and low cost.

  18. In-Field, In Situ, and In Vivo 3-Dimensional Elemental Mapping for Plant Tissue and Soil Analysis Using Laser-Induced Breakdown Spectroscopy.

    Science.gov (United States)

    Zhao, Chunjiang; Dong, Daming; Du, Xiaofan; Zheng, Wengang

    2016-10-22

    Sensing and mapping element distributions in plant tissues and its growth environment has great significance for understanding the uptake, transport, and accumulation of nutrients and harmful elements in plants, as well as for understanding interactions between plants and the environment. In this study, we developed a 3-dimensional elemental mapping system based on laser-induced breakdown spectroscopy that can be deployed in- field to directly measure the distribution of multiple elements in living plants as well as in the soil. Mapping is performed by a fast scanning laser, which ablates a micro volume of a sample to form a plasma. The presence and concentration of specific elements are calculated using the atomic, ionic, and molecular spectral characteristics of the plasma emission spectra. Furthermore, we mapped the pesticide residues in maize leaves after spraying to demonstrate the capacity of this method for trace elemental mapping. We also used the system to quantitatively detect the element concentrations in soil, which can be used to further understand the element transport between plants and soil. We demonstrate that this method has great potential for elemental mapping in plant tissues and soil with the advantages of 3-dimensional and multi-elemental mapping, in situ and in vivo measurement, flexible use, and low cost.

  19. Plant Tissue Culture and Its Application on Fruit Tree%植物组织培养及其在果树上的应用

    Institute of Scientific and Technical Information of China (English)

    刘书荣; 刘静; 吴曼

    2011-01-01

    Plant tissue culture refers to the use of totipotency cells in order to regeneration plant.It has important value with its application on fruit tree which could breeding rapidly,produce virus-free,cultivate new varieties,overcome such hybrids obstacles and save germplasm resources.This paper reviewed the progress of plant tissue culture on fruit tree while suggests references for further study of plant tissue culture.%植物组织培养是指利用细胞全能性培养再生植株的技术,由于其具有快速繁殖、产生脱毒苗、培育新品种、克服杂交不亲和障碍和保存种质资源等应用优点,在果树应用上有重要的价值。通过介绍植物组织培养在果树上的应用,以为今后研究提供参考。

  20. Characterization of plant-growth-promoting effects and concurrent promotion of heavy metal accumulation in the tissues of the plants grown in the polluted soil by Burkholderia strain LD-11.

    Science.gov (United States)

    Huang, Gui-Hai; Tian, Hui-Hui; Liu, Hai-Ying; Fan, Xian-Wei; Liang, Yu; Li, You-Zhi

    2013-01-01

    Plant-growth-promoting (PGP) bacteria especially with the resistance to multiple heavy metals are helpful to phytoremediation. Further development of PGP bacteria is very necessary because of the extreme diversity of plants, soils, and heavy metal pollution. A Burkholderia sp. strain, numbered LD-11, was isolated, which showed resistances to multiple heavy metals and antibiotics. It can produce indole-3-acetic acid, 1-aminocyclopropane-1-carboxylic acid deaminase and siderophores. Inoculation with the LD-11 improved germination of seeds of the investigated vegetable plants in the presence of Cu, promoted elongation of roots and hypocotyledonary axes, enhanced the dry weights of the plants grown in the soils polluted with Cu and/or Pb, and increased activity of the soil urease and the rhizobacteria diversity. Inoculation with the LD-11 significantly enhanced Cu and/or Pb accumulation especially in the roots of the plants grown in the polluted soils. Notably, LD-11 could produce siderophores in the presence of Cu. Conclusively, the PGP effects and concurrent heavy metal accumulation in the plant tissues results from combined effects of the above-mentioned multiple factors. Cu is an important element that represses production of the siderophore by the bacteria. Phytoremediation by synergistic use of the investigated plants and the bacterial strain LD-11 is a phytoextraction process.

  1. The Review of Organic Additives in Plant Tissue Culture%植物组织培养中有机添加物应用研究

    Institute of Scientific and Technical Information of China (English)

    李亮; 张冬敏; 雷华辉; 钟凤林

    2012-01-01

    At plant tissue culture,the inclusion of organic additives in the medium has obvious stimulative effect on cultivator's proliferation and differentiation.In this paper,we review the roles of organic additives and its application in plant tissue culture at present.%植物组织培养中,在培养基内加入有机添加物,对培养物的分化、增殖有明显促进作用。文章综述了有机添加物的作用及其目前在植物组织培养中的应用情况。

  2. Use of co-loaded Fluo-3 and Fura Red fluorescent indicators for studying the cytosolic Ca(2+)concentrations distribution in living plant tissue.

    Science.gov (United States)

    Walczysko, P; Wagner, E; Albrechtová, J T

    2000-07-01

    A method for visualisation of cytosolic [Ca(2+)] distribution was applied to living plant tissue. A mixture of the fluorescent probes Fluo-3 and Fura Red was used. The emitted fluorescence was scanned simultaneously in two channels with a laser-scanning confocal microscope and rationing was performed. The homogeneity of the Fluo-3/Fura Red concentration ratio throughout the tissue after AM-ester loading was proven. In vitro calibration permitted conversion of Fluo-3/Fura Red fluorescence ratios to [Ca(2+)] values. Apparent K(D)of 286 nM, R(min)of 0.43 and R(max)of 18 were calculated. The in vivo determination of extreme ratio values was performed by permeabilizing the plasmalemma for Ca(2+)with a ionophore and manipulating the extracellular [Ca(2+)]. The resultant R(minv)of 1.33 and R(maxv)of 2.69 for vegetative apices, and R(mini)of 1.26 and R(maxi)of 3.45 for apices induced to flowering, suggested incomplete equalization of extra- and intracellular Ca(2+)levels in these experiments. In Chenopodium rubrum, the cytosolic [Ca(2+)] patterns of apical tissue obtained using Fluo-3 and Fura Red were significantly different between vegetative apices and apices after photoperiodic flower induction. This methodological approach may also be helpful for studying cytosolic [Ca(2+)] distribution in other living plant tissues. Copyright 2000 Harcourt Publishers Ltd.

  3. Research on Tissue Culture Technology of Plant Anther%植物花药组织培养技术的研究

    Institute of Scientific and Technical Information of China (English)

    王万奇; 李文龙; 王媛媛; 廖栩; 穆丹

    2015-01-01

    植物花药组织培养技术在育种和基础理论研究中已应用广泛 ,研究阐述了花药组织培养的技术发展现状 ,系统地论述了植物花药组织培养过程中外植体的不同、培养基配方、接种方式、培养条件的不同对组培结果的影响 ,并对花药组织培养技术的应用进行概述 ,使花药组织培养技术研究体系更加清晰系统.%Plant anther tissue culture technology had been widely used in breeding and basic theoretical re-search .The development status of anther culture was interviewed ,the effects of anthers of the plant tissue cul-ture process of explants of different medium formulations ,inoculation on the tissue culture results in different culture conditions systematic exposition were studied ,anther tissue culture application was overviewed .

  4. Effects of micro electric current load during cooling of plant tissues on intracellular ice crystal formation behavior and pH.

    Science.gov (United States)

    Ninagawa, Takako; Kawamura, Yukio; Konishi, Tadashi; Narumi, Akira

    2016-08-01

    Cryopreservation techniques are expected to evolve further to preserve biomaterials and foods in a fresh state for extended periods of time. Long-term cryopreservation of living materials such as food and biological tissue is generally achieved by freezing; thus, intracellular freezing occurs. Intracellular freezing injures the cells and leads to cell death. Therefore, a dream cryopreservation technique would preserve the living materials without internal ice crystal formation at a temperature low enough to prevent bacterial activity. This study was performed to investigate the effect of micro electrical current loading during cooling as a new cryopreservation technique. The behavior of intracellular ice crystal formation in plant tissues with or without an electric current load was evaluated using the degree of supercooling, degree of cell deformation, and grain size and growing rate of intracellular ice crystal. Moreover, the transition of intracellular pH during plant tissue cooling with or without electric current loading was also examined using the fluorescence intensity ratio to comprehend cell activity at lower temperatures. The results indicated that micro electric current load did not only decrease the degree of cell deformation and grain size of intracellular ice crystal but also reduced the decline in intracellular pH due to temperature lowering, compared with tissues subjected to the same cooling rate without an electric current load. Thus, the effect of electric current load on cryopreservation and the potential of a new cryopreservation technique using electric current load were discussed based on these results.

  5. Photochemical studies of a fluorescent chlorophyll catabolite--source of bright blue fluorescence in plant tissue and efficient sensitizer of singlet oxygen.

    Science.gov (United States)

    Jockusch, Steffen; Turro, Nicholas J; Banala, Srinivas; Kräutler, Bernhard

    2014-02-01

    Fluorescent chlorophyll catabolites (FCCs) are fleeting intermediates of chlorophyll breakdown, which is seen as an enzyme controlled detoxification process of the chlorophylls in plants. However, some plants accumulate large amounts of persistent FCCs, such as in senescent leaves and in peels of yellow bananas. The photophysical properties of such a persistent FCC (Me-sFCC) were investigated in detail. FCCs absorb in the near UV spectral region and show blue fluorescence (max at 437 nm). The Me-sFCC fluorescence had a quantum yield of 0.21 (lifetime 1.6 ns). Photoexcited Me-sFCC intersystem crosses into the triplet state (quantum yield 0.6) and generates efficiently singlet oxygen (quantum yield 0.59). The efficient generation of singlet oxygen makes fluorescent chlorophyll catabolites phototoxic, but might also be useful as a (stress) signal and for defense of the plant tissue against infection by pathogens.

  6. Medicinal Plants for the Treatment of Local Tissue Damage Induced by Snake Venoms: An Overview from Traditional Use to Pharmacological Evidence

    Science.gov (United States)

    Félix-Silva, Juliana; Silva-Junior, Arnóbio Antônio; Zucolotto, Silvana Maria

    2017-01-01

    Snakebites are a serious problem in public health due to their high morbimortality. Most of snake venoms produce intense local tissue damage, which could lead to temporary or permanent disability in victims. The available specific treatment is the antivenom serum therapy, whose effectiveness is reduced against these effects. Thus, the search for complementary alternatives for snakebite treatment is relevant. There are several reports of the popular use of medicinal plants against snakebites worldwide. In recent years, many studies have been published giving pharmacological evidence of benefits of several vegetal species against local effects induced by a broad range of snake venoms, including inhibitory potential against hyaluronidase, phospholipase, proteolytic, hemorrhagic, myotoxic, and edematogenic activities. In this context, this review aimed to provide an updated overview of medicinal plants used popularly as antiophidic agents and discuss the main species with pharmacological studies supporting the uses, with emphasis on plants inhibiting local effects of snake envenomation. The present review provides an updated scenario and insights into future research aiming at validation of medicinal plants as antiophidic agents and strengthens the potentiality of ethnopharmacology as a tool for design of potent inhibitors and/or development of herbal medicines against venom toxins, especially local tissue damage. PMID:28904556

  7. Medicinal Plants for the Treatment of Local Tissue Damage Induced by Snake Venoms: An Overview from Traditional Use to Pharmacological Evidence

    Directory of Open Access Journals (Sweden)

    Juliana Félix-Silva

    2017-01-01

    Full Text Available Snakebites are a serious problem in public health due to their high morbimortality. Most of snake venoms produce intense local tissue damage, which could lead to temporary or permanent disability in victims. The available specific treatment is the antivenom serum therapy, whose effectiveness is reduced against these effects. Thus, the search for complementary alternatives for snakebite treatment is relevant. There are several reports of the popular use of medicinal plants against snakebites worldwide. In recent years, many studies have been published giving pharmacological evidence of benefits of several vegetal species against local effects induced by a broad range of snake venoms, including inhibitory potential against hyaluronidase, phospholipase, proteolytic, hemorrhagic, myotoxic, and edematogenic activities. In this context, this review aimed to provide an updated overview of medicinal plants used popularly as antiophidic agents and discuss the main species with pharmacological studies supporting the uses, with emphasis on plants inhibiting local effects of snake envenomation. The present review provides an updated scenario and insights into future research aiming at validation of medicinal plants as antiophidic agents and strengthens the potentiality of ethnopharmacology as a tool for design of potent inhibitors and/or development of herbal medicines against venom toxins, especially local tissue damage.

  8. A rapid and robust assay for detection of S-phase cell cycle progression in plant cells and tissues by using ethynyl deoxyuridine

    Directory of Open Access Journals (Sweden)

    Horváth Gábor V

    2010-01-01

    Full Text Available Abstract Background Progress in plant cell cycle research is highly dependent on reliable methods for detection of cells replicating DNA. Frequency of S-phase cells (cells in DNA synthesis phase is a basic parameter in studies on the control of cell division cycle and the developmental events of plant cells. Here we extend the microscopy and flow cytometry applications of the recently developed EdU (5-ethynyl-2'-deoxyuridine-based S-phase assay to various plant species and tissues. We demonstrate that the presented protocols insure the improved preservation of cell and tissue structure and allow significant reduction in assay duration. In comparison with the frequently used detection of bromodeoxyuridine (BrdU and tritiated-thymidine incorporation, this new methodology offers several advantages as we discuss here. Results Applications of EdU-based S-phase assay in microscopy and flow cytometry are presented by using cultured cells of alfalfa, Arabidopsis, grape, maize, rice and tobacco. We present the advantages of EdU assay as compared to BrdU-based replication assay and demonstrate that EdU assay -which does not require plant cell wall digestion or DNA denaturation steps, offers reduced assay duration and better preservation of cellular, nuclear and chromosomal morphologies. We have also shown that fast and efficient EdU assay can also be an efficient tool for dual parameter flow cytometry analysis and for quantitative assessment of replication in thick root samples of rice. Conclusions In plant cell cycle studies, EdU-based S-phase detection offers a superior alternative to the existing S-phase assays. EdU method is reliable, versatile, fast, simple and non-radioactive and it can be readily applied to many different plant systems.

  9. ABA biosynthesis defective mutants reduce some free amino acids accumulation under drought stress in tomato leaves in comparison with Arabidopsis plants tissues

    Directory of Open Access Journals (Sweden)

    Adnan Ali Al.Asbahi

    2012-05-01

    Full Text Available The ability of plants to tolerate drought conditions is crucial for plant survival and crop production worldwide. The present data confirm previous findings reported existence of a strong relation between abscisic acid (ABA content and amino acid accumulation as response water stress which is one of the most important defense mechanism activated during water stress in many plant species. Therefore, free amino acids were measured to determine any changes in the metabolite pool in relation to ABA content. The ABA defective mutants of Arabidopsis plants were subjected to leaf dehydration for Arabidopsis on Whatman 3 mm filter paper at room temperature while, tomato mutant plants were subjected to drought stresses for tomato plants by withholding water. To understand the signal transduction mechanisms underlying osmotic stress-regulating gene induction and activation of osmoprotectant free amino acid synthesizing genes, we carried out a genetic screen to isolate Arabidopsis mutants defective in ABA biosynthesis under drought stress conditions. The present results revealed an accumulation of specific free amino acid in water stressed tissues in which majority of free amino acids are increased especially those playing an osmoprotectant role such as proline and glycine. Drought stress related Amino acids contents are significantly reduced in the mutants under water stress condition while they are increased significantly in the wild types plants. The exhibited higher accumulation of other amino acids under stressed condition in the mutant plants suggest that, their expressions are regulated in an ABA independent pathways. In addition, free amino acids content changes during water stress condition suggest their contribution in drought toleration as common compatible osmolytes.

  10. 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

  11. Acetone enhances the direct analysis of total condensed tannins in plant tissues by the butanol-HCl-iron assay

    Science.gov (United States)

    The butanol-HCl spectrophotometric assay is widely used to quantify extractable and insoluble forms of condensed tannin (CT, syn. proanthocyanidin) in foods, feeds, and foliage of herbaceous and woody plants. However, this method underestimates total CT content when applied directly to plant materia...

  12. Analysis on Teaching Reform of Plant Tissue Culture in Lfigher Vocational Education%高等职业院校《植物组织培养》教学改革探析

    Institute of Scientific and Technical Information of China (English)

    白美发

    2009-01-01

    By analyzing the teaching reform of "Plant Tissue Culture"s teaching content,teachinf methods,practical training and examination,the teaching model of Plant Tissue Culture was innovated,and the teaching quality of Plant Tissue Culture was improued.%通过对的教学内容、教学方法、手段以及实训、考试等方面进行了教学改革探讨,创新了高职教学模式,提高了的教学质量.

  13. The combination of quantitative PCR and western blot detecting CP4-EPSPS component in Roundup Ready soy plant tissues and commercial soy-related foodstuffs.

    Science.gov (United States)

    Xiao, Xiao; Wu, Honghong; Zhou, Xinghu; Xu, Sheng; He, Jian; Shen, Wenbiao; Zhou, Guanghong; Huang, Ming

    2012-06-01

    With the widespread use of Roundup Ready soy (event 40-3-2) (RRS), the comprehensive detection of genetically modified component in foodstuffs is of significant interest, but few protein-based approaches have been found useful in processed foods. In this report, the combination of quantitative PCR (qPCR) and western blot was used to detect cp4-epsps gene and its protein product in different RRS plant tissues and commercial soy-containing foodstuffs. The foods included those of plant origin produced by different processing procedures and also some products containing both meat and plant protein concentrates. The validity of the 2 methods was confirmed first. We also showed that the CP4-EPSPS protein existed in different RRS plant tissues. In certain cases, the results from the western blot and the qPCR were not consistent. To be specific, at least 2 degraded fragments of CP4-EPSPS protein (35.5 and 24.6 kDa) were observed. For dried bean curd crust and deep-fried bean curd, a degraded protein fragment with the size of 24.6 kDa appeared, while cp4-epsps gene could not be traced by qPCR. In contrast, we found a signal of cp4-epsps DNA in 3 foodstuffs, including soy-containing ham cutlet product, meat ball, and sausage by qPCR, while CP4-EPSPS protein could not be detected by western blot in such samples. Our study therefore concluded that the combination of DNA- and protein-based methods would compensate each other, thus resulting in a more comprehensive detection from nucleic acid and protein levels. The combination of quantitative PCR (qPCR) and western blot was used to detect cp4-epsps gene and its protein product in different Roundup Ready soy (event 40-3-2) plant tissues and commercial soy-containing foodstuffs. The foods included those of plant origin produced by different processing procedures and also some products containing a combination of both meat and plant protein concentrates. This study indicated that the combination of DNA- and protein-based methods

  14. Understanding cross-communication between aboveground and belowground tissues via transcriptome analysis of a sucking insect whitefly-infested pepper plants.

    Science.gov (United States)

    Park, Yong-Soon; Ryu, Choong-Min

    2014-01-03

    Plants have developed defensive machinery to protect themselves against herbivore and pathogen attacks. We previously reported that aboveground whitefly (Bemisia tabaci Genn.) infestation elicited induced resistance in leaves and roots and influenced the modification of the rhizosphere microflora. In this study, to obtain molecular evidence supporting these plant fitness strategies against whitefly infestation, we performed a 300 K pepper microarray analysis using leaf and root tissues of pepper (Capsicum annuum L.) applied with whitefly, benzo-(1,2,3)-thiadiazole-7-carbothioic acid S-methyl ester (BTH), and the combination of BTH+whitefly. We defined differentially expressed genes (DEGs) as genes exhibiting more than 2-fold change (1.0 based on log2 values) in expression in leaves and roots in response to each treatment compared to the control. We identified a total of 16,188 DEGs in leaves and roots. Of these, 6685, 6752, and 4045 DEGs from leaf tissue and 6768, 7705, and 7667 DEGs from root tissue were identified in the BTH, BTH+whitefly, and whitefly treatment groups, respectively. The total number of DEGs was approximately two-times higher in roots than in whitefly-infested leaves subjected to whitefly infestation. Among DEGs, whitefly feeding induced salicylic acid and jasmonic acid/ethylene-dependent signaling pathways in leaves and roots. Several transporters and auxin-responsive genes were upregulated in roots, which can explain why biomass increase is facilitated. Using transcriptome analysis, our study provides new insights into the molecular basis of whitefly-mediated intercommunication between aboveground and belowground plant tissues and provides molecular evidence that may explain the alteration of rhizosphere microflora and root biomass by whitefly infestation.

  15. Deep RNA-Seq profile reveals biodiversity, plant-microbe interactions and a large family of NBS-LRR resistance genes in walnut (Juglans regia) tissues.

    Science.gov (United States)

    Chakraborty, Sandeep; Britton, Monica; Martínez-García, P J; Dandekar, Abhaya M

    2016-03-01

    Deep RNA-Seq profiling, a revolutionary method used for quantifying transcriptional levels, often includes non-specific transcripts from other co-existing organisms in spite of stringent protocols. Using the recently published walnut genome sequence as a filter, we present a broad analysis of the RNA-Seq derived transcriptome profiles obtained from twenty different tissues to extract the biodiversity and possible plant-microbe interactions in the walnut ecosystem in California. Since the residual nature of the transcripts being analyzed does not provide sufficient information to identify the exact strain, inferences made are constrained to the genus level. The presence of the pathogenic oomycete Phytophthora was detected in the root through the presence of a glyceraldehyde-3-phosphate dehydrogenase. Cryptococcus, the causal agent of cryptococcosis, was found in the catkins and vegetative buds, corroborating previous work indicating that the plant surface supported the sexual cycle of this human pathogen. The RNA-Seq profile revealed several species of the endophytic nitrogen fixing Actinobacteria. Another bacterial species implicated in aerobic biodegradation of methyl tert-butyl ether (Methylibium petroleiphilum) is also found in the root. RNA encoding proteins from the pea aphid were found in the leaves and vegetative buds, while a serine protease from mosquito with significant homology to a female reproductive tract protease from Drosophila mojavensis in the vegetative bud suggests egg-laying activities. The comprehensive analysis of RNA-seq data present also unraveled detailed, tissue-specific information of ~400 transcripts encoded by the largest family of resistance (R) genes (NBS-LRR), which possibly rationalizes the resistance of the specific walnut plant to the pathogens detected. Thus, we elucidate the biodiversity and possible plant-microbe interactions in several walnut (Juglans regia) tissues in California using deep RNA-Seq profiling.

  16. Novel use of positively charged nylon transfer membranes for trapping indoleacetic acid or other small anions during efflux from plant tissues

    Science.gov (United States)

    Evans, M. L.; Hangarter, R. P.

    1993-01-01

    Positively charged nylon blotting membranes were used as an anion binding medium to trap [14C]indoleactic acid (IAA) as it exited cells at the basal ends of Coleus blumei L. stem and Zea mays L. coleoptile segments. Autoradiography was used to visualize where the [14C] that moved out of the cut ends was localized on the nylon membrane. Diffusion of [14C]IAA from the initial point of contact with the nylon membrane was minimal. Comparison of the autoradiograms with anatomical tissue prints of the cut ends of the segments was used to determine what tissues participate in IAA movement. The results of these initial studies were consistent with other reports suggesting that [14C]IAA movement was primarily associated with vascular tissues in both C. blumei stems and corn coleoptiles, but the resolution was not sufficient to identify which vascular tissues were involved in IAA transport. With further refinements, this technique could also be used for studying the movement of other small charged molecules through plant tissues.

  17. Analytical and Radio-Histo-Chemical Experiments of Plants and Tissue Culture Cells Treated with Lunar and Terrestrial Materials

    Science.gov (United States)

    Halliwell, R. S.

    1973-01-01

    The nature and mechanisms of the apparent simulation of growth originally observed in plants growing in contact with lunar soil during the Apollo project quarantine are examined. Preliminary experiments employing neutron activated lunar soil indicate uptake of a few elements by plants. It was found that while the preliminary neutron activation technique allowed demonstration of uptake of minerals it presented numerous disadvantages for use in critical experiments directed at elucidating possible mechanisms of stimulation.

  18. Characterization of the soil organic matter and plant tissues in an initial stage of the plant succession and soil development by means of curie-point pyrolysis coupled with GC-MS

    Science.gov (United States)

    Rahmonov, Oimahmad; Kowalski, Witold J.; Bednarek, Renata

    2010-12-01

    The composition of the soil organic matter in soil developing under the influence of vegetation during the primary succession in the poor sandy area was investigated. The Curie-point pyrolysis method coupled with gas chromatographic separation and mass spectrometric identification of pyrolysates was applied during the investigation. A comparison of occurrence and diversity in composition of organic compounds in plant tissues and humus horizon of soils under the communities of coniferous forest series in the initial stages and phases was carried out. A large diversity of organic compounds under Algae-Cyanophyta communities, biological soil crusts and Polytrichum piliferum was noticed. A clear differentiation in the composition of the soil organic matter at different phases of succession under predominating communities with cryptogamous and vascular plants was observed. The analysis of organic compounds in plant tissues was found to facilitate the determination of origin of various groups of organic compounds in the soil. The results obtained from the Curie point pyrograms in the humus horizon (A) under grasses ( Corynephorus canescens, Koeleria glauca) differed from the pyrolysates obtained under Algae-Cyanophyta communities. The polysaccharide derivates are more frequent in the pyrolysis products under algae, grasses ( Corynephorus canescens, Koeleria glauca) and mosses than under Pinus sylvestris. In the beginning of the terminal stage of succession, lignin, phenols and aliphatic substances, prevailed over the polysaccharide derivates in the humus horizon.

  19. Increasing CO[sub 2] concentration inhibits cytochrome c oxidase (cytox) in vitro, cytochrome pathway (cytpath) activity in plant mitochondria and dark respiration in plant tissue

    Energy Technology Data Exchange (ETDEWEB)

    Gonzalez-Meler, M.A.; Drake, B.G.; Jacob, J. (Smithsonian Environmental Research Center, Edgewater, MD (United States)); Ribas-Carbo, M.; Siedow, J.N. (Duke Univ., Durham, NC (United States)); Aranda, X.; Azcon-Bieto, J.; Palet, A. (Universitat Barcelona (Spain))

    1994-06-01

    Dark respiration is inhibited in many plant be exposure to elevated atmospheric CO[sub 2] concentration. The addition of 0.2mM free CO[sub 2] in the reaction medium decreased citpath activity in Pisum sativum and Glycine max mitochondria at pH 7.2, possibly by inhibiting cytox. Under similar conditions, activity of purified cytox from beef heart was also inhibited. Cytox activity extracted from plants grown in elevated CO[sub 2] for 7 years was lower than in those grown in normal ambient. The relationship among these effects and the rate of respiration as well as the role of the alternative pathway in each case will be discussed.

  20. Capacity of the aquatic fern (Salvinia minima Baker) to accumulate high concentrations of nickel in its tissues, and its effect on plant physiological processes

    Energy Technology Data Exchange (ETDEWEB)

    Fuentes, Ignacio I.; Espadas-Gil, Francisco; Talavera-May, Carlos; Fuentes, Gabriela; Santamaría, Jorge M., E-mail: jorgesm@cicy.mx

    2014-10-15

    Highlights: • We document the capacity of an aquatic fern to hyper-accumulate Ni. • Effects of high Ni concentrations uptake on plant performance is documented. • High concentration of Ni in tissues damage photosynthesis. • Damage is related to carboxylation mechanisms than to electron transfer efficiency. • S. minima is a good candidate for remediation of water bodies contaminated with Ni. - Abstract: An experiment was designed to assess the capacity of Salvinia minima Baker to uptake and accumulate nickel in its tissues and to evaluate whether or not this uptake can affect its physiology. Our results suggest that S. minima plants are able to take up high amounts of nickel in its tissues, particularly in roots. In fact, our results support the idea that S. minima might be considered a hyper-accumulator of nickel, as it is able to accumulate 16.3 mg g{sup −1} (whole plant DW basis). Our results also showed a two-steps uptake pattern of nickel, with a fast uptake of nickel at the first 6 to 12 h of being expose to the metal, followed by a slow take up phase until the end of the experiment at 144 h. S. minima thus, may be considered as a fern useful in the phytoremediation of residual water bodies contaminated with this metal. Also from our results, S. minima can tolerate fair concentrations of the metal; however, at concentrations higher than 80 μM Ni (1.5 mg g{sup −1} internal nickel concentration), its physiological performance can be affected. For instance, the integrity of cell membranes was affected as the metal concentration and exposure time increased. The accumulation of high concentrations of internal nickel did also affect photosynthesis, the efficiency of PSII, and the concentration of photosynthetic pigments, although at a lower extent.

  1. Dealing with the problem of non-specific in situ mRNA hybridization signals associated with plant tissues undergoing programmed cell death

    Directory of Open Access Journals (Sweden)

    Jokela Anne

    2010-02-01

    Full Text Available Abstract Background In situ hybridization is a general molecular method typically used for the localization of mRNA transcripts in plants. The method provides a valuable tool to unravel the connection between gene expression and anatomy, especially in species such as pines which show large genome size and shortage of sequence information. Results In the present study, expression of the catalase gene (CAT related to the scavenging of reactive oxygen species (ROS and the polyamine metabolism related genes, diamine oxidase (DAO and arginine decarboxylase (ADC, were localized in developing Scots pine (Pinus sylvestris L. seeds. In addition to specific signals from target mRNAs, the probes continually hybridized non-specifically in the embryo surrounding region (ESR of the megagametophyte tissue, in the remnants of the degenerated suspensors as well as in the cells of the nucellar layers, i.e. tissues exposed to cell death processes and extensive nucleic acid fragmentation during Scots pine seed development. Conclusions In plants, cell death is an integral part of both development and defence, and hence it is a common phenomenon in all stages of the life cycle. Our results suggest that extensive nucleic acid fragmentation during cell death processes can be a considerable source of non-specific signals in traditional in situ mRNA hybridization. Thus, the visualization of potential nucleic acid fragmentation simultaneously with the in situ mRNA hybridization assay may be necessary to ensure the correct interpretation of the signals in the case of non-specific hybridization of probes in plant tissues.

  2. Engineering the cell wall by reducing de-methyl-esterified homogalacturonan improves saccharification of plant tissues for bioconversion

    Science.gov (United States)

    Lionetti, Vincenzo; Francocci, Fedra; Ferrari, Simone; Volpi, Chiara; Bellincampi, Daniela; Galletti, Roberta; D’Ovidio, Renato; De Lorenzo, Giulia; Cervone, Felice

    2010-01-01

    Plant cell walls represent an abundant, renewable source of biofuel and other useful products. The major bottleneck for the industrial scale-up of their conversion to simple sugars (saccharification), to be subsequently converted by microorganisms into ethanol or other products, is their recalcitrance to enzymatic saccharification. We investigated whether the structure of pectin that embeds the cellulose-hemicellulose network affects the exposure of cellulose to enzymes and consequently the process of saccharification. Reduction of de-methyl-esterified homogalacturonan (HGA) in Arabidopsis plants through the expression of a fungal polygalacturonase (PG) or an inhibitor of pectin methylesterase (PMEI) increased the efficiency of enzymatic saccharification. The improved enzymatic saccharification efficiency observed in transformed plants could also reduce the need for acid pretreatment. Similar results were obtained in PG-expressing tobacco plants and in PMEI-expressing wheat plants, indicating that reduction of de-methyl-esterified HGA may be used in crop species to facilitate the process of biomass saccharification. PMID:20080727

  3. Plant growth and metal distribution in tissues of Prosopis juliflora-velutina grown on chromium contaminated soil in the presence of Glomus deserticola

    OpenAIRE

    Arias, Jack A.; Peralta-Videa, Jose R.; Ellzey, Joanne T.; Viveros, Marian N.; Ren, Minghua; Mokgalaka-Matlala, Ntebogeng S.; Castillo-Michel, Hiram; Gardea-Torresdey, Jorge L.

    2010-01-01

    Arbuscular mycorrhizal fungi have been known to increase metal uptake in plants. In this study, mesquite (Prosopis juliflora-velutina) inoculated with Glomus deserticola or amended with EDTA were grown for 30 days in soil containing Cr(III) or Cr(VI) at 0, 40, 80, and 160 mg kg−1. Total amylase activity (TAA) was monitored as a stress indicator. Element concentrations and distribution in tissue were determined using ICP-OES, electron scanning microprobe, and TEM. Inoculated Cr(VI) treated pla...

  4. Leaf and root volatiles produced by tissue cultures of Alpinia zerumbet (pers. Burtt & Smith under the influence of different plant growth regulators

    Directory of Open Access Journals (Sweden)

    Cristiane Pimentel Victório

    2011-01-01

    Full Text Available Volatiles produced by plantlets of Alpinia zerumbet were obtained by means of simultaneous distillation-extraction (SDE. The effects of indole-3-acetic acid, kinetin, thidiazuron and 6-benzylaminopurine on leaf and root volatile composition obtained by tissue cultures were investigated. A higher content of b-pinene and a lower content of sabinene were observed in leaf volatile of plantlets cultured in control, IAA and IAA+ TDZ media, as compared with those of donor plants. In vitro conditions were favorable to increase caryophyllene content. Volatile compounds from the root were characterized mainly by camphene, fenchyl-acetate and bornyl acetate; which constitute about 60% of total volatile.

  5. Determination of trace amounts of molybdenum in plant tissue by solvent extraction-atomic-absorption and direct-current plasma emission spectrometry.

    Science.gov (United States)

    Lajunen, L H; Kubin, A

    1986-03-01

    Methods are presented for determination of molybdenum in plant tissue by flame and graphite-furnace atomic-absorption spectrometry and direct-current argon-plasma emission spectrometry. The samples are digested in HNO(3)-H(2)SO(4)-HC1O(4) mixture, and Mo is separated and concentrated by chelation and extraction. Three organic solvents (methyl isobutyl ketone, di-isobutyl ketone and isoamyl alcohol) and two ligands (8-hydroxyquinoline and toluene-3,4-dithiol) were studied. The procedure were tested on pine needle and birch leaf samples.

  6. Plant growth and metal distribution in tissues of Prosopis juliflora-velutina grown on chromium contaminated soil in the presence of Glomus deserticola.

    Science.gov (United States)

    Arias, Jack A; Peralta-Videa, Jose R; Ellzey, Joanne T; Viveros, Marian N; Ren, Minghua; Mokgalaka-Matlala, Ntebogeng S; Castillo-Michel, Hiram; Gardea-Torresdey, Jorge L

    2010-10-01

    Arbuscular mycorrhizal fungi have been known to increase metal uptake in plants. In this study, mesquite (Prosopis juliflora-velutina) inoculated with Glomus deserticola or amended with EDTA were grown for 30 days in soil containing Cr(III) or Cr(VI) at 0, 40, 80, and 160 mg kg(-1). Total amylase activity (TAA) was monitored as a stress indicator. Element concentrations and distribution in tissue were determined using ICP-OES, electron scanning microprobe, and TEM. Inoculated Cr(VI) treated plants had 21% and 30% more Cr than uninoculated and EDTA treated roots, respectively, at 80 mg Cr kg(-1) treatment. In the case of Cr(III), EDTA produced the highest Cr accumulation in roots. TAA was higher in inoculated plants grown with Cr(III) at 80 and 160 mg kg(-1) and Cr(VI) at 40 and 160 mg kg(-1). The X-ray mapping showed higher metal concentrations in the vascular system of inoculated plants and the TEM micrographs demonstrated the presence of G. deserticola in roots.

  7. Impact of Temperature and Nutrients on Carbon: Nutrient Tissue Stoichiometry of Submerged Aquatic Plants: An Experiment and Meta-Analysis.

    Science.gov (United States)

    Velthuis, Mandy; van Deelen, Emma; van Donk, Ellen; Zhang, Peiyu; Bakker, Elisabeth S

    2017-01-01

    Human activity is currently changing our environment rapidly, with predicted temperature increases of 1-5°C over the coming century and increased nitrogen and phosphorus inputs in aquatic ecosystems. In the shallow parts of these ecosystems, submerged aquatic plants enhance water clarity by resource competition with phytoplankton, provide habitat, and serve as a food source for other organisms. The carbon:nutrient stoichiometry of submerged aquatic plants can be affected by changes in both temperature and nutrient availability. We hypothesized that elevated temperature leads to higher carbon:nutrient ratios through enhanced nutrient-use efficiency, while nutrient addition leads to lower carbon:nutrient ratios by the luxurious uptake of nutrients. We addressed these hypotheses with an experimental and a meta-analytical approach. We performed a full-factorial microcosm experiment with the freshwater plant Elodea nuttallii grown at 10, 15, 20, and 25°C on sediment consisting of pond soil/sand mixtures with 100, 50, 25, and 12.5% pond soil. To address the effect of climatic warming and nutrient addition on the carbon:nutrient stoichiometry of submerged freshwater and marine plants we performed a meta-analysis on experimental studies that elevated temperature and/or added nutrients (nitrogen and phosphorus). In the microcosm experiment, C:N ratios of Elodea nuttallii decreased with increasing temperature, and this effect was most pronounced at intermediate nutrient availability. Furthermore, higher nutrient availability led to decreased aboveground C:P ratios. In the meta-analysis, nutrient addition led to a 25, 22, and 16% reduction in aboveground C:N and C:P ratios and belowground C:N ratios, accompanied with increased N content. No consistent effect of elevated temperature on plant stoichiometry could be observed, as very few studies were found on this topic and contrasting results were reported. We conclude that while nutrient addition consistently leads to

  8. 3D reconstructions with pixel-based images are made possible by digitally clearing plant and animal tissue

    Science.gov (United States)

    Reconstruction of 3D images from a series of 2D images has been restricted by the limited capacity to decrease the opacity of surrounding tissue. Commercial software that allows color-keying and manipulation of 2D images in true 3D space allowed us to produce 3D reconstructions from pixel based imag...

  9. Enhancing Hispanic Minority Undergraduates' Botany Laboratory Experiences: Implementation of an Inquiry-Based Plant Tissue Culture Module Exercise

    Science.gov (United States)

    Siritunga, Dimuth; Navas, Vivian; Diffoot, Nanette

    2012-01-01

    Early involvement of students in hands-on research experiences are known to demystify research and promote the pursuit of careers in science. But in large enrollment departments such opportunities for undergraduates to participate in research are rare. To counteract such lack of opportunities, inquiry-based laboratory module in plant tissue…

  10. 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.

  11. Evaluation of reference genes for quantitative real-time PCR in oil palm elite planting materials propagated by tissue culture.

    Directory of Open Access Journals (Sweden)

    Pek-Lan Chan

    Full Text Available BACKGROUND: The somatic embryogenesis tissue culture process has been utilized to propagate high yielding oil palm. Due to the low callogenesis and embryogenesis rates, molecular studies were initiated to identify genes regulating the process, and their expression levels are usually quantified using reverse transcription quantitative real-time PCR (RT-qPCR. With the recent release of oil palm genome sequences, it is crucial to establish a proper strategy for gene analysis using RT-qPCR. Selection of the most suitable reference genes should be performed for accurate quantification of gene expression levels. RESULTS: In this study, eight candidate reference genes selected from cDNA microarray study and literature review were evaluated comprehensively across 26 tissue culture samples using RT-qPCR. These samples were collected from two tissue culture lines and media treatments, which consisted of leaf explants cultures, callus and embryoids from consecutive developmental stages. Three statistical algorithms (geNorm, NormFinder and BestKeeper confirmed that the expression stability of novel reference genes (pOP-EA01332, PD00380 and PD00569 outperformed classical housekeeping genes (GAPDH, NAD5, TUBULIN, UBIQUITIN and ACTIN. PD00380 and PD00569 were identified as the most stably expressed genes in total samples, MA2 and MA8 tissue culture lines. Their applicability to validate the expression profiles of a putative ethylene-responsive transcription factor 3-like gene demonstrated the importance of using the geometric mean of two genes for normalization. CONCLUSIONS: Systematic selection of the most stably expressed reference genes for RT-qPCR was established in oil palm tissue culture samples. PD00380 and PD00569 were selected for accurate and reliable normalization of gene expression data from RT-qPCR. These data will be valuable to the research associated with the tissue culture process. Also, the method described here will facilitate the selection

  12. Researching and teaching experiment of ornamental plant tissue culture%观赏植物组织培养教学研究与体会

    Institute of Scientific and Technical Information of China (English)

    李青

    2011-01-01

    长期从事观赏植物组织培养本科教学工作并结合多年从事观赏植物组织培养研究工作及实践;在该课程课堂教学和实验教学中对理论和实践教学形式从多方面探索、改进和逐渐积累的经验总结;同时也从中感悟到在以教师为主导的教学中注意激发学生的学习热情,以达到更好的“教”和“学”的效果。%This paper is summary of the author has long been engaged in ornamental plant tissue culture undergraduate teaching and combined with years of practice of studies on ornamental plant tissue culture. In the course of classroom instruction and laboratory teaching, the author explored the theoretical and practical teaching methods in many ways, kept on improving and accumulating it, and also realized that the teacherled instruction should pay attention to stimulate learning enthusiasm of students, in order to achieve better effect of teaching and learning.

  13. Capacity of the aquatic fern (Salvinia minima Baker) to accumulate high concentrations of nickel in its tissues, and its effect on plant physiological processes.

    Science.gov (United States)

    Fuentes, Ignacio I; Espadas-Gil, Francisco; Talavera-May, Carlos; Fuentes, Gabriela; Santamaría, Jorge M

    2014-10-01

    An experiment was designed to assess the capacity of Salvinia minima Baker to uptake and accumulate nickel in its tissues and to evaluate whether or not this uptake can affect its physiology. Our results suggest that S. minima plants are able to take up high amounts of nickel in its tissues, particularly in roots. In fact, our results support the idea that S. minima might be considered a hyper-accumulator of nickel, as it is able to accumulate 16.3 mg g(-1) (whole plant DW basis). Our results also showed a two-steps uptake pattern of nickel, with a fast uptake of nickel at the first 6 to 12h of being expose to the metal, followed by a slow take up phase until the end of the experiment at 144 h. S. minima thus, may be considered as a fern useful in the phytoremediation of residual water bodies contaminated with this metal. Also from our results, S. minima can tolerate fair concentrations of the metal; however, at concentrations higher than 80 μM Ni (1.5 mg g(-1) internal nickel concentration), its physiological performance can be affected. For instance, the integrity of cell membranes was affected as the metal concentration and exposure time increased. The accumulation of high concentrations of internal nickel did also affect photosynthesis, the efficiency of PSII, and the concentration of photosynthetic pigments, although at a lower extent. Copyright © 2014 Elsevier B.V. All rights reserved.

  14. Copper-Containing Amine Oxidases and FAD-Dependent Polyamine Oxidases Are Key Players in Plant Tissue Differentiation and Organ Development.

    Science.gov (United States)

    Tavladoraki, Paraskevi; Cona, Alessandra; Angelini, Riccardo

    2016-01-01

    Plant polyamines are catabolized by two classes of amine oxidases, the copper amine oxidases (CuAOs) and the flavin adenine dinucleotide (FAD)-dependent polyamine oxidases (PAOs). These enzymes differ to each other in substrate specificity, catalytic mechanism and subcellular localization. CuAOs and PAOs contribute to several physiological processes both through the control of polyamine homeostasis and as sources of biologically-active reaction products. CuAOs and PAOs have been found at high level in the cell-wall of several species belonging to Fabaceae and Poaceae families, respectively, especially in tissues fated to undertake extensive wall loosening/stiffening events and/or in cells undergoing programmed cell death (PCD). Apoplastic CuAOs and PAOs have been shown to play a key role as a source of H2O2 in light- or developmentally-regulated differentiation events, thus influencing cell-wall architecture and maturation as well as PCD. Moreover, growing evidence suggests a key role of intracellular CuAOs and PAOs in several facets of plant development. Here, we discuss recent advances in understanding the contribution of different CuAOs/PAOs, as well as their cross-talk with different intracellular and apoplastic metabolic pathways, in tissue differentiation and organ development.

  15. Second harmonic imaging of plants tissues and cell implosion using two-photon process in ZnO nanoparticles.

    Science.gov (United States)

    Urban, Ben E; Neogi, Purnima B; Butler, Sween J; Fujita, Yasuhisa; Neogi, Arup

    2012-03-01

    The optical properties of colloidal ZnO nanoparticle (NP) solutions, with size ranging from several nm to around 200 nm, have been tailored to have high optical nonlinearity for bioimaging with no auto-fluorescence above 750 nm and minimal auto-fluorescence below 750 nm. The high second harmonic conversion efficiency enables selective tissue imaging and cell tracking using tunable near-infrared femtosecond laser source ranging from 750-980 nm. For laser energies exceeding the two-photon energy of the bandgap of ZnO (half of 3.34 eV), the SHG signal greatly decreases and the two-photon emission becomes the dominant signal. The heat generated due to two-photon absorption within the ZnO NPs enable selective cell or localized tissue destruction using excitation wavelength ranging from 710-750 nm. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  16. 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.

  17. Assessment of phosphopeptide enrichment/precipitation method for LC-MS/MS based phosphoproteomic analysis of plant tissue

    DEFF Research Database (Denmark)

    Ye, Juanying; Rudashevskaya, Elena; Hansen, Thomas Aarup

      Introduction Mass spectrometry (MS) is a powerful technology for study of PTMs, including protein phosphorylation. Due to the low abundance of many phosphoproteins and the relatively poor ionization efficiency of phosphopeptides, specific enrichment of phosphopeptides prior to MS analysis is ne...... (MSA) method was used for phosphopeptide fragmentation. The resulting fragment ion spectra were processed with Proteome Discoverer software (Thermo Electron, Bremen, Germany). Results We first investigated the global phosphorylation profile of plant plasma membrane proteins by enriching...

  18. A study of starch gelatinisation behaviour in hydrothermally-processed plant food tissues and implications for in vitro digestibility.

    Science.gov (United States)

    Edwards, Cathrina H; Warren, Frederick J; Campbell, Grant M; Gaisford, Simon; Royall, Paul G; Butterworth, Peter J; Ellis, Peter R

    2015-12-01

    The aim of this study was to investigate the role of the plant food matrix in influencing the extent of starch gelatinisation during hydrothermal processing, and its implications for starch digestibility. Differential scanning calorimetry (DSC) was used to provide a detailed examination of the gelatinisation behaviour of five distinct size fractions (diameters food materials. We observed clear differences in the gelatinisation behaviour of matched size-fractions of chickpeas and durum wheat. In chickpea materials, the TEG values (34-100%) were inversely related to particle size, whereas in durum wheat, no size-dependent limitations on TEG were observed. The TEG values were completely consistent with the extent of starch amylolysis in all size fractions of both durum wheat and chickpea. Microstructural analysis following hydrothermal processing confirmed the presence of some partially gelatinised birefringent starch within intact chickpea cells. Birefringent starch granules were not present in any of the processed fractions of durum wheat. The differences in gelatinisation behaviour of these plant species seem to reflect the individual cell wall properties of these materials. These findings demonstrate the applicability of DSC to real food materials to provide insight into the mechanisms by which the food matrix (particularly the plant cell walls) influences gelatinisation, and consequently, starch amylolysis.

  19. Postharvest application of organic and inorganic salts to control potato (Solanum tuberosum L.) storage soft rot: plant tissue-salt physicochemical interactions.

    Science.gov (United States)

    Yaganza, E S; Tweddell, R J; Arul, J

    2014-09-24

    Soft rot caused by Pectobacterium sp. is a devastating disease affecting stored potato tubers, and there is a lack of effective means of controlling this disease. In this study, 21 organic and inorganic salts were tested for their ability to control soft rot in potato tubers. In the preventive treatment, significant control of soft rot was observed with AlCl3 (≥66%) and Na2S2O3 (≥57%) and to a lesser extent with Al lactate and Na benzoate (≥34%) and K sorbate and Na propionate (≥27%). However, only a moderate control was achieved by curative treatment with AlCl3 and Na2S2O3 (42%) and sodium benzoate (≥33%). Overall, the in vitro inhibitory activity of salts was attenuated in the presence of plant tissue (in vivo) to different degrees. The inhibitory action of the salts in the preventive treatment, whether effective or otherwise, showed an inverse linear relationship with water ionization capacity (pK') of the salt ions, whereas in the curative treatment, only the effective salts showed this inverse linear relationship. Salt-plant tissue interactions appear to play a central role in the attenuated inhibitory activity of salts in potato tuber through reduction in the availability of the inhibitory ions for salt-bacteria interactions. This study demonstrates that AlCl3, Na2S2O3, and Na benzoate have potential in controlling potato tuber soft rot and provides a general basis for understanding of specific salt-tissue interactions.

  20. Establishment of a rapid, inexpensive protocol for extraction of high quality RNA from small amounts of strawberry plant tissues and other recalcitrant fruit crops.

    Science.gov (United States)

    Christou, Anastasis; Georgiadou, Egli C; Filippou, Panagiota; Manganaris, George A; Fotopoulos, Vasileios

    2014-03-01

    Strawberry plant tissues and particularly fruit material are rich in polysaccharides and polyphenolic compounds, thus rendering the isolation of nucleic acids a difficult task. This work describes the successful modification of a total RNA extraction protocol, which enables the isolation of high quantity and quality of total RNA from small amounts of strawberry leaf, root and fruit tissues. Reverse-transcription polymerase chain reaction (RT-PCR) amplification of GAPDH housekeeping gene from isolated RNA further supports the proposed protocol efficiency and its use for downstream molecular applications. This novel procedure was also successfully followed using other fruit tissues, such as olive and kiwifruit. In addition, optional treatment with RNase A following initial nucleic acid extraction can provide sufficient quality and quality of genomic DNA for subsequent PCR analyses, as evidenced from PCR amplification of housekeeping genes using extracted genomic DNA as template. Overall, this optimized protocol allows easy, rapid and economic isolation of high quality RNA from small amounts of an important fruit crop, such as strawberry, with extended applicability to other recalcitrant fruit crops.

  1. An Uncommon Plant Growth Regulator, Diethyl Aminoethyl Hexanoate, Is Highly Effective in Tissue Cultures of the Important Medicinal Plant Purple Coneflower (Echinacea purpurea L.

    Directory of Open Access Journals (Sweden)

    Xiao-Lu Chen

    2013-01-01

    Full Text Available We investigated the effects of various concentrations of diethyl aminoethyl hexanoate (DA-6 on the regeneration and growth of adventitious buds in in vitro purple coneflower cultures. Among the 3 types of explants tested, leaf explants required higher concentrations of DA-6 than petiole and root explants in order to obtain high regeneration rates, while root explants required the lowest concentration of DA-6. Additionally, explants with higher ploidy levels were more sensitive to the addition of DA-6, while explants with lower ploidy levels required higher concentrations of DA-6 to achieve its maximal regeneration rate. Interestingly, the application of a concentration that was conducive to the regeneration of explants with lower ploidy levels was inhibitory to the regeneration of explants with higher ploidy levels. Moreover, during the growth of regenerated buds, DA-6 application significantly improved plant height and weight, root weight, root thickness, root number, primary root length, total root length, and root/top ratio. Differences in the responses of explants to supplementation with DA-6 were also observed among explants with different ploidy levels, with buds having lower ploidy levels responding to lower concentrations of DA-6. Taken together, the results of the present experiments showed that proper application of DA-6 could increase in vitro culture efficiency in purple coneflower.

  2. A study of arsenic speciation in soil, irrigation water and plant tissue: A case study of the broad bean plant, Vicia faba.

    Science.gov (United States)

    Sadee, Bashdar A; Foulkes, Mike E; Hill, Steve J

    2016-11-01

    Samples of soil, the broad bean plant, Vicia faba and irrigation water were collected from the same agricultural site in Dokan, in the Kurdistan region of Iraq. Total arsenic and arsenic speciation were determined in all materials by ICP-MS and HPLC-ICP-MS, respectively. Available arsenic (11%) was also determined within the soil, together with Cd, Cr, Cu, Ni, Zn, Fe and Mn. The concentrations of total arsenic were: soil (5.32μgg(-1)), irrigation water (1.06μgL(-1)), roots (2.065μgg(-1)) and bean (0.133μgg(-1)). Stems, leaves and pods were also measured. Inorganic As(V) dominated soil (90%) and root (78%) samples. However, organo-arsenic (MMA, 48% and DMA, 19%) was the more dominant species in the edible bean. The study provides an insight into the uptake, preferred disposal route, speciation changes and loss mechanism involved for arsenic with this food source. Copyright © 2016. Published by Elsevier Ltd.

  3. An uncommon plant growth regulator, diethyl aminoethyl hexanoate, is highly effective in tissue cultures of the important medicinal plant purple coneflower (Echinacea purpurea L.).

    Science.gov (United States)

    Chen, Xiao-Lu; Zhang, Jun-Jie; Chen, Rong; Li, Qing-Ling; Yang, Yue-Sheng; Wu, Hong

    2013-01-01

    We investigated the effects of various concentrations of diethyl aminoethyl hexanoate (DA-6) on the regeneration and growth of adventitious buds in in vitro purple coneflower cultures. Among the 3 types of explants tested, leaf explants required higher concentrations of DA-6 than petiole and root explants in order to obtain high regeneration rates, while root explants required the lowest concentration of DA-6. Additionally, explants with higher ploidy levels were more sensitive to the addition of DA-6, while explants with lower ploidy levels required higher concentrations of DA-6 to achieve its maximal regeneration rate. Interestingly, the application of a concentration that was conducive to the regeneration of explants with lower ploidy levels was inhibitory to the regeneration of explants with higher ploidy levels. Moreover, during the growth of regenerated buds, DA-6 application significantly improved plant height and weight, root weight, root thickness, root number, primary root length, total root length, and root/top ratio. Differences in the responses of explants to supplementation with DA-6 were also observed among explants with different ploidy levels, with buds having lower ploidy levels responding to lower concentrations of DA-6. Taken together, the results of the present experiments showed that proper application of DA-6 could increase in vitro culture efficiency in purple coneflower.

  4. Effective half-lives of ¹³⁷Cs from persimmon tree tissue parts in Japan after Fukushima Dai-ichi Nuclear Power Plant accident.

    Science.gov (United States)

    Tagami, Keiko; Uchida, Shigeo

    2015-03-01

    To estimate the radiocesium decreasing rates from persimmon trees during a period of about 3 y following the accident at Tokyo Electric Power Company's Fukushima Dai-ichi Nuclear Power Plant (FDNPP), we conducted measurements of tree tissue parts collected in 2011-2013. The sampling was carried out in Chiba, 220 km south of FDNPP; radioactive fallouts discharged from FDNPP had mainly been observed in March-April 2011 on the sampling site. We measured (137)Cs concentrations in the tree tissue parts, i.e., fruits (flesh, skin and seeds), leaves and newly emerged branches, and then the effective half-lives (T(eff)) of (137)Cs were calculated. Leaf samples were classified into two types by sampling months according to the growing stages, that is, immature (April-May) and mature (June-November) leaves. All these parts showed exponential declines in (137)Cs concentration with good adjusted contribution ratios of higher than ca. 0.7. The calculated T(eff) values from all tissue parts were similar with the average of 229 d (range: 216-243 d). From these results, we concluded that each tree tissue was representative for the calculation of Teff. For comparison to these observation results, open source food monitoring data from 2011 to 2013 including (137)Cs data for persimmon fruits collected in Fukushima Prefecture were used to calculate T(eff) for persimmon trees. Values of Teff were obtained for persimmon fruits grown in each local government area in Fukushima Prefecture and they ranged from 303 to 475 d.

  5. Extraction of nucleic acids from yeast cells and plant tissues using ethanol as medium for sample preservation and cell disruption.

    Science.gov (United States)

    Linke, Bettina; Schröder, Kersten; Arter, Juliane; Gasperazzo, Tatiana; Woehlecke, Holger; Ehwald, Rudolf

    2010-09-01

    Here we report that dehydrated ethanol is an excellent medium for both in situ preservation of nucleic acids and cell disruption of plant and yeast cells. Cell disruption was strongly facilitated by prior dehydration of the ethanol using dehydrated zeolite. Following removal of ethanol, nucleic acids were extracted from the homogenate pellet using denaturing buffers. The method provided DNA and RNA of high yield and integrity. Whereas cell wall disruption was essential for extraction of DNA and large RNA molecules, smaller molecules such as tRNAs could be selectively extracted from undisrupted, ethanol-treated yeast cells. Our results demonstrate the utility of absolute ethanol for sample fixation, cell membrane and cell wall disruption, as well as preservation of nucleic acids during sample storage.

  6. In vitro Plant Regeneration from the Mature Tissue of Navel Orange (Citrus sinensis L. Osbeck) by Direct Organogenesis

    Institute of Scientific and Technical Information of China (English)

    HUANG Jia-quan; YIN Li-yan; YANG Xiao-hong; SUN Zhong-hai

    2005-01-01

    An efficient in vitro regeneration system by direct organogenesis from mature nodal and internodal stem segments of Newhall navel orange (Citrus sinensis L. Osbeck) was developed. Illuminating conditions together with plant growth regulators affected the adventitious bud regeneration frequency and efficiency. The initial 15 d darkness inoculation is beneficial for the adventitious bud regeneration. The highest regeneration frequency (85.2%) and bud formation efficiency (3.7 per responsive internodal stem segment) were obtained in the media supplemented with 1.0 mg L-1 BAP and 0.5 mg L-1 NAA. ABA at 0.2 mg L-1 positively affected the bud formation efficiency, which amounted to 8.5 buds per internodal segment in the presence of BAP at 1.0 mg L-1. The adventitious shoots successfully rooted and were transferred to the soil.

  7. Separation and quantification of monothiols and phytochelatins from a wide variety of cell cultures and tissues of trees and other plants using high performance liquid chromatography.

    Science.gov (United States)

    Minocha, Rakesh; Thangavel, P; Dhankher, Om Parkash; Long, Stephanie

    2008-10-17

    The HPLC method presented here for the quantification of metal-binding thiols is considerably shorter than most previously published methods. It is a sensitive and highly reproducible method that separates monobromobimane tagged monothiols (cysteine, glutathione, gamma-glutamylcysteine) along with polythiols (PC(2), PC(3), PC(4) and PC(5)) within 23min from a wide variety of samples. Total run time of the method is 35min. Detection limits for thiols is 33fmol for 10microlL injection. This method will be applicable to study the metal detoxification mechanisms for a wide variety of cell cultures and tissues of plants and trees including algae, Arabidopsis, crambe, rice, and red spruce.

  8. 异体组织埋藏法改善视网膜色素变性%Discuss sight function for the patients with retinitis pigmentosa by planting foreign tissue

    Institute of Scientific and Technical Information of China (English)

    付群; 杨尊之; 毕毅敏

    2001-01-01

    @@Background: Retinitis pigmentosa(RP) mostly harms sensitization organ and pigment epidermis . It is a progress and transmissible eye illness .It has severity harm to patients's eyesight.But now without specially cure. Objective: To introduce planting foreign tissue cure retinitis pigmentosa and appraise sight function.Design: we cure the patients with retinitis pigmentosa by planting foreign tissue. The patients have next one or two clinical symptoms. (1)Moon blindness;(2) Dwindle in vision field;(3)Retinal vas attenuate in evidence. Vas Choroid can been see cirrhosis.(4)The wave and wave's swing debases in retinal electricity chart.(5) Usually by heredity . After ,we value the visual acuity and visual field for treatment and aftertreatment , know that the complexion rejuvenate sight function.At one time, we confirm that planting foreign tissue is a valid method.

  9. Modulation of selenium tissue distribution and selenoprotein expression in Atlantic salmon (Salmo salar L.) fed diets with graded levels of plant ingredients.

    Science.gov (United States)

    Betancor, Monica B; Dam, Thi M C; Walton, James; Morken, Thea; Campbell, Patrick J; Tocher, Douglas R

    2016-04-01

    Increased substitution of marine ingredients by terrestrial plant products in aquafeeds has been proven to be suitable for Atlantic salmon farming. However, a reduction in n-3 long-chain PUFA is a consequence of this substitution. In contrast, relatively little attention has been paid to the effects of fishmeal and oil substitution on levels of micronutrients such as Se, considering fish are major sources of this mineral for human consumers. To evaluate the effects of dietary marine ingredient substitution on tissue Se distribution and the expression of Se metabolism and antioxidant enzyme genes, Atlantic salmons were fed three feeds based on commercial formulations with increasing levels of plant proteins (PP) and vegetable oil. Lipid content in flesh did not vary at any sampling point, but it was higher in the liver of 1 kg of fish fed higher PP. Fatty acid content reflected dietary input and was related to oxidation levels (thiobarbituric acid-reactive substances). Liver had the highest Se levels, followed by head kidney, whereas the lowest contents were found in brain and gill. The Se concentration of flesh decreased considerably with high levels of substitution, reducing the added value of fish consumption. Only the brain showed significant differences in glutathione peroxidase, transfer RNA selenocysteine 1-associated protein 1b and superoxide dismutase expression, whereas no significant regulation of Se-related genes was found in liver. Although Se levels in the diets satisfied the essential requirements of salmon, high PP levels led to a reduction in the supply of this essential micronutrient.

  10. Indirect regeneration from in vitro leaf tissue of periwinkle (Catharanthus roseus L. in response to different treatments of plant growth regulators

    Directory of Open Access Journals (Sweden)

    B.E. Sayed-Tabatabaei

    2012-03-01

    Full Text Available Periwinkle (Catharanthus roseus L. belongs to the Apocynaceae family and accumulates more than 130 terpenoid indole alkaloids (TIAs, of which two dimeric alkaloids Vinblastine and Vincristine have antineoplastic activity and are useful for treatment of various cancers. Therefore, the production of these drugs has been emphasized in plant tissue culture. In this research, 25 treatments of plant growth regulators to produce callus from leaf explants and seven treatments for regeneration of calli were considered. Analysis of variance showed that the effect of different hormonal treatments in the production of callus from leaf explants is significant (P<0.01. Application of 0.1 mg/L BAP and 5 mg/L NAA in nutrient media produced a lot of calli and roots. In addition, nutrient media containing activated charcoal and without it were found to be suitable for production of callus and regeneration, respectively. In conclusion, the results showed that indirect regeneration of leaf explants is not a suitable method for micropropagation of periwinkle due to difficult regeneration of callus, probability of mutation and lower number of produced branches. But this method can be utilized for production of secondary metabolites.

  11. Aequorin-Based Luminescence Imaging Reveals Stimulus- and Tissue-Specific Ca2+ Dynamics in Arabidopsis Plants

    Institute of Scientific and Technical Information of China (English)

    Xiaohong Zhu; Ying Feng; Gaimei Liang; Na Liu; Jian-Kang Zhu

    2013-01-01

    Calcium ion is a versatile second messenger for diverse cell signaling in response to developmental and environmental cues.The specificity of Ca2+-mediated signaling is defined by stimulus-elicited Ca2+ signature and down-stream decoding processes.Here,an Aequorin-based luminescence recording system was developed for monitoring Ca2+ in response to various stimuli in Arabidopsis.With the simple,highly sensitive,and robust Ca2+ recording,this system revealed stimulus-and tissue-specific Ca2+ signatures in seedlings.Cellular Ca2+ dynamics and relationship to Aequorin-based Ca2+ recording were explored using a GFP-based Ca2+ indicator,which suggested that a synchronous cellular Ca2+ signal is responsible for cold-induced Ca2+ response in seedlings,whereas asynchronous Ca2+ oscillation contributes to osmotic stress-induced Ca2+ increase in seedlings.The optimized recording system would be a powerful tool for the identification and characterization of novel components in Ca2+-mediated stress-signaling pathways.

  12. [Plant lectins and embryonic tissue factors as probes for studying the mechanisms of neural induction in amphibians].

    Science.gov (United States)

    Mikhaĭlov, A T; Gorgoliuk, N A

    1992-01-01

    Using various experimental techniques, we have demonstrated that animal pole ectoderm (APE) of Rana temporaria embryos at the stage of early gastrula is a good target tissue for testing the neuralizing (N) factors. In this respect R. temporaria APE is comparable with APE of some other amphibian species. We found that concanavalin A (con A), phytohemagglutinin (PHA) and embryonic brain-derived neuralizing factor (EBDNF; a factor extracted from the chick embryonic brain and partially purified) have a pronounced N-effect on the APE of R. temporaria. In order to analyse possible mechanisms of N-action of these factors, we have cultured APE explants for 3 or 18 h in the medium containing various concentrations of con A, PHA of EBDNF. All these factors could produce neuralization in 50% explants. However, the optimal concentration and time of exposure were different. This is an evidence for different mechanisms of reception and transmission of a N-signal in each particular case. It appears that the APE consists of several cell subpopulations which differ in their threshold sensitivity to the N-effect of studied agents.

  13. Development and function of membrane systems in plant tissue. Annual technical progress report, 15 September 1981-15 August 1982

    Energy Technology Data Exchange (ETDEWEB)

    Hanson, J B

    1982-01-01

    Over the past 11 months we have continued investigation of ion transport mechanisms in corn roots and mitochondria. In mitochondria we find that only citrate and isocitrate are transported by the H/sup +//citrate symporter. However, the in vivo function of this carrier remains in doubt because citrate does not appear to be an effective substrate for corn mitochondria. Studies with roots have been directed to why various types of injury or shock all result in temporary blockage of the H/sup +/-efflux pump in the plasmamembrane. It appears this may be due to an injury-mediated Ca/sup 2 +/ influx into the tissue, which by raising free Ca/sup 2 +/ in the cytosal activates calmodulin (CaM). In turn, the Ca.CaM complex appears to activate protein kinase, phosphorylating membrane proteins. It is possible that one of these phosphorylated proteins is responsible for inactivation of the H/sup +/-ATPase. Future work is planned around the consequences of Ca/sup 2 +/ influx into the root cell subsequent to injury, investigating the recovery of the H/sup +/-ATPase and the initiation of the biosyntheses which lead to augmented ion transport.

  14. Plant tissue culture independent Agrobacterium tumefaciens mediated In-planta transformation strategy for upland cotton (Gossypium hirsutum

    Directory of Open Access Journals (Sweden)

    Bipinchandra B. Kalbande

    2016-06-01

    Full Text Available A new method of transgenic development called “In-planta” transformation method, where Agrobacterium is used to infect the plantlets but the steps of in vitro regeneration of plants is totally avoided. In this study, we have reported a simple In-planta method for efficient transformation of diploid cotton Gossypium hirsutum cv LRK-516 Anjali using Agrobacterium tumefaciens EHA-105 harbouring recombinant binary vector plasmid pBinAR with Arabidopsis At-NPR1 gene. Four day old plantlets were used for transformation. A vertical cut was made at the junction of cotyledonary leaves, moderately bisecting the shoot tip and exposing meristem cells at apical meristem. This site was infected with Agrobacterium inoculum. The transgenic events obtained were tested positive for the presence of At-NPR1 gene with promoter nptII gene. They are also tested negative for vector backbone integration and Agrobacterium contamination in T0 events. With this method a transformation frequency of 6.89% was reported for the cv LRK-516.

  15. Exploration and Practice on Experiment Teaching Reform of Plant Tissue Culture%植物组织培养实验教学改革探索与实践

    Institute of Scientific and Technical Information of China (English)

    尚宏芹

    2011-01-01

    针对植物组织培养实验课程教学中存在的问题,从教学内容、教学方式、教学过程、考核方式等方面对植物组织培养实验教学改革进行了探索.%Aiming at the problems in the experiment teaching of plant tissue culture,exploration was carried on experiment teaching reform of plant tissue culture from teaching content,teaching methods,teaching process and evaluation methods.

  16. Development and analysis of a highly flexible multi-gene expression system for metabolic engineering in Arabidopsis seeds and other plant tissues.

    Science.gov (United States)

    Shockey, Jay; Mason, Catherine; Gilbert, Matthew; Cao, Heping; Li, Xiangjun; Cahoon, Edgar; Dyer, John

    2015-09-01

    Production of novel value-added compounds in transgenic crops has become an increasingly viable approach in recent years. However, in many cases, product yield still falls short of the levels necessary for optimal profitability. Determination of the limiting factors is thus of supreme importance for the long-term viability of this approach. A significant challenge to most metabolic engineering projects is the need for strong coordinated co-expression of multiple transgenes. Strong constitutive promoters have been well-characterized during the >30 years since plant transformation techniques were developed. However, organ- or tissue-specific promoters are poorly characterized in many cases. Oilseeds are one such example. Reports spanning at least 20 years have described the use of certain seed-specific promoters to drive expression of individual transgenes. Multi-gene engineering strategies are often hampered by sub-optimal expression levels or improper tissue-specificity of particular promoters, or rely on the use of multiple copies of the same promoter, which can result in DNA instability or transgene silencing. We describe here a flexible system of plasmids that allows for expression of 1-7 genes per binary plasmid, and up to 18 genes altogether after multiple rounds of transformation or sexual crosses. This vector system includes six seed-specific promoters and two constitutive promoters. Effective constitutive and seed-specific RNA interference gene-suppression cloning vectors were also constructed for silencing of endogenous genes. Taken together, this molecular toolkit allows combinatorial cloning for multiple transgene expression in seeds, vegetative organs, or both simultaneously, while also providing the means to coordinately overexpress some genes while silencing others.

  17. The induction of stromule formation by a plant DNA-virus in epidermal leaf tissues suggests a novel intra- and intercellular macromolecular trafficking route

    Directory of Open Access Journals (Sweden)

    Björn eKrenz

    2012-12-01

    Full Text Available Stromules are dynamic thin protrusions of membrane envelope from plant cell plastids. Despite considerable progress in understanding the importance of certain cytoskeleton elements and motor proteins for stromule maintenance, their function within the cell has yet to be unraveled. Several viruses cause a remodulation of plastid structures and stromule biogenesis within their host plants. For RNA-viruses these interactions were demonstrated to be relevant to the infection process. An involvement of plastids and stromules is assumed in the DNA-virus life cycle as well, but their functional role needs to be determined. Recent findings support a participation of heat shock cognate 70 kDa protein (cpHSC70-1-containing stromules induced by a DNA-virus infection (Abutilon mosaic virus, AbMV, Geminiviridae in intra- and intercellular molecule exchange. The chaperone cpHSC70-1 was shown to interact with the AbMV movement protein (MP. Bimolecular fluorescence complementation confirmed the interaction of cpHSC70-1 and MP, and showed a homo-oligomerization of either protein in planta. The complexes were detected at the cellular margin and co-localized with plastids. In healthy plant tissues cpHSC70-1-oligomers occurred in distinct spots at chloroplasts and in small filaments extending from plastids to the cell periphery. AbMV-infection induced a cpHSC70-1-containing stromule network that exhibits elliptical dilations and transverses whole cells. Silencing of the cpHSC70-gene revealed an impact of cpHSC70 on chloroplast stability and restricted AbMV movement, but not viral DNA accumulation. Based on these data, a model is suggested in which these stromules function in molecule exchange between plastids and other organelles and perhaps other cells. AbMV may utilize cpHSC70-1 for trafficking along plastids and stromules into a neighboring cell or from plastids into the nucleus. Experimental approaches to investigate this hypothesis are discussed.

  18. Influence of subacute treatment of some plant growth regulators on serum marker enzymes and erythrocyte and tissue antioxidant defense and lipid peroxidation in rats.

    Science.gov (United States)

    Celik, Ismail; Tuluce, Yasin; Isik, Ismail

    2006-01-01

    This study aims to investigate the effects of the plant growth regulators (PGRs) (2,3,5-triiodobenzoic acid (TIBA), Naphthaleneacetic acid (NAA), and 2,4-dichlorofenoxyacetic acid (2,4-D)) on serum marker enzymes (aspartate aminotransferase (AST), alanin aminotransferase (ALT), creatine phosphokinase (CPK), and lactate dehydrogenase (LDH)), antioxidant defense systems (reduced glutathione (GSH), glutathione reductase (GR), superoxide dismutase (SOD), glutathione-S-transferase (GST), and catalase (CAT)), and lipid peroxidation content (malondialdehyde = MDA) in various tissues of rats. 50 and 100 ppm of PGRs as drinking water were administered orally to rats (Sprague-Dawley albino) ad libitum for 25 days continuously. The PGRs treatment caused different effects on the serum marker enzymes, antioxidant defense systems, and the MDA content in experimented rats compared to controls. Results showed that TIBA caused a significant decrease in serum AST activity with both the dosage whereas serum CPK was significantly increased with 100 ppm dosage of TIBA. Meanwhile, serum AST, CPK, and LDH activities were significantly increased with both dosage of NAA and 2,4-D. The lipid peroxidation end-product MDA significantly increased in the all tissues treated with both dosages of PGRs without any change in the brain and erythrocyte of rats treated with both the dosages of 2,4-D. The GSH depletion in the kidney and brain tissues of rats treated with both dosages of PGRs was found to be significant. Furthermore, the GSH depletion in the erythrocyte of rats treated with both dosages of PGRs except 50 ppm dosage of 2,4-D was significant too. Also, the GSH level in the liver was significantly depleted with 50 ppm of 2,4-D and NAA, whereas the GSH depletion in the same tissue did not significantly change with the treatment. The activity of antioxidant enzymes was also seriously affected by PGRs; SOD significantly decreased in the liver, heart, kidney, and brain of rats treated with

  19. The spatial distribution of acid phosphatase activity in ectomycorrhizal tissues depends on soil fertility and morphotype, and relates to host plant phosphorus uptake.

    Science.gov (United States)

    Alvarez, Maricel; Huygens, Dries; Díaz, Leila Milena; Villanueva, Claudia Añazco; Heyser, Wolfgang; Boeckx, Pascal

    2012-01-01

    Acid phosphatase (ACP) enzymes are involved in the mobilization of soil phosphorus (P) and polyphosphate accumulated in the fungal tissues of ectomycorrhizal roots, thereby influencing the amounts of P that are stored in the fungus and transferred to the host plant. This study evaluated the effects of ectomycorrhizal morphotype and soil fertility on ACP activity in the extraradical mycelium (ACP(myc)), the mantle (ACP(mantle)) and the Hartig net region (ACP(Hartig)) of ectomycorrhizal Nothofagus obliqua seedlings. ACP activity was quantified in vivo using enzyme-labelled fluorescence-97 (ELF-97) substrate, confocal laser microscopy and digital image processing routines. There was a significant effect of ectomycorrhizal morphotype on ACP(myc), ACP(mantle) and ACP(Hartig), while soil fertility had a significant effect on ACP(myc) and ACP(Hartig). The relative contribution of the mantle and the Hartig net region to the ACP activity on the ectomycorrhizal root was significantly affected by ectomycorrhizal morphotype and soil fertility. A positive correlation between ACP(Hartig) and the shoot P concentration was found, providing evidence that ACP activity at the fungus:root interface is involved in P transfer from the fungus to the host. It is concluded that the spatial distribution of ACP in ectomycorrhizas varies as a function of soil fertility and colonizing fungus.

  20. Quantitative analysis of gibberellins in plant tissues by GPC-HPLC-LC/MS%GPC-HPLC-LC/MS测定植物组织中的赤霉素

    Institute of Scientific and Technical Information of China (English)

    李金克; 邓文红; 陈少良

    2014-01-01

    以胡杨叶片、烟草组培苗及萌发绿豆为材料,研发了利用凝胶渗透色谱( GPC)纯化、液相质谱( LC-MS)定性、高效液相色谱( HPLC)定量测定植物组织中赤霉素( GAS )的方法。植物样品用80%甲醇研磨后4℃浸提过夜,抽滤离心(5000 r/min,20 min)、滤液加2滴浓氨水浓缩至水相,冻融离心,上清液,调pH 2.5~3.0乙酸乙酯萃取,过Sep-pak C18小柱,用含5%甲醇的 CH2 Cl2溶解样品,经0.22μm滤膜过滤后,通过 GPC 纯化、LC-MS 定性,利用HPLC外标曲线法定量。经检测:胡杨叶片中GA3含量为1162.7898 ng/g鲜质量;烟草中GA3含量为920.9067 ng/g鲜质量;萌发绿豆中GA3含量为700.9236 ng/g鲜质量。%We developed a protocol to determine the contents of gibberellins ( GAs) in various plant tissues, such as Populus euphratica leaves, Nicotiana tabacum plantlets, and germinated green beans ( Vigna radiata) . GAs samples were purified with gel permeation chromatography ( GPC) , confirmed by liquid chromatography-mass spectrometry ( LC-MS) , and quantified by high performance liquid chromatography ( HPLC) . Plant samples were ground with 80% methanol and extracted at 4℃ overnight. Crude extract was filtrated and then centrifuged at 5 000 r/min for 20 min. Two drops of ammonia were added to the liquid sample which was concentrated to aqueous phase, then frozen, thawed and centrifuged. Upper liquids were re-extracted with ethyl acetate at pH 2. 5 -3. 0 ( × 3 ) , and purified by Sep-pak C18 columns. The samples were resolved by 5% methanol-CH2 Cl2 and filtrated through 0. 22μm filter. Then samples were purified through GPC and GAs fluid was collected for subsequent quantification. GAs in the GPC-purified samples was quantified with HPLC by means of external standard curves. The gibberellins detected in plant tissues were confirmed with LC-MS. The result showed that the GA3 content was 1 162. 789 8 ng/g FW in poplar leaves, 920. 906 7 ng/g FW in tobacco leaves and 700

  1. 光调控在植物组织培养中的应用研究进展%Research Advances of Photoregulation and Its Utilization in Plant Tissue Culture

    Institute of Scientific and Technical Information of China (English)

    谷艾素; 张欢; 崔瑾

    2011-01-01

    Photoregulation is a new and effective environmental control technology,which is of great significance in plant tissue culture. In this paper,applications of photoregulation in plant tissue culture,that is the effect of light intensity,photoperiod,light quality on plant growth and development,photosynthesis,callus induction,proliferation and differentiation,organogenesis and somatic embryogenesis,physiological characteristics and plant secondary metabolites of plant materials in vitro at home and abroad in recent years were mainly reviewed,so as to provide a reference for plant cell engineering.%光调控是植物组织培养中-种有效的环境控制技术.该文对近年来国内外有关光调控在植物组织培养中的应用,即光照强度、光周期、光质对组培植物的生长发育、光合作用、愈伤组织诱导及其增殖与分化、器官和体细胞胚发生、生理特性及次生代谢物质等方面的影响研究进展进行了综述,为植物细胞工程提供参考.

  2. (Catharanthus roseus) tissue culture

    African Journals Online (AJOL)

    STORAGESEVER

    2008-08-18

    Aug 18, 2008 ... indole alkaloids in plant tissue cultures of C. roseus have. *Corresponding ... alkaloids from C. roseus cell cultures have failed (review- ed by Van der ..... that vinblastine occur in callus culture with differentiated roots. Dimeric ...

  3. Simultaneous determination of 24 or more acidic and alkaline phytohormones in femtomole quantities of plant tissues by high-performance liquid chromatography-electrospray ionization-ion trap mass spectrometry.

    Science.gov (United States)

    Liu, Shichang; Chen, Weiqi; Qu, Long; Gai, Ying; Jiang, Xiangning

    2013-02-01

    Phytohormones act at relatively low concentrations as major regulatory factors of plant growth and development, and cross talk of phytohormones is currently of great interest throughout the plant science community. To meet this demand, a method that is capable of simultaneously analyzing diverse plant hormones is essential. This paper introduces a high-performance liquid chromatographic separation technique coupled with sensitive and selective ion trap mass spectrometry to simultaneously determine 24 or more acidic and alkaline phytohormones, including auxin, cis- and trans-abscisic acid, 11 cytokinins, and 10 gibberellins, in a single injection of sample. A binary solid-phase extraction using Oasis MCX cartridges for cations and Oasis MAX cartridges for anions was used to prepurify more than 24 acidic and alkaline phytohormones from a single plant extract. The method showed good linearity for all 24 phytohormones with R(2) values ranging from 0.9903 to 0.9997. Limits of detection for most of the phytohormones were in the femtomole range with some extending into the sub-femtomole range. This method was applied to hundreds of plant samples comprising different tissues from various plants, including herbaceous, woody climbing, and woody plants to demonstrate feasibility and to validate the methodology.

  4. [Plant hormones, plant growth regulators].

    Science.gov (United States)

    Végvári, György; Vidéki, Edina

    2014-06-29

    Plants seem to be rather defenceless, they are unable to do motion, have no nervous system or immune system unlike animals. Besides this, plants do have hormones, though these substances are produced not in glands. In view of their complexity they lagged behind animals, however, plant organisms show large scale integration in their structure and function. In higher plants, such as in animals, the intercellular communication is fulfilled through chemical messengers. These specific compounds in plants are called phytohormones, or in a wide sense, bioregulators. Even a small quantity of these endogenous organic compounds are able to regulate the operation, growth and development of higher plants, and keep the connection between cells, tissues and synergy between organs. Since they do not have nervous and immume systems, phytohormones play essential role in plants' life.

  5. Preparation of magnetic indole-3-acetic acid imprinted polymer beads with 4-vinylpyridine and β-cyclodextrin as binary monomer via microwave heating initiated polymerization and their application to trace analysis of auxins in plant tissues.

    Science.gov (United States)

    Zhang, Yi; Li, Yuanwen; Hu, Yuling; Li, Gongke; Chen, Yueqin

    2010-11-19

    Auxin is a crucial phytohormone for precise control of growth and development of plants. Due to its low concentration in plant tissues which are rich in interfering substances, the accurate determination of auxins remains a challenge. In this paper, a new strategy for isolation and enrichment of auxins from plant tissues was obtained by the magnetic molecularly imprinted polymer (mag-MIP) beads, which were prepared by microwave heating initiated suspension polymerization using indole-3-acetic acid (IAA) as template. In order to obtain higher selective recognition cavities, an enhanced imprinting method based on binary functional monomers, 4-vinylpyridine (4-VP) and β-cyclodextrin (β-CD), was adopted for IAA imprinting. The morphological and magnetic characteristics of the mag-MIP beads were characterized by scanning electron microscopy, Fourier-transform infrared spectroscopy and vibrating sample magnetometry. A majority of resultant beads were within the size range of 80-150μm. Porous surface morphology and good magnetic property were observed. Furthermore, the mag-MIP beads fabricated with 4-VP and β-CD as binary functional monomers exhibited improved recognition ability to IAA, as compared with the mag-MIP beads prepared with the individual monomer separately. Competitive rebinding experiment results revealed that the mag-MIP beads exhibited a higher specific recognition for the template than the non-imprinted polymer (mag-NIP) beads. An extraction method by mag-MIP beads coupled with high performance liquid chromatography (HPLC) was developed for determination of IAA and indole-3-butyric acid (IBA) in plant tissues. Linear ranges for IAA and IBA were in the range of 7.00-100.0μgL(-1) and 10.0-100.0μgL(-1), and the detection limits were 3.9 and 7.4μgL(-1), respectively. The analytical performance was also estimated by seedlings or immature embryos samples from three different plant tissues, pea, rice and wheat. Recoveries were in the range of 70

  6. Contributions of Chinese Botanists to Plant Tissue Culture in the 20th Century%二十世纪我国植物学家对植物组织培养的贡献

    Institute of Scientific and Technical Information of China (English)

    朱至清

    2002-01-01

    This paper looks back to the development of plant tissue culture in China in the last century. Since 1934, tissue culture studies in China has kept up with the international development in the fields. Progress has been made by Chinese in nearly every branches of tissue culture, including in vitro organogenesis, shoot tip culture, anther culture, ovary culture, endosperm culture, protoplast culture as well as mass cell culture. On the basis of reviewing the articles written by Chinese on plant tissue culture, the internationally recognized contributions are specially mentioned. The applications of plant tissue culture to agriculture and industry in China are also introduced.%回顾了上一世纪我国植物组织培养的发展.1934年以来,我国的植物组织培养研究一直与国际发展同步进行.我国学者在离体器官发生、茎尖培养、花药培养、子房培养、胚乳培养、原生质体培养和细胞大量培养等分支领域都取得重要进展.本文在引证我国研究者发表的植物组织培养论文的基础上,着重评述了那些被国际同行公认的研究成果.此外,还介绍了植物组织培养在我国农业和工业上应用的情况.

  7. Advance on Virus-free Plant Tissue Culture and its Application on Crocus sativus L.%植物组培脱毒技术及其在药用植物藏红花中的应用

    Institute of Scientific and Technical Information of China (English)

    李军; 高广春; 李白; 朱志明

    2014-01-01

    The technique of virus elimination in plant tissue culture can eliminate the virus, rejuvenate, and improve yield and quality of plants. This paper summarized the virus-free methods and its application in the recent years, discussed the virus-free methods used in medicinal plant Crocus sativus L..%植物组培脱毒技术可以脱除患病毒病植株的病毒,起到植株复壮,提高产量及质量的作用。综述了植物组织培养脱毒的技术方法及其近几年的应用情况,同时针对药用植物藏红花脱毒球茎培育中应用的脱毒技术做了总结及探讨。

  8. A microsatellite based method for quantification of fungi in decomposing plant material elucidates the role of Fusarium graminearum DON production in the saprophytic competition with Trichoderma atroviride in maize tissue microcosms.

    Science.gov (United States)

    Naef, Andreas; Senatore, Mauro; Défago, Geneviève

    2006-02-01

    Common PCR assays for quantification of fungi in living plants cannot be used to study saprophytic colonization of fungi because plant decomposition releases PCR-inhibiting substances and saprophytes degrade the plant DNA which could serve as internal standard. The microsatellite PCR assays presented here overcome these problems by spiking samples prior to DNA extraction with mycelium of a reference strain. PCR with fluorescent primers co-amplifies microsatellite fragments of different length from target and reference strains. These fragments were separated in a capillary sequencer with fluorescence detection. The target/reference ratio of fluorescence signal was used to calculate target biomass in the sample. Such PCR assays were developed for the mycotoxin deoxynivalenol (DON)-producing wheat and maize pathogen Fusarium graminearum and the biocontrol agent Trichoderma atroviride, using new microsatellite markers. In contrast to real-time PCR assays, the novel PCR assays showed reliable fungal biomass quantification in samples with differentially decomposed plant tissue. The PCR assays were used to quantify the two fungi after competitive colonization of autoclaved maize leaf tissue in microcosms. Using a DON-producing F. graminearum wild-type strain and its nontoxigenic mutant we found no evidence for a role of DON production in F. graminearum defense against T. atroviride. The presence of T. atroviride resulted in a 36% lower wild-type DON production per biomass.

  9. Inhibition of flower formation by antisense repression of mitochondrial citrate synthase in transgenic potato plants leads to a specific disintegration of the ovary tissues of flowers.

    OpenAIRE

    Landschütze, V; Willmitzer, L.; Müller-Röber, B

    1995-01-01

    The tricarboxylic acid (TCA) cycle constitutes a major component of the mitochondrial metabolism of eucaryotes, including higher plants. To analyze the importance of this pathway, we down-regulated mitochondrial citrate synthase (mCS; EC 4.1.3.7), the first enzyme of the TCA cycle, in transgenic potato plants using an antisense RNA approach. Several transformants were identified with reduced citrate synthase activity (down to approximately 6% of wild-type activity). These plants were indistin...

  10. Tissue culture and plant regeneration of Ardisia crenata%朱砂根的组织培养与植株再生

    Institute of Scientific and Technical Information of China (English)

    马明东; 刘均利; 蒲尚饶

    2009-01-01

    目的:研究药用植物朱砂根组培快繁技术,为其产业化生产提供科学依据.方法:考察不同基本培养基、植物激素、添加物等对腋芽诱导、增殖及植株再生的影响.结果:以朱砂根带芽茎段为外植体,初代培养腋芽诱导的最佳培养基为MS+6-BA 0.5 mg·L~(-1)+NAA 0.1 mg·L~(-1).腋芽增殖的最佳培养基配方为MS+6-BA 2.0 mg·L~(-1)+NAA 0.1 mg·L~(-1)+KT 0.5 mg·L~(-1).生根培养基为1/2MS+IBA 0.2 mg·L~(-1);添加0.2%的活性炭可明显促进根的生长,提高生根率、生根数.最有利于朱砂根无菌苗移栽存活的基质类型为河沙.珍珠岩-蛭石(1:1:1),或蛭石.珍珠岩(1:1),成活率在80%以上.结论:通过腋芽增殖快繁育苗技术可获得完整植株,达到快速繁殖的目的.%Objective: Our research studied the fast-breeding technology of Ardisia crenata sims by using tissue culture and provided the scientific foundation for industry production. Method: The effects of axillary buds and plant regeneration of different basic medium, hormones and additives on induction and multiplication were studied. Result: The best culture medium for the induction of axillary buds, which took the stems of A. crenate were as explants, was MS + 6-BA 0. 5 mg · L~(-1) + NAA 0. 1 mg · L~(-1), and the best medium for multiplication was MS + 6-BA 2. 0 mg · L~(-1) + NAA 0. 1 mg · L~(-1) + KT 0. 5 mg · L~(-1), the best medium for roots generation was 1/2MS + IBA 0. 2 mg · L~(-1). We also found that the roots'generation, roots rate and mean number of roots can be promoted by adding 0. 2% Ac, and the most suitable ground substance was river sand-perlite-vermiculite (1: 1:1 ) or perlite-vermiculite (1:1 ).With axillary buds and plant regeneration methode, more than 80% A. crenata sims could be regenerated integratedly. Conclusion:A. crenata sims can be regenerated integratedly and breeded fast by using axillary bud proliferation technology.

  11. Targeted delivery of 1,25-dihydroxyvitamin D3 to colon tissue and identification of a major 1,25-dihydroxyvitamin D3 glycoside from Solanumglaucophyllum plant leaves.

    Science.gov (United States)

    Zimmerman, Duane R; Koszewski, Nicholas J; Hoy, Derrel A; Goff, Jesse P; Horst, Ronald L

    2015-04-01

    Leaves of the Solanum glaucophyllum (Sg) plant, indigenous to South America, have long been known for their calcinogenic toxicity in ruminant animals. It was determined the leaves contained glycosidic derivatives of 1,25-dihydroxyvitamin D3 (1,25D3) and liberation of the free hormone by rumen bacterial populations elicited a hypercalcemic response. Our interest in the leaves is predicated on the concept that the glycoside forms of 1,25D3 would target release of the active hormone in the lower gut of non-ruminant mammals. This would provide a means of delivering 1,25D3 directly to the colon, where the hormone has been shown to have beneficial effects in models of inflammatory bowel disease (IBD) and colon cancer. We fed mice for 10 days with variable amounts of Sg leaf. Feeding 7-333μg leaf/day produced no changes in plasma Ca(2+) and 1,25D3 concentrations, and only at ≥1000μg leaf/day did these values become significantly elevated compared to controls. Gene expression studies from colon tissue indicated a linear relationship between the amount of leaf consumed and expression of the Cyp24a1 gene. In contrast, Cyp24a1 gene expression in the duodenums and ileums of these mice was unchanged compared to controls. One of the major 1,25D3-glycosides was isolated from leaves following extraction and purification by Sep-Pak cartridges and HPLC fractionation. Ultraviolet absorbance was consistent with modification of the 1-hydroxyl group, and positive ion ESI mass spectrometry indicated a diglycoside of 1,25D3. 2-Dimensional NMR analyses were carried out and established the C1 proton of the A-ring was interacting with a C1' sugar proton, while the C3 proton of the A-ring was linked with a second C1' sugar proton. The structure of the isolated compound is therefore consistent with a β-linked 1,3-diglycoside of 1,25D3. Thus, Sg leaf administered to mice at up to 333 ug/day can elicit colon-specific enhancement of Cyp24a1 gene expression without inducing hypercalcemia, and

  12. Early evaluation and on field conditions of resistance to Mycosphaerella fijiensis Morelet of plants from Grande naine (AAA cultivar, obtained through out tissue culture and mutations induction

    Directory of Open Access Journals (Sweden)

    Lourdes R. García

    2003-04-01

    Full Text Available The present work was carried out in the Plants Biotechnology Institute of the Central University of Las Villas. The plant material from the cv. Grande Naine (AAA was treated with physical mutagenic agents(gamma radiation 60Co source to induce genetic variability. The behaviour of the population to the black Sigatoka was evaluated. A somaclone was selected by its disease resistance and was in vitro multiplied and the plants were acclimatized to evaluate its behaviour facing the disease on greenhouse conditions and in a second cycle of multiplication in the field. The results showed that in the majority of the plants were not found differences respect cv Grande Naine, just one presented similar reaction to cv. ‘FHIA 18’ (AAAB (partially resistant as for the variable evaluated, being obtained a frequency of 0.018% for this character. This plant was named IBP 446. After 60 days of application of the mycelial homogenized of M. fijiensis in micropropagated plants of this somaclone, differences in the respect affectation states were found at susceptible witness in greenhouse conditions. When plants of the IBP 446 were evaluated in a second cycle of multiplication differences were found with the susceptible control only at flowering, while they behaved similar at susceptible control in the crop. Key words: early detection, breeding, mutation, Black Sigatoka

  13. Use of tissue culture techniques for producing virus-free plant in garlic and their identification through real-time PCR.

    Science.gov (United States)

    Taşkın, Hatıra; Baktemur, Gökhan; Kurul, Mehmet; Büyükalaca, Saadet

    2013-01-01

    This study was performed for comparison of meristem culture technique with shoot tip culture technique for obtaining virus-free plant, comparison of micropropagation success of two different nutrient media, and determination of effectiveness of real-time PCR assay for the detection of viruses. Two different garlic species (Allium sativum and Allium tuncelianum) and two different nutrient media were used in this experiment. Results showed that Medium 2 was more successful compared to Medium 1 for both A. tuncelianum and A. sativum (Kastamonu garlic clone). In vitro plants obtained via meristem and shoot tip cultures were tested for determination of onion yellow dwarf virus (OYDV) and leek yellow stripe virus (LYSV) through real-time PCR assay. In garlic plants propagated via meristem culture, we could not detect any virus. OYDV and LYSV viruses were detected in plants obtained via shoot tip culture. OYDV virus was observed in amount of 80% and 73% of tested plants for A. tuncelianum and A. sativum, respectively. LYSV virus was found in amount of 67% of tested plants of A. tuncelianum and in amount of 87% of tested plants of A. sativum in this study.

  14. Use of Tissue Culture Techniques for Producing Virus-Free Plant in Garlic and Their Identification through Real-Time PCR

    Directory of Open Access Journals (Sweden)

    Hatıra Taşkın

    2013-01-01

    Full Text Available This study was performed for comparison of meristem culture technique with shoot tip culture technique for obtaining virus-free plant, comparison of micropropagation success of two different nutrient media, and determination of effectiveness of real-time PCR assay for the detection of viruses. Two different garlic species (Allium sativum and Allium tuncelianum and two different nutrient media were used in this experiment. Results showed that Medium 2 was more successful compared to Medium 1 for both A. tuncelianum and A. sativum (Kastamonu garlic clone. In vitro plants obtained via meristem and shoot tip cultures were tested for determination of onion yellow dwarf virus (OYDV and leek yellow stripe virus (LYSV through real-time PCR assay. In garlic plants propagated via meristem culture, we could not detect any virus. OYDV and LYSV viruses were detected in plants obtained via shoot tip culture. OYDV virus was observed in amount of 80% and 73% of tested plants for A. tuncelianum and A. sativum, respectively. LYSV virus was found in amount of 67% of tested plants of A. tuncelianum and in amount of 87% of tested plants of A. sativum in this study.

  15. Review on tissue culture of A ralia plants%楤木属植物组织培养研究综述

    Institute of Scientific and Technical Information of China (English)

    程莹; 李根有; 夏国华; 黄晌决; 黄宇锋

    2011-01-01

    This paper reviews the main research progress on tissue culture of Aralia. The explants, medium and plant growth regulators (PCRs) have important impacts on Aralia's callus culture, adventitious bud and somatic embryogenesis. Leaf blade, leaf stalk, young stem, apical bud, cotyledon and inflorescence can all be used as explants in primary culture. Leaf blade and young stem are the most commonly used explants. Aralia is usually cultured in solid media and liquid media. Liquid media is commonly used for somatic embryos culture. MS (Murashige-Skoog) is a common basic medium for in vitro culture. The types, concentrations of PCRs, and their combinations in the medium are key factors for Aralia in vitro culture. Thereinto, auxin is the main factor for callus induction, ABA is used for synchronization for somatic embryogenesis. Rooting is easy to plantlets in vitro and plantlets adapted well to cultural substrate under normal acclimatization and transplants. Research on molecular regulation of somatic embryogenesis of Aralia is still in a preliminary stage. [Ch, 1 tab, 26 ref.]%阐述了国内外楤木属A ralia植物组织培养研究的现状与进展.外植体、培养基、植物生长调节物质对楤木属植物愈合组织、不定芽和体细胞胚诱导均产生重要影响,叶片、叶柄、嫩茎、顶芽、子叶、花序等均可用作初代培养的外植体,其中叶片和嫩茎是最常用的外植体;培养方式有固体培养和液体培养,其中液体培养通常用于体细胞胚的培养;MS(Murashige and Skoog)是最常用的基本培养基,培养基中植物生长调节剂种类、浓度及其配比是楤木属植物离体培养成功与否的关键因素,其中生长素对愈合组织诱导起到主要作用,脱落酸(ABA)可以有效调控体细胞胚同步化;楤木属植物生根容易,采用常规的炼苗移栽,成活率较高.楤木属植物体细胞胚胎发生调控的分子机制的研究尚处于初步阶段.

  16. The Plant Cell Surface

    Institute of Scientific and Technical Information of China (English)

    Anne-Mie C.Emons; Kurt V.Fagerstedt

    2010-01-01

    @@ Multicellular organization and tissue construction has evolved along essentially different lines in plants and animals. Since plants do not run away, but are anchored in the soil, their tissues are more or less firm and stiff. This strength stems from the cell walls, which encase the fragile cytoplasm, and protect it.

  17. PLANT BIOPRINTING: NOVEL PERSPECTIVE FOR PLANT BIOTECHNOLOGY

    Directory of Open Access Journals (Sweden)

    Adhityo WICAKSONO

    2015-12-01

    Full Text Available Bioprinting is a technical innovation that has revolutionized tissue engineering. Using conventional printer cartridges filled with cells as well as a suitable scaffold, major advances have been made in the biomedical field, and it is now possible to print skin, bones, blood vessels, and even organs. Unlike animal systems, the application of bioprinting in simple plant tissue cells is still in a nascent phase and has yet to be studied. One major advantage of plants is that all living parts are reprogrammable in the form of totipotent cells. Plant bioprinting may improve scientists’understanding of plant shape and morphogenesis, and could serve for the mass production of desired tissues or plants, or even the production of plant-based biomaterial for industrial uses. This perspectives paper explores these possibilities using knowledge on what is known about bioprinting in other biosystems.

  18. The Development of Plant Biotechnology.

    Science.gov (United States)

    Torrey, John G.

    1985-01-01

    Examines major lines of thought leading to what is meant by plant biotechnology, namely, the application of existing techniques of plant organ, tissue, and cell culture, plant molecular biology, and genetic engineering to the improvement of plants and of plant productivity for the benefit of man. (JN)

  19. 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.

  20. Research Methods in Study on Optimum Medium in Plant Tissue Culture%植物组织培养寻求最佳培养基的方法研究

    Institute of Scientific and Technical Information of China (English)

    邓正正

    2015-01-01

    Over the past few decades, plant tissue culture has evolved into one of the major research tools in many fields such as ontogeny, organogenesis and physiological metabolism, etc. In practice, it has reached a level of sophistication where, for some species, its adaptation to large-scale industrial use has become possible, especially for quick reproducing elite seedlings. Though great progress has been achieved most of successful culturing procedures in vitro were base on experiences rather than on theories and concepts. This paper reviewed recent advances in search of media that would be suitable to plant regenerations in plant tissue culture. With focusing on the methods of optimizing plant growth regulate, macro elements and nutrition components at different culturing stages, it is hope that improving the research means for media would make steps from traditional experiments into more intentional and help to development of seeking optimal medium components.%植物组织培养技术在植物的个体发育、 器官再生和生理代谢等理论研究中已成为一种重要手段, 最佳培养基的寻找, 是组培成败的关键. 从基本培养基和植物生长调节剂入手, 植物组织培养最佳培养基的研究方法由传统的定性研究转向现代的定量研究, 加快寻求最佳培养基的步伐.

  1. Bicarbonate as tracer for plant assimilated C and homogeneity of 14C and 15N distribution in ryegrass and white clover tissue by alternative labeling approaches

    DEFF Research Database (Denmark)

    Rasmussen, Jim; Kusliene, Gedrime; Jacobsen, Ole Stig;

    2013-01-01

    Aims: Application of carbon (C) and nitrogen (N) isotopes is an essential tool to study C and N flows in plant-soil-microorganisms systems. When targeting single plants in a community the tracers need to be added via e.g., leaf-labeling or stem-feeding approaches. In this study we: (i) investigated...... parts with high and low 14C activity were separated and analyzed for 15N enrichment. Results Bicarbonate applied by leaf-labeling efficiently introduced 14C into both white clover and ryegrass, although the 14C activity in particular for white clover was found predominantly in the labeled leaf. Using 14...... that 15N also had a heterogeneous distribution (up to two orders of magnitude). Conclusion Bicarbonate can efficiently be used to introduce 14C or 13C into plant via the leaf-labeling method. Both 14C and 15N showed heterogeneous distribution in the plant, although the distribution of 15N was more even...

  2. Contribution of Wastewater Treatment Plants to Concentrations of PBDEs, PFCs, PCBs, DDT and Synthetic Musks in Fish Tissue from U.S. Urban Waters

    Science.gov (United States)

    Wastewater treatment plants (WWTPs) are tasked with removing a wide variety of contaminants from influents, including BOD and nutrients from human waste as well as any and all other compounds that emanate from homes and commercial facilities in the communities they serve. Traces ...

  3. Study of the comparative dynamics of the incorporation of tissue free-water tritium (TFWT) in bulrushes (Typha latifolia) and carp (Cyprinus carpio) in the Almaraz nuclear power plant cooling reservoir

    Energy Technology Data Exchange (ETDEWEB)

    Baeza, A. [Department of Applied Physics, Faculty of Veterinary, University of Extremadura, Avda de la Universidad s/n, 10071 Caceres (Spain)], E-mail: ymiralle@unex.es; Garcia, E. [Department of Applied Physics, Technical Forest Engineering School, University of Extremadura, 10600 (Plasencia) Caceres (Spain); Paniagua, J.M. [Department of Applied Physics, Polytechnic School, University of Extremadura, Avda de la Universidad s/n, 10071 Caceres (Spain); Rodriguez, A. [Department of Applied Physics, Faculty of Veterinary, University of Extremadura, Avda de la Universidad s/n, 10071 Caceres (Spain)

    2009-03-15

    The Almaraz nuclear power plant (Spain) uses the water of Arrocampo reservoir for cooling, and consequently raises the radioactive levels of the aquatic ecosystem of this reservoir. From July 2002 to June 2005, monthly samples of surface water, bulrushes (Typha latifolia) and carp (Cyprinus carpio) were collected from this reservoir. They were analyzed to determine the temporal evolution of the levels of {sup 3}H in surface water and of its transfer from the surface water to free-water in the tissues (TFWT) of the aforementioned two organisms. The tritium levels in the surface water oscillate with a biannual period, with their values in the study period ranging between 53 and 433 Bq/L. The incorporation of tritium to bulrushes and carp was fairly similar, the respective mean concentration factors being 0.74 and 0.8 (unitless, as Bq/L tissue water per Bq/L reservoir water). The temporal evolution of the levels fairly closely followed that observed for the surface water tritium, although detailed analysis showed the dominant periodicity for the bulrushes to be annual. This difference reflects the influence on the incorporation of tritium to bulrushes of diverse environmental and metabolic factors, especially evapotranspiration and the seasonal growth of this plant.

  4. Study of the comparative dynamics of the incorporation of tissue free-water tritium (TFWT) in bulrushes (Typha latifolia) and carp (Cyprinus carpio) in the Almaraz nuclear power plant cooling reservoir.

    Science.gov (United States)

    Baeza, A; García, E; Paniagua, J M; Rodríguez, A

    2009-03-01

    The Almaraz nuclear power plant (Spain) uses the water of Arrocampo reservoir for cooling, and consequently raises the radioactive levels of the aquatic ecosystem of this reservoir. From July 2002 to June 2005, monthly samples of surface water, bulrushes (Typha latifolia) and carp (Cyprinus carpio) were collected from this reservoir. They were analyzed to determine the temporal evolution of the levels of (3)H in surface water and of its transfer from the surface water to free-water in the tissues (TFWT) of the aforementioned two organisms. The tritium levels in the surface water oscillate with a biannual period, with their values in the study period ranging between 53 and 433 Bq/L. The incorporation of tritium to bulrushes and carp was fairly similar, the respective mean concentration factors being 0.74 and 0.8 (unitless, as Bq/L tissue water per Bq/L reservoir water). The temporal evolution of the levels fairly closely followed that observed for the surface water tritium, although detailed analysis showed the dominant periodicity for the bulrushes to be annual. This difference reflects the influence on the incorporation of tritium to bulrushes of diverse environmental and metabolic factors, especially evapotranspiration and the seasonal growth of this plant.

  5. Plant Tissue Culture in the Process of Pollution and its Control%植物组织培养过程中的污染及其防治方法

    Institute of Scientific and Technical Information of China (English)

    刘玲玲

    2012-01-01

    Contamination of plant tissue culture is a common problem,It seriously affected the normal development of training materials and growth,and to the research and production resulted in varying degrees of damage.This paper appears in plant tissue culture analysis of the causes of pollution,and proposed pollution control and reduce the rate of effective methods for research and production of a certain reference value.%污染是植物组织培养过程中的常见问题之一,它严重影响了培养材料的正常发育和生长,并给科研和生产造成了不同程度的损失.主要对植物组织培养中出现的污染原因进行了分析,并提出了控制和降低污染率的有效方法,为科研和生产提供了一定的参考价值.

  6. Avaliação por RAPD de plantas de abacaxizeiro cultivar Smooth Cayenne derivadas do seccionamento do talo e cultura de tecidos Evaluation of pineapple plants cultivar Smooth Cayenne from peduncle division and tissue culture by RAPD

    Directory of Open Access Journals (Sweden)

    MARIA VITÓRIA CECCHETTI GOTTARDI

    2002-04-01

    Full Text Available Foram coletadas, em área comercial da fazenda Córrego dos Bois, município de Canápolis -- MG, plantas de abacaxizeiro cultivar Smooth Cayenne, para serem avaliadas quanto à propagação pelo método do seccionamento do talo e cultura de tecidos, bem como análise por RAPD das mudas decorrentes destes dois processos de propagação. A propagação pelo seccionamento do talo foi eficiente na produção de mudas, tanto em quantidade como em qualidade, em um curto espaço de tempo, além de apresentar a mesma característica genotípica (análise por RAPD das plantas-matrizes de origem. Já no processo de produção de mudas por cultura de tecidos, não foi obtida uma quantidade suficiente de mudas que comprovasse a utilização de uma metodologia mais sofisticada. Além da perda por contaminação em laboratório de 70% do material em estudo, foi necessária a utilização de um longo período, aproximadamente 18 meses, para a obtenção das mudas. Na análise por RAPD das plantas decorrentes deste processo de propagação, foram observados padrões de bandas diferentes em algumas amostras, as quais podem estar relacionadas com uma possível variação somaclonal.Plants of pineapple Smooth Cayenne cultivar were collected from a commercial area, Córrego dos Bois farm, in Canápolis, state of Minas Gerais, to evaluate two different processes of propagation, peduncle division and tissue culture. These same plants were characterized by RAPD analysis. According to the two propagation method, the peduncle division was efficient in seedlings propagation, in quantity as well as in quality, at a short period of time, resulting the same phenotype and genetic characteristics (RAPD analysis from the matrix descent plants. In the tissue culture method, the quantity of seedlings obtained were not good enough that could prove the use of a sophisticated methodology. Besides loosing 70% of the material, it was necessary to expend a long period of time, at

  7. Pretreatment and enzymatic hydrolysis of wheat straw (Triticum aestivum L.)--the impact of lignin relocation and plant tissues on enzymatic accessibility.

    Science.gov (United States)

    Hansen, Mads A T; Kristensen, Jan Bach; Felby, Claus; Jørgensen, Henning

    2011-02-01

    Wheat straw is a potential feedstock for bioethanol production. This paper investigates tissues from whole internode sections subjected to hydrothermal pretreatment at 185°C and subsequent enzymatic hydrolysis up to 144 h. Analyses revealed an increase in surface lignin as hydrolysis progressed, which could be coupled to the gradual decrease in hydrolysis rate over time. The data support the hypothesis of lignin extraction from the cell wall matrix during pretreatment and deposition as droplets upon cooling. These droplets are assumed to accumulate during enzymatic hydrolysis. Additionally, after 144 h of enzymatic hydrolysis the cortex had vanished, exposing the heavier lignified vascular tissue. Accumulation of lignin droplets and exposure of residual lignin could be part of the explanation for the decreasing hydrolysis rate. Flattening of macrofibrils after pretreatment together with more indentations on the surfaces was also observed, possibly caused by a proposed synergistic effect of cellobiohydrolases and endoglucanases. Copyright © 2010 Elsevier Ltd. All rights reserved.

  8. Synthesis of functional materials by radiation and qualification testing of organic materials in nuclear power plant; evaluation of tissue compatibility of hydrogels

    Energy Technology Data Exchange (ETDEWEB)

    Jeon, H. J.; Kim, K. C.; Lee, Z. S.; Kim, J. Y.; Yoo, K. E. [Catholic University, Seoul (Korea)

    2002-04-01

    The hydrogels, owing to their adaptive physicochemical properties to the soft connective tissue, are known to have potentials to be applied as skin prosthetics. But hydrogels currently used are prepared by chemical crosslinking procedures. A prosthetics prepared by chemical crosslinking may damage the skin due to eluted crosslinking agent and/or other chemicals involved. That is the drawback of the skin substitutes made of typical hydrogels. The use of the irradiation crosslinking technique may supercede the demerits of conventionally used hydrogels, since this crossolinking technique entirely depends on physical method. The objective of this study is to prepare a series of hydrogels prepared by irradiation crosslinking procedure and to evaluate physical and tissue compatible profiles of these. In vitro and in vivo animal tests were employed to evaluate the properties as well as the comparibility of the samples in order to figure out the possibilities of the use as skin substitutes. 7 refs., 4 figs., 4 tabs. (Author)

  9. The plant Extracts of Momordica Charantia and Trigonella Foenum Graecum Have Antioxidant and Anti-Hyperglycemic Properties for Cardiac Tissue During Diabetes Mellitus

    Directory of Open Access Journals (Sweden)

    Uma Nath Tripathi

    2009-01-01

    Full Text Available Oxidative stress is currently suggested to play a major role in the development of diabetes mellitus. There is an increasing demand of natural anti-diabetic agents, as continuous administration of existing drugs and insulin are associated with many side effects and toxicity. The present study was aimed to investigate the effect of Momordica charantia (MC and Trigonella foenum graecum (TFG extracts (aqueous on antioxidant status and lipid peroxidation in heart tissue of normal and alloxan induced diabetic rats. In a 30 days treatment, rats were divided into six groups (I-VI of five animals in each, experiments were repeated thrice. Administration of MC (13.33 g pulp/kg body weight/day and TFG (9 g seeds powder/kg body weight/day extracts in diabetic rats has remarkably improved the elevated levels of fasting blood glucose. A significant decrease in lipid peroxidation (p<0.001 and significant increase in the activities of key antioxidant enzymes such as superoxide dismutase (SOD, catalase (CAT, glutathione-s-transferase (GST and reduced glutathione (GSH contents in heart tissue of diabetic rats were observed (group V and VI upon MC and TFG treatment. Our studies demonstrate the anti-hyperglycemic and anti-oxidative potential of Momordica charantia and Trigonella foenum graecum, which could exert beneficial effects against the diabetes and associated free radicals complications in heart tissue.

  10. Characterization of eight terpenoids from tissue cultures of the Chinese herbal plant, Tripterygium wilfordii, by high-performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry.

    Science.gov (United States)

    Su, Ping; Cheng, Qiqing; Wang, Xiujuan; Cheng, Xiaoqing; Zhang, Meng; Tong, Yuru; Li, Fei; Gao, Wei; Huang, Luqi

    2014-09-01

    In this study, a reliable method for analysis and identification of eight terpenoids in tissue cultures of Tripterygium wilfordii has been established using high-performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry (HPLC-ESI-MS). Our study indicated that sterile seedlings, callus cultures and cell-suspension cultures can rapidly increase the amount of biological materials. HPLC-ESI-MS was used to identify terpenoids from the extracts of these tissue cultures. Triptolide, triptophenolide, celastrol and wilforlide A were unambiguously determined by comparing the retention times, UV spectral data, and mass fragmentation behaviors with those of the reference compounds. Another four compounds were tentatively identified as triptonoterpenol, triptonoterpene, 22β-hydroxy-3-oxoolean-12-en-29-oic acid and wilforlide B, based on their UV and mass spectrometry spectra. The quantitative analysis showed that all three materials contain triptolide, triptophenolide, celastrol, wilforlide A, and the contents of the four compounds in the cell-suspension cultures were 53.1, 240, 129 and 964 µg/g, respectively, which were at least 2.0-fold higher than these in the sterile seedlings and callus cultures. Considering the known pharmacological activity of triptolide and celastrol, we recommend the cell-suspension cultures as biological materials for future studies, such as clinical and toxicological studies. The developed method was validated by the evaluation of its precision, linearity, detection limits and recovery, and it was successfully used to identify and quantify the terpenoids in the tissue cultures.

  11. Thallium speciation in plant tissues-Tl(III) found in Sinapis alba L. grown in soil polluted with tailing sediment containing thallium minerals.

    Science.gov (United States)

    Krasnodębska-Ostręga, Beata; Sadowska, Monika; Ostrowska, Sylwia

    2012-05-15

    Besides the dominant species in plants-Tl(I), noticeable amounts of Tl(III) (about 10% of total Tl content) were found in extracts of plants cultivated in the presence of tailing sediments, which are the main source of anthropogenic thallium already present in the environment. It is an important step of gaining knowledge about the detoxification mechanisms developed by Sinapis alba. This plant species is highly tolerant to Tl and it is able to cumulate high amounts of Tl and transport it into the above-ground organs. For more adequate estimation of accumulating abilities of S. alba, the elements' bioavailability was taken into consideration. The obtained bioconcentration factors of Cd (AF=0.6) and Zn (AF=1-2) were significantly lower than of Tl (AF=100-200). The biomass production was similar to the biomass of control cultivation. The results were based on ICP MS measurements of total elements' content and HPLC ICP MS for speciation analysis. The quality of obtained results was evaluated based on the intermethod comparison with voltammetry as a reference method. Comparison of data obtained using ICP MS and electrochemical methods (after a proper chemical treatment) was also used for indication of Tl(III) presence and for proving that Tl(I) was not transferred into Tl(III) during analytical procedures. Copyright © 2012 Elsevier B.V. All rights reserved.

  12. 濒危植物翅果油树组织培养研究进展%Advance of New Researches on Tissue Culture of the Rare and Endangered Plant Elaeagnus mollis

    Institute of Scientific and Technical Information of China (English)

    刘庚伟; 陈惠

    2011-01-01

    This review summarized the recent research achievements on the process of tissue culture of the rare and endangered plant Elaeagnus mollis, including sterilizing protocols of different explants derived from adult plants and seeds for tube germination, two types of plants regeneration(organ type and organ genesis type),methods of rooting and transplanting, factors affecting culture, as well as browning and vitrification phenomena and avoiding measures. And the further biotechnology research fields of E. mollis were prospected.%综述了濒危植物翅果油树组织培养过程中成年植株与试管萌发种子苗取材的外植体的种类与消毒程序、植株再生的2条途径(器官型与器官发生型)、生根与移栽、影响培养的因素及培养过程中出现的褐化和玻璃化现象的防除等,并提出了翅果油树生物技术研究今后的发展方向.

  13. Pretreatment and enzymatic hydrolysis of wheat straw (Triticum aestivum L.) – The impact of lignin relocation and plant tissues on enzymatic accessibility

    DEFF Research Database (Denmark)

    Hansen, Mads Anders Tengstedt; Kristensen, Jan Bach; Felby, Claus

    2011-01-01

    Wheat straw is a potential feedstock for bioethanol production. This paper investigates tissues from whole internode sections subjected to hydrothermal pretreatment at 185 °C and subsequent enzymatic hydrolysis up to 144 h. Analyses revealed an increase in surface lignin as hydrolysis progressed......, which could be coupled to the gradual decrease in hydrolysis rate over time. The data support the hypothesis of lignin extraction from the cell wall matrix during pretreatment and deposition as droplets upon cooling. These droplets are assumed to accumulate during enzymatic hydrolysis. Additionally...

  14. Phenylphenalenones Accumulate in Plant Tissues of Two Banana Cultivars in Response to Herbivory by the Banana Weevil and Banana Stem Weevil

    Science.gov (United States)

    Hölscher, Dirk; Buerkert, Andreas; Schneider, Bernd

    2016-01-01

    Phenylphenalenone-type compounds accumulated in the tissues of two banana cultivars—Musa acuminata cv. “Grande Naine” (AAA) and Musa acuminata × balbisiana Colla cv. “Bluggoe” (ABB)—when these were fed on by the banana weevil (Cosmopolites sordidus (Germ.) (Coleoptera: Curculionidae)) and the banana stem weevil (Odoiporus longicollis (Oliver) (Coleoptera: Curculionidae)). The chemical constituents of the banana material were separated by means of chromatographic techniques and identified by NMR spectroscopy. One new compound, 2-methoxy-4-phenylphenalen-1-one, was found exclusively in the corm material of “Bluggoe” that had been fed on by the weevils. PMID:27571112

  15. 8种园林彩叶植物叶片显色部位分析%Coloration tissue analysis of leaf color in eight garden colored-leaf plants

    Institute of Scientific and Technical Information of China (English)

    邱东萍; 赵云

    2012-01-01

    [Objective]The purpose of this experiment was to study leaf structure and coloration tissue location of he-liophyte including Hibiscus rosa-sinensis var. variegata, Codiaeum variegatum var. pictum, Brassica oleracea var. acephala and Loropetalum chinense Olive var. rubrum Yieh.and shade plants including Syngonium podophyllum cv. ' White Butterfly' , Rhoeo discolor Hance, Stromanthe sanguinea Sond and Excoecaria cochinchinensis Lou. [ Method ] After fixed with FAA, the fresh leaves were made into temporary slices to observe under microscope and photographed. [ Result ]The coloration tissue of Excoecaria cochinchinensis Lour, Hibiscus rosa-sinensis var. variegata, Brassica oleracea var. acephala, Rhoeo discolor Hance and Stromanthe sanguinea Sond existed in leaf epidermis, and Codiaeum variegatum var. pictum, Loropetalum chinense Olive var. rubrum Yieh, and Syngonium podophyllum cv. 'White Butter-fly' existed in palisade tissue or spongy tissue of mesophyll. [Conclusion]In summary, heliophyte and shade plants had similar coloration tissue with pigment in leaf epidermis or mesophyll tissue. Ecological characteristics were not the sole indicator for color expression in the color-leafed plants.%[目的]研究阳生植物羽衣甘蓝、花叶扶桑、红花檵木、细叶变叶木及阴生植物紫背万年青、紫背竹芋、红背桂和白蝶合果芋共8种彩叶植物的叶片结构及其显色部位,为探索园林彩叶植物的显色特性和呈色机理提供理论依据.[方法]取上述8种植物的新鲜叶片作为材料,用FAA固定后制作叶片结构的临时切片,显微观察摄影并分析.[结果]红背桂、花叶扶桑、羽衣甘蓝、紫背万年青、紫背竹芋这5种植物的显色部位在叶表皮中;细叶变叶木、红花檵木和白蝶合果芋这3种植物的显色部位在叶肉的栅栏组织或海绵组织中.[结论]阴生植物和阳生植物的显色部位类似,色素存在于叶表皮或叶肉组织中,因此不能以生态

  16. Detection of transgenic and endogenous plant DNA fragments and proteins in the digesta, blood, tissues, and eggs of laying hens fed with phytase transgenic corn.

    Directory of Open Access Journals (Sweden)

    Qiugang Ma

    Full Text Available The trials were conducted to assess the effects of long-term feeding with phytase transgenic corn (PTC to hens on laying performance and egg quality, and investigate the fate of transgenic DNA and protein in digesta, blood, tissues, and eggs. Fifty-week old laying hens (n = 144 were fed with a diet containing 62.4% PTC or non-transgenic isogenic control corn (CC for 16 weeks. We observed that feeding PTC to laying hens had no adverse effect on laying performance or egg quality (P>0.05 except on yolk color (P<0.05. Transgenic phyA2 gene and protein were rapidly degraded in the digestive tract and were not detected in blood, heart, liver, spleen, kidney, breast muscle, and eggs of laying hens fed with diet containing PTC. It was concluded that performance of hens fed diets containing PTC, as measured by egg production and egg quality, was similar to that of hens fed diets formulated with CC. There was no evidence of phyA2 gene or protein translocation to the blood, tissues, and eggs of laying hens.

  17. Detection of transgenic and endogenous plant DNA fragments and proteins in the digesta, blood, tissues, and eggs of laying hens fed with phytase transgenic corn.

    Science.gov (United States)

    Ma, Qiugang; Gao, Chunqi; Zhang, Jianyun; Zhao, Lihong; Hao, Wenbo; Ji, Cheng

    2013-01-01

    The trials were conducted to assess the effects of long-term feeding with phytase transgenic corn (PTC) to hens on laying performance and egg quality, and investigate the fate of transgenic DNA and protein in digesta, blood, tissues, and eggs. Fifty-week old laying hens (n = 144) were fed with a diet containing 62.4% PTC or non-transgenic isogenic control corn (CC) for 16 weeks. We observed that feeding PTC to laying hens had no adverse effect on laying performance or egg quality (P>0.05) except on yolk color (PTransgenic phyA2 gene and protein were rapidly degraded in the digestive tract and were not detected in blood, heart, liver, spleen, kidney, breast muscle, and eggs of laying hens fed with diet containing PTC. It was concluded that performance of hens fed diets containing PTC, as measured by egg production and egg quality, was similar to that of hens fed diets formulated with CC. There was no evidence of phyA2 gene or protein translocation to the blood, tissues, and eggs of laying hens.

  18. Effect of inulin extracts or inulin-containing plant supplement on blood lipid indices and fatty acid profile in fattener tissues.

    Science.gov (United States)

    Grela, E R; Sobolewska, S; Roziński, T

    2014-01-01

    The objective of the study was to evaluate the effect of inulin or dandelion, chicory and Jerusalem artichoke powder on lipid indices and fatty acid profile in fattener tissues. The experiment involved 120 crossbred pigs (PL x PLW) x Duroc with an initial body weight of 25.0 +/- 0.5 kg. Animals were assigned into 6 groups. A diet for group I (control) did not comprise an inulin additive, group II and III received 2% inulin (water or water-alcohol extraction of inulin from chicory root, respectively), while the mixtures for group IV, V and VI contained 4% root powder from Jerusalem artichoke (topinambur), chicory or dandelion, respectively. The animals were slaughtered at 115 kg body weight. The samples of blood, liver and the muscle longissimus dorsi tissues as well as whole hearts were collected for analysis. Fatty acid profile, some lipid indices and crude fat and cholesterol content were evaluated. Dietary supplement of 40 g dandelion powder resulted in preferable significant changes in the blood lipid indices and fatty acids composition (increased PUFA share and decreased n-6/n-3 ratio).

  19. The plant extracts of Momordica charantia and Trigonella foenum graecum have antioxidant and anti-hyperglycemic properties for cardiac tissue during diabetes mellitus

    Science.gov (United States)

    Tripathi, Uma Nath

    2009-01-01

    Oxidative stress is currently suggested to play a major role in the development of diabetes mellitus. There is an increasing demand of natural anti-diabetic agents, as continuous administration of existing drugs and insulin are associated with many side effects and toxicity. The present study was aimed to investigate the effect of Momordica charantia (MC) and Trigonella foenum graecum (TFG) extracts (aqueous) on antioxidant status and lipid peroxidation in heart tissue of normal and alloxan induced diabetic rats. In a 30 days treatment, rats were divided into six groups (I-VI) of five animals in each, experiments were repeated thrice. Administration of MC (13.33 g pulp/kg body weight/day) and TFG (9 g seeds powder/kg body weight/day) extracts in diabetic rats has remarkably improved the elevated levels of fasting blood glucose. A significant decrease in lipid peroxidation (pMomordica charantia and Trigonella foenum graecum, which could exert beneficial effects against the diabetes and associated free radicals complications in heart tissue. PMID:20716916

  20. 植物组培过程中污染微生物种类及其季节性的变化%Varieties and seasonal characters of pollution microflora in plant tissue culture

    Institute of Scientific and Technical Information of China (English)

    方丽; 王连平; 茹水江; 王汉荣

    2013-01-01

    为研究植物组培过程中污染微生物种类及其季节性变化规律,以浙江省杭州和宁波的植物组培生产车间中的污染物为监测对象,利用NA、PDA等培养基对污染物进行分离纯化和培养.结果发现:植物组培污染物主要为细菌和真菌,其中细菌占52.13%,分布于9个种,主要为芽孢杆菌属和假单胞菌属;真菌占43.87%,分为18个种,芽枝霉(Cladosporium spp.)、黑曲霉(Aspergillus niger)、青霉(Penicillum spp.)和酵母为主要污染真菌.污染物的数量及真菌与细菌的污染比例呈现季节性变化,夏秋季是污染物的高发期,而春冬季节污染发生相对较少.%To study the varieties and seasonal characters of pollution microflora in plant tissue culture, the pollutant in Hangzhou and Ningbo plant tissue production workshops in Zhejiang province were monitored. The microbes were separated and purified through culturing on the NA and PDA media. The results showed that bacteria and fungi were the major pollutants. Bacteria accounted for 52. 13% covering 9 species, and were mainly from the genus Bacillus spp. and Pseudomonas spp. Fungi accounted for 43. 87% covering 18 species, and were mainly from Aspergillus ni-ger, Cladosporium spp. , Penicillium spp. and yeast. The total pollutants showed a seasonal change, so did fungal and bacterial contamination ratio in plant tissue production. The peak period of pollutant occurrence was in summer and autumn, while in winter and spring pollution occurred rarely.

  1. Protocol: An improved high-throughput method for generating tissue samples in 96-well format for plant genotyping (Ice-Cap 2.0

    Directory of Open Access Journals (Sweden)

    Krysan Patrick J

    2007-06-01

    Full Text Available Abstract Background We previously developed a high-throughput system called 'Ice-Cap' for growing Arabidopsis seedlings in a 96-well format and rapidly collecting tissue for subsequent DNA extraction and genotyping. While the originally described Ice-Cap method is an effective tool for high-throughput genotyping, one shortcoming of the first version of Ice-Cap is that optimal seedling growth is highly dependent on specific environmental conditions. Here we describe several technical improvements to the Ice-Cap method that make it much more robust and provide a detailed protocol for implementing the method. Results The key innovation underlying Ice-Cap 2.0 is the development of a continuous watering system. The addition of the watering system allows the seedling growth plates to be incubated without a lid for the duration of the growth period, which in turn allows for much more uniform and robust seedling growth than was observed using the original method. We also determined that inserting wooden skewers between the upper and lower plates prior to tissue harvest made it easier to separate the plates following freezing. Seedlings grown using the Ice-Cap 2.0 method remain viable in the Ice-Cap plates twice as long as seedlings grown using the original method. Conclusion The continuous watering system that we have developed provides an effective solution to the problem of sub-optimal seedling growth that can be encountered when using the originally described Ice-Cap system. This novel watering system and several additional modifications to the Ice-Cap procedure have improved the robustness and utility of the method.

  2. An industrialized technique for raising tissue culture plant of Acacia cincinnata%卷荚相思组培工厂化育苗技术

    Institute of Scientific and Technical Information of China (English)

    张月娇

    2012-01-01

    通过对卷荚相思组培工厂化育苗技术研究的阐述,总结了卷荚相思组培工厂化育苗最优培养基配方为:诱导培养基为改良MS+ 6-BA 0.3 mg/L+ KT 0.5 mg/L+ NAA 0.3 mg/L+蔗糖30 g/L+卡拉胶8 g/L,继代增殖培养基为改良MS+ 6-BA 0.8 mg/L+ KT 0.3 mg/L+ NAA 0.6 mg/L+蔗糖30 g/L+卡拉胶8 g/L,生根培养基为1/2改良MS+ NAA 0.5 mg/L+蔗糖20 g/L+卡拉胶8 g/L,pH值为5.8.用此培养基配方进行卷荚相思组培工厂化育苗其诱导率可达70%,有效芽增殖倍数可达3倍以上,生根率可达95%以上.%A research on tissue culture technique of Acacia cincinnata was summarized and the optimal mediums were proposed in this paper. The medium of improved MS +6-BA 0. 3mg/L + KT 0. 5mg/L + NAA 0. 3mg/L + sucrose 30 g/L + carrageenan 8g/L was optimal for inducement, the medium of improved MS + 6-BA 0. 8mg/L + KT 0. 3mg/L + NAA 0. 6mg/L + sucrose 30 g/L + carrageenan 8 g/L was optimal for subculture. The medium of 1/2 improved MS + NAA 0. 5mg/L + sucrose 20 g/L + carrageenan 8 g/L was optimal for rooting, and the Ph was 5. 8. Using these mediums, the inducement rate of industrialized tissue culture of Acacia cincinnata could be up to 70% , effective bud multiplication was more than three times, the rooting rate was more than 95%.

  3. Standardized production of Phyllanthus tenellus Roxb. by plant tissue culture Produção padronizada de Phyllanthus tenellus Roxb. por cultura de tecidos vegetais

    Directory of Open Access Journals (Sweden)

    Cristiane Pimentel Victório

    2010-06-01

    Full Text Available Exigencies as ethic plant raw material are part of the needs of modern phytotherapy. Micropropagation offers opportunities to obtain mass propagation of superior genotypes in short time. This study aimed to develop a protocol of direct and indirect organogenesis of Phyllanthus tenellus Roxb. Nodal segments from plantlets obtained by in vitro germination were subcultured in modified Murashige and Skoog medium added with different plant growth regulators: IAA (indole-3-acetic acid, IBA (indole-3-butyric acid, GA3 (3-giberelic acid and KIN (kinetin. The highest proliferation rate was obtained using the combinations: IBA, KIN + GA3 (3.5 mg L-1 and IBA + KIN (2.4 mg L-1. Rooting was intensified after 40 days, reaching 100% for all media with indole-3-butyric acid. Addition of 2,4 dichlorophenoxyacetic acid (2,4D provided the best results for production of friable calli. Acclimatization was 100% effective for plantlets cultured in control medium, with decrease in survival rate in grown plantlets from media added with growth regulators.Exigências como matérias-primas vegetais fidedignas fazem parte das necessidades da fitoterapia moderna. A propagação in vitro permite a obtenção em massa de genótipos superiores, em curto espaço de tempo. Este estudo teve como objetivo desenvolver um protocolo de organogênese direta e indireta de Phyllanthus tenellus Roxb. Segmentos nodais de plântulas obtidas por germinação in vitro foram introduzidos em meio Murashige e Skoog adicionado de diferentes reguladores de crescimento vegetal: AIA (ácido indol-3-acético, AIB (ácido indol-3-butírico, AG3 (ácido giberélico-3 e CIN (cinetina. A maior taxa de proliferação foi obtida utilizando as combinações: AIB, CIN + AG3 (3,5 mg L-1 e AIB + CIN (2,4 mg L-1. A rizogênese foi intensificada após 40 dias de cultivo, atingindo 100% para todos os meios contendo AIB. A adição de ácido 2,4 diclorofenoxiacético (2,4D proporcionou os melhores resultados para

  4. Physicochemical characteristics and fatty acid profiles of muscle tissues from Hanwoo steers fed a total mixed ration supplied with medicinal plant by-products.

    Science.gov (United States)

    Lee, Shin Ja; Kim, Do Hyung; Yang, Han Sul; Nam, Ki Chang; Ahn, Seung Kyu; Park, Sung Kwon; Choi, Chang Weon; Lee, Sung Sill

    2017-10-01

    Using medicinal plant by-products (MPBP) as feed additives may be an eco-friendly option as substitutes for feedstuffs and may assist in reducing the improper disposal of MPBP. Therefore, this study was conducted to evaluate the influences of MPBP on the meat quality of Hanwoo steers fed a total mixed ration (TMR). Twenty seven steers (body weight = 573±57 kg) were randomly divided into three treatments with a control group and two tested groups as follows: control, 1,000 g/kg TMR; treatment 1 (MPBP30), 970 g/kg TMR and 30 g/kg MPBP; treatment 2 (MPBP50), 950 g/kg TMR and 50 g/kg MPBP. Average daily gain, feed conversion ratio and the Commission Internationale de l'Eclairage L* of muscle were improved (p<0.05, respectively) by MPBP30. Stearic acid (C18:0) was decreased (linear effect, p = 0.012), while oleic acid (C18:1) was increased (linear effect, p = 0.055) by MPBP level. Saturated fatty acid (SFA) and polyunsaturated fatty acid (PUFA) were decreased for MPBP50 while unsaturated fatty acid (USFA) and monounsaturated fatty acid (MUFA) were increased for MPBP 50. USFA and SFA ratio was increased for MPBP50 as well. These results indicated that MPBP supplementation in Hanwoo steers fed a TMR increased feed efficiency and meat color (lightness) with altering fatty acid proportions. Therefore, MPBP may be successfully used in ruminant feeding.

  5. Microwave dissolution of plant tissue and the subsequent determination of trace lanthanide and actinide elements by inductively coupled plasma-mass spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Alvarado, J.S.; Neal, T.J.; Smith, L.L.; Erickson, M.D.

    1997-08-01

    Recently there has been much concern with the ability of plants to uptake heavy metals from their surroundings. With the development of instrumental techniques with low detection limits such as inductively coupled plasma-mass spectrometry (ICP-MS), attention is shifting toward achieving faster and more elegant ways of oxidizing the organic material inherent in environmental samples. Closed-vessel microwave dissolution was compared with conventional methods for the determination of concentrations of cerium, samarium, europium, terbium, uranium and thorium in a series of samples from the National Institute of Standards and Technology and from fields in Idaho. The ICP-MS technique exhibited detection limits in parts-per-trillion and linear calibration plots over three orders of magnitude for the elements under study. The results obtained by using nitric acid and hydrogen peroxide in a microwave digestion system for the analysis of reference materials showed close agreement with the accepted values. These values were compared with results obtained from dry- and wet-ashing procedures. The findings from an experiment comparing radiometric techniques for the determination of actinide elements to ICP-MS are reported.

  6. Physiological indexese macro- and micronutrients in plant tissue and essential oil of Mentha piperita L. grown in nutrient solution with variation in N, P, K and Mg levels

    Directory of Open Access Journals (Sweden)

    E.F.S. David

    2014-03-01

    Full Text Available Mentha piperita L. is an aromatic and medicinal species of the family Lamiaceae, known as mint or peppermint, and its leaves and branches produce essential oil rich in menthol. This study aimed to evaluate physiological indexes, macro- and micronutrients inthe shootsand essential oil of Mentha piperita L. grown in nutrient solution number 2 of Hoagland and Arnon (1950 with different N, P, K and Mg levels. Shoot length, dry mass of the different organs, total dry mass, leaf area, essential oil yield and composition, and macronutrient (N, P, K, Mg, Ca, S and micronutrient (Mn, Cu, Fe, Zn contents in the shoot were evaluated. Plants treated with 65%N/50%P/25%K/100%Mg had a tendency towards longer shoot, greaterroot and leaf blade dry masses, higher essential oil yield, higher menthol levels and lower menthone levels. The results showed that Mentha can be grown in nutrient solution by reducing 65% N, 50% P, 25% K and 100% Mg. This solution had better development compared to the other tested treatments. Therefore,we recommendMentha piperita L. to be grown with such nutrient levels.

  7. Identification, Cloning, and Potential Application of Genes Related to Somatic Embryogenesis in Plant Tissue Culture%植物组织培养再生相关基因鉴定、克隆和应用研究进展

    Institute of Scientific and Technical Information of China (English)

    叶兴国; 佘茂云; 王轲; 杜丽璞; 徐惠君

    2012-01-01

    collectively described. They are somatic embryogenesis receptor-like kinase, arabinogalactan-proteins, nitrite reductase, auxin binding protein, and antioxidant enzyme. Regeneration associated genes are prospected to be potentially used in plant genetic breeding, whose applications will be involved in the improvement of plant regeneration efficiency and transformation efficiency, also in obtaining transgenic plants with bio-safety. However, main candidate genes related to regeneration might vary in different plants or tissues, or function through different pathways. Therefore, cloning and characterization of some important genes related to somatic embryogenesis or organogenesis should be strengthened in future.

  8. Hydrolyzable Tannins of Tamaricaceous Plants. 7.1 Structures and Cytotoxic Properties of Oligomeric Ellagitannins from Leaves of Tamarix nilotica and Cultured Tissues of Tamarix tetrandra.

    Science.gov (United States)

    Orabi, Mohamed A A; Taniguchi, Shoko; Sakagami, Hiroshi; Yoshimura, Morio; Amakura, Yoshiaki; Hatano, Tsutomu

    2016-04-22

    Partially unacylated new oligomeric hydrolyzable tannins, nilotinin T2 (1, trimer) and nilotinin Q1 (2, tetramer), together with four known trimers, nilotinin T1 (3) and hirtellins T1-T3 (4-6), and a dimer, tamarixinin B (7), were isolated from the aqueous acetone extracts of leaves of Tamarix nilotica. Among them, the new trimer 1 and the known trimers 4 and 6, in addition to the partially unacylated new trimer nilotinin T3 (8), the known dimers nilotinin D3 (9) and tamarixinin C (10), and the monomer tellimagrandin I (11), were isolated from the cultured shoots of Tamarix tetrandra. The structures of the new hydrolyzable tannins were established by chromatographic analyses and extensive 1D and 2D NMR, HRESI-TOFMS, and ECD spectroscopic experiments. Among the new oligomeric tannins, the particular unacylated position of a glucose core is attributed to a possible biosynthetic route. Isolation of the same oligomeric tannins from cultured shoots of T. tetrandra emphasizes the unique biogenetic ability of the obtained cultures on production of the structurally and biologically characteristic tamaricaceous tannins commonly produced by the intact Tamarix plants. Additionally, tannins obtained in the present study together with gemin D (12) and 1,3-di-O-galloyl-4,6-O-(aS)-hexahydroxydiphenoyl-β-d-glucose (13), from our previous investigation of the leaves of T. nilotica, exhibited variable tumor-specific cytotoxic effects. The ellagitannin trimers 4, 6, and 8 and the dimer 9 exerted predominant tumor-selective cytotoxic effects with high specificity toward human promyelocytic leukemia cells.

  9. Enzymes that control the thiamine diphosphate pool in plant tissues. Properties of thiamine pyrophosphokinase and thiamine-(di)phosphate phosphatase purified from Zea mays seedlings.

    Science.gov (United States)

    Rapala-Kozik, Maria; Gołda, Anna; Kujda, Marta

    2009-04-01

    The pool of thiamine diphosphate (TDP), available for TDP-dependent enzymes involved in the major carbohydrate metabolic pathways, is controlled by two enzyme systems that act in the opposite directions. The thiamine pyrophosphokinase (TPK) activates thiamine into TDP and the numerous phosphatases perform the reverse two-step dephosphorylation of TDP to thiamine monophosphate (TMP) and then to free thiamine. Properties and a possible cooperation of those enzymes in higher plants have not been extensively studied. In this work, we characterize highly purified preparations of TPK and a TDP/TMP phosphatase isolated from 6-day Zea mays seedlings. TPK was the 29-kDa monomeric protein, with the optimal activity at pH 9.0, the K(m) values of 12.4microM and 4.7mM for thiamine and ATP, respectively, and the V(max) value of 360pmol TDPmin(-1)mg(-1) protein. The enzyme required magnesium ions, and the best phosphate donor was GTP. The purified phosphatase was the dimer of 24kDa subunits, showed the optimal activity at pH 5.0 and had a rather broad substrate specificity, although TDP, but not TMP, was one of the preferable substrates. The K(m) values for TDP and TMP were 36microM and 49microM, respectively, and the V(max) value for TDP was significantly higher than for TMP (164 versus 60nmolesmin(-1)mg(-1) protein). The total activities of TPK and TDP phosphatases were similarly decreased when the seedlings were grown under the illumination, suggesting a coordinated regulation of both enzymes to stabilize the pool of the essential coenzyme.

  10. Tissue types (image)

    Science.gov (United States)

    There are 4 basic types of tissue: connective tissue, epithelial tissue, muscle tissue, and nervous tissue. Connective tissue ... and binds them together (bone, blood, and lymph tissues). Epithelial tissue provides a covering (skin, the linings of ...

  11. Conditional sterility in plants

    Science.gov (United States)

    Meagher, Richard B.; McKinney, Elizabeth; Kim, Tehryung

    2010-02-23

    The present disclosure provides methods, recombinant DNA molecules, recombinant host cells containing the DNA molecules, and transgenic plant cells, plant tissue and plants which contain and express at least one antisense or interference RNA specific for a thiamine biosynthetic coding sequence or a thiamine binding protein or a thiamine-degrading protein, wherein the RNA or thiamine binding protein is expressed under the regulatory control of a transcription regulatory sequence which directs expression in male and/or female reproductive tissue. These transgenic plants are conditionally sterile; i.e., they are fertile only in the presence of exogenous thiamine. Such plants are especially appropriate for use in the seed industry or in the environment, for example, for use in revegetation of contaminated soils or phytoremediation, especially when those transgenic plants also contain and express one or more chimeric genes which confer resistance to contaminants.

  12. Plant vascular development

    NARCIS (Netherlands)

    Rybel, De Bert; Mähönen, Ari Pekka; Helariutta, Yrjö; Weijers, Dolf

    2016-01-01

    Vascular tissues in plants are crucial to provide physical support and to transport water, sugars and hormones and other small signalling molecules throughout the plant. Recent genetic and molecular studies have identified interconnections among some of the major signalling networks that regulate

  13. 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...

  14. 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...

  15. The influence of freezing and tissue porosity on the material properties of vegetable tissues

    CERN Document Server

    Ralfs, J D

    2002-01-01

    Tissue porosity and fluid flow have been shown to be important parameters affecting the mechanical and sensorial behaviour of edible plant tissues. The quantity of fluid and the manner with which it was released on compression of the plant tissue were also important regarding the sensory perception and a good indication of any structural damage resulting from freezing, for example. Potato, carrot and Chinese water chestnut were used to study the effects freezing has on model plant tissues. Mechanical and structural measurements of the plant tissue were correlated with sensory analysis. Conventional freezing was shown to cause severe structural damage predominantly in the form of cavities between or through cells, resulting in decreases in mechanical strength and stiffness, and samples that were perceived in the mouth as 'soft' and 'wet'. The location and size of the cavities formed from ice crystals, depended on the particular plant tissue being frozen, the processing it was subjected to prior to freezing, th...

  16. Tissue Classification

    DEFF Research Database (Denmark)

    Van Leemput, Koen; Puonti, Oula

    2015-01-01

    Computational methods for automatically segmenting magnetic resonance images of the brain have seen tremendous advances in recent years. So-called tissue classification techniques, aimed at extracting the three main brain tissue classes (white matter, gray matter, and cerebrospinal fluid), are now...... well established. In their simplest form, these methods classify voxels independently based on their intensity alone, although much more sophisticated models are typically used in practice. This article aims to give an overview of often-used computational techniques for brain tissue classification...

  17. Methods of saccharification of polysaccharides in plants

    Science.gov (United States)

    Howard, John; Fake, Gina

    2014-04-29

    Saccharification of polysaccharides of plants is provided, where release of fermentable sugars from cellulose is obtained by adding plant tissue composition. Production of glucose is obtained without the need to add additional .beta.-glucosidase. Adding plant tissue composition to a process using a cellulose degrading composition to degrade cellulose results in an increase in the production of fermentable sugars compared to a process in which plant tissue composition is not added. Using plant tissue composition in a process using a cellulose degrading enzyme composition to degrade cellulose results in decrease in the amount of cellulose degrading enzyme composition or exogenously applied cellulase required to produce fermentable sugars.

  18. Plant Transporter Identification

    DEFF Research Database (Denmark)

    Larsen, Bo

    Membrane transport proteins (transporters) play a critical role for numerous biological processes, by controlling the movements of ions and molecules in and out of cells. In plants, transporters thus function as gatekeepers between the plant and its surrounding environment and between organs......, tissues, cells and intracellular compartments. Since plants are highly compartmentalized organisms with complex transportation infrastructures, they consequently have many transporters. However, the vast majority of predicted transporters have not yet been experimentally verified to have transport...... activity. This project contains a review of the implemented methods, which have led to plant transporter identification, and present our progress on creating a high-throughput functional genomics transporter identification platform....

  19. Plant Transporter Identification

    DEFF Research Database (Denmark)

    Larsen, Bo

    Membrane transport proteins (transporters) play a critical role for numerous biological processes, by controlling the movements of ions and molecules in and out of cells. In plants, transporters thus function as gatekeepers between the plant and its surrounding environment and between organs......, tissues, cells and intracellular compartments. Since plants are highly compartmentalized organisms with complex transportation infrastructures, they consequently have many transporters. However, the vast majority of predicted transporters have not yet been experimentally verified to have transport...... activity. This project contains a review of the implemented methods, which have led to plant transporter identification, and present our progress on creating a high-throughput functional genomics transporter identification platform....

  20. Genetically engineered plants with increased vegetative oil content

    Science.gov (United States)

    Benning, Christoph

    2017-05-23

    The invention relates to genetically modified agricultural plants with increased oil content in vegetative tissues, as well as to expression systems, plant cells, seeds and vegetative tissues related thereto.

  1. Cloning: plants – micropropagation/tissue culture

    Science.gov (United States)

    Clonal micropropagation is the multiplication of the buds and shoots that occur in leaf axils on a defined nutrient medium in an aseptic in vitro environment. The resulting shoots are either subdivided for continued multiplication or rooted and acclimatized to the greenhouse or field. Micropropagati...

  2. Plant Tissue Cultures of Juniperus virginiana.

    Science.gov (United States)

    Kašparová, Marie; Spilková, Jirina; Cvak, Ladislav; Siatka, Tomáš; Martin, Jan

    2016-05-01

    Callus cultures of Juniperus virginiana L. (varieties 'Hetzii', 'Glauca', 'Grey Owl') were derived from fresh leaves of garden-grown trees on Schenk and Hildebrandt medium supplemented with 3.0 mg/L of α-naphthaleneacetic acid, 0.2 mg/L of kinetin and 15 mg/L of ascorbic acid. The growth characteristics of one-year-old and two-years-old cultures were determined. The maximum biomass in all varieties was achieved on the 35th day of the cultivation period. The increase in fresh weights of two-years-old callus cultures, when compared with one-year-old callus cultures, was as follows: variety 'Hetzii' by 25%, variety 'Glauca' by 29% and variety 'Grey Owl' by 49%. J. virginiana suspension cultures (varieties 'Hetzii', 'Glauca', 'Grey Owl') were derived from two-years-old callus cultures on Schenk and Hildebrandt medium supplemented with 3.0 mg/L of α-naphthaleneacetic acid, 0.2 mg/L of kinetin and 15 mg/L of ascorbic acid. The maximum biomass of all varieties was found on the 21st day of the cultivation period. These results indicate that a sub-cultivation interval of 35 days for callus cultures and of 21st days for suspension cultures can be recommended. The callus and suspension cultures of J. virginiana of the variety 'Glauca' have the best survivability and thus provide the most biomass.

  3. Tissue Classification

    Energy Technology Data Exchange (ETDEWEB)

    Robinson, David Gerald [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States)

    2015-01-01

    The project began as a e ort to support InLight and Lumidigm. With the sale of the companies to a non-New Mexico entity, the project then focused on supporting a new company Medici Technologies. The Small Business (SB) is attempting to quantify glucose in tissue using a series of short interferometer scans of the nger. Each scan is produced from a novel presentation of the nger to the device. The intent of the project is to identify and, if possible, implement improved methods for classi cation, feature selection, and training to improve the performance of predictive algorithms used for tissue classi cation.

  4. 卓越农林人才培养视野下植物组织培养实验双语教学的探索与实践%Exploration and Practice of Bilingual Teaching in Plant tissue Culture from the Perspective of the Cultivation of Excellent Talents in Agriculture and Forestry

    Institute of Scientific and Technical Information of China (English)

    洪森荣

    2016-01-01

    卓越农林人才培养是地方本科院校向应用技术类型高校转型发展的需要。植物组织培养实验是一门实践性的课程,传统的植物组织培养实验教学能教会学生常规的实验操作,但学生毕业工作后查阅、参考并使用植物组织培养实验方面的英文资料往往会遇到一些困难。针对上饶师范学院生命科学学院植物组织培养实验教学的实际情况,作者改变了原来的中文授课方式,基于卓越农林人才的培养,开展了植物组织培养实验的双语教学,取得了较好的效果,并提出了植物组织培养实验双语教学中存在的问题及其对策。%The cultivation of excellent talents in agriculture and forestry is the need for the transformation and development of local colleges and universities to those of applied technology type .Plant tissue culture experi‐ment is a practical course ,the traditional plant tissue culture experiment teaching can teach students the con‐ventional experimental operation ,but after obtaining employment ,the students often encounter some difficul‐ties when referring to English materials on plant tissue culture experiment .Aimed at plant tissue culture ex‐periment teaching actual situation in College of Life Sciences in Shangrao Normal University ,the original Chi‐nese teaching method was changed ,the bilingual teaching of plant tissue culture experiment based on the culti‐vation of outstanding talents in agriculture and forestry was carried out and achieved better effects .At the same time ,and the problems and countermeasures existing in the bilingual teaching of plant tissue culture ex‐periment were put forward .

  5. Initiation of somatic embryos and regeneration of plants from primordial shoots of 10-year-old somatic white spruce and expression profiles of 11 genes followed during the tissue culture process.

    Science.gov (United States)

    Klimaszewska, Krystyna; Overton, Catherine; Stewart, Don; Rutledge, Robert G

    2011-03-01

    Adult conifers are notoriously recalcitrant in vegetative propagation and micropropagation that would result in the regeneration of juvenile propagules through somatic embryogenesis (SE) has not been demonstrated to date. Because SE-derived material is more amenable in subsequent tissue culture experiments compared with seed-derived material, a multi-year study was conducted to investigate induction of SE from primordial shoot (PS) explants that were excised from shoot buds of somatic embryo-derived white spruce. The SE induction experiments were carried out first with greenhouse-grown and later with field-grown trees each year from 2002 (2-year-old) to 2010 (10-year-old). Of the four genotypes tested, 893-2 and 893-12 never responded, 893-1 responded up to year 4 and 893-6 consistently responded every year. In 2010, for the first time, three of the 17 893-6 clonal trees produced male strobili as well as SE from cultured PS explants. SE induction was associated with formation of a nodule on the surface of an elongated needle primordium or in callus. Early somatic embryos were detectable after about 3 weeks of culture. Of 11 genes whose expression profiles were followed during the PS cultures, CHAP3A, VP1, WOX2 and SAP2C were expressed exclusively in the early stages of SE, and could potentially be used as markers of embryogenecity. Mature somatic embryos and plants were produced from the explants of responding genotype. Implication of these results for future research on adult conifer recalcitrance in micropropagation is discussed.

  6. Annual Plant Reviews

    DEFF Research Database (Denmark)

    , three dimensional structures and functions of each protein in a biological system. In plant science, the number of proteome studies is rapidly expanding after the completion of the Arabidopsis thaliana genome sequence, and proteome analyses of other important or emerging model systems and crop plants...... are in progress or are being initiated. Proteome analysis in plants is subject to the same obstacles and limitations as in other organisms, but the nature of plant tissues, with their rigid cell walls and complex variety of secondary metabolites, means that extra challenges are involved that may not be faced when...... analysing other organisms. This volume aims to highlight the ways in which proteome analysis has been used to probe the complexities of plant biochemistry and physiology. It is aimed at researchers in plant biochemistry, genomics, transcriptomics and metabolomics who wish to gain an up-to-date insight...

  7. Microbially produced phytotoxins and plant disease management ...

    African Journals Online (AJOL)

    Microbially produced phytotoxins and plant disease management. ... African Journal of Biotechnology ... Pathogenic fungi and bacteria often damage their host (plants) tissues by producing toxic metabolites, which induced various symptoms ...

  8. Memristors in plants.

    Science.gov (United States)

    Volkov, Alexander G; Tucket, Clayton; Reedus, Jada; Volkova, Maya I; Markin, Vladislav S; Chua, Leon

    2014-01-01

    We investigated electrical circuitry of the Venus flytrap, Mimosa pudica and Aloe vera. The goal was to discover if these plants might have a new electrical component--a resistor with memory. This element was postulated recently and the researchers were looking for its presence in different systems. The analysis was based on cyclic current-voltage characteristic where the resistor with memory should manifest itself. We found that the electrostimulation of plants by bipolar sinusoidal or triangle periodic waves induces electrical responses in the Venus flytrap, Mimosa pudica and Aloe vera with fingerprints of memristors. Tetraethylammonium chloride, an inhibitor of voltage gated K(+) channels, transforms a memristor to a resistor in plant tissue. Our results demonstrate that a voltage gated K(+) channel in the excitable tissue of plants has properties of a memristor. This study can be a starting point for understanding mechanisms of memory, learning, circadian rhythms, and biological clocks.

  9. Eggplant (Solanum melongena L.: tissue culture, genetic transformation and use as an alternative model plant Berinjela (Solanum melongena L.: cultura de tecidos, transformação genética e uso como planta modelo

    Directory of Open Access Journals (Sweden)

    Claudia Magioli

    2005-03-01

    Full Text Available Eggplant is an agronomically important non-tuberous solanaceous crop grown primarily for its large oval fruit. In popular medicine, eggplant is indicated for the treatment of several diseases, including diabetes, arthritis, asthma and bronchitis. Eggplant is susceptible to a number of diseases and pests capable of causing serious crop losses. This problem has been addressed by hybridizing eggplant with wild resistant Solanum species, which present a wide genetic diversity and are source of useful agronomic traits. The application of in vitro methodologies to eggplant has resulted in considerable success. Eggplant tissues present a high morphogenetic potential that is useful for developmental studies as well as for establishing biotechnological approaches to produce improved varieties, such as embryo rescue, in vitro selection, somatic hybridization and genetic transformation. Taken together, these characteristics also make eggplant a complete model for studies on different areas of plant science, including control of gene expression and assessment of genetic stability of somaclones derived from different morphogenetic processes. In the present study, important factors that affect the efficiency of in vitro regeneration through organogenesis and embryogenesis as well as genetic transformation are analyzed. The potential of this species as a model plant for studying various aspects of plant genetics and physiology is also discussed.A berinjela é uma espécie solanácea não tuberosa de importância agronômica, cultivada principalmente por seus frutos. Na medicina popular, a berinjela é indicada para o tratamento de várias doenças, incluindo diabetes, artrite, asma e bronquite. A berinjela é suscetível a várias doenças e pragas que causam perdas econômicas significativas. Esse problema tem sido abordado com técnicas convencionais de melhoramento, utilizando espécies silvestres resistentes de Solanum, que possuem uma grande diversidade

  10. 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,...

  11. 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,...

  12. Imprinting in plants

    Institute of Scientific and Technical Information of China (English)

    GUTIERREZ-MARCOS Jose

    2009-01-01

    Genomic imprinting leads to the differential expression of parental alleles after fertilization. Imprinting appears to have evolved independently in mammals and flowering plants to regulate the development of nutrient-transfer placental tissues. In addition, the regulation of imprinting in both mammals and flowering plants involves changes in DNA methylation and histone methylation, thus suggesting that the epigenetic signals that regulate imprinting have been co-opted in these distantly related species.

  13. 76 FR 31171 - Importation of Plants for Planting; Establishing a Category of Plants for Planting Not Authorized...

    Science.gov (United States)

    2011-05-27

    ... allowed that it may be possible to safely move small amounts of tissue-cultured plants that have been... agriculture, supporting livestock grazing and meat production and providing habitat for native pollinators...

  14. Walnut tissue culture: research and field applications

    Science.gov (United States)

    Jose M. Lopez

    2004-01-01

    Vitrotech Biotecnologia Vegetal began researching propagating Juglans regia (English walnut) and various Juglans hybrids by tissue culture in 1993 and has operated on a commercial scale since 1996. Since this time, more than one and a half million walnuts of different species have been propagated and field planted. Tissue cultured...

  15. Tissue culture of Cecropia glaziovii Sneth (urticaceae: vegetative micropropagation and plant regeneration from callus Cultura de tecidos de Cecropia glaziovii Sneth (Urticaceae: micropropagação vegetativa e regeneração de plantas via calos

    Directory of Open Access Journals (Sweden)

    Marcos Nopper Alves

    2010-10-01

    Full Text Available Cecropia glaziovii is a tree with used in Brazilian popular medicine. Methods allowing the clonal propagation of this species are of great interest for superior genotype multiplication and perpetuation. For this reason, we examined the effect of different culture media and different types of explants on adventitious shoot regeneration from callus and buds of C. glaziovii. Leaves, petioles and stipules obtained from aseptically grown seedlings or from pre-sterilized plants were used to initiate cultures. Adventitious shoot regeneration was achieved when apical and axillary buds were inoculated on gelled Murashige & Skoog (MS medium supplemented with 6-benzylaminopurine alone (BAP (1.0, 5.0 or 10.0 mg L-1 or combined with -naphthalene acetic acid (NAA (1.0 or 2.0 mg L-1, after 40 days of culture. Best callus production was obtained after 30 days of petioles' culture on gelled MS medium with 2,4 dichlorophenoxyacetic acid (2,4-D (5.0 mg L-1 combined with BAP (1.0 mg L-1. Successful shoot regeneration from callus was achieved when MS medium supplemented with zeatin (ZEA (0.1 mg L-1 alone or combined with 2,4-D (1.0 or 5.0 mg L-1 was inoculated with friable callus obtained from petioles. All shoots were rooted by inoculation on MS medium supplemented with indole-3-acetic acid (IAA (1.0 mg L-1. Rooted plants transferred to potting soil were successfully established. All in vitro regenerated plantlets showed to be normal, without morphological variations, being also identical to the source plant. Our study has shown that C. glaziovii can be propagated by tissue culture methods, allowing large scale multiplication of superior plants for pharmacological purposes.Cecropia glaziovii é uma planta lenhosa, popularmente usada no Brasil como medicinal. Métodos que visem a sua propagação clonal podem ser de grande utilidade na preservação de seus genótipos de elite. Foram examinados efeitos de diferentes reguladores de crescimento e explantes na forma

  16. Immersion freezing by plant nanocelloluse and plant phytolith particles

    Science.gov (United States)

    Rudich, Yinon; Reicher, Naama

    2017-04-01

    Using a new microfluidics setup (WISDOM), we examined the immersion freezing abilities of two types of airborne particles from biogenic sources: Plant nano crystalline cellulose particles and plant phytoliths. Plant opal phytoliths (POP) form in tissues of living plants during their growth, and can be found in soils following plants decay and in biomass burning plumes. Several measurements have identified these micro-sized particles in the atmosphere, but their ice nucleation properties have not yet been studied. We will present the new WISDOM device and first results on the efficiency of these biogenic particles to act as ice nuclei in the atmosphere under mixed clouds conditions.

  17. Cellulose metabolism in plants.

    Science.gov (United States)

    Hayashi, Takahisa; Yoshida, Kouki; Park, Yong Woo; Konishi, Teruko; Baba, Kei'ichi

    2005-01-01

    Many bacterial genomes contain a cellulose synthase operon together with a cellulase gene, indicating that cellulase is required for cellulose biosynthesis. In higher plants, there is evidence that cell growth is enhanced by the overexpression of cellulase and prevented by its suppression. Cellulase overexpression could modify cell walls not only by trimming off the paracrystalline sites of cellulose microfibrils, but also by releasing xyloglucan tethers between the microfibrils. Mutants for membrane-anchored cellulase (Korrigan) also show a typical phenotype of prevention of cellulose biosynthesis in tissues. All plant cellulases belong to family 9, which endohydrolyzes cellulose, but are not strong enough to cause the bulk degradation of cellulose microfibrils in a plant body. It is hypothesized that cellulase participates primarily in repairing or arranging cellulose microfibrils during cellulose biosynthesis in plants. A scheme for the roles of plant cellulose and cellulases is proposed.

  18. Terrestrial plant methane production

    DEFF Research Database (Denmark)

    Mikkelsen, Teis Nørgaard; Bruhn, Dan; Møller, Ian M.

    We evaluate all experimental work published on the phenomenon of aerobic methane (CH4) generation in terrestrial plants. We conclude that the phenomenon is true. Four stimulating factors have been observed to induce aerobic plant CH4 production, i.e. cutting injuries, increasing temperature......, ultraviolet radiation and reactive oxygen species. Further, we analyze rates of measured emission of aerobically produced CH4 in pectin and in plant tissues from different studies and argue that pectin is very far from the sole contributing precursor. Hence, scaling up of aerobic CH4 emission needs to take...... the aerobic methane emission in plants. Future work is needed for establishing the relative contribution of several proven potential CH4 precursors in plant material....

  19. Plant Biotech Lab Manual.

    Science.gov (United States)

    Tant, Carl

    This book provides laboratory experiments to enhance any food science/botany curriculum. Chapter 1, "Introduction," presents a survey of the techniques used in plant biotechnology laboratory procedures. Chapter 2, "Micronutrition," discusses media and nutritional requirements for tissue culture studies. Chapter 3, "Sterile Seeds," focuses on the…

  20. Mechanisms in Plant Development

    Energy Technology Data Exchange (ETDEWEB)

    Hake, Sarah [USDA ARS Plant Gene Expression Center

    2013-08-21

    This meeting has been held every other year for the past twenty-two years and is the only regularly held meeting focused specifically on plant development. Topics covered included: patterning in developing tissues; short and long distance signaling; differentiation of cell types; the role of epigenetics in development; evolution; growth.

  1. Plant Biotech Lab Manual.

    Science.gov (United States)

    Tant, Carl

    This book provides laboratory experiments to enhance any food science/botany curriculum. Chapter 1, "Introduction," presents a survey of the techniques used in plant biotechnology laboratory procedures. Chapter 2, "Micronutrition," discusses media and nutritional requirements for tissue culture studies. Chapter 3, "Sterile Seeds," focuses on the…

  2. Photochemical tissue bonding

    Science.gov (United States)

    Redmond, Robert W.; Kochevar, Irene E.

    2012-01-10

    Photochemical tissue bonding methods include the application of a photosensitizer to a tissue and/or tissue graft, followed by irradiation with electromagnetic energy to produce a tissue seal. The methods are useful for tissue adhesion, such as in wound closure, tissue grafting, skin grafting, musculoskeletal tissue repair, ligament or tendon repair and corneal repair.

  3. Biogeographical diversity of plant associated microbes in arcto-alpine plants

    NARCIS (Netherlands)

    Kumar, Manoj Gopala Krishnan

    2016-01-01

    Terrestrial plants and microbes have co-evolved since the emergence of the former on Earth. Associations with microorganisms can be either beneficial or detrimental for plants. Microbes can be found in the soil surrounding the plant roots, but also in all plant tissues, including seeds. In arcto-alp

  4. Tissue Photolithography

    Science.gov (United States)

    Wade, Lawrence A.; Kartalov, Emil; Shibata, Darryl; Taylor, Clive

    2011-01-01

    Tissue lithography will enable physicians and researchers to obtain macromolecules with high purity (greater than 90 percent) from desired cells in conventionally processed, clinical tissues by simply annotating the desired cells on a computer screen. After identifying the desired cells, a suitable lithography mask will be generated to protect the contents of the desired cells while allowing destruction of all undesired cells by irradiation with ultraviolet light. The DNA from the protected cells can be used in a number of downstream applications including DNA sequencing. The purity (i.e., macromolecules isolated form specific cell types) of such specimens will greatly enhance the value and information of downstream applications. In this method, the specific cells are isolated on a microscope slide using photolithography, which will be faster, more specific, and less expensive than current methods. It relies on the fact that many biological molecules such as DNA are photosensitive and can be destroyed by ultraviolet irradiation. Therefore, it is possible to protect the contents of desired cells, yet destroy undesired cells. This approach leverages the technologies of the microelectronics industry, which can make features smaller than 1 micrometer with photolithography. A variety of ways has been created to achieve identification of the desired cell, and also to designate the other cells for destruction. This can be accomplished through chrome masks, direct laser writing, and also active masking using dynamic arrays. Image recognition is envisioned as one method for identifying cell nuclei and cell membranes. The pathologist can identify the cells of interest using a microscopic computerized image of the slide, and appropriate custom software. In one of the approaches described in this work, the software converts the selection into a digital mask that can be fed into a direct laser writer, e.g. the Heidelberg DWL66. Such a machine uses a metalized glass plate (with

  5. Aeroponics for the culture of organisms, tissues and cells.

    Science.gov (United States)

    Weathers, P J; Zobel, R W

    1992-01-01

    Characteristics of aeroponics are discussed. Contrast is made, where appropriate, with hydroponics and aero-hydroponics as applies to research and commercial applications of nutrient mist technology. Topics include whole plants, plant tissue cultures, cell and microbial cultures, and animal tissue cultures with regard to operational considerations (moisture, temperature, minerals, gaseous atmosphere) and design of apparati.

  6. Aloe Vera for Tissue Engineering Applications.

    Science.gov (United States)

    Rahman, Shekh; Carter, Princeton; Bhattarai, Narayan

    2017-02-14

    Aloe vera, also referred as Aloe barbadensis Miller, is a succulent plant widely used for biomedical, pharmaceutical and cosmetic applications. Aloe vera has been used for thousands of years. However, recent significant advances have been made in the development of aloe vera for tissue engineering applications. Aloe vera has received considerable attention in tissue engineering due to its biodegradability, biocompatibility, and low toxicity properties. Aloe vera has been reported to have many biologically active components. The bioactive components of aloe vera have effective antibacterial, anti-inflammatory, antioxidant, and immune-modulatory effects that promote both tissue regeneration and growth. The aloe vera plant, its bioactive components, extraction and processing, and tissue engineering prospects are reviewed in this article. The use of aloe vera as tissue engineering scaffolds, gels, and films is discussed, with a special focus on electrospun nanofibers.

  7. Aloe Vera for Tissue Engineering Applications

    Directory of Open Access Journals (Sweden)

    Shekh Rahman

    2017-02-01

    Full Text Available Aloe vera, also referred as Aloe barbadensis Miller, is a succulent plant widely used for biomedical, pharmaceutical and cosmetic applications. Aloe vera has been used for thousands of years. However, recent significant advances have been made in the development of aloe vera for tissue engineering applications. Aloe vera has received considerable attention in tissue engineering due to its biodegradability, biocompatibility, and low toxicity properties. Aloe vera has been reported to have many biologically active components. The bioactive components of aloe vera have effective antibacterial, anti-inflammatory, antioxidant, and immune-modulatory effects that promote both tissue regeneration and growth. The aloe vera plant, its bioactive components, extraction and processing, and tissue engineering prospects are reviewed in this article. The use of aloe vera as tissue engineering scaffolds, gels, and films is discussed, with a special focus on electrospun nanofibers.

  8. Study Progress on Tissue Culture of Maize Mature Embryo

    Science.gov (United States)

    Wang, Hongzhen; Cheng, Jun; Cheng, Yanping; Zhou, Xioafu

    It has been paid more and more attention on maize tissue culture as it is a basic work in maize genetic transformation, especially huge breakthrough has been made in maize tissue culture utilizing mature embryos as explants in the recent years. This paper reviewed the study progress on maize tissue culture and plant regeneration utilizing mature embryos as explants from callus induction, subculture, plant regeneration and browning reduction and so on.

  9. Mathematical Models Light Up Plant Signaling

    NARCIS (Netherlands)

    Chew, Y.H.; Smith, R.W.; Jones, H.J.; Seaton, D.D.; Grima, R.; Halliday, K.J.

    2014-01-01

    Plants respond to changes in the environment by triggering a suite of regulatory networks that control and synchronize molecular signaling in different tissues, organs, and the whole plant. Molecular studies through genetic and environmental perturbations, particularly in the model plant Arabidopsis

  10. Mathematical Models Light Up Plant Signaling

    NARCIS (Netherlands)

    Chew, Y.H.; Smith, R.W.; Jones, H.J.; Seaton, D.D.; Grima, R.; Halliday, K.J.

    2014-01-01

    Plants respond to changes in the environment by triggering a suite of regulatory networks that control and synchronize molecular signaling in different tissues, organs, and the whole plant. Molecular studies through genetic and environmental perturbations, particularly in the model plant Arabidopsis

  11. Plant Sex Determination.

    Science.gov (United States)

    Pannell, John R

    2017-03-06

    Sex determination is as important for the fitness of plants as it is for animals, but its mechanisms appear to vary much more among plants than among animals, and the expression of gender in plants differs in important respects from that in most animals. In this Minireview, I provide an overview of the broad variety of ways in which plants determine sex. I suggest that several important peculiarities of plant sex determination can be understood by recognising that: plants show an alternation of generations between sporophytic and gametophytic phases (either of which may take control of sex determination); plants are modular in structure and lack a germ line (allowing for a quantitative expression of gender that is not common in animals); and separate sexes in plants have ultimately evolved from hermaphroditic ancestors. Most theorising about sex determination in plants has focused on dioecious species, but we have much to learn from monecious or hermaphroditic species, where sex is determined at the level of modules, tissues or cells. Because of the fundamental modularity of plant development and potentially important evolutionary links between monoecy and dioecy, it may be useful to relax the distinction often made between 'developmental sex determination' (which underpins the development of male versus female flowers in monoecious species) and 'genetic sex determination' (which underpins the separation of males and females in dioecious species, often mediated by a genetic polymorphism and sex chromosomes). I also argue for relaxing the distinction between sex determination involving a genetic polymorphism and that involving responses to environmental or hormonal cues, because non-genetic cues might easily be converted into genetic switches.

  12. Mycoplasma infections of plants.

    Science.gov (United States)

    Bove, J M

    1981-07-01

    Plants can be infected by two types of wall-less procaryotes, spiroplasmas and mycoplasma-like organisms (MLO), both located intracellularly in the phloem tissues of affected plants. Spiroplasmas have been cultured, characterized and shown to be true members of the class Mollicutes. MLO have not yet been cultured or characterized; they are thought to be mycoplasma-like on the basis of their ultrastructure as seen in situ, their sensitivity to tetracycline and resistance to penicillin. Mycoplasmas can also be found on the surface of plants. These extracellularly located organisms are members of the following genera: Spiroplasma. Mycoplasma and Acholeplasma. The presence of such surface mycoplasmas must not be overlooked when attempts to culture MLO from affected plants are undertaken. Sensitive serological techniques such as the enzyme-linked immunosorbent assay (ELISA) can successfully be used to compare the MLO located in the phloem of affected plants with those eventually cultured from the same plants. In California and Morocco periwinkles naturally infected with both Spiroplasma citri and MLO have been reported. With such doubly infected plants, the symptom expression has been that characteristic of the MLO disease (phyllody or stolbur), not that given by S. citri. Only S. citri can be cultured from such plants, but this does not indicate that S. citri is the causal agent of the disease expressed by the plant. In California many nonrutaceous plants have been found to be infected with S. citri. Stubborn affected citrus trees represent an important reservoir of S. citri, and Circulifer tenellus is an active leafhopper vector of S. citri. Hence, it is not surprising that in California MLO-infected fruit trees could also become infected with S. citri but it would not mean that S. citri is the causal agent of the disease. Criteria are discussed that are helpful in distinguishing between MLO infections and S. citri infections.

  13. Tissue culture of Sophora tonkinensis Gapnep. and its quality evaluation.

    Science.gov (United States)

    Kun-Hua, Wei; Lin-Xuan, Li; Yong-Cai, Huang; Mei-Ying, Wang; Cui, Li; Jian-Hua, Miao

    2013-10-01

    Sophora tonkinensis Gapnep. is an important rare medicinal plant in China. There were only a few papers on the rapid propagation of S. tonkinensis through in vitro tissue culture, and still no report focuses on the quality analysis of in vitro tissue culture plantlets. The different concentrations of 6-benzylaminopurine (BAP), kinetin (KT), and indole-3-acetic acid (IAA) were used to establish and screen the optimal rapid propagation technology of S. tonkinensis by orthogonal test; the different concentrations of a-naphthalene acetic acid (NAA), indole-3-butyric acid (IBA), and ABT rooting power (ABT) were used to screen the optimal rooting technology. For quality evaluation of tissue culture plants, three different sites were chose to finish planting experiment. The leaf characteristics, radix ex rhizoma yield, and contents of matrine and oxymatrine were evaluated, respectively, to provide evidence of high yield and good qualities of tissue culture plants. A large number of buds could be induced directly from epicotyl and hypocotyl explants on the Murashige and Skoog (MS) medium supplemented with 1.5 mg/l BAP, 0.5 mg/l IAA, and 0.5 mg/l KT; the best root induction medium was solid MS medium at half the macronutrient concentration supplemented with 1.0 mg/l NAA, 0.4 mg/l IBA, and 0.1 mg/l ABT. The rooting rate was 98%. All tissue culture plants showed normal leaf characteristics. Tissue culture plants from two sites possessed higher radix ex rhizoma yield and overall productivity of matrine and oxymatrine than those of seed plants. Tissue culture is a rapid, effective, and convenient propagation method for S. tonkinensis, and the quality of S. tonkinensis tissue culture plants meets the requirement of quality standard of China Pharmacopoeia (edition 2010), the crude drug from S. tonkinensis tissue culture plants will be suitable for substituting the crude drug from seed plants.

  14. Tissue culture of ornamental cacti

    Directory of Open Access Journals (Sweden)

    Eugenio Pérez-Molphe-Balch

    2015-12-01

    Full Text Available Cacti species are plants that are well adapted to growing in arid and semiarid regions where the main problem is water availability. Cacti have developed a series of adaptations to cope with water scarcity, such as reduced leaf surface via morphological modifications including spines, cereous cuticles, extended root systems and stem tissue modifications to increase water storage, and crassulacean acid metabolism to reduce transpiration and water loss. Furthermore, seeds of these plants very often exhibit dormancy, a phenomenon that helps to prevent germination when the availability of water is reduced. In general, cactus species exhibit a low growth rate that makes their rapid propagation difficult. Cacti are much appreciated as ornamental plants due to their great variety and diversity of forms and their beautiful short-life flowers; however, due to difficulties in propagating them rapidly to meet market demand, they are very often over-collected in their natural habitats, which leads to numerous species being threatened, endangered or becoming extinct. Therefore, plant tissue culture techniques may facilitate their propagation over a shorter time period than conventional techniques used for commercial purposes; or may help to recover populations of endangered or threatened species for their re-introduction in the wild; or may also be of value to the preservation and conservation of the genetic resources of this important family. Herein we present the state-of-the-art of tissue culture techniques used for ornamental cacti and selected suggestions for solving a number of the problems faced by members of the Cactaceae family.

  15. Overcoming DNA extraction problems from carnivorous plants

    OpenAIRE

    Fleischmann, Andreas; Heubl, Günther

    2009-01-01

    We tested previously published protocols for DNA isolation from plants with high contents of polyphenols and polysaccharides for several taxa of carnivorous plants. However, we did not get satisfying results with fresh or silica dried leaf tissue obtained from field collected or greenhouse grown plants, nor from herbarium specimens. Therefore, we have developed a simple modified protocol of the commercially available Macherey- Nagel NucleoSpin® Plant kit for rapid, effective and reproducible ...

  16. 75 FR 36060 - Notice of Request for Extension of Approval of an Information Collection; Blood and Tissue...

    Science.gov (United States)

    2010-06-24

    ... Collection; Blood and Tissue Collection at Slaughtering and Rendering Establishments AGENCY: Animal and Plant... collection associated with regulations for blood and tissue collection at slaughtering and rendering... FURTHER INFORMATION CONTACT: For information on regulations for blood and tissue collection...

  17. Poisonous Plants

    Science.gov (United States)

    ... Publications and Products Programs Contact NIOSH NIOSH POISONOUS PLANTS Recommend on Facebook Tweet Share Compartir Photo courtesy ... U.S. Department of Agriculture Many native and exotic plants are poisonous to humans when ingested or if ...

  18. Undifferentiated Connective Tissue Disease

    Science.gov (United States)

    ... Home Conditions Undifferentiated Connective Tissue Disease (UCTD) Undifferentiated Connective Tissue Disease (UCTD) Make an Appointment Find a Doctor ... L. Goldstein, MD, MMSc (February 01, 2016) Undifferentiated connective tissue disease (UCTD) is a systemic autoimmune disease. This ...

  19. Aquatic plants

    DEFF Research Database (Denmark)

    Madsen, T. V.; Sand-Jensen, K.

    2006-01-01

    Aquatic fl owering plants form a relatively young plant group on an evolutionary timescale. The group has developed over the past 80 million years from terrestrial fl owering plants that re-colonised the aquatic environment after 60-100 million years on land. The exchange of species between...... terrestrial and aquatic environments continues today and is very intensive along stream banks. In this chapter we describe the physical and chemical barriers to the exchange of plants between land and water....

  20. Manufacturing Plants

    Institute of Scientific and Technical Information of China (English)

    TANG YUANKAI

    2010-01-01

    @@ Sunshine, air and soil are indispensable for green plants. This might be axi-omatic but not in a plant factory. By creating a plant factory, scientists are trying to grow plants where natural elements are deficient or absent, such as deserts,islands, water surfaces, South and North poles and space, as well as in human habi-tats such as skyscrapers in modern cities.

  1. Manufacturing Plants

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    China starts to produce vegetables and fruits in a factory sunshine,air and soil are indispensable for green plants. This might be axiomatic but not in a plant factory. By creating a plant factory,scientists are trying to grow plants where natural elements are deficient or absent,such as deserts, islands,water surfaces,South and North poles and space,as well as in human habitats such as skyscrapers in modern cities.

  2. Cultivable endophytic bacteria from leaf bases of Agave tequilana and their role as plant growth promoters

    OpenAIRE

    Martínez-Rodríguez,Julia del C.; Marcela De la Mora-Amutio; Luis A. Plascencia-Correa; Esmeralda Audelo-Regalado; Guardado, Francisco R.; Elías Hernández-Sánchez; Peña-Ramírez, Yuri J.; Adelfo Escalante; Miguel J. Beltrán-García; Tetsuya Ogura

    2015-01-01

    Agave tequilana Weber var. 'Azul' is grown for the production of tequila, inulin and syrup. Diverse bacteria inhabit plant tissues and play a crucial role for plant health and growth. In this study culturable endophytic bacteria were extracted from leaf bases of 100 healthy Agave tequilana plants. In plant tissue bacteria occurred at mean population densities of 3 million CFU/g of fresh plant tissue. Three hundred endophytic strains were isolated and 16s rDNA sequences grouped the bacteria in...

  3. Medicinal Plants.

    Science.gov (United States)

    Phillipson, J. David

    1997-01-01

    Highlights the demand for medicinal plants as pharmaceuticals and the demand for health care treatments worldwide and the issues that arise from this. Discusses new drugs from plants, anticancer drugs, antiviral drugs, antimalarial drugs, herbal remedies, quality, safety, efficacy, and conservation of plants. Contains 30 references. (JRH)

  4. Plant growth-promoting bacterial endophytes.

    Science.gov (United States)

    Santoyo, Gustavo; Moreno-Hagelsieb, Gabriel; Orozco-Mosqueda, Ma del Carmen; Glick, Bernard R

    2016-02-01

    Bacterial endophytes ubiquitously colonize the internal tissues of plants, being found in nearly every plant worldwide. Some endophytes are able to promote the growth of plants. For those strains the mechanisms of plant growth-promotion known to be employed by bacterial endophytes are similar to the mechanisms used by rhizospheric bacteria, e.g., the acquisition of resources needed for plant growth and modulation of plant growth and development. Similar to rhizospheric plant growth-promoting bacteria, endophytic plant growth-promoting bacteria can act to facilitate plant growth in agriculture, horticulture and silviculture as well as in strategies for environmental cleanup (i.e., phytoremediation). Genome comparisons between bacterial endophytes and the genomes of rhizospheric plant growth-promoting bacteria are starting to unveil potential genetic factors involved in an endophytic lifestyle, which should facilitate a better understanding of the functioning of bacterial endophytes.

  5. 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.

  6. Alkaloid production by callous tissue cultures of Cereus peruvianus (Cactaceae).

    Science.gov (United States)

    de Oliveira, Arildo José Braz; Machado, Maria Fátima Pires da Silva

    2003-02-01

    The morphologically undifferentiated cells of nonregenerant callous tissue of Cereus peruvianus cultured in the original medium and in medium supplemented with tyrosine were used as an alkaloid source. Comparison of alkaloid production by C. peruvianus plants and by callous tissues indicated that alkaloid levels were almost twice as high in callous tissues as in shoots of C. peruvianus plants. The ratio of alkaloid concentration between mature plant and morphologically undifferentiated cells of callous tissue was 1:1.7. A relationship between culture medium containing tyrosine and alkaloid production was also observed in the callous tissues of C. peruvianus. Since increased alkaloid production may be induced by additional factors such as tyrosine, increasing levels of tyrosine or other conditions of the culture medium may be considered factors for inducing higher alkaloid production by C. peruvianus callous tissues.

  7. Plant injury induced by ozone

    Energy Technology Data Exchange (ETDEWEB)

    Hill, A.C.; Pack, M.R.; Treshow, M.; Downs, R.J.; Transtrum, L.G.

    1961-06-01

    Phytotoxicity of ozone to 34 plant species was studied in controlled-atmosphere greenhouses. Plants were subjected at various stages of growth to 0.13-0.72 ppm ozone for 2-hour periods. Injury symptoms developed on 28 species. Some of the most sensitive species were small grains, alfalfa, spinach, and tobacco. There was a general tendency for sensitivity to increase with maturity of tissue. Palisade cells were most readily injured by ozone. On plants with adaxial palisade parenchyma, chlorotic spots and bleached necrotic areas developed on the upper leaf surface. Injury was equally apparent from either leaf surface of plants with undifferentiated mesophyll. Necrotic spots extending completely through the leaf developed on plants with either mesophyll structure when injury was severe. Ozone caused conspicuous tumors to develop on broccoli leaves. Symptoms similar to those produced by ozone fumigations have been observed on a wide range of plant species growing near several large metropolitan centers. 18 references, 8 figures, 2 tables.

  8. Hyaluronidase, phospholipase A2 and protease inhibitory activity of plants used in traditional treatment of snakebite-induced tissue necrosis in Mali, DR Congo and South Africa

    DEFF Research Database (Denmark)

    Schmidt, Marianne Molander; Nielsen, Line Hagner; Søgaard, Søren Vinter

    2014-01-01

    ETHNOPHARMACOLOGICAL RELEVANCE: Snakebite envenomation, every year, causes estimated 5-10,000 mortalities and results in more than 5-15,000 amputations in sub-Saharan Africa alone. Antiserum is not easily accessible in these regions or doctors are simply not available, thus more than 80% of all p...... patients seek traditional practitioners as first-choice. Therefore it is important to investigate whether the plants used in traditional medicine systems contain compounds against the necrosis-inducing enzymes of snake venom....

  9. Alteration of plant meristem function by manipulation of the Retinoblastoma-like plant RRB gene

    Science.gov (United States)

    Durfee, Tim; Feiler, Heidi; Gruissem, Wilhelm; Jenkins, Susan; Roe, Judith; Zambryski, Patricia

    2007-01-16

    This invention provides methods and compositions for altering the growth, organization, and differentiation of plant tissues. The invention is based on the discovery that, in plants, genetically altering the levels of Retinoblastoma-related gene (RRB) activity produces dramatic effects on the growth, proliferation, organization, and differentiation of plant meristem.

  10. Alteration of plant meristem function by manipulation of the Retinoblastoma-like plant RRB gene

    Energy Technology Data Exchange (ETDEWEB)

    Durfee, Tim (Madison, WI); Feiler, Heidi (Albany, CA); Gruissem, Wilhelm (Forch, CH); Jenkins, Susan (Martinez, CA); Roe, Judith (Manhattan, KS); Zambryski, Patricia (Berkeley, CA)

    2007-01-16

    This invention provides methods and compositions for altering the growth, organization, and differentiation of plant tissues. The invention is based on the discovery that, in plants, genetically altering the levels of Retinoblastoma-related gene (RRB) activity produces dramatic effects on the growth, proliferation, organization, and differentiation of plant meristem.

  11. Lipid hydroperoxides in plants.

    Science.gov (United States)

    Griffiths, G; Leverentz, M; Silkowski, H; Gill, N; Sánchez-Serrano, J J

    2000-12-01

    Hydroperoxides are the primary oxygenated products of polyunsaturated fatty acids and were determined spectrophotometrically based on their reaction with an excess of Fe2+ at low pH in the presence of the dye Xylenol Orange. Triphenylphosphine-mediated hydroxide formation was used to authenticate the signal generated by the hydroperoxides. The method readily detected lipid peroxidation in a range of plant tissues including Phaseolus hypocotyls (26 +/- 5 nmol.g of fresh weight(-1); mean +/- S.D.), Alstroemeria floral tissues (sepals, 66+/-13 nmol.g of fresh weight(-1); petals, 49+/-6 nmol.g of fresh weight(-1)), potato leaves (334+/-75 nmol.g of fresh weight(-1)), broccoli florets (568+/-68 nmol.g of fresh weight(-1)) and Chlamydomonas cells (602+/-40 nmol.g of wet weight(-1)). Relative to the total fatty acid content of the tissues, the percentage hydroperoxide content was within the range of 0.6-1.7% for all tissue types (photosynthetic and non-photosynthetic) and represents the basal oxidation level of membrane fatty acids in plant cells. Leaves of transgenic potato with the fatty acid hydroperoxide lyase enzyme expressed in the antisense orientation were elevated by 38%, indicating a role for this enzyme in the maintenance of cellular levels of lipid hydroperoxides.

  12. Biodiversity effects on plant stoichiometry.

    Science.gov (United States)

    Abbas, Maike; Ebeling, Anne; Oelmann, Yvonne; Ptacnik, Robert; Roscher, Christiane; Weigelt, Alexandra; Weisser, Wolfgang W; Wilcke, Wolfgang; Hillebrand, Helmut

    2013-01-01

    In the course of the biodiversity-ecosystem functioning debate, the issue of multifunctionality of species communities has recently become a major focus. Elemental stoichiometry is related to a variety of processes reflecting multiple plant responses to the biotic and abiotic environment. It can thus be expected that the diversity of a plant assemblage alters community level plant tissue chemistry. We explored elemental stoichiometry in aboveground plant tissue (ratios of carbon, nitrogen, phosphorus, and potassium) and its relationship to plant diversity in a 5-year study in a large grassland biodiversity experiment (Jena Experiment). Species richness and functional group richness affected community stoichiometry, especially by increasing C:P and N:P ratios. The primacy of either species or functional group richness effects depended on the sequence of testing these terms, indicating that both aspects of richness were congruent and complementary to expected strong effects of legume presence and grass presence on plant chemical composition. Legumes and grasses had antagonistic effects on C:N (-27.7% in the presence of legumes, +32.7% in the presence of grasses). In addition to diversity effects on mean ratios, higher species richness consistently decreased the variance of chemical composition for all elemental ratios. The diversity effects on plant stoichiometry has several non-exclusive explanations: The reduction in variance can reflect a statistical averaging effect of species with different chemical composition or a optimization of nutrient uptake at high diversity, leading to converging ratios at high diversity. The shifts in mean ratios potentially reflect higher allocation to stem tissue as plants grew taller at higher richness. By showing a first link between plant diversity and stoichiometry in a multiyear experiment, our results indicate that losing plant species from grassland ecosystems will lead to less reliable chemical composition of forage for

  13. Autoluminescent plants.

    Directory of Open Access Journals (Sweden)

    Alexander Krichevsky

    Full Text Available Prospects of obtaining plants glowing in the dark have captivated the imagination of scientists and layman alike. While light emission has been developed into a useful marker of gene expression, bioluminescence in plants remained dependent on externally supplied substrate. Evolutionary conservation of the prokaryotic gene expression machinery enabled expression of the six genes of the lux operon in chloroplasts yielding plants that are capable of autonomous light emission. This work demonstrates that complex metabolic pathways of prokaryotes can be reconstructed and function in plant chloroplasts and that transplastomic plants can emit light that is visible by naked eye.

  14. Plant volatiles.

    Science.gov (United States)

    Baldwin, Ian T

    2010-05-11

    Plant volatiles are the metabolites that plants release into the air. The quantities released are not trivial. Almost one-fifth of the atmospheric CO2 fixed by land plants is released back into the air each day as volatiles. Plants are champion synthetic chemists; they take advantage of their anabolic prowess to produce volatiles, which they use to protect themselves against biotic and abiotic stresses and to provide information - and potentially disinformation - to mutualists and competitors alike. As transferors of information, volatiles have provided plants with solutions to the challenges associated with being rooted in the ground and immobile.

  15. Plant defense against insect herbivores

    DEFF Research Database (Denmark)

    Fürstenberg-Hägg, Joel; Zagrobelny, Mika; Bak, Søren

    2013-01-01

    have adapted to resist plant defenses, and in some cases even sequester the compounds and reuse them in their own defense. Both plant defense and insect adaptation involve metabolic costs, so most plant-insect interactions reach a stand-off, where both host and herbivore survive although......Plants have been interacting with insects for several hundred million years, leading to complex defense approaches against various insect feeding strategies. Some defenses are constitutive while others are induced, although the insecticidal defense compound or protein classes are often similar....... Insect herbivory induce several internal signals from the wounded tissues, including calcium ion fluxes, phosphorylation cascades and systemic- and jasmonate signaling. These are perceived in undamaged tissues, which thereafter reinforce their defense by producing different, mostly low molecular weight...

  16. Exploring interactions of plant microbiomes

    Directory of Open Access Journals (Sweden)

    Fernando Dini Andreote

    2014-12-01

    Full Text Available A plethora of microbial cells is present in every gram of soil, and microbes are found extensively in plant and animal tissues. The mechanisms governed by microorganisms in the regulation of physiological processes of their hosts have been extensively studied in the light of recent findings on microbiomes. In plants, the components of these microbiomes may form distinct communities, such as those inhabiting the plant rhizosphere, the endosphere and the phyllosphere. In each of these niches, the "microbial tissue" is established by, and responds to, specific selective pressures. Although there is no clear picture of the overall role of the plant microbiome, there is substantial evidence that these communities are involved in disease control, enhance nutrient acquisition, and affect stress tolerance. In this review, we first summarize features of microbial communities that compose the plant microbiome and further present a series of studies describing the underpinning factors that shape the phylogenetic and functional plant-associated communities. We advocate the idea that understanding the mechanisms by which plants select and interact with their microbiomes may have a direct effect on plant development and health, and further lead to the establishment of novel microbiome-driven strategies, that can cope with the development of a more sustainable agriculture.

  17. Tissue bionics: examples in biomimetic tissue engineering.

    Science.gov (United States)

    Green, David W

    2008-09-01

    Many important lessons can be learnt from the study of biological form and the functional design of organisms as design criteria for the development of tissue engineering products. This merging of biomimetics and regenerative medicine is termed 'tissue bionics'. Clinically useful analogues can be generated by appropriating, modifying and mimicking structures from a diversity of natural biomatrices ranging from marine plankton shells to sea urchin spines. Methods in biomimetic materials chemistry can also be used to fabricate tissue engineering scaffolds with added functional utility that promise human tissues fit for the clinic.

  18. Comparative proteomics and metallomics studies in Arabidopsis thaliana leaf tissues: evaluation of the selenium addition in transgenic and nontransgenic plants using two-dimensional difference gel electrophoresis and laser ablation imaging.

    Science.gov (United States)

    Maciel, Bruna C M; Barbosa, Herbert S; Pessôa, Gustavo S; Salazar, Marcela M; Pereira, Gonçalo A G; Gonçalves, Danieli C; Ramos, Carlos H I; Arruda, Marco A Z

    2014-04-01

    The main goal of this work is to evaluate some differential protein species in transgenic (T) and nontransgenic (NT) Arabidopsis thaliana plants after their cultivation in the presence or absence of sodium selenite. The transgenic line was obtained through insertion of CaMV 35S controlling nptII gene. Comparative proteomics through 2D-DIGE is carried out in four different groups (NT × T; NT × Se-NT (where Se is selenium); Se-NT × Se-T, and T × Se-T). Although no differential proteins are achieved in the T × Se-T group, for the others, 68 differential proteins (by applying a regulation factor ≥1.5) are achieved, and 27 of them accurately characterized by ESI-MS/MS. These proteins are classified into metabolism, energy, signal transduction, disease/defense categories, and some of them are involved in the glycolysis pathway-Photosystems I and II and ROS combat. Additionally, laser ablation imaging is used for evaluating the Se and sulfur distribution in leaves of different groups, corroborating some results obtained and related to proteins involved in the glycolysis pathway. From these results, it is possible to conclude that the genetic modification also confers to the plant resistance to oxidative stress.

  19. Electronic plants

    Science.gov (United States)

    Stavrinidou, Eleni; Gabrielsson, Roger; Gomez, Eliot; Crispin, Xavier; Nilsson, Ove; Simon, Daniel T.; Berggren, Magnus

    2015-01-01

    The roots, stems, leaves, and vascular circuitry of higher plants are responsible for conveying the chemical signals that regulate growth and functions. From a certain perspective, these features are analogous to the contacts, interconnections, devices, and wires of discrete and integrated electronic circuits. Although many attempts have been made to augment plant function with electroactive materials, plants’ “circuitry” has never been directly merged with electronics. We report analog and digital organic electronic circuits and devices manufactured in living plants. The four key components of a circuit have been achieved using the xylem, leaves, veins, and signals of the plant as the template and integral part of the circuit elements and functions. With integrated and distributed electronics in plants, one can envisage a range of applications including precision recording and regulation of physiology, energy harvesting from photosynthesis, and alternatives to genetic modification for plant optimization. PMID:26702448

  20. Concentrações de carboidratos em tecidos de pessegueiro (Prunus persica (L. Batsch cv. jubileu em plantas com ou sem sintomas de morte-precoce durante o período de dormência Carbohydrates content in peach trees tissues (Prunus persica (L. Batsch cv. jubileu in plants with and without peach-tree-short-life symptoms during the dormant period

    Directory of Open Access Journals (Sweden)

    Anderson Carlos Marafon

    2007-04-01

    Full Text Available O objetivo deste trabalho foi determinar as concentrações de amido e açúcares solúveis em tecidos de gemas e base de gemas de plantas de pessegueiro (Prunus persica (L. Batsch cultivar Jubileu, com e sem sintomas da Morte-Precoce-do-Pessegueiro, durante o período de repouso. Os ramos foram coletados em quatro épocas durante a dormência (11-06, 11-07, 29-07 e 05-08, no inverno de 2003. Foram conduzidos dois experimentos separadamente, um para cada pomar. A elevada concentração de amido nos tecidos das plantas sem sintomas de morte-precoce na fase da saída de dormência propiciou um adequado suprimento energético para que a brotação e a floração destas plantas ocorressem de maneira uniforme e regular. Por outro lado, a antecipação na quebra de dormência das plantas com sintomas da morte-precoce, provocada pelo desencadeamento da síndrome, intensificou a degradação do amido e do sorbitol em ambos os tecidos na saída de dormência, possivelmente, para o fornecimento de glicose e frutose.The aim of this study was to quantify the carbohydrates content (starch and soluble sugars in bud tissues and bud base of peach trees (Prunus persica (L. Batsch cultivar Jubileu in plants with and without Peach-tree-short-life (PTSL symptoms during the dormant period. The branches were collected in four different times during the dormant period (June, 11th, July, 11th, July, 29th, August, 05th in the winter of 2003. Two experiments were conducted separately, one for each orchard. The higher concentration of starch in plnt tissues without PTSL symptoms in the final of the dormant period provided a suitable energetic supply , leading to a regular and uniform budbreak of the plants. On the other hand, the budbreak anticipation of the plants with PTSL symptoms, which was provoked by the syndrome starting, intensified the starch and sorbitol breakdown in both tissues in the final of the dormant period, possibly producing glucose and fructose.

  1. Application of plant impedance for diagnosing plant disease

    Science.gov (United States)

    Xu, Huirong; Jiang, Xuesong; Zhu, Shengpan; Ying, Yibin

    2006-10-01

    Biological cells have components acting as electrical elements that maintain the health of the cell by regulation of the electrical charge content. Plant impedance is decided by the state of plant physiology and pathology. Plant physiology and pathology can be studies by measuring plant impedance. The effect of Cucumber Mosaic Virus red bean isolate (CMV-RB) on electrical resistance of tomato leaves was studied by the method of impedance measurement. It was found that the value of resistance of tomato leaves infected with CMV-RB was smaller than that in sound plant leaves. This decrease of impedances in leaf tissue was occurred with increased severity of disease. The decrease of resistance of tomato leaves infected with CMV-RB could be detected by electrical resistance detecting within 4 days after inoculation even though significant visible differences between the control and the infected plants were not noted, so that the technique for measurement of tomato leaf tissue impedance is a rapid, clever, simple method on diagnosis of plant disease.

  2. Acúmulo de matéria seca e exigências nutricionais de plantas de alho provenientes de cultura de tecidos e de propagação convencional Dry matter accumulation and nutritional requirements of garlic planted with cloves obtained by tissue culture or produced directly on the field

    Directory of Open Access Journals (Sweden)

    Francisco Vilela Resende

    1999-11-01

    by tissue culture and plants originated from cloves produced directly in the field. A randomized block design with four replications was used in the split-plot scheme. The treatments consisted of garlic plants obtained from tissue culture (meristem-tip culture and from cloves produced in the field, evaluated 30, 50, 70, 90, 110, 130, and 150 days after planting. At each period, six plants/plot were collected and dry matter of both aerial part and bulb were evaluated. Accumulated amounts of N, P, K, Ca, Mg, S, B, Zn, Cu, Mn and Fe in the garlic plants were also determined. In general, the amount of nutrient uptake corresponded to the development of the plants in both propagation system. Nutrient uptake was more intense between 70 and 110 days in the aerial part of the plants, and 90 and 150 days in the bulb. Differences in nutrient uptake between these two multiplication systems were verified only at the maximum development of both aerial and bulb portions. Plants originated from field propagation required more nitrogen than potassium when compared to those previously propagated by tissue culture. Plants originated from tissue culture accumulated more nutrients than those obtained from cloves produced directty on the field. These differences in nutrient accumulation were observed at the harvesting time and were expressed as percentage: Ca - 83.2%, K - 77.8%, S - 70.0%, Mg - 62.7%, P - 55.5%, N - 16.0%, Fe - 116.6%, Mn - 94.5%, Cu - 64.7%, Zn - 62.7%, B - 57.3%.

  3. Compatible plant-aphid interactions: how aphids manipulate plant responses.

    Science.gov (United States)

    Giordanengo, Philippe; Brunissen, Laurence; Rusterucci, Christine; Vincent, Charles; van Bel, Aart; Dinant, Sylvie; Girousse, Christine; Faucher, Mireille; Bonnemain, Jean-Louis

    2010-01-01

    To access phloem sap, aphids have developed a furtive strategy, their stylets progressing towards sieve tubes mainly through the apoplasmic compartment. Aphid feeding requires that they overcome a number of plant responses, ranging from sieve tube occlusion and activation of phytohormone-signalling pathways to expression of anti-insect molecules. In addition to bypassing plant defences, aphids have been shown to affect plant primary metabolism, which could be a strategy to improve phloem sap composition in nutrients required for their growth. During compatible interactions, leading to successful feeding and reproduction, aphids cause alterations in their host plant, including morphological changes, modified resource allocation and various local as well as systemic symptoms. Repeated salivary secretions injected from the first probe in the epidermal tissue up to ingestion of sieve-tube sap may play a crucial role in the compatibility between the aphid and the plant.

  4. Growth variability in a tissue governed by stress dependent growth

    Science.gov (United States)

    Alim, Karen; Boudaoud, Arezki

    2012-02-01

    Cell wall mechanics lie at the heart of plant cell growth and tissue morphogenesis. Conversely, mechanical forces generated at tissue level can feedback on cellular dynamics. Differential growth of neighboring cells is one eminent origin of mechanical forces and stresses in tissues where cells adhere to each other. How can stresses arising from differential growth orchestrate large scale tissue growth? We show that cell growth coupled to the cell's main stress can reduce or increase tissue growth variability. Employing a cell-based two dimensional tissue model we investigate the dynamics of a tissue with stress depending growth dynamics. We find that the exact cell division rule strongly affects not only the tissue geometry and topology but also its growth dynamics. Our results should enable to infer underlying growth dynamics from live tissue statistics.

  5. Microscopia electrônica de microrganismos do tipo micoplasma nos tecidos de milho afetado pelo enfezamento e nos órgãos da cigarrinha vectora portadora Electron microscopy of mycoplasma-like organisms in corn stunt - Infected plant tissues and in the organs of the leafhopper vector

    Directory of Open Access Journals (Sweden)

    E. W. Kitajima

    1972-01-01

    Full Text Available Exames electrono-microscópicos de tecido foliar ou radicular de milho (Zea maysL. afetado pelo enfezamento do milho, tanto a forma vermelha como a pálida, e de órgãos da cigarrinha vectora, Dalbulus maidisDe L. & W., portadora, demonstraram a ocorrência de corpúsculos pleomórficos, do tipo micoplasma, consistentemente associados a essa anomalia. Não se notou diferença na morfologia entre os microrganismos do tipo micoplasma associados às duas formas do enfezamento da planta nem tampouco entre estes e os presentes no inseto vector. Verificou-se também que na planta esses corpúsculos do tipo micoplasma ocorriam somente nos vasos crivados, e que no inseto eles se achavam presentes em diversos tipos de tecidos de diferentes órgãos - tubo digestivo, músculo, túbulos de Malphigi, epiderme, gânglio nervoso, glândula salivar, tecido adiposo. Nas células do inseto, os corpúsculos ocorriam dispersos no citoplasma ou em cavidades limitadas por membrana.Corn stunt in São Paulo State is usually of minor importance during normal summer crops but might induce significant losses if corn is planted late in the season. It is transmitted by leaf - hopper Dalbulus maidisDe L. & W. and two different forms of this disease have been recognized the chlorotic, similar to the Rio Grande type of the U.S., and the red, resembling the Mesa Central type of Mexico. Electron microscopic examination of tissues from both affected plants or leafhopper reared on diseased plants, demonstrated the presence of pleomorphic, mycoplasma-like bodies, consistently associated with the corn stunt, which probably represent the causal agent the latter. No significant difference could be observed in the morphology of these mycoplasma-like bodies associated with the two disease types or with the insect vector. In the affected plants, these corpuscles were only found in the sieve tubes; in the insect body they were detected in several types of tissues of different organs

  6. Sample preparation for SEM of plant surfaces

    OpenAIRE

    A.K. Pathan; Bond, J.; R.E. Gaskin

    2010-01-01

    Plant tissues must be dehydrated for observation in most electron microscopes. Although a number of sample processing techniques have been developed for preserving plant tissues in their original form and structure, none of them are guaranteed artefact-free. The current paper reviews common scanning electron microscopy techniques and the sample preparation methods employed for visualisation of leaves under specific types of electron microscopes. Common artefacts introduced by specific techniq...

  7. Uptake by plants of radionuclides from FUSRAP waste materials

    Energy Technology Data Exchange (ETDEWEB)

    Knight, M.J.

    1983-04-01

    Radionuclides from FUSRAP wastes potentially may be taken up by plants during remedial action activities and permanent near-surface burial of contaminated materials. In order to better understand the propensity of radionuclides to accumulate in plant tissue, soil and plant factors influencing the uptake and accumulation of radionuclides by plants are reviewed. In addition, data describing the uptake of the principal radionuclides present in FUSRAP wastes (uranium-238, thorium-230, radium-226, lead-210, and polonium-210) are summarized. All five radionuclides can accumulate in plant root tissue to some extent, and there is potential for the translocation and accumulation of these radionuclides in plant shoot tissue. Of these five radionuclides, radium-226 appears to have the greatest potential for translocation and accumulation in plant shoot tissue. 28 references, 1 figure, 3 tables.

  8. Plant Defense against Insect Herbivores

    Science.gov (United States)

    Fürstenberg-Hägg, Joel; Zagrobelny, Mika; Bak, Søren

    2013-01-01

    Plants have been interacting with insects for several hundred million years, leading to complex defense approaches against various insect feeding strategies. Some defenses are constitutive while others are induced, although the insecticidal defense compound or protein classes are often similar. Insect herbivory induce several internal signals from the wounded tissues, including calcium ion fluxes, phosphorylation cascades and systemic- and jasmonate signaling. These are perceived in undamaged tissues, which thereafter reinforce their defense by producing different, mostly low molecular weight, defense compounds. These bioactive specialized plant defense compounds may repel or intoxicate insects, while defense proteins often interfere with their digestion. Volatiles are released upon herbivory to repel herbivores, attract predators or for communication between leaves or plants, and to induce defense responses. Plants also apply morphological features like waxes, trichomes and latices to make the feeding more difficult for the insects. Extrafloral nectar, food bodies and nesting or refuge sites are produced to accommodate and feed the predators of the herbivores. Meanwhile, herbivorous insects have adapted to resist plant defenses, and in some cases even sequester the compounds and reuse them in their own defense. Both plant defense and insect adaptation involve metabolic costs, so most plant-insect interactions reach a stand-off, where both host and herbivore survive although their development is suboptimal. PMID:23681010

  9. Electroanalysis of Plant Thiols

    Directory of Open Access Journals (Sweden)

    Rene Kizek

    2007-06-01

    Full Text Available Due to unique physico-chemical properties of –SH moiety thiols comprise widegroup of biologically important compounds. A review devoted to biological functions ofglutathione and phytochelatins with literature survey of methods used to analysis of thesecompounds and their interactions with cadmium(II ions and Murashige-Skoog medium ispresented. For these purposes electrochemical techniques are used. Moreover, we revealedthe effect of three different cadmium concentrations (0, 10 and 100 μM on cadmiumuptake and thiols content in maize plants during 192 hours long experiments usingdifferential pulse anodic stripping voltammetry to detect cadmium(II ions and highperformance liquid chromatography with electrochemical detection to determineglutathione. Cadmium concentration determined in tissues of the plants cultivated innutrient solution containing 10 μM Cd was very low up to 96 hours long exposition andthen the concentration of Cd markedly increased. On the contrary, the addition of 100 μMCd caused an immediate sharp increase in all maize plant parts to 96 hours Cd expositionbut subsequently the Cd concentration increased more slowly. A high performance liquidchromatography with electrochemical detection was used for glutathione determination intreated maize plants after 96 and 192 hours of treatment. The highest total content of glutathione per one plant was 6 μg (96 h, 10 μM Cd in comparison with non-treated plant (control where glutathione content was 1.5 μg. It can be concluded that electrochemical techniques have proved to be useful to analyse plant thiols.

  10. The Research Development in Determination Method of Sugar Organic Compounds in Plant Tissue%植物组织中糖化合物测定方法的研究进展

    Institute of Scientific and Technical Information of China (English)

    孙艳涛; 由欣

    2011-01-01

    糖是植物生命现象中重要的生物高分子化合物。本文研究了近十年植物组织中糖类化合物的测定方法,为植物组织中的糖化合物的测定提供理论依据。%Sugar is plant life phenomenon in the important biological macromolecular compounds.This paper studies the measurement method of the saccharine compounds during nearly ten years,including the paper sulfuric acid phenol,chromatography,thin layer chromatography,gas chromatography,and high performance capillary electrophoresis method.

  11. Gene-enzyme relationships in somatic cells and their organismal derivatives in higher plants. Progress report. [In vitro cultivation of Nicotiana tissues and enzymological studies of gene expression at the cell level

    Energy Technology Data Exchange (ETDEWEB)

    None

    1979-01-01

    Progress over the first 9 months of the project has been substantial along several avenues. We have focussed on Nicotiana sylvestris for intensive study for the reasons specified. The individual characteristics of this organism dictate the need to adapt cell culture techniques to the particular requirements of this species. We have devoted considerable effort to optimization of our system through largely empirical experimentation. Methodological advances have been made to improve techniques for isolating enzyme substrates (mainly pretyrosine) that are not commercially available and for refining analytical techniques for the qualitative assay of the new enzyme activities of aromatic biosynthesis recently found by our group. Enzymological studies have been carried out in organismal plant material as a part of the ultimate goal of defining gene expression at the organismal level in relationship to expression at the cell culture level.

  12. Selection of bacterial wilt-resistant tomato through tissue culture.

    Science.gov (United States)

    Toyoda, H; Shimizu, K; Chatani, K; Kita, N; Matsuda, Y; Ouchi, S

    1989-06-01

    Bacterial wilt-resistant plants were obtained using a tomato tissue culture system. A virulent strain ofPseudomonas solanacearum secreted some toxic substances into the culture medium. Leaf explant-derived callus tissues which were resistant to these toxic substances in the culture filtrate were selectedin vitro and regenerated into plants. These plants expressed bacterial wilt resistance at the early infection stage to suppress or delay the growth of the inoculated bacteria. On the other hand, complete resistance was obtained in self-pollinated progeny of regenerants derived from non-selected callus tissues. These plants showed a high resistance when inoculated with this strain, and were also resistant when planted in a field infested with a different strain of the pathogen.

  13. Plant response to polluted air

    Energy Technology Data Exchange (ETDEWEB)

    Kendrick, J.B. Jr.; Darley, E.F.; Middleton, J.T.; Paulus, A.O.

    1956-08-01

    Field observations and controlled fumigation experiments have shown that plants differ in their response to atmospheric contamination by ethylene, herbicides, fluorides, sulfur dioxide, and smog, or oxidized hydrocarbons. Controlled experiments have also shown that plant response to air pollution varies with species and variety of plant, age of plant tissue, soil fertility levels, soil moisture, air temperatures during the prefumigation growth period, and presence of certain agricultural chemicals on leaves. The leaves of many plants; such as tomato, African marigold, fuchsia, pepper, and potato, become curved and malformed in the presence of ethylene, while those of cantaloupe, China aster, gardenia, Cattleya orchid, and snapdragon do not. Ethylene may cause serious damage to the sepals of orchids without injury to the petals or leaves.

  14. Measuring tissue oxygenation

    Science.gov (United States)

    Soyemi, Olusola O. (Inventor); Soller, Babs R. (Inventor); Yang, Ye (Inventor)

    2009-01-01

    Methods and systems for calculating tissue oxygenation, e.g., oxygen saturation, in a target tissue are disclosed. In some embodiments, the methods include: (a) directing incident radiation to a target tissue and determining reflectance spectra of the target tissue by measuring intensities of reflected radiation from the target tissue at a plurality of radiation wavelengths; (b) correcting the measured intensities of the reflectance spectra to reduce contributions thereto from skin and fat layers through which the incident radiation propagates; (c) determining oxygen saturation in the target tissue based on the corrected reflectance spectra; and (d) outputting the determined value of oxygen saturation.

  15. Plant Behavior

    Science.gov (United States)

    Liu, Dennis W. C.

    2014-01-01

    Plants are a huge and diverse group of organisms, ranging from microscopic marine phytoplankton to enormous terrestrial trees epitomized by the giant sequoia: 300 feet tall, living 3000 years, and weighing as much as 3000 tons. For this plant issue of "CBE-Life Sciences Education," the author focuses on a botanical topic that most…

  16. Plant Behavior

    Science.gov (United States)

    Liu, Dennis W. C.

    2014-01-01

    Plants are a huge and diverse group of organisms, ranging from microscopic marine phytoplankton to enormous terrestrial trees epitomized by the giant sequoia: 300 feet tall, living 3000 years, and weighing as much as 3000 tons. For this plant issue of "CBE-Life Sciences Education," the author focuses on a botanical topic that most…

  17. Microwave-assisted solvent extraction and analysis of shikimic acid from plant tissues Extração por solvente assistida por micro-ondas e análise do ácido chiquímico dos tecidos de plantas

    Directory of Open Access Journals (Sweden)

    M.B Matallo

    2009-12-01

    Full Text Available A better method for determination of shikimate in plant tissues is needed to monitor exposure of plants to the herbicide glyphosate [N-(phosphonomethylglycine] and to screen the plant kingdom for high levels of this valuable phytochemical precursor to the pharmaceutical oseltamivir. A simple, rapid, and efficient method using microwave-assisted extraction (MWAE with water as the extraction solvent was developed for the determination of shikimic acid in plant tissues. High performance liquid chromatography was used for the separation of shikimic acid, and chromatographic data were acquired using photodiode array detection. This MWAE technique was successful in recovering shikimic acid from a series of fortified plant tissues at more than 90% efficiency with an interference-free chromatogram. This allowed the use of lower amounts of reagents and organic solvents, reducing the use of toxic and/or hazardous chemicals, as compared to currently used methodologies. The method was used to determine the level of endogenous shikimic acid in several species of Brachiaria and sugarcane (Saccharum officinarum and on B. decumbens and soybean (Glycine max after treatment with glyphosate. The method was sensitive, rapid and reliable in all cases.Um método melhor para a determinação do chiquimato em tecidos de plantas é necessário para monitorar a exposição destas ao herbicida glyphosate N-(fosfonometilglicina] e selecionar o gênero da planta para altos níveis desse valioso precursor fitoquímico ao fármaco oseltamivir. Um método simples, rápido e eficiente, que usa a extração assistida por micro-ondas (MWAE com água como solvente de extração, foi desenvolvido para a determinação do ácido chiquímico em tecidos de plantas. Foi empregada cromatografia líquida de alto desempenho para a separação do ácido chiquímico, e os dados cromatográficos foram obtidos utilizando arranjo de fotodiodo. Essa técnica de MWAE obteve sucesso na recupera

  18. Plant macro- and micronutrient minerals

    Science.gov (United States)

    All plants must obtain a number of inorganic mineral elements from their environment to ensure successful growth and development of both vegetative and reproductive tissues. A total of fourteen mineral nutrients are considered to be essential. Several other elements have been shown to have beneficia...

  19. Plant minichromosomes.

    Science.gov (United States)

    Birchler, James A; Graham, Nathaniel D; Swyers, Nathan C; Cody, Jon P; McCaw, Morgan E

    2016-02-01

    Plant minichromosomes have the potential for stacking multiple traits on a separate entity from the remainder of the genome. Transgenes carried on an independent chromosome would facilitate conferring many new properties to plants and using minichromosomes as genetic tools. The favored method for producing plant minichromosomes is telomere-mediated chromosomal truncation because the epigenetic nature of centromere function prevents using centromere sequences to confer the ability to organize a kinetochore when reintroduced into plant cells. Because haploid induction procedures are not always complete in eliminating one parental genome, chromosomes from the inducer lines are often present in plants that are otherwise haploid. This fact suggests that minichromosomes could be combined with doubled haploid breeding to transfer stacked traits more easily to multiple lines and to use minichromosomes for massive scale genome editing.

  20. Multiplexed lasing in tissues

    Science.gov (United States)

    Chen, Yu-Cheng; Chen, Qiushu; Fan, Xudong

    2017-02-01

    Biolasers are an emerging technology for next generation biochemical detection and clinical applications. Progress has recently been made to achieve lasing from biomolecules and single living cells. Tissues, which consist of cells embedded in extracellular matrix, mimic more closely the actual complex biological environment in a living body and therefore are of more practical significance. Here, we developed a highly versatile tissue laser platform, in which tissues stained with fluorophores are sandwiched in a high-Q Fabry-Pérot microcavity. Distinct lasing emissions from muscle and adipose tissues stained respectively with fluorescein isothiocyanate (FITC) and boron-dipyrromethene (BODIPY), and hybrid muscle/adipose tissue with dual-staining were achieved with a threshold of only 10 μJ/mm2. Additionally, we investigated how tissue structure/geometry, tissue thickness, and staining dye concentration affect the tissue laser. It is further found that, despite large fluorescence spectral overlap between FITC and BODIPY in tissues, their lasing emissions could be clearly distinguished and controlled due to their narrow lasing bands and different lasing thresholds, thus enabling highly multiplexed detection. Our tissue laser platform can be broadly applicable to various types of tissues/diseases. It provides a new tool for a wide range of biological and biomedical applications, such as diagnostics/screening of tissues and identification/monitoring of biological transformations in tissue engineering.