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Differences in plant function in phosphorus- and nitrogen-limited mangrove ecosystems.

Mangrove ecosystems can be either nitrogen (N) or phosphorus (P) limited and are therefore vulnerable to nutrient pollution. Nutrient enrichment with either N or P may have differing effects on ecosystems because of underlying differences in plant physiological responses to these nutrients in either N- or P-limited settings. Using a common mangrove species, Avicennia germinans, in sites where growth was either N or P limited, we investigated differing physiological responses to N and P limitation and fertilization. We tested the hypothesis that water uptake and transport, and hydraulic architecture, were the main processes limiting productivity at the P-limited site, but that this was not the case at the N-limited site. We found that plants at the P-deficient site had lower leaf water potential, stomatal conductance and photosynthetic carbon-assimilation rates, and less conductive xylem, than those at the N-limited site. These differences were greatly reduced with P fertilization at the P-limited site. By contrast, fertilization with N at the N-limited site had little effect on either photosynthetic or hydraulic traits. We conclude that growth in N- and P-limited sites differentially affect the hydraulic pathways of mangroves. Plants experiencing P limitation appear to be water deficient and undergo more pronounced changes in structure and function with relief of nutrient deficiency than those in N-limited ecosystems.

Belize↗

Structural motifs of syringyl peroxidases predate not only the gymnosperm-angiosperm divergence but also the radiation of tracheophytes.

* The most distinctive variation in the monomer composition of lignins in vascular land plants is that found between the two main groups of seed plants. Thus, while gymnosperm lignins are typically composed of guaiacyl (G) units, angiosperm lignins are largely composed of similar levels of G and syringyl (S) units. * However, and contrary to what might be expected, peroxidases isolated from basal (Cycadales and Ginkgoales) and differentially evolved (Coniferales and Gnetales) gymnosperms are also able to oxidize S moieties, and this ability is independent of the presence or absence of S-type units in their lignins. * The results obtained led us to look at the protein database to search for homologies between gymnosperm peroxidases and true eudicot S-peroxidases, such as the Zinnia elegans peroxidase. * The findings showed that certain structural motifs characteristic of eudicot S-peroxidases (certain amino acid sequences and beta-sheet secondary structures) predate the gymnosperm-angiosperm divergence and the radiation of tracheophytes, since they are found not only in peroxidases from basal gymnosperms, ferns and lycopods, but also in peroxidases from the moss Physcomitrella patens (Bryopsida) and the liverwort Marchantia polymorpha (Marchantiopsida), which, as typical of bryophytes, do not have xylem tissue nor lignins.

Amino Acid Motifs↗

The distribution of acetylcholine in the Malayan jack-fruit plant, Artocarpus integra.

The distribution of acetylcholine in the seeds and leaves of the Malayan Jack-fruit plant, Artocarpus integra, has been studied with the view to obtaining evidence for the site of its formation. The terminal growing leaves on the side branches had a very high concentration of acetylcholine (770 mug./g.), while the acetylcholine content of the other leaves on the same branch progressively decreased with age. The total amount of acetylcholine stored in the terminal growing leaves was only 42 mug., but in the second leaves which had grown nearly to their full size it was 540 mug. From the third leaves, the amount of acetylcholine stored gradually decreased. The midribs and the secondary veins of the leaves when combined had a higher concentration of acetylcholine than had the blades. The acetylcholine concentration of the pith of the stem was 4.2 times higher than that of the cortex-phloem layer while that of the xylem layer was the lowest; in the root the pith had a value only one-seventh of the cortex. The younger part of the pith and the cortex-phloem layers of the stem contained more acetylcholine than the older parts. These findings support the view that the acetylcholine is synthesized in the growing leaves. An unusual lenticel-like structure in the cortex layer of the root contained more acetylcholine than the surrounding tissue.

Acetylcholine↗

Protein extraction from woody plants.

In this chapter we present a protocol for total protein extraction optimized for wood-forming tissue (differentiating secondary xylem). The protocol is then used for a series of other organs (root, leaf, pollen, bud, flower, cambium, and phloem) in broadleaf (oak and poplar) and conifer (pine) species. Proteins are first extracted from tissue powdered in liquid nitrogen using the TCA-acetone method and then solubilized in an optimized buffer. The resulting 2D gels can be viewed at http://cbi.labri.fr/outils/protic/index.php.

Chemical Precipitation↗

Effects of total dietary nitrogen and nitrogen form on the development of xylophagous leafhoppers

The influence of primary nutrients on the development of the leafhopper Homalodisca coagulata (Say) was examined using Glycine max (L.) Merrill subjected to treatments using different forms of nitrogen fertilization (urea and nitrate) and inoculation with Rhizobium bacteria. Concentrations of amino acids, organic acids, carbohydrates, and ureides in xylem fluid varied with treatment; differences were most pronounced in young plants (6 weeks after planting), and subsided as plants aged. Plants receiving urea fertilization had lower concentrations of organic nitrogen and carbon, but more balanced profiles of amino acids. Leafhoppers on urea-fertilized plants had much higher success in development (62 to 87% survival to adult stage) than H. coagulata developing on nitrate-fertilized plants (31-42%). The sex ratio of successfully developed adults varied greatly with treatments, indicating that female survival was much higher on urea-fertilized plants. Males and females utlilized diets differently as indicated by the carbon and nitrogen content of adult carcasses. Consumption rates and nitrogen assimilation efficiencies were higher for young (third instar) leafhoppers on urea-fertilized plants. Instar duration and total time of development were not affected by treatment. Highest rates of success in development on diets providing the lowest (yet most balanced) concentrations of total nitrogen, amino acids, and organic compounds underscored the importance of nutrient balance for leafhopper development. Arch. Copyright 1999 Wiley-Liss, Inc.

Journal Article↗

Featured organism: pathogen special: Vibrio cholerae, Pseudomonas aeruginosa and Xylella fastidiosa.

One could almost say that it is the latest fashion to sequence a bacterial genome. However, this would belittle the efforts of those working on these important organisms, whose data will greatly help those working on the prevention of disease in the fields of medicine and agriculture. In this feature we present a guided tour of the latest additions to the 'sequenced microbes' club. Vibrio cholerae is the causative agent of cholera, which is still a threat in countries with poor sanitation and unsafe drinking water. Pseudomonas aeruginosa is responsible for a large proportion of opportunistic human infections, typically infecting those with compromised immune systems, particularly cystic fibrosis patients, those patients on respirators and burn victims. Xylella fastidiosa is a plant pathogen that attacks citrus fruits by blocking the xylem, resulting in juiceless fruits of no commercial value.

Gammaproteobacteria↗

Isoxaflutole: the background to its discovery and the basis of its herbicidal properties.

This paper reviews the discovery of isoxaflutole (IFT), focusing on the chemical and physicochemical properties which contribute to the herbicidal behaviour of this new herbicide. IFT (5-cyclopropyl-1,2-isoxazol-4-yl alpha alpha alpha-trifluoro-2-mesyl-p-tolyl ketone) is a novel herbicide for pre-emergence control of a wide range of important broadleaf and grass weeds in corn and sugarcane. The first benzoyl isoxazole lead was synthesised in 1989 and IFT in 1990, and the herbicidal potential of the latter was identified in 1991. The decision to develop the molecule was taken after two years of field testing in North America. The biochemical target of IFT is 4-hydroxyphenylpyruvate dioxygenase (HPPD), inhibition of which leads to a characteristic bleaching of susceptible species. The inhibitor of HPPD is the diketonitrile derivative of IFT formed from opening of the isoxazole ring. The diketonitrile (DKN) is formed rapidly in plants following root and shoot uptake. The DKN is both xylem and phloem mobile leading to high systemicity. IFT also undergoes conversion to the DKN in the soil. The soil half-life of IFT ranges from 12 h to 3 days under laboratory conditions and is dependent on several factors such as soil type, pH and moisture. The log P of IFT is 2.19 and the water solubility is 6.2 mg litre-1, whereas the corresponding values for the DKN are 0.4 and 326 mg litre-1, respectively. These properties restrict the mobility of IFT, which is retained at the soil surface where it can be taken up by surface-germinating weed seeds. The DKN, which has a laboratory soil half-life of 20-30 days, is more mobile and is taken up by the roots. In addition to influencing the soil behaviour of IFT and DKN, the greater lipophilicity of IFT leads to greater uptake by seed, shoot and root tissues. In both plants and soil, the DKN is converted to the herbicidally inactive benzoic acid. This degradation is more rapid in maize than in susceptible weed species and this contributes to the mechanism of selectivity, together with the greater sowing depth of the crop.

4-Hydroxyphenylpyruvate Dioxygenase↗

Genetic analysis and epigenetic silencing of At4CL1 and At4CL2 expression in transgenic Arabidopsis.

4-coumarate::CoA ligase (4CL) gene family members are involved in channeling carbon flow into branch pathways of phenylpropanoid metabolism. Transgenic Arabidopsis plants containing the At4CL1 or At4CL2 promoter fused to the beta-glucuronidase (GUS) reporter gene show developmentally regulated GUS expression in the xylem tissues of the root and shoot. To identify regulatory genes involved in the developmental regulation of At4CL and other phenylpropanoid-specific genes, we generated ethyl methyl sulfate mutagenized populations of At4CL1::GUS and At4CL2::GUS transgenic lines and screened approximately 16,000 progeny for reduced or altered GUS expression. Several lines with reproducible patterns of reduced GUS expression were identified. However, the GUS-expression phenotype segregated in a non-Mendelian manner in all of the identified lines. Also, GUS expression was restored by 5-azacytidine (aza) treatment, suggesting inhibitory DNA methylation of the transgene. Southern analysis confirmed DNA methylation of the proximal promoter sequences of the transgene only in the mutant lines. In addition, retransformation of At4CL::GUS lines with further At4CL promoter constructs enhanced the GUS-silencing phenotype. Taken together, these results suggest that the isolated mutants are epimutants. Apparently, two different modes of silencing were engaged in the At4CL1::GUS and At4CL2::GUS silenced lines. While silencing in the seedlings of the At4CL1::GUS lines was root specific in seedlings, it affected all organs in the At4CL2::GUS lines. Also, At4CL1::GUS transgene silencing was confined to the transgene but At4CL2::GUS silencing extended to the endogenous At4CL2 gene. Organ-specific silencing of the At4CL1::GUS transgene cannot be explained by current models in the literature.

Arabidopsis↗

Electrophoretic analysis of coniferyl alcohol oxidase and related laccases.

Gradient gel electrophoretic methods enabled a distinction to be made between coniferyl alcohol oxidase (CAO) of lignifying cell walls and a pI approximately 9 pine "laccase" recently implicated in lignification (Science 1993 260, 672). Following treatment of a partially purified protein mixture from developing xylem of Pinus strobus with 2-[N-morpholine]ethanesulfonic acid (MES) buffer, isoelectric focusing and sodium dodecyl sulphate-polyacrylamide gel electrophoresis indicated that CAO had been selectively precipitated by MES and thereby purified to electrophoretic homogeneity. Purified CAO was determined to be a cell-wall-bound glycoprotein (38% glycan), M(r) 107,500, pI 7.6, pH and temperature optima 6.3 and 30 degrees C, respectively. By graphite-furnace atomic-absorption analysis, CAO contained one copper atom per protein molecule. Proteins obtained from lignifying cambial derivatives of conifers (family Pinaceae) and from Rhus typhina bark were compared with CAO and the pI approximately 9 pine "laccase" following electrophoresis and Western blotting. For Abies balsamea, Larix laricina, Picea rubens, Pinus banksiana, Pinus taeda, and R. typhina, the isoelectric points of oxidatively active bands were identical to those of purified CAO. In addition, for all species only the pI 7.6 band was immunoreactive with antibodies against periodate-deglycosylated CAO.

Alcohol Oxidoreductases↗

Toxicity of cotton phytoalexins to zoopathogenic fungi.

The sesquiterpenoid phytoalexins desoxyhemigossypol, desoxymethoxyhemigossypol, and hemigossypolone formed in cotton (Gossypium hirsutum and G. barbadense) stem xylem infected with Verticillium dahliae were shown to be highly toxic to zoopathogenic fungi. This appears to be the first study of the toxicity of terpenoid phytoalexins to zoopathogenic fungi. The toxicities of the phytoalexins expressed as MIC (micrograms ml-1) values were 8 to 128 against four isolates of Candida albicans and one isolate of Cryptococcus neoformans. These highly toxic compounds or their derivatives may prove useful for the treatment of animal mycoses.

Antifungal Agents↗

Translocation and translaminar bioavailability of two neonicotinoid insecticides after foliar application to cabbage and cotton.

A laboratory study was undertaken to investigate the leaf systemic properties and the translaminar aphicidal activity of two commercialised neonicotinoid (chloronicotinyl) insecticides. For that purpose [14C]imidacloprid was subjected to uptake and translocation studies in cabbage and cotton after foliar application. Foliar penetration and short-term translocation patterns of imidacloprid were similar in both plant species. Nevertheless imidacloprid penetrated twice as much into cabbage leaves as it did into cotton leaves. It showed a comparable translaminar behaviour and was entirely translocated acropetally, indicating its well-known xylem mobility. The translaminar and acropetal movement of imidacloprid and acetamiprid were quantified by simple laboratory bioassays using the green peach aphid, Myzus persicae (Sulzer), and the cotton aphid, Aphis gossypii (Glover), as typical homopteran pests for cabbage and cotton, respectively. A single dose (7.5 micrograms AI per leaf) applied to the upper leaf surface of cabbage and cotton was tested against aphids feeding on the lower leaf surface both close to and distant from the site of application 1, 5 and 12 days after treatment. The translaminar residual activity of imidacloprid on cabbage leaves was superior to that of acetamiprid, whereas its translaminar efficacy against A gossypii on cotton was inferior to that of acetamiprid. However, oral ingestion bioassays using an artificial double membrane feeding system revealed no significant differences in intrinsic activity between the two neonicotinoids tested.

Anabasine↗

Spatial and temporal distribution of imidacloprid and thiamethoxam in citrus and impact on Homalodisca coagulata populations.

Titers of two systemic neonicotinoid insecticides in citrus trees were measured in conjunction with conventional evaluations of their impact on glassy-winged sharpshooter (Homalodisca coagulata (Say); GWSS) populations. Xylem fluid samples were collected at regular intervals and from multiple locations within field-grown citrus trees to determine imidacloprid and thiamethoxam concentrations using commercial ELISA kits. Uptake profiles varied considerably with peak mean titers of imidacloprid occurring 6-8 weeks after application compared with 2 weeks for thiamethoxam. The persistence of each compound also varied as near-peak levels of imidacloprid were sustained for another 6-10 weeks before gradually declining. In contrast, thiamethoxam titers declined more rapidly after the initial peak, possibly reflecting an application rate only one-quarter of that used for imidacloprid. Within-tree distributions were more similar for the two compounds, with no significant effect due to height of the sample (upper or lower half) or to the quadrant location within the tree, with the exception of one quadrant in the thiamethoxam-treated trees. Substantial reductions in GWSS nymphs and adults were observed in imidacloprid-treated trees during the 2001 trial and were sustained for 4-5 months after treatment. Treatment effects on nymphs were not as well pronounced in the 2002 trial, when overall GWSS infestations were much reduced from the previous year. However, consistently lower adult infestations were still observed in 2002 for both treatments compared with untreated trees. Information on the spatial and temporal profiles in citrus trees was obtained for both compounds to complement field impact data and improve understanding of their pest management potential.

Animals↗

Monoterpene synthases of loblolly pine (Pinus taeda) produce pinene isomers and enantiomers.

The turpentine fraction of conifer oleoresin is a complex mixture of monoterpene olefins and plays important roles in defense and in the mediation of chemical communication between conifer hosts and insect predators. The stereochemistry of the turpentine monoterpenes is critical in these interactions, influencing host recognition, toxicity, and potency of derived pheromones, and the stereochemical composition of these compounds lends insight into their biogenetic origin, with implications for the numbers and types of enzymes responsible and their corresponding genes. Analysis of the oleoresin from several tissues of loblolly pine (Pinus taeda) showed the derived turpentine to consist mainly of (+)-(3R:5R)-alpha-pinene and (-)-(3S:5S)-beta-pinene. Cell-free extracts from xylem tissue yielded three monoterpene synthases which together account for the monoterpene isomer and enantiomer content of the turpentine of this tissue. The major products of these enzymes, produced from the universal precursor of monoterpenes, geranyl diphosphate, were shown to be (+)-alpha-pinene, (-)-alpha-pinene, and (-)-beta-pinene, respectively. In most properties (molecular mass of approximately 60 kDa, K(m) for geranyl diphosphate of 3 microM, requirement for monovalent and divalent cations), these enzymes resemble other monoterpene synthases from conifer species.

Animals↗

Anatomical features of pepper plants (Capsicum annuum L.) grown under red light-emitting diodes supplemented with blue or far-red light.

Pepper plants (Capsicum annuum L. cv., Hungarian Wax) were grown under metal halide (MH) lamps or light-emitting diode (LED) arrays with different spectra to determine the effects of light quality on plant anatomy of leaves and stems. One LED (660) array supplied 90% red light at 660 nm (25nm band-width at half-peak height) and 1% far-red light between 700-800nm. A second LED (660/735) array supplied 83% red light at 660nm and 17% far-red light at 735nm (25nm band-width at half-peak height). A third LED (660/blue) array supplied 98% red light at 660nm, 1% blue light between 350-550nm, and 1% far-red light between 700-800nm. Control plants were grown under broad spectrum metal halide lamps. Plants were gron at a mean photon flux (300-800nm) of 330 micromol m-2 s-1 under a 12 h day-night photoperiod. Significant anatomical changes in stem and leaf morphologies were observed in plants grown under the LED arrays compared to plants grown under the broad-spectrum MH lamp. Cross-sectional areas of pepper stems, thickness of secondary xylem, numbers of intraxylary phloem bundles in the periphery of stem pith tissues, leaf thickness, numbers of choloplasts per palisade mesophyll cell, and thickness of palisade and spongy mesophyll tissues were greatest in peppers grown under MH lamps, intermediate in plants grown under the 660/blue LED array, and lowest in peppers grown under the 660 or 660/735 LED arrays. Most anatomical features of pepper stems and leaves were similar among plants grown under 660 or 660/735 LED arrays. The effects of spectral quality on anatomical changes in stem and leaf tissues of peppers generally correlate to the amount of blue light present in the primary light source.

Calcium Oxalate↗

Effects of elevated selenium and salinity concentrations in root zone on selenium and salt secretion in saltgrass (Distichlis spicata L.).

The effects of elevated selenium (Se) and salinity concentrations in the root zone on Se and salt secretion and accumulation were studied for an inland (Kesterson) and a coastal (Bodega Bay) saltgrass in sand culture and under greenhouse conditions. The results of this study indicate that the secretory mechanism of the saltgrass exhibits high ion specificity. Plants of both ecotypes were more efficient at Cl- and Na+ secretion than at SO4(2-) and Se secretion, suggesting that the saltgrass secretion mechanism is adapted primarily to saline environments high in NaCl. The saltgrass plants of the Kesterson ecotype secreted more Se than the Bodega Bay plants when treated with Se alone. However, the Bodega Bay plants secreted more Se than the Kesterson plants when the plants were treated with Se+NaCl. These differences reflect a ecotypical difference in which the Kesterson plants are more adaptive to the seleniferous soil, and the Bodega Bay plants are more adaptive to a coastal saline soil high in NaCl. Sulfate availability inhibited both Se accumulation and Se secretion in the plants of both ecotypes by approximately 98%. Ion secretion molar ratios of Na:Cl were calculated and the results suggest that Na+ secretion is dependent on the availability of Cl-. Selenium was taken up by plants with little discrimination, and thus it may be regarded as a master of chemical mimicry sharing similar physical and chemical properties with sulfur (S). Selenium and salt accumulation indices and secretion efficiency indices were calculated and found that the accumulation indices were higher for Se than for S, suggesting that Se uptake may be more passive and less regulated by active transport than S. Secretion efficiency was much higher for Na+ and Cl- than for Se and SO4(2-), but the efficiency indices between ecotypes were comparable, suggesting that the secretion mechanism in this species is designed mainly for adaptation of high NaCl concentrations. About 85% of the secreted Se was found in selenate form. This finding suggests that the mechanism of Se translocation and Se secretion in the saltgrass resembles the movement of selenate in xylem found in nonhalophytic plants. Less than 15% of organic Se was detected in the secretions and only trace amount of selenite was detected. Overall, the Se secretion mechanism in saltgrass does not seem to contribute substantial Se speciation in the plant soil system.

Poaceae↗

Structure of native and expanded sobemoviruses by electron cryo-microscopy and image reconstruction.

Rice yellow mottle virus (RYMV) and southern bean mosaic virus, cowpea strain (SCPMV) are members of the Sobemovirus genus of RNA-containing viruses. We used electron cryo-microscopy (cryo-EM) and icosahedral image analysis to examine the native structures of these two viruses at 25 A resolution. Both viruses have a single tightly packed capsid layer with 180 subunits assembled on a T=3 icosahedral lattice. Distinctive crown-like pentamers emanate from the 12 5-fold axes of symmetry. The exterior face of SCPMV displays deep valleys along the 2-fold axes and protrusions at the quasi-3-fold axes. While having a similar topography, the surface of RYMV is comparatively smooth. Two concentric shells of density reside beneath the capsid layer of RYMV and SCPMV, which we interpret as ordered regions of genomic RNA. In the presence of divalent cations, SCPMV particles swell and fracture, whereas the expanded form of RYMV is stable. We previously proposed that the cell-to-cell movement of RYMV in xylem involves chelation of Ca(2+) from pit membranes of infected cells, thereby stabilizing the capsid shells and allowing a pathway for spread of RYMV through destabilized membranes. In the context of this model, we propose that the expanded form of RYMV is an intermediate in the in vivo assembly of virions.

Amino Acid Sequence↗

Fast NMR flow measurements in plants using FLASH imaging.

A fast method for quantitative NMR imaging of flow velocities in intact plants is described. The purpose of this method is to observe dynamic changes of flow velocity in the xylem of plants after fast changes of environmental conditions. The spatial image resolution is 47 x 188 micrometer(2) in-plane. The method applies a fast gradient echo sequence (FLASH). Compared to other flow NMR imaging sequences, the imaging time was reduced by a factor of 6 with comparable signal-to-noise ratio. A complete flow measurement consists of a set of 8 different flow weighted images with a total acquisition time of 3.5 min.

Magnetic Resonance Spectroscopy↗

Microscopic imaging of slow flow and diffusion: a pulsed field gradient stimulated echo sequence combined with turbo spin echo imaging.

In this paper we present a pulse sequence that combines a displacement-encoded stimulated echo with rapid sampling of k-space by means of turbo spin echo imaging. The stimulated echo enables the use of long observation times between the two pulsed field gradients that sample q-space completely. Propagators, constructed with long observation times, could discriminate slowly flowing protons from diffusing protons, as shown in a phantom in which a plug flow with linear velocity of 50microm/s could clearly be distinguished from stationary water. As a biological application the apparent diffusion constant in longitudinal direction of a transverse image of a maize plant stem had been measured as a function of observation time. Increasing contrast in the apparent diffusion constant image with increasing observation times were caused by differences in plant tissue: although the plant stem did not take up any water, the vascular bundles, concentrated in the outer ring of the stem, could still be discerned because of their longer unrestricted diffusional pathways for water in the longitudinal direction compared to cells in the parenchymal tissue. In the xylem region of a tomato pedicel flowing water could be distinguished from a large amount of stationary water. Linear flow velocities up to 0.67 mm/s were measured with an observation time of 180 ms.

Algorithms↗