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PubMedDate not supplied

Localization of cauliflower mosaic virus in the cell nucleus of Brassica pekinensis L.

Cauliflower mosaic virus (CaMV) particles were observed in the nuclei of xylem parenchyma cells in Brassica pekinensis L. doubly infected by CaMV and turnip mosaic virus (TuMV). CaMV particles were aggregated in the nucleoplasm but not embedded in viroplasms. This phenomenon was not detected in cell nuclei of mesophyll tissue. Typical features associated with infection by either CaMV or TuMV normally occurred in the cytoplasm of cells of both tissues: two types of viroplasms with embedded CaMV particles and cylindrical inclusions induced by TuMV. Among the main hypotheses which could explain this particular CaMV localization, we checked that it was not the result of the coinfection with TuMV, since we also found CaMV particles in the same place in single infected plants. Other explanations are discussed, including a possible particularity of the infection in xylem tissue or a specific property of an unusual CaMV isolate.

Brassica↗
PubMed2005

Colonization of cantaloupe roots by Monosporascus cannonballus.

Penetration of Monosporascus cannonballus into and growth within cantaloupe roots was studied using light and electron microcopy. Germ tubes penetrated the epidermis, and hyphae grew, without branching, almost directly to the xylem. The hyphae traversed the endodermis into protoxylem cells, and then grew extensively within the lumen of metaxylem vessels. Eventually, the hyphae grew back out into the cortical cells. A relatively low percentage of cells within both the cortex and xylem of lesions contained hyphae. The hyphae were generally localized within the lesion and could rarely be isolated more than 2 mm away from the margin of the lesion. Regardless of tissue type, hyphae were predominately intracellular. M. cannonballus appeared to be most similar to vascular wilt pathogens in its mode of parasitism, but does not spread via the vascular system to above-ground plant tissues.

California↗
PubMed2002

Tracing the history of nuclear releases: determination of 129I in tree rings.

Concentrations of the long-lived radioisotope 1291 were measured in dated tree rings in order to determine whether the distribution of this isotope reflects the history of nuclear deposition. 129I concentrations and 129I/127I ratios were analyzed in tree rings and bark samples from four trees at West Valley, NY, and from one tree at Rochester, NY. West Valley was the site of short-lived nuclear fuel reprocessing activities (1966-1972), while Rochester, located 115 km to the northeast, provided a regional control site for the study. The selected trees reflect different modes of fluid and nutrient transport in trees, with three species of ring-porous trees (elm, oak, and locust), one semidiffuse (cherry), and one diffuse-porous tree (maple). The results show that 1291 levels in ring-porous trees, in which xylem or hydrologic tissue is localized in the outermost growth ring, are generally well correlated with the expected 1291 deposition pattern for the region. In contrast, tree rings of the more common semidiffuse to diffuse-porous wood, where xylem is disseminated throughout the trunk, show a less well developed 129I signal, probably due to the transport of iodine ions across annual rings. Iodine concentrations in the tree rings range from 0.04 to 2 mg/kg, 129I/127I ratios from 6 x 10(-10) to 3.8 x 10(-6). Tree bark and the outermost rings show significantly higher 129I concentrations than the wood of the trunk. The 129I/127I ratios for bark are very similar to values obtained for surface soil and water at the two localities, while inner rings have ratios similar to those in deeper layers of the soil, reflecting different pathways for 129I uptake and the differences in ambient 1291 levels between the atmosphere and deep soil. Although ring porous trees preserve the depositional pattern of nuclear releases, rings older than or close to the onset of the nuclear age have 129I/127I ratios significantly above the preanthropogenic level, suggesting that even in these trees some redistribution of 129I occurs throughout the trunk. Our results indicate that growth rings from ring-porous wood are useful in time-series analyses of regional 129I deposition, yielding reliable information on relative changes in 129I concentrations but requiring caution in the reconstruction of absolute ambient concentrations during any given time.

Air Movements↗
PubMed2003

TCE diffusion to the atmosphere in phytoremediation applications.

The phytoremediation of trichloroethylene (TCE) and other chlorinated compounds has been studied over the past decade, and full-scale systems are in place. The results regarding TCE fates and removal pathways are inconclusive and conflicting, particularly the results regarding volatilization to the atmosphere. Research presented here demonstrates that TCE is taken up by trees and volatilized to the atmosphere. TCE diffusion along the transpiration pathway is shown to be the primary process for TCE volatilization, although volatilization can occur from both stems and leaves. Two concurrent processes influence the eventual fate: transport with transpiration stream through xylem tissues and diffusion from transpiration stream to atmosphere. TCE diffusion flux invariably decreased with height for trees planted in soil or grown hydroponically. In both laboratory experiments and field sampling, TCE concentrations in the transpiration stream (e.g., xylem tissues) decreased with elevation. In field samples, TCE concentrations also decreased in the radial direction, providing fundamental evidence for diffusion. The TCE concentrations in tissues responded linearly to the exposure concentrations at the roots, while TCE diffusion from tree stems was influenced by concentration and transpiration rates.

Biodegradation, Environmental↗
PubMed2004

Modeling of TCE diffusion to the atmosphere and distribution in plant stems.

Fate of chlorinated solvents in phytoremediation has been delineated by many discoveries made in recent years. Plant uptake, metabolism, rhizosphere degradation, accumulation, and volatilization were shown to occur to differing degrees for many organic contaminants including chlorinated solvents. Among these mechanistic findings, recent research confirmed that volatile organic compounds (VOCs) volatilize from stems and that the resulting diffusive flux to the atmosphere is related to exposure concentration and to height up the stem. A comprehensive model was developed based upon all identified fate and transport mechanisms for VOCs, including translocation in the xylem flow and diffusion. The dispersion and diffusion in the radial direction were considered as one process (effective diffusion) as the two could not be investigated individually. The mechanism-based model mathematically indicates an exponential decrease of concentrations with height. While an analytic solution for the comprehensive model was not attained, it can serve as a starting point for other modeling efforts. The comprehensive model was simplified in this work for practical application to experimentally obtained data on trichloroethylene (TCE) fate. Model output correlated well with experimental results, and effective diffusivities for TCE in plant tissues were obtained through the model calibrations. The simplified model approximated TCE concentrations in the transpiration stream as well as TCE volatilization to the atmosphere. Xylem transport, including advection, dispersion, and diffusion through cell walls with subsequent volatilization to the atmosphere, is a major fate for VOCs in phytoremediation.

Atmosphere↗
PubMed2006

Fractionation mechanisms of rare earth elements (REEs) in hydroponic wheat: an application for metal accumulation by plants.

Fractionations of rare earth elements (REEs) in wheat (Triticum aestivum L.) were observed through application of exogenous mixed REEs under hydroponic conditions. Middle REE (MREE), light REE (LREE), and heavy REE (HREE) enrichments were found in roots, stems, and leaves, respectively, accompanied by the tetrad effect (an effect that can cause a split of REE patterns into four consecutive segments) in these organs. Investigations into REE speciation in roots and in the xylem sap with X-ray absorption spectroscopy (XAS) and nanometer-sized TiO2 adsorption techniques, associated with other controlled experiments, demonstrated that REE fractionations in wheat were caused by the combined effects of chemical precipitation, cell wall absorption, and solution complexation by organic ligands in the xylem vessels. REE fractionations in wheat, which were derived from the small differences of chemical properties across REE series, may reflect a sensitive internal chemical environment that influences plant accumulation for REEs and their analogues actinide radionuclides.

Adsorption↗
PubMed2004

Metabolic profiling: a new tool in the study of wood formation.

In the realm of plant genomics, metabolic profiling has become a valuable tool with which to assess the effect of genetic and/or environmental factors on plant development. This paper reports the first application of metabolic profiling on differentiating xylem tissue of loblolly pine. A protocol is presented for the analysis of loblolly pine xylem tissue. The effects of sample preparation, extraction, and derivatization on the corresponding metabolite profiles and yields have been investigated and are reported. Gas chromatography-mass spectroscopy has been used to quantify >60 polar and lipophilic metabolites from wood-forming tissue. It was possible to assign chemical structures to approximately half of these compounds. Comparison of six loblolly pine genotypes, three high cellulose (50-52%) and three medium (45-48%) cellulose, showed distinct metabolic profiles. Principal component analysis enabled the assignment of metabolic phenotypes using these large data sets. Metabolic phenotype clustering occurred in which the three high-cellulose genotypes were segregated from the medium-cellulose genotypes. These results demonstrate the use of metabolic profiling for the study of wood-forming tissue and as a tool in functional genomics.

Cellulose↗
PubMed2005

Relationship of carotenoids and tocopherols in a sample of carrot root-color accessions and carrot germplasm carrying Rp and rp alleles.

Carotenoids and tocopherols are powerful antioxidants synthesized in plants from a common precursor. They may offer significant health benefits to humans. Seed oils have been shown to possess high levels of tocopherols, but little is known about their levels in the edible portions of most vegetable crops. A two-year field experiment was conducted at two locations to assess levels of major carotenoids and tocopherols in carrot (Daucus carota) root and leaf tissue. Levels of compounds in root tissue reported on a dry weight basis were as follows: alpha-tocopherol, 0.04-0.18 ppm; lycopene, 0.00-52.94 ppm; alpha-carotene, 10.63-1504.76 ppm; and beta-carotene, 26.69-1673.76 ppm. Higher levels of all carotenoids were measured in phloem tissue than in xylem. Leaf tissue levels of tocopherols measured on a dry weight basis ranged from 0.02 to 0.85 ppm, whereas levels of carotenoids ranged from 12.81 to 411.66 ppm. In xylem tissue, alpha-tocopherol was significantly (P < or =0.001) positively correlated with alpha-carotene (r = 0.65) and with beta-carotene (r = 0.52). This positive correlation indicates it may be possible to select for both increased alpha-tocopherol and carotenoids in carrot. The reduced pigment (rp) mutation of carrot exhibited a 96% reduction in levels of alpha- and beta-carotene and a 25-43% reduction in alpha-tocopherol when compared to a near-isogenic line. In plants homozygous for rp, a substantial increase was observed in phytoene, a precursor to carotenoids, suggesting the location of the rp lesion in the carotenoid synthesis pathway.

Alleles↗
PubMed1997

Characterization of a gene encoding a DNA-binding protein that interacts in vitro with vascular specific cis elements of the phenylalanine ammonia-lyase promoter.

A study of the expression of a bean phenylalanine ammonia-lyase (PAL) promoter/beta-glucuronidase gene fusion in transgenic tobacco has shown that the PAL2 promoter has a modular organization. Expression of the PAL2 promoter in the vascular system involves positive and negative regulatory cis elements. Among these elements is an AC-rich motif implicated in xylem expression and a suppressing cis element for phloem expression. Using radiolabelled complementary oligonucleotides bearing the AC-rich motif, a cDNA clone encoding a DNA-binding protein has been isolated from a tobacco lambda gt11 expression library. This factor, named AC-rich binding factor (ACBF), showed binding specificity to the AC-rich region. The specificity of ACBF for the AC-rich region was also shown using a gel retardation assay with an ACBF recombinant protein extract. The deduced amino acid sequence from ACBF contains a long repeat of glutamine residues as found in well characterized transcription factors. Interestingly, ACBF shared sequence similarity to conserved amino acid motifs found in RNA-binding proteins. Genomic gel blot analysis indicated the presence of a small gene family of sequences related to ACBF within the tobacco nuclear genome. Analysis of tobacco mRNA using the ACBF cDNA as probe showed that while ACBF mRNA was present in all tissues examined, the highest transcript accumulation occurred in stem tissues. The functional characteristics of the AC-rich sequence were examined in transgenic tobacco. A heptamer of the AC-rich sequence, in front of a minimal 35S promoter from cauliflower mosaic virus (-46 to +4), conferred specific expression in xylem.

Amino Acid Sequence↗
PubMed1998

A novel protein associated with citrus blight has sequence similarities to expansin.

A protein associated with citrus blight (CB), a disease of unknown cause, was partially characterized. The 12 kDa protein, designated p12, is diagnostic of CB and is present in leaves and xylem fluid from roots and stems of CB-affected trees. The protein, and up to six other CB-specific proteins, are readily detected by SDS-PAGE of xylem fluid from CB-affected trees. The partial N-terminal amino acid sequence of p12 was found to be unique based on database searches. A cDNA library from CB-affected root cambium was screened with a 60 bp fragment, obtained by PCR amplification of cDNA with degenerate primers designed using the amino acid sequence of p12, and two clones were selected. These clones were sequenced revealing a 674 nucleotide cDNA with a 393 nt ORF which included sequence predicted by the N-terminal amino acid sequence of p12. The amino acid sequence based on the p12 ORF was found to be up to 49% similar and 31% identical to expansins. Bacterial expression of the cloned ORF, which encodes an 11.8 kDa protein plus an N-terminal hydrophobic signal peptide, produced an immunoreactive protein of the expected size. By northern blot analysis, it was determined that p12 transcripts are present in root and stem cambium, but not in leaves of CB-affected trees, suggesting transport of the protein to leaves. Southern hybridization analysis of citrus genomic DNA indicated that p12 is a citrus encoded protein.

Amino Acid Sequence↗
PubMed1999

cDNA cloning and heterologous expression of coniferin beta-glucosidase.

Coniferin beta-glucosidase (CBG) catalyzes the hydrolysis of monolignol glucosides to release the cinnamyl alcohols for oxidative polymerization to lignin. Utilizing the N-terminal amino acid sequence of the purified enzyme, the corresponding full-length cDNA sequence was isolated from a Pinus contorta xylem-specific library. The isolated 1909 nucleotide cDNA was confirmed to be that of CBG on the basis of its high homology to family 1 glycosyl hydrolases, the sequence identity with the N-terminal amino acid residues of the purified enzyme, and the coniferin hydrolytic activity and substrate specificity profile displayed by the recombinant protein when expressed in Escherichia coli. The presence of a 23 amino acid N-terminal signal peptide in the deduced 513 amino acid enzyme suggests that CBG is a secretory protein targeted to the ER. The isolation of CBG cDNA will facilitate the evaluation of the importance of this enzyme in the ultimate stages of lignin biosynthesis and could be a valuable tool in manipulating lignin levels in xylem cell walls.

Amino Acid Sequence↗
PubMed2003

Abnormal 'wrinkled' cell walls and retarded development of transgenic Arabidopsis thaliana plants expressing endo-1,4-beta-glucanase (cell) antisense.

Transgenic Arabidopsis thaliana plants expressing cel1 antisense exhibit reduced levels of cel1 mRNA and protein compared with wild-type plants. The former display significant alterations in their phenotype. cel1 antisense plants have shorter stems and roots and are mechanically weaker than their wild-type counterparts. In cel1 antisense plants, the cell wall structure is markedly disrupted: both fluorescent confocal microscopy and scanning electron microscopy revealed 'wrinkled' cell walls, thus indicating that CEL1 plays an important role in cell wall relaxation during cell growth and expansion. In cel1 antisense plants, the number of xylem elements per bundle is smaller than in the wild-type. In addition, both xylem elements and interfascicular fibers are significantly less lignified in the former. It is suggested that in A. thaliana, abnormal cell wall deposition affected by CEL1 depletion is associated not only with cell growth, but also with the differentiation process in the vascular and supporting tissues.

Arabidopsis↗
PubMed2003

Regulated expression of Arabidopsis shaker K+ channel genes involved in K+ uptake and distribution in the plant.

Potassium is the most abundant cation in the cytosol, where it plays a role in basal functions. Rapid uptake and distribution of K+ is therefore required for plant growth. Three members of the so-called Shaker K+ channel gene family (nine genes identified in Arabidopsis) play a role in these transports: AKT1, SKOR and AKT2. The encoded proteins are involved in K+ uptake by the root, K+ secretion into the xylem sap and K+ transport in the phloem tissues, respectively. Using the GUS reporter strategy, we have found that another Shaker channel gene, AtKC1, is expressed in epidermal and cortical cells in roots (supporting the hypothesis of a role in K+ uptake from the soil, together with AKT1), and in trichomes and hydathodes in leaves. These four genes were selected for expression studies, and two-hybrid experiments were performed for channels displaying overlapping expression patterns. The data support the hypothesis that physical interactions could occur in planta between AtKC1 and AKT1, and between AKT1 and AKT2. Potassium deficiency, salt stress and hormonal treatments (ABA, BA, 2,4-D) were found to differentially affect channel mRNA levels, each channel displaying its own regulation pattern. The most prominent effects were (1) a strong induction of AtKC1 transcript accumulation in leaves (hydathodes, trichomes and leaf epidermis) in response to NaCl treatment, suggesting a key role of the protein in adaptation to saline conditions, and (2) a strong decrease in SKOR transcript levels by hormones, supporting the hypothesis that K+ secretion into the xylem sap is under tight hormonal control.

2,4-Dichlorophenoxyacetic Acid↗
PubMed2004

Morphogenetic processes: application to cambial growth dynamics.

Both the physiological and the pathological morphogenetic processes that we can meet in embryogenesis, neogenesis and degenerative dysgenesis present common features: they are ruled by three different kinds of mechanisms, one related to cell migration, the second to cell differentiation and the third to cell proliferation. We deal here with an application to the cambial growth which essentially involves the third type of mechanism. Woody plants produce secondary tissue (secondary xylem and phloem) from a meristematic tissue called vascular cambium, responsible for the radial growth of a tree. This paper focuses on the formation of secondary xylem, considered in two dimensions in a cross-section framework. A new discrete modelling approach is used, based on the cellular scale, in order to attain a more accurate understanding of how the elementary microscopic behaviour of each cell takes part in the macroscopic morphogenesis. The mathematical model essentially uses an occurrence method simulating the main features of radial growth with simple geometric rules, such as Thom's division rule (Thom,1972)to account for the cell proliferation. The study applies to concrete instances in which the changes made in the geometrical cellular patterns of the vascular cambium clearly affect the shape of the tree, as in Pinus radiata (D. Don.).

Morphogenesis↗
PubMed2001

Genes encoding chitinase-antifreeze proteins are regulated by cold and expressed by all cell types in winter rye shoots.

One group of antifreeze proteins (AFPs) is composed of two chitinases that accumulate in the apoplast of winter rye leaves during cold acclimation. In this study, the 28- and 35-kDa chitinase-AFPs were localized in nonacclimated and cold-acclimated rye leaves by immunoelectron microscopy with an antiserum produced against the purified winter rye 35-kDa chitinase-AFP. In cold-acclimated winter rye leaves, labelled chitinase-AFPs were abundant in the walls of epidermal, parenchymal sheath and mesophyll cells and xylem vessels, while less label was present in walls of vascular parenchyma cells. In contrast, chitinase labelling was essentially absent in the nonacclimated cells except in xylem vessels. As shown by RNA blotting, the transcripts of chitinase-AFPs accumulated to a high level in rye leaves during cold acclimation, to a lesser extent in crowns and were not detectable in roots. mRNA transcripts of the 28-kDa chitinase-AFP were localized in rye leaves by in situ hybridization. The chitinase-AFP transcripts were found in the same cell types as the protein itself. We conclude that all metabolically active cell types in cold-acclimated winter rye leaves and crowns are able to synthesize chitinase-AFPs and secrete them into adjacent cell walls, where they may interact with ice to delay its propagation through the plant and modify its growth.

Journal Article↗
PubMed2001

Caffeoyl-coenzyme A 3-O-methyltransferase enzyme activity, protein and transcript accumulation in flax (Linum usitatissimum) stem during development.

Flax (Linum usitatissimum) is a commercially important fiber crop in Europe. Lignification of its phloem fibers, although weak, causes a decrease in their commercial quality. In flax, fiber lignin mainly consists of guaiacyl (G) units in contrast to the mixed guaiacyl-syringyl (G-S) lignin type occurring in xylem fibers. G lignins are reported as more condensed polymers due to a higher frequency of 5-5 linkages, whereas the deposition of syringyl end groups in lignins increases the proportion of alky-aryl ether linkages as beta-O-4-bonds. The type of linkages within a lignin polymer depends on the methylation of either the 3-hydroxyl groups or both 3-OH and 5-OH groups, which is controlled by two enzymes: caffeate 3-O-methyltransferase (COMT) and caffeoyl-coenzyme A 3-O-methyltransferase (CCoAOMT). First, we measured the in vitro activity of both OMTs in the flax stem tissues during stem development. CCoAOMT activity varied in the same way as COMT, i.e. increased gradually with stem maturity, from the top to the bottom of the stem, was maximum at the flowering stage and was lower, but still scorable, in the outer fiber-bearing tissues than in the xylem cells. In a second step, we focused our studies on the characterization of CCoAOMT in order to understand the implication of this enzyme in the lignification of flax fiber cells. CCoAOMT activity appeared to be associated with the accumulation of an acidic 33-kDa polypeptide identified as a CCoAOMT after immunological cross-reactivity with a poplar CCoAOMT and microsequencing. The differential accumulation of the CCoAOMT protein was confirmed by immunolocalization on tissue prints and correlated with that of the transcripts, suggesting a transcriptional regulation of CCoAOMT in the flax stem.

Journal Article↗
PubMed2001

Overlapping expression of cytosolic glutamine synthetase and phenylalanine ammonia-lyase in immature leaf blades of rice.

In order to estimate whether cytosolic glutamine synthetase (GS1; EC 6.3.1.2) is partly coupled to the reaction of phenylalanine ammonia-lyase (PAL; EC 4.3.1.5) in developing organs of rice (Oryza sativa L.), we compared the expression pattern of transcripts and proteins for GS1 and PAL in the tissue sections from leaf blades at various stages of development. In immature vascular bundles of unexpanded leaf blades, GS1 mRNA was mainly detected in xylem parenchyma cells, mestome-sheath cells, and sclerenchyma cells. PAL transcripts were also accumulated in these cell types. Vascular bundles in midribs of immature leaf blades contained mRNAs and proteins for both GS1 and PAL abundantly in sclerenchyma cells, although distribution of these two proteins was not completely overlapped. In immature vascular bundles in midribs, lignin deposition was observed in cell walls of xylem parenchyma cells, mestome-sheath cells and sclerenchyma cells. These results implied that a part of GS1 in unexpanded leaf blades is possibly involved in reassimilation of ammonia released from PAL reaction during the lignin production.

Journal Article↗
PubMed2002

Water entry into detached root systems saturates with increasing externally applied pressure; a result inconsistent with models of simple passive diffusion.

The most widely accepted model of radial water entry from the soil into the xylem of roots is based on principles of ordinary passive diffusion. However, long-standing problems with this model remain unresolved, which concern variable intrinsic properties of conductivity, Lp, changing reflection coefficients, sigma, and inaccurate resolution of osmotic differentials between the soil and xylem. Our study re-examined pressure flow relationships in isolated roots of tomato (Lycopersicon esculentum Mill. cv. Montfavet), pea (Pisum sativum cv. Baccara) and soybean (Glycine max L. Merryl cv. Essor) manipulated in a pressure chamber. In addition to problems previously recognized with the simple passive diffusion model, a new conflict, flow saturation, was observed at high pressures. Experiments revealed that the plateau in flow, Jmax seen at high pressures followed natural rhythms diurnally and developmentally, and was not due to root damage or unnatural flow restriction. Near the end of the photoperiod, Jmax closely correlated with root dry mass. The above inconsistencies between observations in pressure-flow kinetics and ordinary passive diffusion indicate that either the current model should be adjusted or a new model should be proposed.

Journal Article↗