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PubMed2002

Detection of root mucilage using an anti-fucose antibody.

Plant root mucilage is known to enhance soil quality by contributing towards the soil carbon pool, soil aggregation, detoxification of heavy metal ions and interactions with rhizospheric microflora. Mucilage consists of many monosaccharide units, including fucose which can be used as an indicator for plant root based polysaccharides. This is the first report of an immunological technique developed to use anti-fucose antibodies as markers for probing and localizing fucosyl residues in mucilage polysaccharide and, in turn, for localization of plant root mucilage. Fucose was complexed with bovine serum albumin to raise antibodies against fucose. A fucose-directed antibody was shown to cross-react with root cap mucilages from grasses. This antibody was used to localize root mucilage polysaccharide in maize and wheat root caps using immunogold electron microscopy. Abundant labelling could be localized on the cell wall, and in the intercellular matrix and vesicles of the peripheral root cap cells. Labelling was less intense in cells towards the centre of the root cap tissue. Control experiments confirmed that immunogold localization of fucose was specific and reliable.

Animals↗
PubMed2006

Abrupt tropical climate change: past and present.

Three lines of evidence for abrupt tropical climate change, both past and present, are presented. First, annually and decadally averaged delta(18)O and net mass-balance histories for the last 400 and 2,000 yr, respectively, demonstrate that the current warming at high elevations in the mid- to low latitudes is unprecedented for at least the last 2 millennia. Second, the continuing retreat of most mid- to low-latitude glaciers, many having persisted for thousands of years, signals a recent and abrupt change in the Earth's climate system. Finally, rooted, soft-bodied wetland plants, now exposed along the margins as the Quelccaya ice cap (Peru) retreats, have been radiocarbon dated and, when coupled with other widespread proxy evidence, provide strong evidence for an abrupt mid-Holocene climate event that marked the transition from early Holocene (pre-5,000-yr-B.P.) conditions to cooler, late Holocene (post-5,000-yr-B.P.) conditions. This abrupt event, approximately 5,200 yr ago, was widespread and spatially coherent through much of the tropics and was coincident with structural changes in several civilizations. These three lines of evidence argue that the present warming and associated glacier retreat are unprecedented in some areas for at least 5,200 yr. The ongoing global-scale, rapid retreat of mountain glaciers is not only contributing to global sea-level rise but also threatening freshwater supplies in many of the world's most populous regions.

Greenhouse Effect↗
PubMed1994

Immunolocalization of an annexin-like protein in corn.

Although calcium has been proposed to be an important regulatory element in plant gravitropic growth, as yet no specific function of Ca2+ in growth regulation has been discovered. Our recent studies on a Ca(2+)-binding protein in pea seedlings called p35 indicate that it is a member of the annexin family of proteins and may play a key role in growth regulation through its function in delivering polysaccharides needed for wall construction. We previously reported the isolation of p35 from pea plumules and the production of polyclonal antibodies to it. Immunolocalizaton analyses of p35 in pea tissues revealed high levels of staining in secretory cell types such as developing vascular cells and outer root cap cells. To test how general was the occurrence and distribution of this annexin-like protein in plant cells we initiated an analysis of annexins in the monocot corn using immunological techniques. Our results indicate the immunochemical properties and localization of corn annexins are very similar to those reported for pea. They are consistent with the postulate that annexins may play a general role in the regulation of the secretion of wall polysaccharides needed for growth, and thus could be an important target of calcium action during gravitropic growth.

Annexins↗
PubMed2002

Developmental anatomy and branching of roots of four Zeylanidium species (podostemaceae), with implications for evolution of foliose roots.

Podostemaceae have markedly specialized and diverse roots that are adapted to extreme habitats, such as seasonally submerged or exposed rocks in waterfalls and rapids. This paper describes the developmental anatomy of roots of four species of Zeylanidium, with emphasis on the unusual association between root branching and root-borne adventitious shoots. In Z. subulatum and Z. lichenoides with subcylindrical or ribbon-like roots, the apical meristem distal (exterior) to a shoot that is initiated within the meristem area reduces and loses meristematic activity. This results in a splitting into two meristems that separate the parental root and lateral root (anisotomous dichotomy). In Z. olivaceum with lobed foliose roots, shoots are initiated in the innermost zone of the marginal meristem, and similar, but delayed, meristem reduction usually occurs, producing a parenchyma exterior to shoots located between root lobes. In some extreme cases, due to meristem recovery, root lobing does not occur, so the margin is entire. In Z. maheshwarii with foliose roots, shoots are initiated proximal to the marginal meristem and there is no shoot-root lobe association. Results suggest that during evolution from subcylindrical or ribbon-like roots to foliose roots, reduction of meristem exterior to a shoot was delayed and then arrested as a result of inward shifting of the sites of shoot initiation. The evolutionary reappearance of a protective tissue or root cap in Z. olivaceum and Z. maheshwarii in the Zeylanidium clade is implied, taking into account the reported molecular phylogeny and root-cap development in Hydrobryum.

Biological Evolution↗
PubMed1996

Apoptosis: A Functional Paradigm for Programmed Plant Cell Death Induced by a Host-Selective Phytotoxin and Invoked during Development.

The host-selective AAL toxins secreted by Alternaria alternata f sp lycopersici are primary chemical determinants in the Alternaria stem canker disease of tomato. The AAL toxins are members of a new class of sphinganine analog mycotoxins that cause cell death in both animals and plants. Here, we report detection of stereotypic hallmarks of apoptosis during cell death induced by these toxins in tomato. DNA ladders were observed during cell death in toxin-treated tomato protoplasts and leaflets. The intensity of the DNA ladders was enhanced by Ca2+ and inhibited by Zn2+. The progressive delineation of fragmented DNA into distinct bodies, coincident with the appearance of DNA ladders, also was observed during death of toxin-treated tomato protoplasts. In situ analysis of cells dying during development in both onion root caps and tomato leaf tracheary elements revealed DNA fragmentation localized to the dying cells as well as the additional formation of apoptotic-like bodies in sloughing root cap cells. We conclude that the fundamental elements of apoptosis, as characterized in animals, are conserved in plants. The apoptotic process may be expressed during some developmental transitions and is the functional process by which symptomatic lesions are formed in the Alternaria stem canker disease of tomato. Sphinganine analog mycotoxins may be used to characterize further signaling pathways leading to apoptosis in plants.

Journal Article↗
PubMed1985

Growth and gravireaction of maize roots treated with a phytotropin.

Effects of the phytotropin 1-(2'-carboxyphenyl)-3-phenylpropane-1,3-dione (CPD) on growth and gravireaction of intact roots and apical root segments of Zea mays L. cv LG-11 were analysed. It is concluded that the compound acts through sites of action in the extension zone and/or the root cap. The nature of the effects is consistent with the proposal that phytotropins may inhibit the gravitropic response by interfering in some way with the movement or action of the growth substance(s) emanating from the root cap.

Darkness↗
PubMed1990

Gravity-induced changes in intracellular potentials in elongating cortical cells of mung bean roots.

Gravity-induced changes in intracellular potentials in primary roots of 2-day-old mung bean (Vigna mungo L. cv. black matpe) seedlings were investigated using glass microelectrodes held by 3-dimensional hydraulic micro-drives. The electrodes were inserted into outer cortical cells within the elongation zone. Intracellular potentials, angle of root orientation with respect to gravity, and position within the root of the impaled cortical cell were measured simultaneously. Gravistimulation caused intracellular potential changes in cortical cells of the elongation zone. When the roots were oriented vertically, the intracellular potentials of the outer cortical cells (2 mm behind the root apex) were approximately - 115 mV. When the roots were placed horizontally cortical cells on the upper side hyperpolarized to - 154 mV within 30 s while cortical cells on the lower side depolarized to about - 62 mV. This electrical asymmetry did not occur in cells of the maturation zone. Because attempts to insert the electrode into cells of the root cap were unsuccessful, these cells were not measured. The hyperpolarization of cortical cells on the upper side was greatly reduced upon application of N,N'-dicyclohexylcarbodiimide (DCCD), an inhibitor of respiratory energy coupling. When stimulated roots were returned to the vertical, the degree of hyperpolarization of cortical cells on the previous upper side decreased within 30 s and approached that of cortical cells in non-stimulated roots. This cycle of hyperpolarization/loss of hyperpolarization was repeatable at least ten times by alternately turning the root from the vertical to the horizontal and back again. The very short (<30 s) lag period of these electrical changes indicates that they may result from stimulus-perception and transduction within the elongation zone rather than from transmission of a signal from the root cap.

Cell Differentiation↗
PubMed1999

Agravitropic mutant for the study of hydrotropism in seedling roots.

Roots have been shown to respond to a moisture gradient by positive hydrotropism. Agravitropic mutant plants are useful for the study of the hydrotropism in roots because on Earth hydrotropism is obviously altered by the gravity response in the roots of normally gravitropic plants. The roots are able to sense water potential gradient as small as 0.5 MPa mm(-1). The root cap includes the sensing apparatus that causes a differential growth at the elongation region of roots. A gradient in apoplastic calcium and calcium influx through plasmamembrane in the root cap is somehow involved in the signal transduction mechanism in hydrotropism, which may cause a differential change in cell wall extensibility at the elongation region. We have isolated an endoxy loglucan transferase (EXGT) gene that is strongly expressed in pea roots and appears to be involved in the differential growth in hydrotropically responding roots. Thus, it is now possible to study hydrotropism in roots by comparing with or separate from gravitropism. These results also imply that microgravity conditions in space are useful for the study of hydrotropism and its interaction with gravitropism.

Calcium Channels↗
PubMed1986

Calmodulin: localization in plant tissues.

Calmodulin was purified from bovine brain by preparative SDS-polyacrylamide gel electrophoresis. The denatured, purified calmodulin was used to immunize rabbits to produce antiserum. This antiserum was used to study the distribution of calmodulin in plant tissues by indirect immunohistochemistry. The root tips from corn seeds, oat seeds, peanuts, spaghetti squash seeds, and the terminal buds of spinach were investigated. A method for plant tissue sectioning and inhibition of endogenous peroxide activity was developed. In the corn root section, reaction product from anti-calmodulin was found mainly in the root cap cells. Lesser but significant amounts of calmodulin were localized in metaxylem elements, in some stele cells surrounding metaxylem elements, in apical initials, and in the cortical cells. Similar findings were also observed in other root tips from oat seeds, peanuts, and spaghetti squash seeds. In the terminal buds of the spinach, calmodulin-stained cells were highly concentrated in the apical meristem and leaf primordium. These findings suggest that the high concentration of calmodulin in the root cap may be important in relation to gravitropism and growth development.

Calmodulin↗
PubMed2004

A stress-inducible plasma membrane protein 3 (AcPMP3) in a monocotyledonous halophyte, Aneurolepidium chinense, regulates cellular Na(+) and K(+) accumulation under salt stress.

Regulation of ion homeostasis is fundamental to physiological activities in plants. Here, we report on the functional characterization of AcPMP3 [Aneurolepidium chinense (a monocotyledonous halophyte) plasma membrane protein 3] under salt stress. Expression of AcPMP3-1 and AcPMP3-2 genes was highly induced by various abiotic stresses, such as salt, cold and drought. Furthermore, abscisic acid, H(2)O(2) and salicylic acid also triggered expression of AcPMP3 genes. In the Deltanha1 Deltapmr2 Deltapmp3 yeast mutant, which lacks the major Na(+) efflux systems (Na(+)/H(+) antiporter and Na(+)-ATPase), its salt-sensitive phenotype was restored by expressing the AcPMP3-1 gene, and the transformants accumulated lesser amounts of Na(+) and K(+) than mutant cells under 50 mM NaCl and 500 mM KCl conditions, respectively. These results suggested that AcPMP3-1 plays a role as a regulator of both Na(+) and K(+) accumulation in the cells. In situ hybridization showed that the AcPMP3-1 transcript was localized in cells of the root cap and root epidermis, which strongly suggested that AcPMP3-1 is essential for regulating Na(+)/K(+) transportation between plant roots and the outer environment under salt stress.

Amino Acid Sequence↗
PubMed1998

Phytepsin, a barley vacuolar aspartic proteinase, is highly expressed during autolysis of developing tracheary elements and sieve cells.

Vacuolarisation, formation of autophagocytotic vacuoles and tonoplast disruption have been reported in plant cells undergoing developmentally regulated programmed cell death (PCD), but little is known about the vacuolar proteins involved. In HeLa cells, cathepsin D, a lysosomal aspartic proteinase has been shown to mediate PCD. Based on immunohistochemical staining of barley roots, we show here that the previously well characterised barley vacuolar aspartic proteinase (phytepsin), a plant homologue to cathepsin D, is highly expressed both during formation of tracheary elements and during partial autolysis of sieve cells. In serial transverse sections of the vascular cylinder, starting from the root tip, phytepsin is expressed in root cap cells, in the tracheary elements of early and late metaxylem, and in the sieve cells of the protophloem and metaphloem. Aleurain, a barley vacuolar cysteine proteinase, is expressed similarly in root cap cells but differently in the tracheary elements of protoxylem and early metaxylem. This is the first evidence that a vacuolar aspartic proteinase, in analogy to cathepsin D in animals, may play a role in the active autolysis of plant cells.

Aspartic Acid Endopeptidases↗
PubMed2000

The roles of three functional sulphate transporters involved in uptake and translocation of sulphate in Arabidopsis thaliana.

To investigate the uptake and long-distance translocation of sulphate in plants, we have characterized three cell-type-specific sulphate transporters, Sultr1;1, Sultr2;1 and Sultr2;2 in Arabidopsis thaliana. Heterologous expression in the yeast sulphate transporter mutant indicated that Sultr1;1 encodes a high-affinity sulphate transporter (Km for sulphate 3.6 +/- 0.6 microM), whereas Sultr2;1 and Sultr2;2 encode low-affinity sulphate transporters (Km for sulphate 0.41 +/- 0.07 mM and >/= 1.2 mM, respectively). In Arabidopsis plants expressing the fusion gene construct of the Sultr1;1 promoter and green fluorescent protein (GFP), GFP was localized in the lateral root cap, root hairs, epidermis and cortex of roots. beta-glucuronidase (GUS) expressed with the Sultr2;1 promoter was specifically accumulated in the xylem parenchyma cells of roots and leaves, and in the root pericycles and leaf phloem. Expression of the Sultr2;2 promoter-GFP fusion gene showed specific localization of GFP in the root phloem and leaf vascular bundle sheath cells. Plants continuously grown with low sulphate concentrations accumulated high levels of Sultr1;1 and Sultr2;1 mRNA in roots and Sultr2;2 mRNA in leaves. The abundance of Sultr1;1 and Sultr2;1 mRNA was increased remarkably in roots by short-term stress caused by withdrawal of sulphate. Addition of selenate in the sulphate-sufficient medium increased the sulphate uptake capacity, tissue sulphate content and the abundance of Sultr1;1 and Sultr2;1 mRNA in roots. Concomitant decrease of the tissue thiol content after selenate treatment was consistent with the suggested role of glutathione (GSH) as a repressive effector for the expression of sulphate transporter genes.

Arabidopsis↗
PubMed1980

Dictyosome-endoplasmic reticulum associations in higher plant cells? A serial-section analysis.

Serial sections through permanganate and glutaraldehyde/osmium fixed dictyosomes from suspension cultured sycamore cells and maize root cap cells have been analyzed. The results may be summarized as follows. For both sycamore and maize dictyosomes are: 1. transition vesicles between ER and dictyosome, and therefore a "forming face" to the dictyosome, not present. 2. direct, tubular ER-cisternal connections doubtful, if at all present; 3. "Maturing faces" to dictyosomes, i.e. a polarity in terms of the pinching-off of secretory vesicles, not a typical feature; 4. two types of vesicle associated with the dictyosome: large and small (in part "coated"); 5. cisternae present as a flattened, continuous central portion at whose periphery an extensive, anastomosing tubule system exists; 6. cisternae in the dictyosome stack not concentrically arranged, but lie somewhat displaced to one another. A characteristic feature of maize dictyosomes, in addition, is a bi- or unilateral distribution of hypertrophied slime vesicles. The kidney-shaped slime vesicles are arranged opposite to one another at the tubular periphery of a cisternae; the region in between is occupied with smaller, occasionally coated, vesicles. It would appear from these results that the endomembrane concept sensu strictu [58] is not applicable to higher plant cells whose main secretory product is polysaccharide rather than protein. Consequences and alternatives in relation to membrane-flow in such cells are discussed.

Endoplasmic Reticulum↗
PubMed2000

Diversity of cell lengths in terminal portions of roots: implications to cell proliferation.

Terminal meristems are responsible for all primary growth of roots. It has been asserted that all cells of root meristems are actively dividing (no cells cycle slowly or arrest in the cycle) and stem cell populations expand exponentially. Because cells do not slide relative to each other in roots, relative cell lengths may be used to determine relative cell cycle durations and/or proportions of cells actively dividing in root tissues. If all cells are cycling, no interphase cells should be longer than critical length (length of longest mitotic cell in the meristem) and cells should exhibit an exponential cell-age distribution. Lengths of all cells were obtained radially across entire median longitudinal root sections at 0.5, 1.0, 1.5, 2.0, 2.5 and 3.0 mm from the founder cell/root cap boundary for five plant species to estimate percentages of cells longer than critical length. For example, up to 15 and 90% of all interphase cells were longer than critical length in 0.5 and 2.0 mm tissues, respectively, indicating that slow-cycling and/or non-proliferative cells are present in such tissues. In order to determine if the distribution of cell lengths in 0.5 mm segments approximated an exponential cell-age distribution, lengths of interphase cells less than critical length were determined. Such interphase cells were placed into ten groups according to cell length and percentages of cells in each group were compared with percentages of cells in groups calculated from an exponential cell-age distribution. Percentages of cells were significantly different from predicted percentages of between 6 and 9 out of ten groups - cell lengths were not distributed exponentially. Because there are significant numbers of interphase cells longer than critical length and since lengths of interphase cells shorter than critical length do not resemble an exponential cell-age distribution, it must be concluded that not all cells in root segments from 0.5 to 3.0 mm root segments are actively dividing. Heretofore, no databases of cell lengths have been used to test these assertions.

Journal Article↗
PubMed2003

Redox-related peroxidative responses evoked by methyl-jasmonate in axenically cultured aeroponic sunflower (Helianthus annuus L.) seedling roots.

Methyl-jasmonate (MeJA) has been proposed to be involved in the evocation of defense reactions, as the oxidative burst in plants, substituting the elicitors or enhancing their effect. 48 h dark- and sterilely cultured (axenic) aeroponic sunflower seedling roots excised and treated with different concentrations of MeJA showed a strong and quick depression of the H(+) efflux rate, 1.80 microM MeJA totally stopping it for approximately 90 min and then reinitiating it again at a lower rate than controls. These results were wholly similar to those obtained with nonsterilely cultured roots and have been interpreted as mainly based on H(+) consumption for O(2)(*-) dismutation to H(2)O(2). Also K(+) influx was strongly depressed by MeJA, even transitorily reverting to K(+) efflux. These results were consistent with those associated to the oxidative burst in plants. MeJA induced massive H(2)O(2) accumulation in the middle lamella and intercellular spaces of both the root cap cells and the inside tissues of the roots. The native acidic extracellular peroxidase activity of the intact (nonexcised) seedling roots showed a sudden enhancement (by about 52%) after 5 min of MeJA addition, maintained for approximately 15 min and then decaying again to control rates. O(2) uptake by roots gave similar results. These and other results for additions of H(2)O(2) or horseradish peroxidase, diphenylene iodonium, and sodium diethyldithiocarbamate trihydrate to the reaction mixture with roots were all consistent with the hypothesis that MeJA induced an oxidative burst, with the generation of H(2)O(2) being necessary for peroxidase activity. Results with peroxidase activity of the apoplastic fluid were in accordance with those of the whole root. Finally, MeJA enhanced NADH oxidation and inhibited hexacyanoferrate(III) reduction by axenic roots, and diphenylene iodonium cancelled out these effects. Redox activities by CN(-)- preincubated roots were also studied. All these results are consistent with the hypothesis that MeJA enhanced the NAD(P)H oxidase of a redox chain linked to the oxidative burst, so enhancing the generation of O(2)(*-) and H(2)O(2), O(2) uptake, and peroxidase activity by roots.

Acetates↗