ATP-dependent regulation of nuclear Ca(2+) levels in plant cells.
(17/200)
Localised alterations in cytoplasmic Ca(2+) levels are an integral part of the response of eukaryotic cells to a plethora of external stimuli. Due to the large size of nuclear pores, it has generally been assumed that intranuclear Ca(2+) levels reflect the prevailing cytoplasmic Ca(2+) levels. Using nuclei prepared from carrot (Daucus carota L.) cells, we now show that Ca(2+) can be transported across nuclear membranes in an ATP-dependent manner and that over 95% of Ca(2+) is accumulated into a pool releasable by the Ca(2+) ionophore A.23187. ATP-dependent nuclear Ca(2+) uptake did not occur in the presence of ADP or ADPgammaS and was abolished by orthovanadate. Confocal microscopy of nuclei loaded with dextran-linked Indo-1 showed that the initial ATP-induced rise in [Ca(2+)] occurs in the nuclear periphery. The occurrence of ATP-dependent Ca(2+) uptake in plant nuclei suggests that alterations of intranuclear Ca(2+) levels may occur independently of cytoplasmic [Ca(2+)] changes. (+info)
Spatial and temporal distribution of solutes in the developing carrot taproot measured at single-cell resolution.
(18/200)
The time-course and spatial distribution of sugars and ions in carrot (Daucus carota L.) was studied at fine resolution using single cell (SiCSA) and tissue analysis. Four phases of osmolyte accumulation in the taproot were identified: an amino acid (germination) phase, when internal sources of amino acids provide seedlings with osmotica; an ion phase, when inorganic and organic ions were the main solutes; a hexose phase, when concentrations of glucose and fructose sharply increased and reached their maximum; and a sucrose phase, when sucrose became the major solute. Spatial distribution of sugar in taproot cells showed a general trend of highest concentration on both sides of the vascular cambium (some 200 mM sucrose, 150 mM glucose) and a minimum in the pith (some 100 mM sucrose, 60 mM glucose) and in periderm. Electrolytes (e.g. potassium) followed a distribution generally reciprocal to that of sugars; minimum in the tissue adjacent to the cambium (some 10 mM) and maximum in the pith and periderm (some 60-100 mM). The cambial cells contained unexpectedly low concentrations of sugars and potassium. These spatial and temporal patterns indicate that amino acids, other electrolytes and sugars are interchangeable in the tissue osmotic balance. The nature of the solute is developmentally determined both temporally and spatially. During the accumulation of electrolytes following the initial amino acid phase, osmotic pressure to 420 mosmol kg-1 rises and then remains constant despite large changes in the concentration of individual solutes. This indicates that osmotic pressure is regulated independently of the individual concentrations of solutes. (+info)
Effects on the growth of carrots (Daucus carota L.), cabbage (Brassica oleracea var. capitata L.) and onion (Allium cepa L.) of restricting the ability of the plants to intercept resources.
(19/200)
The objective of this paper is to assess the size and penetration of edge effects in carrot, cabbage and onion field crops and the extent to which these edge effects are modified by the presence of aerial or soil competition between the crop rows. In all three crops, large weight differences developed between the plants in the edge rows and those in the central rows. There was no indication of plant weight fluctuating between large and small values with each successive row in from the edge, as suggested by others. In carrot and onion, edge effects were greatly reduced by the presence of either white reflective aerial partitions or soil partitions, indicating that these species competed for both light and soil resources in UK field conditions. In cabbage, the mere presence of clear aerial partitions between rows reduced edge effects and there was little effect of soil partitions. This indicates the predominance of shoot over root competition in this species. The differences between species are possibly related to the architectural flexibility of their shoots. These results suggest that, within crops, carrot and onion plants compete for light over a distance of about 20 cm in each direction and for below-ground resources over a distance of about 50 cm in each direction. For cabbage, interactions between plants appeared to be dominated by the requirement for sufficient space to deploy the shoots for efficient light interception. (+info)
KDC1, a novel carrot root hair K+ channel. Cloning, characterization, and expression in mammalian cells.
(20/200)
Potassium is an essential nutrient which plays an important role in many aspects of plant growth and development. Plants have developed a number of highly specific mechanisms to take up potassium from the soil; these include the expression of K(+) transporters and potassium channels in root cells. Despite the fact that root epidermal and hair cells are in direct contact with the soil, the role of these tissues in K(+) uptake is not well understood. Here we report the molecular cloning and functional characterization of a novel potassium channel KDC1 which forms part of a new subfamily of plant K(in) channels. Kdc1 was isolated from carrot root RNA and in situ hybridization experiments show Kdc1 to be highly expressed in root hair cells. Expressing the KDC1 protein in Chinese hamster ovary cells identified it as a voltage and pH-dependent inwardly rectifying potassium channel. An electrophysiological analysis of carrot root hair protoplasts confirmed the biophysical properties of the Kdc1 gene product (KDC1) in the heterologous expression system. KDC1 thus represents a major K(+) uptake channel in carrot root hair cells. (+info)
Inositol(1,4,5)trisphosphate production in plant cells: an early response to salinity and hyperosmotic stress.
(21/200)
Salinity and hyperosmotic stress are environmental factors that severely affect the growth and development of plants. Adaptation to these stresses is known to be a complex multistep process, but a rise in cytoplasmic Ca(2+) and increased polyphosphoinositide turnover have now been identified as being amongst the early events leading to the development of tolerance. To determine whether a causal link exists between these two events we have investigated the effects of several salts and osmotic agents on levels of inositol(1, 4,5)trisphosphate (Ins(1,4,5)P(3)) in plant cells. Our data show that salts as well as osmotic agents induce a rapid and up to 15-fold increase in cellular Ins(1,4,5)P(3) levels. The increase in Ins(1,4,5)P(3) occurs in a dose-dependent manner and levels remain elevated for at least 10 min. These data indicate that increased Ins(1,4,5)P(3) production is a common response to salt and hyperosmotic stresses in plants and that it may play an important role in the processes leading to stress tolerance. (+info)
In vitro nuclear reconstitution could be induced in a plant cell-free system.
(22/200)
A cell-free system derived from carrot cell cytosol extract has been developed for reassembling nuclear structure around the added demembranated sperm chromatin of Xenopus. Morphological evidence suggests that reassembled nuclei display the typical characteristics of normal eukaryotic nuclei, such as double-layered nuclear membrane and nuclear pores. Micrococcal nuclease treatment indicates that remodeling of the demembranated sperm chromatin has occurred and the structure of nucleosome is formed during nuclear reconstitution. These data indicate that the nuclear reconstitution can be induced in cell-free systems from plants, and the self-assembly of the nucleus is ubiquitous in both animal and plant cells. (+info)
Carrot cells contain two top1 genes having the coding capacity for two distinct DNA topoisomerases I.
(23/200)
Five DNA topoisomerase I cDNA clones were isolated from a carrot (Daucus carota) cDNA library and two classes of nucleotide sequences were found. One component of the first class, pTop9, perfectly matches the open reading frame of pTop28, a truncated top1 cDNA previously described, and extended it by 594 nucleotides (top1alpha). A member of the second class, pTop11, contains an open reading frame 2727 bp long (top1ss) with a coding capacity for a second putative DNA topoisomerase I of 101 kDa. Both pTop9 and pTop11 clones are full length cDNAs. The two deduced amino acid sequences share a relevant similarity (89%) only at the C-terminal domain, whereas the similarity is reduced to 32% in the N-terminal region. Southern blot analysis and PCR amplification of genomic DNAs from carrot pure lines suggested the presence of two distinct loci. Northern blot analysis revealed the presence of two distinct transcripts of 3.0 and 3.2 kb in both cycling and starved cell populations. Three fusion peptides corresponding to the N-terminal domain of the alpha and ss forms and from the common C-terminal domain of carrot topoisomerases I were overexpressed in E. coli cells and used to raise antibodies in rabbit. Immunolocalization seems to suggest the presence of two topoisomerases I in carrot nuclei. (+info)
Distinct patterns of expression but similar biochemical properties of protein L-isoaspartyl methyltransferase in higher plants.
(24/200)
Protein L-isoaspartyl methyltransferase is a widely distributed repair enzyme that initiates the conversion of abnormal L-isoaspartyl residues to their normal L-aspartyl forms. Here we show that this activity is expressed in developing corn (Zea mays) and carrot (Daucus carota var. Danvers Half Long) plants in patterns distinct from those previously seen in winter wheat (Triticum aestivum cv Augusta) and thale cress (Arabidopsis thaliana), whereas the pattern of expression observed in rice (Oryza sativa) is similar to that of winter wheat. Although high levels of activity are found in the seeds of all of these plants, relatively high levels of activity in vegetative tissues are only found in corn and carrot. The activity in leaves was found to decrease with aging, an unexpected finding given the postulated role of this enzyme in repairing age-damaged proteins. In contrast with the situation in wheat and Arabidopsis, we found that osmotic or salt stress could increase the methyltransferase activity in newly germinated seeds (but not in seeds or seedlings), whereas abscisic acid had no effect. We found that the corn, rice, and carrot enzymes have comparable affinity for methyl-accepting substrates and similar optimal temperatures for activity of 45 degrees C to 55 degrees C as the wheat and Arabidopsis enzymes. These experiments suggest that this enzyme may have specific roles in different plant tissues despite a common catalytic function. (+info)