Salt Marsh Claviceps purpurea in Native and Invaded Spartina Marshes in Northern California.

The fungal pathogen Claviceps purpurea (subgroup G3) has a worldwide distribution on salt marsh Spartina spp. In Northern California (United States), native Spartina foliosa sustains high rates of infection by G3 C. purpurea in marshes north of the San Francisco Estuary. Invasive populations of S. alterniflora and S. alterniflora × foliosa hybrids are virtually disease free in the same estuary, although S. alterniflora is host to G3 C. purpurea in its native range (Atlantic Coast of the United States). Greenhouse inoculation experiments showed no differences in susceptibility among S. foliosa, S. alterniflora, and Spartina hybrids. Under field conditions, S. foliosa sustained a higher incidence of disease in coastal marshes than in marshes within the bay. This geographic effect may be attributable to environmental differences between the coast and the bay proper, with the former being more conducive to infection by C. purpurea. Seed set of S. foliosa spikelets was 40 to 70% lower on infected than on uninfected inflorescences, but seed germination was not affected. The C. purpurea epidemic on S. foliosa on the coast north of the San Francisco Estuary further reduces the meager competitive ability of this declining native plant species.

3,7-Dichloroquinolinecarboxylic Acid Inhibits Cell-Wall Biosynthesis in Maize Roots.

The mode of action of the herbicide 3,7-dichloroquinolinecar-boxylic acid (quinclorac) was examined by measuring incorporation of [14C]glucose, [14C]acetate, [3H]thymidine, and [3H]uridine into maize (Zea mays) root cell walls, fatty acids, DNA, and RNA, respectively. Among the precursors examined, 10 [mu]M quinclorac inhibited [14C]glucose incorporation into the cell wall within 3 h. Fatty acid and DNA biosynthesis were subsequently inhibited, whereas RNA biosynthesis was unaffected. In contrast to the cellulose synthesis inhibitor 2,6-dichlorobenzonitrile, quinclorac strongly inhibited cellulose and a hemicellulose fraction presumed to be glucuronoarabinoxylan. However, the synthesis of (1->3),(1->4)-[beta]-D-glucans was only slightly inhibited. The degree of inhibition was time- and dose-dependent. By 4 h after treatment, the concentration that inhibited [14C]glucose incorporation into the cell wall, cellulose, and the sensitive hemicellulose fraction by 50% was about 15, 5, and 20 [mu]M, respectively. Concomitant with an inhibition of [14C]glucose incorporation into the cell wall, quinclorac treatment led to a marked accumulation of radioactivity in the cytosol. The increased radioactivity was found mostly in glucose and fructose. However, total levels of glucose, fructose, and uridine diphosphate-glucose were not changed greatly by quinclorac. These data suggest that quinclorac acts primarily as a cell-wall biosynthesis inhibitor in a susceptible grass by a mechanism that is different from that of 2,6-dichlorobenzonitrile.

Effects of Acetyl-Coenzyme A Carboxylase Inhibitors on Root Cell Transmembrane Electric Potentials in Graminicide-Tolerant and -Susceptible Corn (Zea mays L.).

Herbicidal activity of aryloxyphenoxypropionate and cyclohexanedione herbicides (graminicides) has been proposed to involve two mechanisms: inhibition of acetyl-coenzyme A carboxylase (ACCase) and depolarization of cell membrane potential. We examined the effect of aryloxyphenoxypropionates (diclofop and haloxyfop) and cyclohexanediones (sethoxydim and clethodim) on root cortical cell membrane potential of graminicide-susceptible and -tolerant corn (Zea mays L.) lines. The graminicide-tolerant corn line contained a herbicide-insensitive form of ACCase. The effect of the herbicides on membrane potential was similar in both corn lines. At a concentration of 50 [mu]M, the cyclohexanediones had little or no effect on the membrane potential of root cells. At pH 6, 50 [mu]M diclofop, but not haloxyfop, depolarized membrane potential, whereas both herbicides (50 [mu]M) dramatically depolarized membrane potential at pH 5. Repolarization of membrane potential after removal of haloxyfop and diclofop from the treatment solution was incomplete at pH 5. However, at pH 6 nearly complete repolarization of membrane potential occurred after removal of diclofop. In graminicide-susceptible corn, root growth was significantly inhibited by a 24-h exposure to 1 [mu]M haloxyfop or sethoxydim, but cell membrane potential was unaffected. In gramincide-tolerant corn, sethoxydim treatment (1 [mu]M, 48 h) had no effect on root growth, whereas haloxyfop (1 [mu]M, 48 h) inhibited root growth by 78%. However, membrane potential was the same in roots treated with 1 [mu]M haloxyfop or sethoxydim. The results of this study indicate that graminicide tolerance in the corn line used in this investigation is not related to an altered response at the cell membrane level as has been demonstrated with other resistant species.

Characterization of Paraquat Transport in Protoplasts from Maize (Zea mays L.) Suspension Cells.

Protoplasts isolated from maize (Zea mays L.) suspension cells were used to study transport of paraquat. [14C]Paraquat uptake was measured in 400-[mu]L centrifuge tubes using silicon oil centrifugation techniques. Approximately 50% of accumulation from a 100 [mu]M paraquat solution occurred in the first 10 s, and net accumulation reached a maximum after about 10 min. Membrane binding accounted for about 30% of apparent accumulation. Concentration-dependent uptake kinetics were characterized by a non-saturating curve, which was resolved into a linear and a saturable component. The Km of the saturable component was 132 [mu]M, and the Vmax was 0.512 nmol [mu]L of protoplasts-1 min-1. In the absence of sucrose, the Vmax of the saturable component was reduced by 52%, suggesting that paraquat uptake across the plasmalemma is energy dependent. Measurement of concentration-dependent binding of paraquat to burst protoplasts showed a linear response. This suggests that the linear component from intact protoplast concentration kinetics represented paraquat binding to the plasmalemma surface. Calcium inhibited the saturable component, and this inhibition was shown by Lineweaver-Burk analysis to be noncompetitive. Putrescine, a divalent cationic polyamine with a charge distribution similar to that of paraquat, competitively inhibited paraquat uptake. These results show that paraquat transport characteristics at the plasmalemma of maize protoplasts are similar to those reported earlier for paraquat transport in roots of intact maize seedlings.

Problems associated with the use of common names in the identification of poisonous plants.

Barriers in communication among professional botanists, physicians and the general populous developed from a disparity in the use of botanical nomenclature. Universally accepted scientific names from plants affords a common ground of understanding among botanists throughout the world. A general phobia with unfamiliar latinized scientific nomenclature has proliferated the use of common or trivial names. These names, however, can vary with geographic region, language, or individual preference often and thus, can lead to misidentification or a delay in the proper identification of a toxic plant.

Membrane Response to Diclofop Acid Is pH Dependent and Is Regulated by the Protonated Form of the Herbicide in Roots of Pea and Resistant and Susceptible Rigid Ryegrass.

Electrophysiological studies in roots of pea (Pisum sativum L.) and rigid ryegrass (Lolium rigidum Gaud.) seedlings were conducted to elucidate the mechanism involved in the membrane response to the herbicide diclofop. In pea, a dicotyledonous plant insensitive to diclofop, membrane depolarization at varying pH values and herbicide concentrations increased at higher concentrations of the protonated form of diclofop acid (pKa 3.57). In unbuffered nutrient solution (pH 5.7), diclofop acid (50 [mu]M) depolarized the membrane potential (Em) in roots of both resistant and susceptible biotypes of rigid ryegrass, whereas recovery of Em occurred only in the resistant biotype following removal of the herbicide. This differential response was correlated with an increase (450%) in the rate of acidification of the external solution by the susceptible biotype, and the Em differences between biotypes were eliminated in solutions buffered at pH 5.0 or 6.0. In addition, p-chloromercuribenzene-sulfonic acid did not prevent the depolarization of Em by 50 [mu]M diclofop acid. It is concluded that the differential membrane response to diclofop acid in herbicide-resistant and -susceptible biotypes of rigid ryegrass is due to pH differences at the cell wall/plasmalemma interface. Although the membrane response is probably not involved in the primary inhibitory effect of diclofop on plant growth, it could reduce the concentration of the permeant protonated form of the herbicide and possibly could contribute to increased tolerance to diclofop and other weak acid herbicides.

Compartmentation Analysis of Paraquat Fluxes in Maize Roots as a Means of Estimating the Rate of Vacuolar Accumulation and Translocation to Shoots.

Efflux analysis conducted after five loading periods of various lengths (2, 6, 12, 18, or 24 h) was used to investigate uptake, compartmentation, and translocation of [14C]paraquat in maize (Zea mays L.) seedlings. The time course for net paraquat uptake (paraquat concentration in uptake solution = 25[mu]M) into maize roots was linear (56.7 nmol g-1 root fresh weight h-1) for 24 h. Estimates of changes in paraquat content in the vacuole, cytoplasm, and cell wall after 2-, 6-, 12-, 18-, and 24-h loading periods indicated that the cell wall saturated rapidly, whereas accumulation of paraquat into the vacuole increased linearly (12.4 nmol g-1 root fresh weight h-1) over 24 h. In contrast to vacuolar accumulation, cytoplasmic paraquat content appeared to approach saturation. The half-time for paraquat efflux from the cell wall (16.6 min [plus or minus] 1.2 SD) and cytoplasm (58.8 min [plus or minus] 8.9 SD remained relatively constant regardless of the length of the loading period, whereas the half-time for efflux from the vacuole was considerably longer and increased linearly with increased loading time (6.1-18.7 h). The time course for paraquat translocation to the shoot was linear within a 24-h exposure to radiolabeled herbicide, but translocation did not begin until 5 h after initiation of treatment. The experimental approach used in these experiments provides a valuable method for examining the movement of paraquat in maize seedlings. Results indicate that the herbicide slowly accumulates in the vacuole of root cells but is also translocated to the shoot.

Transport Interactions between Paraquat and Polyamines in Roots of Intact Maize Seedlings.

Interactions between absorption of paraquat and the polyamines putrescine, cadaverine, and spermine in roots of intact maize (Zea mays L. cv 3377 Pioneer) seedlings were examined. Concentration-dependent kinetics for paraquat and putrescine influx were similar and both kinetic curves could be resolved into a linear and a saturable component. The linear component was previously shown to represent cell wall/membrane binding. The saturable components for paraquat and putrescine uptake, which represent influx across the plasmalemma, had K(m) values of 98 and 120 micromolar, respectively, and V(max) values of 445 and 456 nanomoles per gram fresh weight per hour, respectively. Lineweaver-Burk transformation of the saturable component of paraquat influx in the presence of varying concentrations of putrescine indicated that the diamine competitively inhibited the saturable component of paraquat uptake. Reciprocal experiments similarly demonstrated that paraquat competitively inhibited the saturable component of putrescine uptake. Competitive inhibition of both paraquat and putrescine influx could also be demonstrated with the diamine cadaverine, which has a charge distribution similar to that of paraquat and putrescine. In contrast, the larger, tetravalent polyamine spermine appeared to noncompetitively inhibit the influx of paraquat and putrescine. These results strongly suggest that paraquat enters maize root cells via a carrier system that normally functions in the transport of diamines with a charge distribution similar to that of paraquat.

Effect of inorganic cations and metabolic inhibitors on putrescine transport in roots of intact maize seedlings.

The specificity and regulation of putrescine transport was investigated in roots of intact maize (Zea mays L.) seedlings. In concentration-dependent transport studies, the kinetics for putrescine uptake could be resolved into a single saturable component that was noncompetitively inhibited by increasing concentrations of Ca(2+) (50 micromolar to 5 millimolar). Similarly, other polyvalent cations, including Mg(2+) (1.8 millimolar) and La(3+) (200 micromolar), almost completely abolished the saturable component for putrescine uptake. This suggests that putrescine does not share a common transport system with other divalent or polyvalent inorganic cations. Further characterization of the putrescine transport system indicated that 0.3 millimolar N-ethyl-maleimide had no effect on putrescine uptake, and 2 millimolar p-chloromercuribenzene sulfonic acid only partially inhibited transport of the diamine (39% inhibition). Metabolic inhibitors, including carbonylcyanide-m-chlorphenylhydrazone (20 micromolar) and KCN (0.5 millimolar), also partially inhibited the saturable component for putrescine uptake (V(max) reduced 48-60%). Increasing the time of exposure to carbonylcyanide-m-chlorphenylhydrazone from 30 minutes to 2 hours did not significantly increase the inhibition of putrescine uptake. Electrophysiological evidence indicates that the inhibitory effect on putrescine uptake by these inhibitors is correlated to a depolarization of the membrane potential, suggesting that the driving force for putrescine uptake is the transmembrane electrical potential across the plasmalemma.

Transport kinetics and metabolism of exogenously applied putrescine in roots of intact maize seedlings.

Putrescine metabolism, uptake, and compartmentation were studied in roots of hydroponically grown intact maize (Zea mays L.) seedlings. In vivo analysis of exogenously applied putrescine indicated that the diamine is primarily metabolized by a cell wall-localized diamine oxidase. Time-dependent kinetics for putrescine uptake could be resolved into a rapid phase of uptake and binding within the root apoplasm, followed by transport across the plasma membrane that was linear for 30 to 40 minutes. Concentration-dependent kinetics for putrescine uptake (between 0.05 and 1.0 millimolar putrescine) appeared to be nonsaturating but could be resolved into a saturable (V(max) 0.397 micromoles per gram fresh weight per hour; K(m) 120 micromolar) and a linear component. The linear component was determined to be cell wall-bound putrescine that was not removed during the desorption period following uptake of [(3)H]putrescine. These results suggest that a portion of the exogenously applied putrescine can be metabolized in maize root cell walls by diamine oxidase activity, but the bulk of the putrescine is transported across the plasmalemma by a carrier-mediated process, similar to that proposed for animal systems.

Effects of diclofop and diclofop-methyl on membrane potentials in roots of intact oat, maize, and pea seedlings.

Growth and electrophysiological studies in roots of intact diclofop-methyl susceptible and resistant seedlings were conducted to test the hypothesis that the herbicide acts primarily as a proton ionophore. The ester formulation of diclofop, at 0.2 micromolar, completely inhibited root growth in herbicide-susceptible oat (Avena sativa L.) after a 96 hour treatment, but induced only a delayed transient depolarization of the membrane potential in oat root cortical cells. Root growth in susceptible maize (Zea mays L.) seedlings was dramatically reduced by exposure to 0.8 micromolar diclofop-methyl, while the same diclofop-methyl exposure hyperpolarized the membrane potential within 48 hours after treatment. Furthermore, exposure of maize roots to the protonophore, carbonyl cyanide m-chlorophenylhydrazone (CCCP) (50 nanomolar), inhibited growth by only 31%, 96 hours after treatment, while the same CCCP exposure depolarized the resting potential by an average of 32 millivolts. Thus, the protonophore hypothesis cannot account for a differential membrane response to phytotoxic levels of diclofop-methyl in two susceptible species. From the results of others, much of the evidence to support the protonophore hypothesis was obtained using high concentrations of diclofop acid (100 micromolar). At a similar concentration, we also report a rapid (3 minute) diclofop-induced depolarization of the membrane potential in roots of susceptible oat and maize, moderately tolerant barley (Hordeum vulgare L.), and resistant pea (Pisum sativum L.) seedlings. Moreover, 100 micromolar diclofop acid inhibited growth in excised cultured pea roots. In contrast, 100 micromolar diclofop-methyl did not inhibit root growth. Since the membrane response to 100 micromolar diclofop acid does not correspond to differential herbicide sensitivity under field conditions, results obtained with very high levels of diclofop acid are probably physiologically irrelevant. The results of this study suggest that the effect of diclofop-methyl on the membrane potentials of susceptible species is probably unrelated to the primary inhibitory effect of the herbicide on plant growth.

Putrescine-induced wounding and its effects on membrane integrity and ion transport processes in roots of intact corn seedlings.

Interactions between putrescine and membrane function were examined with the use of a recently developed microelectrode system that enables us simultaneously to quantify membrane potentials and net K(+) fluxes associated with individual cells at the root surface of an intact corn (Zea mays L.) seedling. In contrast to the results of others, our analyses indicate that exogenous putrescine (0.5 millimolar), in the absence of calcium, does not maintain membrane stability. In addition, putrescine caused a wound response characterized by a gradual depolarization of the membrane potential and a considerable net efflux of K(+) from the root. In the presence of calcium, both short term (20 minutes) and long term (24 hours) exposure to a high concentration of exogenous putrescine (5 millimolar) also caused a reduction in the resting membrane potential and a significant K(+) efflux. However, preincubating corn roots in a solution containing the antioxidant ascorbate ameliorated the wounding effects of putrescine and slightly increased potassium uptake. A similar preincubation in the absence of calcium did not protect membranes against putrescine-induced damage. The ameliorating effect of ascorbate on putrescine-induced membrane damage suggests that the wounding response of high putrescine levels in corn roots involves the catabolism of the polyamine by a cell wall diamine oxidase, with the concomitant production of hydrogen peroxide and free radicals resulting in peroxidative damage of the plasmalemma.