Effects of diet on titratable acid-base excretion in grasshoppers

Despite the potential for diet to affect organismal acid-base status, especially in herbivores, little is known about the effects of diet on acid-base loading and excretion. We tested the effects of diet on acid-base loading and excretion in grasshoppers by (a) comparing the fecal acid-base content of 15 grasshopper species collected from the field and (b) comparing fecal acid-base excretion rates of Schistocerca americana grasshoppers fed vegetable diets that differed in their ashed and raw acid-base contents. The field experiments indicated that grass-feeding species excrete fairly neutral fecal pellets, while forb/mixed-feeding species vary widely in their fecal acid-base contents. In the laboratory experiment, acid-base excretion rates were positively correlated with dietary ashed base intake rates but were not correlated with the acid-base content of raw, unashed diet or feeding rate. These experiments suggest that some diets could strongly challenge the acid-base homeostasis of herbivores; in some grasshoppers, dietary acid-base loads could produce certainly lethal 1-unit changes in average body pH within 6 h if they were not excreted.     

Field studies of the relationship between herbivore damage and tannin concentration in bracken (Pteridium aquilinum Kuhn)

Summary The acceptance of secondary plant metabolites as herbivore deterrents rests primarily on their deleterious effects on herbivores. Efforts to demonstrate differential fitness in natural plant populations with varying concentrations of tannin have failed, since coevolved plant predators may physiologically or behaviorally circumvent the defense, which results in apparently equal amounts of damage to defended and undefended individuals. In this study, two approaches were used to overcome this difficulty. 1) Theoretically, more energy should be allocated to the defense of parts which contribute more heavily to the plant's fitness. Bracken fern clones produce fronds throughout the growing season. Fronds which are produced early should be more heavily defended than late-emerging fronds which will return less photosynthate per unit cost of production. The results of this study do not support this prediction; it appears that the production of tannin is more closely linked to environmental factors such as water stress than to date of frond emergence. Fronds which emerged in August contained as much tannin as fronds which emerged in May. 2) By recording the temporal occurrence of herbivore damage in bracken ferns, it was found that in fronds which escaped attack until after reaching maturity there was a significant negative correlation between tannin concentration in the frond and the amount of damage experienced. This result supports the generally accepted assumption that herbivory has been a selective force in the evolution of tannin as a defensive substance.     

Development of a toxic bait for control of eastern lubber grasshopper (Orthoptera: Acrididae)

This study assessed baits for eastern lubber grasshopper, Romalea guttata (Houttuyn). When offered a choice among several grain-based baits (rolled oats, wheat bran, oat bran, yeast, corn meal, cornflakes) and vegetable oils (canola, corn, peanut, soybean), eastern lubber grasshopper adults preferred bait consisting of wheat bran carrier with corn oil as an added phagostimulant. Other carriers were accepted but consumed less frequently. Discrimination by eastern lubber grasshoppers among oils was poor. Similarly, addition of flavorings (peppermint, anise, lemon, banana) resulted in few significant effects. The carbaryl, wheat bran, and oil bait developed in this study was effective at causing eastern lubber grasshopper mortality in field-cage studies. Significant mortality occurred even though grasshoppers had to locate dishes of bait in a large cage, and could feed on daylilies, or grass growing through the bottom of the cage, rather than on the bran flakes. Consumption of as little as a single carbaryl-treated bran flake could induce mortality, although individuals varied greatly in their susceptibility. The bait matrix developed in this study was readily consumed when in the presence of some plant species. We expect that wheat bran and corn oil bait would be most effective as protection for less preferred plants (tomato, pepper, eggplant, leek, parsley, fennel, daylily, lily of the Nile, and canna lily) because baits were readily consumed in the presence of these plants. Plants that are readily consumed in the presence of bait (preferred plants) included butter crunch lettuce, carrot, yellow squash, cauliflower, collards, green onion, chive, cucumber, cabbage, cantalope, endive, red leaf lettuce, society garlic, caladium, and amaryllis. Baits are likely to be less effective in the presence of such plants. On average, vegetables in Solanaceae (i.e., tomato, pepper, and eggplant) and Apiaceae (i.e., fennel and parsley) elicited high levels of bait-feeding activity, indicating that these vegetables were not highly preferred. The plants tested from Liliaceae, Cucurbitaceae, Asteraceae, and Brassicaceae elicited an intermediate-to-low level of bait feeding.     

Characteristics of Arsenic Accumulation by Pteris and non-Pteris Ferns

This research was conducted to understand the mechanisms of arsenic hyperaccumulation in Pteris vittata by comparing the characteristics of arsenic accumulation in Pteris and non-Pteris ferns. Seven Pteris (P.vittata, P. Cretica Rowerii, P. Cretica Parkerii, P. Cretica Albo-lineata, P. Quadriavrita, P. Ensiformis and P. Dentata) and six non-Pteris (Arachnoides simplicor, Didymochlaena truncatula, Dryopteris atrata, Dryopteris erythrosora, Cyrtomium falcatum, and Adiantum hispidulum) ferns were exposed to 0, 1 and 10 mgL⁻¹ arsenic as sodium arsenate for 14-d in hydroponic systems. As a group, the Pteris ferns were more efficient in arsenic accumulation than the non-Pteris ferns, with P. vittata being the most efficient followed by P. cretica. When exposed to 10 mg L⁻¹ As, arsenic concentrations in the fronds and roots of P. vittata were 1748 and 503 mg kg⁻¹. Though not all Pteris ferns were efficient in accumulating arsenic, none of the non-Pteris ferns was an efficient As accumulator (the highest concentration being 452 mg kg⁻¹). The fact that frond arsenic concentrations in the control were highly correlated with those exposed to As (r ² = 0.76–0.87) may suggest that they may be used as a preliminary tool to screen potential arsenic hyperaccumulators. Our research confirms that the ability of P. vittata to translocate arsenic from the roots to the fronds (73–77% As in the fronds), reduce arsenate to arsenite in the fronds (>50% AsIII in the fronds), and maintain high concentrations of phosphate in the roots (48–53% in the roots) all contributed to its arsenic tolerance and hyperaccumulation.     

Comparison of root absorption, translocation and tolerance of arsenic in the hyperaccumulator Pteris vittata and the nonhyperaccumulator Pteris tremula

* Several fern species can hyperaccumulate arsenic, although the mechanisms are not fully understood. Here we investigate the roles of root absorption, translocation and tolerance in As hyperaccumulation by comparing the hyperaccumulator Pteris vittata and the nonhyperaccumulator Pteris tremula. * The two species were grown in a pot experiment with 0-500 mg As kg-1 added as arsenate, and in a short-term (8 h) uptake experiment with 5 microM arsenate under phosphorus-sufficient conditions. * In the pot experiment, P. vittata accumulated up to 2500 mg As kg-1 frond d. wt and suffered no phytotoxicity. P. tremula accumulated<100 mg As kg-1 frond d. wt and suffered severe phytotoxicity with additions of >or=25 mg As kg-1. In the short-term uptake experiment, P. vittata had a 2.2-fold higher rate of arsenate uptake than P. tremula, and distributed more As taken up to the fronds (76%) than did P. tremula (9%). * Our results show that enhanced root uptake, efficient root-to-shoot translocation, and a much elevated tolerance through internal detoxification all contribute to As hyperaccumulation in P. vittata.     

An arsenic hyperaccumulating fern,Pteris vittata L. (Pteridaceae) broadly affects terrestrial invertebrate abundance

1. The Chinese brake fern (Pteris vittata L.; Pteridaceae) can accumulate up to 27 000 mg kg^?1 dry wt. of arsenic (As) from the soil into its above‐ground biomass. They may use this As to deter invertebrate threats. 2. This study explored how As concentrations [As] in the fern, and in soil associated with the fern, influenced the abundance and composition of various invertebrates. 3. Populations of P. vittata were identified in the field. Soils from the base of the fern and from 3 m away of each plant were collected and pitfall traps were installed. Soil and fern arsenic concentrations ([As]) were measured via inductively coupled plasma mass spectrometry and invertebrates were identified to order and classified by feeding guild. 4. Increased [As] did not affect all feeding guilds and orders equally. For example, individual herbivore abundance did not decrease as [As] increased, but predator abundance did. In many cases, the impact of soil [As] on invertebrates depended on the distance from the fern. Fern [As] also influenced components of the community, but only at 3 m away from the fern. Furthermore, the abundances of many invertebrate groups were higher beneath the fern, where [As] was higher. 5. These results suggest that hyperaccumulated As can impact the invertebrate community, but the defensive benefits of hyperaccumulation are more complex than have been previously described. The authors advocate that future studies examining the potential defensive benefits of hyperaccumulation should do so in a natural setting that incorporates this complexity and invertebrate richness.     

The role of the host-specific grasshopper Cornops aquaticum (Orthoptera: Acrididae) as consumer of native Eichhornia crassipes (Pontederiaceae) floating meadows

Cornops aquaticum is a widely distributed semiaquatic grasshopper in the Neotropics. The development, feeding and oviposition of C. aquaticum take place on Pontederiaceae, especially on species of Eichhornia. Several aspects of the feeding of C. aquaticum are studied because is one of the most important herbivores of the highly invasive floating Eichhornia crassipes in native areas. The aims of this paper were: (1) to quantify the amount of E. crassipes consumed by C. aquaticum, (2) to determine the growth rate and the conversion efficiency of food ingested by this grasshopper, and (3) to determine the possible effect of consumption on E. crassipes productivity. Thirty individuals from each specific age class were used in the experiment: nymphs A, nymphs B, adult males and adult females. Insects were individually confined in plastic pots with a leaf of E. crassipes. We estimated feeding by individual, consumption index (CI), relative growth rate (GR) and efficiency of conversion of ingested food to body substance (ECI). The impact of C. aquaticum consumption on E. crassipes floating meadows was assessed with the abundance of the grasshopper, and the available data on primary production of the host plant at the study site. Food intake of C. aquaticum was 11.23% of plant productivity. Food consumption, growth rate and food conversion efficiency of this grasshopper varied according to the specific age classes. Damage caused by C. aquaticum is high in comparison with the damage caused by other semiaquatic and grassland grasshoppers, however it is not enough to prevent the growth and coverage of native E. crassipes floating meadows because abundance of grasshoppers are realtively low and the growth rate and productivity of the host plant is high.     

Selenium accumulation protects plants from herbivory by Orthoptera via toxicity and deterrence

To investigate whether selenium (Se) accumulation in plants provides a chemical defense against generalist insect herbivores, the feeding preference and performance of a mix of orthopteran species were investigated. The selenium hyperaccumulator Stanleya pinnata and accumulator Brassica juncea were used in herbivory studies in the laboratory, and S. pinnata was also used in a manipulative field experiment. In laboratory studies, both crickets and grasshoppers avoided plants pretreated with selenate, while those given no choice died after eating leaves with elevated Se (447 +/- 68 and 230 +/- 68 microg Se g(-1) DW, respectively). B. juncea has previously been shown to accumulate selenate, while S. pinnata hyperaccumulates methyl-selenocysteine. Thus, these findings demonstrate that both inorganic and organic forms of selenium protect plants from herbivory. Grasshoppers fed S. pinnata contained methylselenocysteine in their midgut and absorbed this form into surrounding tissues. In a manipulative field experiment, methylselenocysteine protected S. pinnata from invertebrate herbivory and increased its long-term survival rate over an entire growth season. * In native habitats of selenium hyperaccumulators, orthopterans represent a major group of insect herbivores. Protection offered by organic selenium accumulation against these herbivores may have promoted the evolution of selenium hyperaccumulation in plants.     

Arsenic hyperaccumulation in gametophytes of Pteris vittata. A new model system for analysis of arsenic hyperaccumulation

The sporophyte of the fern Pteris vittata is known to hyperaccumulate arsenic (As) in its fronds to >1% of its dry weight. Hyperaccumulation of As by plants has been identified as a valuable trait for the development of a practical phytoremediation processes for removal of this potentially toxic trace element from the environment. However, because the sporophyte of P. vittata is a slow growing perennial plant, with a large genome and no developed genetics tools, it is not ideal for investigations into the basic mechanisms underlying As hyperaccumulation in plants. However, like other homosporous ferns, P. vittata produces and releases abundant haploid spores from the parent sporophyte plant which upon germination develop as free-living, autotrophic haploid gametophyte consisting of a small (<1 mm) single-layered sheet of cells. Its small size, rapid growth rate, ease of culture, and haploid genome make the gametophyte a potentially ideal system for the application of both forward and reverse genetics for the study of As hyperaccumulation. Here we report that gametophytes of P. vittata hyperaccumulate As in a similar manner to that previously observed in the sporophyte. Gametophytes are able to grow normally in medium containing 20 mm arsenate and accumulate >2.5% of their dry weight as As. This contrasts with gametophytes of the related nonaccumulating fern Ceratopteris richardii, which die at even low (0.1 mm) As concentrations. Interestingly, gametophytes of the related As accumulator Pityrogramma calomelanos appear to tolerate and accumulate As to intermediate levels compared to P. vittata and C. richardii. Analysis of gametophyte populations from 40 different P. vittata sporophyte plants collected at different sites in Florida also revealed the existence of natural variability in As tolerance but not accumulation. Such observations should open the door to the application of new and powerful genetic tools for the dissection of the molecular mechanisms involved in As hyperaccumulation in P. vittata using gametophytes as an easily manipulated model system.     

The ecological significance of nickel hyperaccumulation: a plant chemical defense

Nickel hyperaccumulating plants have more than 1000 mg Ni kg^–1 dry weight when grown on nickel-bearing soils. We hypothesized that Ni hyperaccumulation could serve as a chemical defense against herbivores In feeding experiments with potential insect herbivores and Ni hyperaccumulating plants, only those inseets fed leaves from plants grown on non-nickel-bearing soil survived or showed a weight gain. Among chemical parameters measured, only Ni content of plants was sufficient to explain this result. When subjected to herbivory by lepidopteran larvae, plants grown on Ni-amended soil showed greater survival and yield than plants on unamended soil. Ni hyperaccumulation may be an effective plant chemical defense against herbivores because of its high lethality, apparent low cost, and broad spectrum of toxicity.     

Stability of AtVSP in the insect digestive canal determines its defensive capability

We have previously demonstrated that Arabidopsis vegetative storage protein (AtVSP) is an acid phosphatase that has anti-insect activity in in vitro feeding assays [Liu et al., 2005. Plant Physiology 139, 1545-1556]. To investigate the functionality of AtVSP in planta as an anti-insect defense protein, we produced AtVSP-overexpressing as well as AtVSP-silenced transgenic Arabidopsis lines, and evaluated impact on the polyphagous American grasshopper Schistocerca americana. Grasshoppers showed no significant difference in weight gain and growth rate when feeding on wild type, overexpressing, or silenced lines, respectively. In addition, AtVSP protein was undetectable in either the midgut or frass of grasshoppers reared on transgenic plants suggesting that AtVSP was unable to withstand proteolytic degradation. To determine the stability of the AtVSP protein in grasshopper digestive canal, midgut extracts from various nymphal stages were incubated with bacterially expressed AtVSP for different periods of time. AtVSP was hydrolyzed rapidly by grasshopper midgut extract, in stark contrast with its fate when incubated with cowpea bruchid midgut extract. Multiple proteases have been detected in the midgut of grasshoppers, which may play important roles in determining the insect response to AtVSP. Results indicate that stability of an anti-insect protein in insect guts is a crucial property integral to the defense protein.     

Insect herbivory and vertebrate grazing impact food limitation and grasshopper populations during a severe outbreak

1. Competition between herbivores often plays an important role in population ecology and appears strongest when densities are high or plant production is low. Phytophagous insects are often highly abundant, but relatively few experiments have examined competition between vertebrates and phytophagous insects. 2. In grassland systems worldwide, grasshoppers are often the dominant phytophagous insect, and livestock grazing is a dominant land use. For this study, a novel experiment was conducted examining competition between vertebrates and invertebrates, where both grasshopper densities and sheep grazing were manipulated inside 10‐m² caged mesocosms during a grasshopper outbreak. We examined how grasshopper densities and the timing of vertebrate herbivory affected grasshopper densities, if the effects of vertebrates on survival and reproduction changed with grasshopper density, and how a naturally occurring grasshopper outbreak affected grasshopper populations in the following year. 3. Densities of grasshoppers at the site peaked at 130 m^–2. Food‐limited competition was stronger in treatments with higher grasshopper densities and repeated or late livestock herbivory, leading to reduced survival, femur length, and functional ovarioles, a measure of future reproduction. Strong food‐limited density‐dependent reproduction and survival led to reduced hatching densities in 2001. 4. As competition was typically stronger with high grasshopper densities than with livestock grazing, competition from vertebrates could be relatively less important for phytophagous insect population dynamics during outbreaks. The experiment provides insights into how competition between insect and vertebrate herbivores influences insect population dynamics, and indicates that severe outbreaks can rapidly subside with strong competition from vertebrate and insect herbivores.     

An arsenate-activated glutaredoxin from the arsenic hyperaccumulator fern Pteris vittata L. regulates intracellular arsenite

To elucidate the mechanisms of arsenic resistance in the arsenic hyperaccumulator fern Pteris vittata L., a cDNA for a glutaredoxin (Grx) Pv5-6 was isolated from a frond expression cDNA library based on the ability of the cDNA to increase arsenic resistance in Escherichia coli. The deduced amino acid sequence of Pv5-6 showed high homology with an Arabidopsis chloroplastic Grx and contained two CXXS putative catalytic motifs. Purified recombinant Pv5-6 exhibited glutaredoxin activity that was increased 1.6-fold by 10 mm arsenate. Site-specific mutation of Cys(67) to Ala(67) resulted in the loss of both GRX activity and arsenic resistance. PvGrx5 was expressed in E. coli mutants in which the arsenic resistance genes of the ars operon were deleted (strain AW3110), a deletion of the gene for the ArsC arsenate reductase (strain WC3110), and a strain in which the ars operon was deleted and the gene for the GlpF aquaglyceroporin was disrupted (strain OSBR1). Expression of PvGrx5 increased arsenic tolerance in strains AW3110 and WC3110, but not in OSBR1, suggesting that PvGrx5 had a role in cellular arsenic resistance independent of the ars operon genes but dependent on GlpF. AW3110 cells expressing PvGrx5 had significantly lower levels of arsenite when compared with vector controls when cultured in medium containing 2.5 mm arsenate. Our results are consistent with PvGrx5 having a role in regulating intracellular arsenite levels, by either directly or indirectly modulating the aquaglyceroporin. To our knowledge, PvGrx5 is the first plant Grx implicated in arsenic metabolism.     

Induced Resistance in Wild Tobacco with Clipped Sagebrush Neighbors: The Role of Herbivore Behavior

Previous experiments showed that wild tobacco plants with experimentally clipped sagebrush neighbors experienced less damage by grasshoppers than tobacco plants with unclipped sagebrush neighbors. This result could have been caused by grasshoppers preferring not to feed near clipped sagebrush. This hypothesis was tested in field choice experiments using six grasshopper species feeding on an unresponsive and uniformly palatable food. When offered food that was either close to clipped sagebrush or close to unclipped sagebrush, grasshoppers showed no preference. When offered food that was either close to sagebrush (3 cm) or far from sagebrush (30 cm), grasshoppers preferred to feed far from sagebrush. However, this preference was similar whether or not the sagebrush had been clipped. Avoidance of feeding near clipped sagebrush, independent of changes in tobacco, was not found to contribute to our earlier result that tobacco near clipped sagebrush suffered less herbivory than tobacco near unclipped sagebrush.     

Assessing density–damage relationships between water hyacinth and its grasshopper herbivore

Plants are variable in their responses to insect herbivory. Experimental increases in densities of phytophagous insects can reveal the type of plant response to herbivory in terms of impact and compensatory ability. The relationship between insect density and plant damage of a grasshopper, Cornops aquaticum Brüner (Orthoptera: Acrididae: Tetrataeniini), a candidate biological control agent, and an invasive aquatic plant, water hyacinth, Eichhornia crassipes Mart. Solms‐Laubach (Pontederiaceae), was investigated to assess potential damage to the weed. The impact of different densities of male and female grasshoppers on E. crassipes growth parameters was determined in a quarantine glasshouse experiment. Damage curves indicated that the relationship between plant biomass reduction and insect density was curvilinear whereas leaf production was linear. Female C. aquaticum were more damaging than males, causing high rates of plant mortality before the end of the trial at densities of three and four per plant. Feeding by C. aquaticum significantly reduced the total plant biomass and the number of leaves produced, and female grasshoppers caused a greater reduction in the number of leaves produced by water hyacinth plants than males. Grasshopper herbivory suppressed vegetative reproduction in E. crassipes, suggesting C. aquaticum could contribute to a reduction in the density and spread of E. crassipes infestations. The results showed that E. crassipes vigour and productivity decreases with an increase in feeding intensity by the grasshopper. Cornops aquaticum should therefore be considered for release in South Africa based on its host specificity and potential impact on E. crassipes.     

Behavioral responses of Schistocerca americana (Orthoptera: Acrididae) to Azadirex (neem)-treated host plants

Azadirex (azadirachtin and other biologically active extracts from neem trees) has been shown to have considerable potential to be used in integrated pest management systems based on its growth regulator/insecticide properties. Less well known are the antifeedant properties. The feeding-deterrent properties of a commercial azadirex formulation (Azatrol EC) were evaluated using both no-choice and choice tests, the American grasshopper, Schistocerca americana (Drury), and four host plants [savoy cabbage, Brassica oleracea variety capitata L.; cos (romaine) lettuce, Lactuca sativa variety longifolia Lam.; sweet orange, Citrus sinensis variety Hamlin L.; and peregrina, Jatropha integerrima Jacq.]. These studies demonstrated that azadirex application can significantly affect the feeding behavior of grasshoppers. Some degree of protection can be afforded to plants that differ markedly in their innate attractiveness to the insect, although the level of protection varies among hosts. The tendency of grasshoppers to sometimes feed on azadirex-treated foliage suggests that it will be difficult to prevent damage from occurring at all times, on all hosts. No evidence of rapid habituation to azadirex was detected. Rapid loss of efficacy was observed under field conditions, suggesting that daily retreatment might be necessary to maintain protection of plants from feeding.     

EFFECTS OF NITROGEN AND PHOSPHORUS LEVELS,AND FROND-HARVESTING ON ABSORPTION,TRANSLOCATION AND ACCUMULATION OF ARSENIC BY CHINESE BRAKE FERN (PTERIS VITTATA L.)

This hydroponic experiment was conducted to determine the effects of nitrogen (N) and phosphorus (P) levels and frond-harvesting on the effectiveness of arsenic (As)-hyperaccumulator Chinese brake fern (Pteris vittata L.) to remove As from contaminated groundwater collected from south Florida. Three-month old ferns were grown in 38-L plastic tanks (two ferns per tank) containing 30-L of As-contaminated water (130 μg·L^?1 As), which was amended with modified 0.25 strength Hoagland's solution #2. Two N (26 or 52 mg·L^?1) and two P levels (1.2 and 2.4 mg·L^?1) were tested in one experiment, whereas the effect of frond-harvesting was tested in a separate experiment. Initially, N had little effect on plant As removal whereas low P treatment was more effective than high P and As was reduced to <5 μg·L^?1 in 28 d compared to 35 d. For well-established ferns, N and P levels had little effect. Reused fern, with or without harvesting the As-rich fronds, took up arsenic more rapidly so the As concentration in the groundwater declined faster (130 to ～10 μg·L^?1 in 8 h). Regardless of the treatments, most As (85–93%) was located in the aboveground tissue (rhizomes and fronds). Frond As concentrations were higher for non-harvested ferns than for ferns where fronds were partially harvested prior to treatment. Conversely, rhizomes accumulated more arsenic in ferns where fronds had been partially harvested. Low-P treatment coupled with reuse of more established ferns with or without harvesting fronds can be used to effectively remove arsenic from contaminated water using P. vittata     

Mechanisms of arsenic hyperaccumulation in Pteris vittata. Uptake kinetics,interactions with phosphate,and arsenic speciation

The mechanisms of arsenic (As) hyperaccumulation in Pteris vittata, the first identified As hyperaccumulator, are unknown. We investigated the interactions of arsenate and phosphate on the uptake and distribution of As and phosphorus (P), and As speciation in P. vittata. In an 18-d hydroponic experiment with varying concentrations of arsenate and phosphate, P. vittata accumulated As in the fronds up to 27,000 mg As kg(-1) dry weight, and the frond As to root As concentration ratio varied between 1.3 and 6.7. Increasing phosphate supply decreased As uptake markedly, with the effect being greater on root As concentration than on shoot As concentration. Increasing arsenate supply decreased the P concentration in the roots, but not in the fronds. Presence of phosphate in the uptake solution decreased arsenate influx markedly, whereas P starvation for 8 d increased the maximum net influx by 2.5-fold. The rate of arsenite uptake was 10% of that for arsenate in the absence of phosphate. Neither P starvation nor the presence of phosphate affected arsenite uptake. Within 8 h, 50% to 78% of the As taken up was distributed to the fronds, with a higher translocation efficiency for arsenite than for arsenate. In fronds, 49% to 94% of the As was extracted with a phosphate buffer (pH 5.6). Speciation analysis using high-performance liquid chromatography-inductively coupled plasma mass spectroscopy showed that >85% of the extracted As was in the form of arsenite, and the remaining mostly as arsenate. We conclude that arsenate is taken up by P. vittata via the phosphate transporters, reduced to arsenite, and sequestered in the fronds primarily as As(III).     

Characterization of glutathione reductase and catalase in the fronds of two Pteris ferns upon arsenic exposure

To better understand the mechanisms of plant tolerance to high concentration of arsenic, we characterized two antioxidant enzymes, glutathione reductase (GR) and catalase (CAT), in the fronds of Pteris vittata, an arsenic-hyperaccumulating fern, and Pteris ensiformis, an arsenic-sensitive fern. The induction, activation and apparent kinetics of GR and CAT in the plants upon arsenic exposure were investigated. Under arsenic exposure (sodium arsenate), CAT activity in P. vittata was increased by 1.5-fold, but GR activity was unchanged. Further, GR was not inhibited or activated by the arsenic in assays. No significant differences in K_m and V_max values of GR or CAT were observed between the two ferns. However, CAT activity in P. vittata was activated by 200 μM arsenate up to 300% compared to the control. Similar but much smaller increases were observed for P. ensiformis and purified bovine liver catalase (133% and 120%, respectively). This research reports, for the first time, the activation of CAT by arsenic in P. vittata. The increased CAT activities may allow P. vittata to more efficiently mediate arsenic-induced stress by preparing the fern for the impeding production of reactive oxygen species resulting from arsenate reduction to arsenite in the fronds.