Calling dynamics and call synchronization in a local group of unison bout callers.

In many species of chorusing frogs, callers can rapidly adjust their call timing with reference to neighboring callers so as to maintain call rate while minimizing acoustic interference. The rules governing the interactions, in particular, who is listening to whom are largely unknown, presumably influenced by distance between callers, caller density, and intensities of interfering calls. We report vocal interactions in a unison bout caller, the green tree frog (Hyla cinerea). Using a microphone array, we monitored bouts from a local group of six callers embedded in a larger chorus. Data were analyzed in a 21-min segment at the peak of the chorus. Callers within this group were localized and their voices were separated for analysis of spatio-temporal interactions. We show that callers in this group: (1) synchronize with one another, (2) prefer to time their calls antiphonally, almost exactly at one-third and two-thirds of the call intervals of their neighbors, (3) tolerate call collision when antiphonal calling is not possible, and (4) perform discrete phase-hopping between three preferred phases when tracking other callers. Further, call collision increases and phase-locking decreases, with increasing inter-caller spacing. We conclude that the precise phase-positioning, phase-tracking, and phase-hopping minimizes acoustic jamming while maintaining chorus synchrony.

Runoff transport of pyrethroids from a residential lawn in central California.

An irrigation runoff study on a residential lawn was conducted in California, northeast of Sacramento, during the summer and fall of 2008 to investigate the contribution of turf uses of pyrethroids to residues in Californian urban creek sediments. This study examined how over irrigation (i.e., irrigation that produces runoff) in the summer season may transport recently applied pyrethroids. The study included liquid and granular applications of both bifenthrin [(2-methyl-3-phenyl-phenyl) methyl 3-(2-chloro-3,3,3-trifluoro-prop-1-enyl)-2,2-dimethyl-cyclopropane-1-carboxylate] and beta-cyfluthrin [Cyano(4-fluoro-3-phenoxyphenyl)methyl 3-(2,2-dichloroethenyl)-2,2-dimethyl-cyclopropanecarboxylate]. Generally, runoff did not occur at irrigation rates of 2.03 cm/h (0.8 in/h) but did occur when the irrigation rates were increased to about 3.81 cm/h (1.5 in/h), generating chemical losses in the first runoff event of up to 0.58 and 0.08% of applied for beta-cyfluthrin and bifenthrin, respectively. Chemical runoff losses dropped significantly between over-irrigation events with the third over-irrigation event chemical runoff losses representing 0.026 and 0.015% of applied for beta-cyfluthrin and bifenthrin, respectively. Runoff losses were generally less for liquid formulations than granular formulations but within a factor of three. Additionally, the study included a simulated winter rainstorm 8 wk after application. The low runoff losses from turf seen in this study suggest that other sources could be contributing to observed residues in urban streams. Other sources could include pyrethroids ending up on impervious surfaces, such as concrete driveways from off-target applications to turf, spills, and other poor handling practices, or pyrechroids applied directly to impervious surfaces for insect control.

Hydrogen sulfide alleviates aluminum toxicity in germinating wheat seedlings.

Protective role of hydrogen sulfide (H(2)S) on seed germination and seedling growth was studied in wheat (Triticum) seeds subjected to aluminum (Al(3+)) stress. We show that germination and seedling growth of wheat is inhibited by high concentrations of AlCl(3). At 30 mmol/L AlCl(3) germination is reduced by about 50% and seedling growth is more dramatically inhibited by this treatment. Pre-incubation of wheat seeds in the H(2)S donor NaHS alleviates AlCl(3)-induced stress in a dose-dependant manner at an optimal concentration of 0.3 mmol/L. We verified that the role of NaHS in alleviating Al(3+) stress could be attributed to H(2)S/HS(-) by showing that the level of endogenous H(2)S increased following NaHS treatment. Furthermore, other sodium salts containing sulfur were ineffective in alleviating Al(3+) stress. NaHS pretreatment significantly increased the activities of amylases and esterases and sustained much lower levels of MDA and H(2)O(2) in germinating seeds under Al(3+) stress. Moreover, NaHS pretreatment increased the activities of guaiacol peroxidase, ascorbate peroxidase, superoxide dismutase and catalase and decreased that of lipoxygenase. NaHS pretreatment also decreased the uptake of Al(3+) in AlCl(3)-treated seed. Taken together these results suggest that H(2)S could increase antioxidant capability in wheat seeds leading to the alleviation of Al(3+) stress.

Summary of potable well monitoring conducted for aldicarb and its metabolites in the United States in 2005.

United States potable well monitoring in the Pacific Northwest, California, Texas, Mississippi delta, and the Southeast was conducted in 2005 to provide estimates of dietary exposure to aldicarb carbamate residues (aldicarb and its two carbamate metabolites) in potentially vulnerable potable wells. Samples were analyzed from 1,673 drinking water wells in nine major use areas that were within 300 m of fields treated at least once with aldicarb between 2002 and 2005. Analyses were performed with a high-performance liquid chromatography/tandem mass spectrometry analytical method with limits of quantitation of 0.021 microg/L for aldicarb and aldicarb sulfone and 0.027 microg/L for aldicarb sulfoxide. The method detection limits were 0.0070 microg/L for aldicarb and aldicarb sulfone and 0.0090 microg/L for aldicarb sulfoxide. Samples from 1,513 of the 1,673 wells contained no aldicarb carbamate residues. Only 10 wells had total aldicarb carbamate residues above 1 microg/L, the maximum being 2.9 microg/L. All residues were below the U.S. Environmental Protection Agency Health Advisory Limit (HAL) of 10 microg/L. Given the number of samples collected in this study, there is a 95% confidence level that the maximum concentration in this study exceeds the 99.8th percentile of concentrations of all potable wells located within 300 m of fields recently treated with aldicarb. Therefore, the data from this study show that the restrictions and management practices on the current product label continue to be effective in preventing residues in excess of the 10 microg/L HAL in potable wells in these use areas.

Effects of formulation on transport of pyrethroids in residential settings.

Washoff of 17 pyrethroid products resulting from a 1-h, 25.4-mm rainfall occurring 24 h after application was measured in indoor studies with concrete slabs. These products included different pyrethroid active ingredients and a range of formulation types. Based on this replicated study, 5 product pairs with contrasting washoff behaviors were chosen for an outdoor study using 6 full-scale house fronts in central California. Products in 4 of these pairs were applied once to different rectangular areas on the driveway (1 product in each pair to 3 house lots and the other to the remaining 3 house lots). The products in the fifth pair were applied 3 times at 2-mo intervals to vertical stucco walls above the driveway. All house lots received natural and simulated rainfall over 7 mo. Indoor studies showed differences up to 170-fold between paired products, whereas the maximum difference between paired products in the field was only 5-fold. In the pair applied to the wall, 1 product had 91 times the washoff of the other in the indoor study, whereas in the field the same product had 15% lower washoff. These results show that, although the formulation may influence washoff under actual use conditions, its influence is complex and not always as predicted by indoor experiments. Because the formulation also affects insect control, washoff research needs to be conducted together with efficacy testing.

Comparison of regulatory estimates of drinking water concentrations with monitoring data.

Currently, regulatory practice in the United States is to estimate potential concentrations in drinking water from surface water by using an index reservoir scenario. This approach extrapolates results from the modeling of a single field with maximum application rates to the watershed scale, based on a percent crop area estimate. Since 1998, Bayer CropScience and its predecessor companies have conducted drinking water monitoring studies with a number of different compounds. The results from these studies show that the index reservoir scenario greatly overpredicts residues in surface water. The most important factor is the overestimation of use within a watershed. Other factors contributing to the overestimation of concentrations are the conservative procedures used to obtain the chemical fate related input parameters and the simplified hydrology. A new procedure based on the USGS WARP model, being developed by a group of scientists from the EPA, USGS, USDA, and industry, will provide more realistic estimates of concentrations of pesticides and their metabolites in drinking water.

Protein secretion in plants.

Protein secretion is an ubiquitous but poorly understood process in plants. Secreted proteins are synthesized on the membranes of the rough endoplasmic reticulum and transported to the cell surface by secretary vesicles formed at the Golgi apparatus. Whereas many of the structural details of this process are known the mechanisms underlying secretion are just beginning to be understood, in this article we review some of the recent developments in this field, and we compare the progress made with animal and plant cells. CONTENTS Summary 567 I. Introduction 568 II. Proteins secreted by plants 568 III. Synthesis and post-translational modification of secreted proteins 571 IV. Molecular requirements for secretion 576 V. Vehicles of secretory transport 581 VI. Regulation of secretion 585 VII. Conclusions and Perspective 587 Acknowledgements 588 References 588.

Signalling in the cereal aleurone: hormones, reactive oxygen and cell death.

The cereal aleurone is widely used as a model system to study hormonal signalling. Abscisic acid (ABA) and gibberellins (GAs) elicit distinct responses in aleurone cells, ranging from those occurring within minutes of hormone addition to those that require several hours or days to complete. Programmed cell death is an example of a response in aleurone layers that is hormonally regulated. GAs promote cell death and cells in intact aleurone layers begin to die 24 h after GA treatment, whereas cell death of aleurone protoplasts begins 4 d after GA treatment. ABA prevents aleurone cell death and addition of ABA to cells pretreated with GA can delay cell death. Aleurone cells do not follow the apoptotic route of programmed cell death. Cells treated with GA, but not ABA, develop large, acidic vacuoles containing a spectrum of hydrolases typical of lytic compartments. Enzymes that metabolize reactive oxygen species are also present in aleurone cells, but ascorbate peroxidase, catalase and superoxide dismutase become less abundant after treatment with GA; activity of these enzymes increases or remains unchanged in ABA-treated cells. We propose a model whereby reactive oxygen species accumulate in GA-treated cells and lead to peroxidation of membrane lipids and plasma membrane rupture. ABBREVIATIONS: RO, reactive oxygen species; HR, hypersensitive response; PSV, protein storage vacuole; PCD, programmed cell death; CAT, catalase; SOD, superoxide dismutase; APX, ascorbate peroxidase.

Comparison of pesticide root zone model 3.12: runoff predictions with field data.

As part of a process to improve confidence in the results of regulatory modeling, predictions of pesticide root zone model (PRZM) 3.12 were compared with measured data collected in nine different runoff field studies. This comparison shows that PRZM 3.12 provides a reasonable estimate of chemical runoff at the edge of a field. Simulations based on the best choices for input parameters (no conservatism built into input parameters) are generally within an order of magnitude of measured data, with better agreement observed both for larger events and for cumulative values over the study period. When the model input parameters are calibrated to improve the hydrology, the fit between predicted and observed data improves (results are usually within a factor of three). When conservatism is deliberately introduced into the input pesticide parameters, substantial overprediction of runoff losses occur. Recommendations for future work to improve regulatory models include implementation of more sophisticated evapotranspiration routines, allowing for seasonal variation of various model parameters (such as curve numbers, crop cover, and Manning's surface roughness coefficients), better procedures for estimating site-specific degradation rates in surface and subsoils, and improved sorption routines.

Review of the validation of models used in Federal Insecticide, Fungicide, and Rodenticide Act environmental exposure assessments.

The first activity of the Federal Insecticide. Fungicide, and Rodenticide Act (FIFRA) Environmental Model Validation Task Force, established to increase confidence in the use of environmental models used in regulatory assessments, was to review the literature information on validation of the pesticide root zone model (PRZM) and the groundwater loading effects of agricultural management systems (GLEAMS). This literature information indicates that these models generally predict the same or greater leaching than observed in actual field measurements, suggesting that these models are suitable for use in regulatory assessments. However, additional validation research conducted using the newest versions of the models would help improve confidence in runoff and leaching predictions because significant revisions have been made in models over the years, few of the literature studies focused on runoff losses, the number of studies having quantitative validation results is minimal, and modelers were aware of the field results in most of the literature studies. Areas for special consideration in conducting model validation research include improving the process for selecting input parameters, developing recommendations for performing calibration simulations, devising appropriate procedures for keeping results of field studies from modelers performing simulations to validate model predictions while providing access for calibration simulations, and developing quantitative statistical procedures for comparing model predictions with experimental results.

Comparison of pesticide root zone model 3.12: leaching predictions with field data.

As part of a process to improve confidence in the results of regulatory modeling, predictions of the pesticide root zone model (PRZM) 3.12 were compared with measured data collected in nine different field leaching studies. Reasonable estimates of leaching were obtained with PRZM 3.12 in homogeneous soils where preferential flow is not significant. The PRZM 3.12 usually did a good job of predicting movement of bromide in soil (soil and soil pore-water concentrations were generally within a factor of two of predicted values). For simulations based on the best choices for input parameters, predictions of soil pore-water concentrations for pesticides were usually within a factor of three and soil pore-water estimates within a factor of 11. When the model input parameters were calibrated to improve the simulation of hydrology, predicted pesticide concentrations in soil pore water were usually within a factor of two of measured concentrations. Because of the sensitivity of leaching to degradation rate, the most accurate predictions were obtained with pesticides with relatively slow degradation rates. When conservative assumptions were used to define input pesticide parameters, predictions of pesticide concentrations were usually a factor of two greater than when using the best estimate of input parameters without any built-in conservatism.

Predicted impact of transgenic, herbicidetolerant corn on drinking water quality in vulnerable watersheds of the mid-western USA.

In the intensely farmed corn-growing regions of the mid-western USA, surface waters have often been contaminated by herbicides, principally as a result of rainfall runoff occurring shortly after application of these to corn and other crops. In some vulnerable watersheds, water quality criteria for chronic human exposure through drinking water are occasionally exceeded. We selected three settings representative of vulnerable corn-region watersheds, and used the PRZM-EXAMS model with the Index Reservoir scenario to predict corn herbicide concentrations in the reservoirs as a function of herbicide properties and use pattern, site characteristics and weather in the watersheds. We compared herbicide application scenarios, including broadcast surface pre-plant atrazine and alachlor applications with a glyphosate pre-plant application, scenarios in which losses of herbicides were mitigated by incorporation or banding, and scenarios in which only glyphosate or glufosinate post-emergent herbicides were used with corn genetically modified to be resistant to them. In the absence of drift, in almost all years a single runoff event dominates the input into the reservoir. As a result, annual average pesticide concentrations are highly correlated with annual maximum daily values. The modeled concentrations were generally higher than those derived from monitoring data, even for no-drift model scenarios. Because of their lower post-emergent application rates and greater soil sorptivity, glyphosate and glufosinate loads in runoff were generally one-fifth to one-tenth those of atrazine and alachlor. These model results indicate that the replacement of pre-emergent corn herbicides with the post-emergent herbicides allowed by genetic modification of crops would dramatically reduce herbicide concentrations in vulnerable watersheds. Given the significantly lower chronic mammalian toxicity of these compounds, and their vulnerability to breakdown in the drinking water treatment process, risks to human populations through drinking water would also be reduced.

Major transport mechanisms of pyrethroids in residential settings and effects of mitigation measures.

The major pathways for transport of pyrethroids were determined in runoff studies conducted at a full-scale test facility in central California, USA. The 6 replicate house lots were typical of front lawns and house fronts of California residential developments and consisted of stucco walls, garage doors, driveways, and residential lawn irrigation sprinkler systems. Each of the 6 lots also included a rainfall simulator to generate artificial rainfall events. Different pyrethroids were applied to 5 surfaces­driveway, garage door and adjacent walls, lawn, lawn perimeter (grass near the house walls), and house walls above grass. The volume of runoff water from each house lot was measured, sampled, and analyzed to determine the amount of pyrethroid mass lost from each surface. Applications to 3 of the house lots were made using the application practices typically used prior to recent label changes, and applications were made to the other 3 house lots according to the revised application procedures. Results from the house lots using the historic application procedures showed that losses of the compounds applied to the driveway and garage door (including the adjacent walls) were 99.75% of total measured runoff losses. The greatest losses were associated with significant rainfall events rather than lawn irrigation events. However, runoff losses were 40 times less using the revised application procedures recently specified on pyrethroid labels.

Laboratory degradation rates of 11 pyrethroids under aerobic and anaerobic conditions.

Degradation of 11 pyrethroids was measured over approximately 100 days in three sediment/water systems under aerobic and anaerobic conditions at 25 °C in the dark. The three California sediments represented a range of textures and organic matter. Test compounds were bifenthrin, cypermethrin, ζ-cypermethrin, cyfluthrin, ß-cyfluthrin, deltamethrin, esfenvalerate, fenpropathrin, γ-cyhalothrin, λ-cyhalothrin, and permethrin. A non-standard design was employed to keep conditions essentially the same for all compounds. The test compounds were applied as two test mixtures (six active ingredients per mixture, with bifenthrin common to both) at approximately 50 µg of test compound/kg of sediment (dry weight). Extracts of sediment/water were cleaned up by solid-phase extraction, concentrated, and analyzed by gas chromatography/mass spectrometry (except deltamethrin) against matrix-matched standards, with cyfluthrin-d6 as an internal standard. Deltamethrin was analyzed by liquid chromatography/tandem mass spectrometry using deltamethrin-phenoxy-(13)C6 as an internal standard. Similar degradation rates of bifenthrin and for related isomeric compounds (e.g., cyfluthrin and ß-cyfluthrin) were generally measured in both mixtures for each sediment. First-order half-lives under aerobic conditions ranged from 2.9 to greater than 200 days, with a median value of 18 days. Under anaerobic conditions, the range was from 20 to greater than 200 days, with a median value of 70 days.

Nitric oxide methods in seed biology.

The ubiquitous signaling molecule nitric oxide (NO) plays an important role in seed biology. Experiments with this biologically important gas require special provisions because NO in aerobic environments is readily converted into other oxides of nitrogen. In this chapter, we describe methods for the application of NO as a gas, and through the use of NO-donor compounds. We included information on the removal or reduction of NO with NO scavengers. Methods for detecting NO using NO-reactive fluorescent probes, and an apparatus incorporating an oxidizer column are also described.

Hydrogen sulfide stimulates ß-amylase activity during early stages of wheat grain germination.

We recently reported that H 2S could significantly promote the germination of wheat grains subjected to aluminum (Al(3+)) stress.1 In these experiments seeds were pretreated with the H 2S donor NaHS for 12 h prior to Al(3+) stress. During this pre-incubation period we observed that H2S increased the activity of grain amylase in the absence of Al(3+). Using embryoless half grains of wheat we now show that H2S preferentially affects the activity of endosperm ß-amylase and that α-amylase synthesis and activity is unaffected by this treatment.

Measuring NO production by plant tissues and suspension cultured cells.

We describe an inexpensive and reliable detector for measuring NO emitted in the gas phase from plants. The method relies on the use of a strong oxidizer to convert NO to NO2 and subsequent capture of NO2 by a Griess reagent trap. The set-up approaches the sensitivity for NO comparable to that of instruments based on chemiluminescence and photoacoustic detectors. We demonstrate the utility of our set-up by measuring NO produced by a variety of well established plant sources. NO produced by nitrate reductase (NR) in tobacco leaves and barley aleurone was readily detected, as was the production of NO from nitrite by the incubation medium of barley aleurone. Arabidopsis mutants that overproduce NO or lack NO-synthase (AtNOS1) also displayed the expected NO synthesis phenotype when assayed by our set-up. We could also measure NO production from elicitor-treated suspension cultured cells using this set-up. Further, we have focused on the detection of NO by a widely used fluorescent probe 4-amino-5-methylamino-2',7'-difluorofluorescein (DAF-FM). Our work points to the pitfalls that must be avoided when using DAF-FM to detect the production of NO by plant tissues. In addition to the dramatic effects that pH can have on fluorescence from DAF-FM, the widely used NO scavengers 2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (PTIO) and 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (cPTIO) can produce anomalous and unexpected results. Perhaps the most serious drawback of DAF-FM is its ability to bind to dead cells and remain NO-sensitive.

The Arabidopsis aleurone layer responds to nitric oxide, gibberellin, and abscisic acid and is sufficient and necessary for seed dormancy.

Seed dormancy is a common phase of the plant life cycle, and several parts of the seed can contribute to dormancy. Whole seeds, seeds lacking the testa, embryos, and isolated aleurone layers of Arabidopsis (Arabidopsis thaliana) were used in experiments designed to identify components of the Arabidopsis seed that contribute to seed dormancy and to learn more about how dormancy and germination are regulated in this species. The aleurone layer was found to be the primary determinant of seed dormancy. Embryos from dormant seeds, however, had a lesser growth potential than those from nondormant seeds. Arabidopsis aleurone cells were examined by light and electron microscopy, and cell ultrastructure was similar to that of cereal aleurone cells. Arabidopsis aleurone cells responded to nitric oxide (NO), gibberellin (GA), and abscisic acid, with NO being upstream of GA in a signaling pathway that leads to vacuolation of protein storage vacuoles and abscisic acid inhibiting vacuolation. Molecular changes that occurred in embryos and aleurone layers prior to germination were measured, and these data show that both the aleurone layer and the embryo expressed the NO-associated gene AtNOS1, but only the embryo expressed genes for the GA biosynthetic enzyme GA3 oxidase.

Nitric oxide reduces seed dormancy in Arabidopsis.

Dormancy is a property of many mature seeds, and experimentation over the past century has identified numerous chemical treatments that will reduce seed dormancy. Nitrogen-containing compounds including nitrate, nitrite, and cyanide break seed dormancy in a range of species. Experiments are described here that were carried out to further our understanding of the mechanism whereby these and other compounds, such as the nitric oxide (NO) donor sodium nitroprusside (SNP), bring about a reduction in seed dormancy of Arabidopsis thaliana. A simple method was devised for applying the products of SNP photolysis through the gas phase. Using this approach it was shown that SNP, as well as potassium ferricyanide (Fe(III)CN) and potassium ferrocyanide (Fe(II)CN), reduced dormancy of Arabidopsis seeds by generating cyanide (CN). The effects of potassium cyanide (KCN) on dormant seeds were tested and it was confirmed that cyanide vapours were sufficient to break Arabidopsis seed dormancy. Nitrate and nitrite also reduced Arabidopsis seed dormancy and resulted in substantial rates of germination. The effects of CN, nitrite, and nitrate on dormancy were prevented by the NO scavenger c-PTIO. It was confirmed that NO plays a role in reducing seed dormancy by using purified NO gas, and a model to explain how nitrogen-containing compounds may break dormancy in Arabidopsis is presented.

Sodium nitroprusside, cyanide, nitrite, and nitrate break Arabidopsis seed dormancy in a nitric oxide-dependent manner.

The seeds of many plant species are dormant at maturity and dormancy loss is a prerequisite for germination. Numerous environmental and chemical treatments are known to lessen or remove seed dormancy, but the biochemical changes that occur during this change of state are poorly understood. Several lines of research have implicated nitric oxide (NO) as a participant in this process. Here, we show that dormant seeds of Arabidopsis thaliana (L.) Heynh. will germinate following treatment with the NO donor sodium nitroprusside (SNP), cyanide (CN), nitrite or nitrate. In all cases, the NO scavenger c-PTIO effectively promotes the maintenance of seed dormancy. c-PTIO does not, however, inhibit germination of fully after-ripened seeds, and c-PTIO does not interact directly with nitrite, nitrate or CN. We also show that volatile CN effectively breaks dormancy of Arabidopsis seeds, and that CN is the volatile compound in SNP that promotes dormancy loss. Our data support the hypothesis that NO is a signaling molecule that plays an important role in the loss of seed dormancy.