Probing mode of action in plant cell cycle by the herbicide endothall, a protein phosphatase inhibitor

The mode of action of endothall, an herbicide which was reported to inhibit plant protein phosphatases 1 (PP1) and 2A (PP2A), was investigated. For initial characterization, a series of bioassays was used for comprehensive physiological profiling of endothall effects which suggested a phytotoxic mode of action similar to mitotic disrupter herbicides. Unlike known microtubule disrupters, endothall did not inhibit soybean tubulin polymerization in vitro. As shown in meristematic corn root tips, endothall distorted the orientation of cell division plane and microtubule spindle structures which led to cell cycle arrest in prometaphase. In tobacco BY-2 cells, malformed spindles together with prometaphase arrest of nuclei and abnormal perinuclear microtubule patterns were detected as early as 4 h of endothall treatment. These effects were also observed after treatment with other protein phosphatase inhibitors, cantharidin and okadaic acid, which phenocopied the mitotic changes described in tonneau1 (ton1) and tonneau2 (ton2) Arabidopsis mutants. These mutants are defective in TONNEAU2 (TON2) protein, a regulatory subunit of PP2A, which governs cell division plane and microtubule orientation. Therefore, PP2A/TON2 phosphatase complex is suggested to be an in planta molecular target of endothall. However, in BY-2 cells, additional effects of endothall, including inhibition of S-phase initiation and DNA synthesis, detected by 5-ethynyl-2′-deoxyuridine (EdU) incorporation, and condensed nuclei arrested in late mitosis were observed which were not reported in Arabidopsiston1 and ton2 mutants. This result indicates that two additional checkpoints in cell cycle were blocked by endothall which are probably not associated with TON2-pathway inhibition. Possibly, inhibition of PP1 and/or other PP2A protein phosphatases are involved in the regulation of these cell cycle phenomena.     

Dynamic plant uptake model applied for drip irrigation of an insecticide to pepper fruit plants

BACKGROUND: Drip application of insecticides is an effective way to deliver the chemical to the plant that avoids off‐site movement via spray drift and minimizes applicator exposure. The aim of this paper is to present a cascade model for the uptake of pesticide into plants following drip irrigation, its application for a soil‐applied insecticide and a sensitivity analysis of the model parameters. RESULTS: The model predicted the measured increase and decline of residues following two soil applications of an insecticide to peppers, with an absolute error between model and measurement ranging from 0.002 to 0.034 mg kg fw^?1. Maximum measured concentrations in pepper fruit were approximately 0.22 mg kg fw^?1. Temperature was the most sensitive component for predicting the peak and final concentration in pepper fruit, through its influence on soil and plant degradation rates. CONCLUSION: Repeated simulations of pulse inputs with the cascade model adequately describe soil pesticide applications to an actual cropped system and reasonably mimic it. The model has the potential to be used for the optimization of practical features, such as application rates and waiting times between applications and before harvest, through the integrated accounting of soil, plant and environmental influences. Copyright © 2011 Society of Chemical Industry     

Mixing of different mineral soil layers by endogeic earthworms affects carbon and nitrogen mineralization

The effect of the endogeic earthworm species Octolasion tyrtaeum (Savigny) on decomposition of uniformly ¹⁴C-labelled lignin (lignocellulose) was studied in microcosms with upper mineral soil (Ah-horizon) from two forests on limestone, representing different stages of succession, a beech- and an ash-tree-dominated forest. Microcosms with and without lower mineral soil (Bw-horizon) were set-up; one O. tyrtaeum was added to half of them. It was hypothesised that endogeic earthworms stabilise lignin and the organic matter of the upper mineral soil by mixing with lower mineral soil of low C content. Cumulative C mineralization was increased by earthworms and by the addition of lower mineral soil. Effects of the lower mineral soil were more pronounced in the beech than in the ash forest. Cumulative mineralization of lignin was strongly increased by earthworms, but only in the beech soil (+24.6%). Earthworms predominantly colonized the upper mineral soil; mixing of the upper and lower mineral soils was low. The presence of lower mineral soil did not reduce the rates of decomposition of organic matter and lignin; however, the earthworm-mediated increase in mineralization was less pronounced in treatments with (+8.6%) than in those without (+14.1%) lower mineral soil. These results indicate that the mixing of organic matter with C-unsaturated lower mineral soil by endogeic earthworms reduced microbial decomposition of organic matter in earthworm casts.     

Changes in the succession and diversity of protozoan and microbial populations in soil spiked with a range of copper concentrations

We studied microbial and protozoan activity, diversity and abundance as affected by Cu²⁺ amendments ranging from 0 to 1000 μg g⁻¹ over a 70-day period. At the end of the experiment the microbial population size, as indicated by substrate-induced respiration, had normalized for all Cu²⁺ concentrations, but 1000 μg g⁻¹. Protozoan abundance was negatively affected by Cu²⁺, although, only in the first few weeks. A more detailed analysis of the individual components that make up the microbial and micro-faunal populations (phospholipid fatty acid (PLFA) profile and protozoan morphotypes), however, yielded a somewhat more complex picture. For the three highest Cu²⁺ amendments (160, 400 and 1000 μg g⁻¹), there still was a significant reduction in number of differentiable protozoan morphotypes. The bacterial PLFA pattern suggested a shift from Gram-negative towards Gram-positive bacteria for the high amendments, a process where protozoan grazing most likely played a significant role. The ratio of the trans/cis isomers of the 16:1ω7 fatty acid indicated that Cu²⁺, even at low and medium concentrations, induced physiological changes in the microbial population. The relatively slight changes in total microbial and micro-faunal abundance and activity, also at the highest Cu²⁺ concentrations, probably reflected the ability of the community to compensate for loss of taxa by functional substitution.     

Quantitative Mineralbestimmung in der Schlufffraktion von Böden auf der Grundlage der chemischen Analyse und unter Anwendung der Karl-Fischer-Titration. I. Verfahren

Quantitative estimation of the mineralogical composition of silt fractions of soils based on both, chemical analysis and application of Karl-Fischer-titration. I. Method This paper describes a method for calculating the mineralogical composition of the silt fractions of soils. The chemical analysis and the temperature dependence of water release are used for determining micas and feldspars, each in three components in a relatively short procedure. The release of water was measured by Karl-Fischer-titration at controlled temperatures. At temperatures above 550°C the water release of the particle size fractions correlates with their content of micas. The quantity of micas can be obtained from the amount of released water and the results of the chemical analysis. The influence of kaolinite, amphibole and the fluoride content on the calculation is described. By considering the potassium content of the mica fraction results of the chemical analysis are used to quantify feldspars.     

Statistical Modeling of the Partitioning of Nonylphenol in Soil

Partition coefficients K _P of nonylphenol (NP) in soil were determined for 193 soil samples which differed widely in content of soil organic carbon (SOC), hydrogen activity, clay content, and in the content of dissolved organic carbon (DOC). By means of multiple linear regression analysis (MLR), pedotransfer functions were derived to predict partition coefficients from soil data. SOC and pH affected the sorption, though the latter was in a range significantly below the pK_a of NP. Quality of soil organic matter presumably plays an important but yet not quantified role in sorption of NP. For soil samples with SOC values less than 3 g kg^?1, model prediction became uncertain with this linear approach. We suggest that using only SOC and pH data results in good prediction of NP sorption in soils with SOC higher than 3 g kg^?1. Considering the varying validity of the linear model for different ranges of the most sensitive parameter SOC, a more flexible, nonlinear approach was tested. The application of an artificial neuronal network (ANN) to predict sorption of NP in soils showed a sigmoidal relation between K _P and SOC. The nonlinear ANN approach provided good results compared to the MLR approach and represents an alternative tool for prediction of NP partitioning coefficients.     

Comparison of the chemical composition of extracts from Scutellaria lateriflora using accelerated solvent extraction and supercritical fluid extraction versus standard hot water or 70% ethanol extraction

The aqueous extract of American skullcap (Scutellaria lateriflora L. (S. lateriflora), Lamiaceae) has been traditionally used by North American Indians as a nerve tonic and for its sedative and diuretic properties. Recent reports stated that flavonoids and possibly amino acids are responsible for the anxiolytic activity. As a part of our search for environmentally friendly solvents to extract the active components from medicinal plants, we used S. lateriflora in a comparison of accelerated solvent extraction (ASE) using water, and supercritical fluid extraction (SFE) using CO2 and 10% EtOH as modifier, at different temperatures. Flavonoids and amino acids were quantified by HPLC-UV and HPLC-MS, respectively. The flavonoid content was compared with conventional extraction methods (hot water extraction and 70% ethanol). The use of ASE at 85 degrees C with water as solvent gave the best results for flavonoid glycosides and amino acids, whereas SFE gave higher yields of flavonoid aglycones. However, the results obtained for total flavonoids were not significatively superior to hot water extraction or 70% aqueous EtOH extract.     

Combined influence of NaCl and sucrose on heat-induced gelation of bovine serum albumin

The combined influence of a strongly interacting cosolvent (NaCl) and a weakly interacting cosolvent (sucrose) on the heat-induced gelation of bovine serum albumin (BSA) was studied. The dynamic shear rheology of 4 wt % BSA solutions containing 0 or 20 wt % sucrose and 0-200 mM NaCl was monitored as they were heated from 30 to 90 degrees C at 1.5 degrees C min(-)(1), held at 90 degrees C for 120 min, and then cooled back to 30 degrees C at -1.5 degrees C min(-)(1). The turbidity of the same solutions was monitored as they were heated from 30 to 95 degrees C at 1.5 degrees C min(-)(1) or held isothermally at 90 degrees C for 10 min. NaCl had a similar effect on BSA solutions that contained 0 or 20 wt % sucrose, with the gelation temperature decreasing and the final gel strength increasing with increasing salt concentration and the greatest changes occurring between 25 and 100 mM NaCl. Nevertheless, the presence of sucrose did lead to an increase in the gelation temperature and final gel strength and a decrease in the final gel turbidity. The impact of NaCl on gel characteristics was attributed primarily to its ability to screen electrostatic interactions between charged protein surfaces, whereas the impact of sucrose was attributed mainly to its ability to increase protein thermal stability and strengthen the attractive forces between proteins through a preferential interaction mechanism.     

Seasonal variation and fire effects on CH4, N2O and CO2 exchange in savanna soils of northern Australia

Tropical savanna ecosystems are a major contributor to global CO₂, CH₄ and N₂O greenhouse gas exchange. Savanna fire events represent large, discrete C emissions but the importance of ongoing soil-atmosphere gas exchange is less well understood. Seasonal rainfall and fire events are likely to impact upon savanna soil microbial processes involved in N₂O and CH₄ exchange. We measured soil CO₂, CH₄ and N₂O fluxes in savanna woodland (Eucalyptus tetrodonta/Eucalyptus miniata trees above sorghum grass) at Howard Springs, Australia over a 16 month period from October 2007 to January 2009 using manual chambers and a field-based gas chromatograph connected to automated chambers. The effect of fire on soil gas exchange was investigated through two controlled burns and protected unburnt areas. Fire is a frequent natural and management action in these savanna (every 1-2 years). There was no seasonal change and no fire effect upon soil N₂O exchange. Soil N₂O fluxes were very low, generally between −1.0 and 1.0 μg N m⁻² h⁻¹, and often below the minimum detection limit. There was an increase in soil NH₄⁺ in the months after the 2008 fire event, but no change in soil NO₃⁻. There was considerable nitrification in the early wet season but minimal nitrification at all other times.Savanna soil was generally a net CH₄ sink that equated to between −2.0 and −1.6 kg CH₄ ha⁻¹ y⁻¹ with no clear seasonal pattern in response to changing soil moisture conditions. Irrigation in the dry season significantly reduced soil gas diffusion and as a consequence soil CH₄ uptake. There were short periods of soil CH₄ emission, up to 20 μg C m⁻² h⁻¹, likely to have been caused by termite activity in, or beneath, automated chambers. Soil CO₂ fluxes showed a strong bimodal seasonal pattern, increasing fivefold from the dry into the wet season. Soil moisture showed a weak relationship with soil CH₄ fluxes, but a much stronger relationship with soil CO₂ fluxes, explaining up to 70% of the variation in unburnt treatments. Australian savanna soils are a small N₂O source, and possibly even a sink. Annual soil CH₄ flux measurements suggest that the 1.9 million km² of Australian savanna soils may provide a C sink of between −7.7 and −9.4 Tg CO₂-e per year. This sink estimate would offset potentially 10% of Australian transport related CO₂-e emissions. This CH₄ sink estimate does not include concurrent CH₄ emissions from termite mounds or ephemeral wetlands in Australian savannas.