Theoretically, the activity of AB-type toxin molecules such as the insecticidal toxin (Cry toxin) from B. thuringiensis, which have one active site and two binding site, is improved in parallel with the binding affinity to its receptor. In this experiment, we tried to devise a method for the directed evolution of Cry toxins to increase the binding affinity to the insect receptor. Using a commercial T7 phage-display system, we expressed Cry1Aa toxin on the phage surface as fusions with the capsid protein 10B. These recombinant phages bound to a cadherin-like protein that is one of the Cry1Aa toxin receptors in the model target insect Bombyx mori. The apparent affinity of Cry1Aa-expressing phage for the receptor was higher than that of Cry1Ab-expressing phage. Phages expressing Cry1Aa were isolated from a mixed suspension of phages expressing Cry1Ab and concentrated by up to 130,000-fold. Finally, random mutations were made in amino acid residues 369-375 in domain 2 of Cry1Aa toxin, the mutant toxins were expressed on phages, and the resulting phage library was screened with cadherin-like protein-coated beads. As a result, phages expressing abnormal or low-affinity mutant toxins were excluded, and phages with high-affinity mutant toxins were selected. These results indicate that a method combining T7 phage display with selection using cadherin-like protein-coated magnetic beads can be used to increase the activity of easily obtained, low-activity Cry toxins from bacteria.

The dissipation behavior of water-extractable pesticides in soils is important when assessing the phytoavailability of pesticides in soils. This process is less understood than pesticide extraction with organic solvents. To elucidate the dissipation behavior of water-extractable pesticides in soils, we conducted an incubation study using 27 pesticides and five Japanese soils. The rate of decrease of the level of pesticides in water extracts was faster in soils than that of total extracts (water extracts and acetone extracts). This suggests that time-dependent sorption contributed to the difference in the dissipation between the pesticides in water and total extracts from soils. Increased apparent sorption coefficients (Kd,app) with time were positively and significantly correlated with Kd,app values of a 0 day incubation [Kd,app(t0)]. This empirical relationship suggests that Kd,app(t0) values can predict the time-dependent increase in Kd,app and the dissipation of water-extractable pesticides in soils.

We analyzed the factors influencing pesticide sorption to soils by testing 17 pesticides with different polarity and chemical structures in 8 soils, including major types in Japan. The soil-water distribution coefficients (K d ) were significantly positively correlated with organic carbon (OC) content. However, the OC-normalized sorption coefficients (K oc ) of many pesticides exhibited high variability among soils. Solid-state 13 C nuclear magnetic resonance (NMR) was performed to elucidate the effect of OC quality on K oc variability. The NMR results indicated that the aromatic carbon content in soil was positively correlated with the K oc values of pesticides. The sorption pattern of pesticides to soils containing abundant aromatic carbon was influenced by the differences in the molecular structures of pesticides, similar to that to activated carbon and graphite. The results indicate the aromatic carbon in soils, particularly black carbon, is an important factor influencing the sorption of pesticides to soils. © Pesticide Science Society of Japan

The relationship between pesticide concentrations in Komatsuna (Brassica rapa var. perviridis) shoots and the extractable concentrations in soils by a sequential soil-extraction method was investigated in 8 pesticides and in 4 soils. Concentrations of many pesticides in Komatsuna shoots showed higher positive correlation with water-extractable than with total-extractable soil concentrations. We also examined the effects of the soil-aging periods (SAPs) between pesticide applications and sowing on pesticide concentrations in Komatsuna shoots. Pesticide concentrations in Komatsuna shoots and soil-water extracts decreased with increasing SAPs. These results imply that estimating the pesticide concentrations in crops based on water-extractable concentrations in soil and setting the appropriate SAPs prevent contamination of crops by pesticide residues in soil. However, the correlation between pesticide concentrations in Komatsuna shoots and water extracts in soils, and the effectiveness of pesticide reduction in Komatsuna shoots by setting long SAPs differed according to the pesticide type. Hence, the applicability of the water-extraction method should be verified for each pesticide.

We performed uptake experiments with 12 different organic chemicals using 16 plant species and determined differences in the ability of plant species to take up and translocate these chemicals. There were differences among the plant species in the shoot and root concentrations of each organic chemical. The root concentration factor values increased with an increasing log of the noctanol-water partition coefficient (log K OW) of organic chemicals. Thus, the concentrations in roots may be predicted to a certain extent because the root concentration factor values were related to the log K OW. The root-to-shoot translocation was related to the log K OW because the shoot-to-root concentration ratio decreased with an increasing log K OW ; however, there was no clear relationship between the shoot concentration factor value and the log K OW , and this differed among plant species.

We cultivated Brassica rapa var. perviridis in soil mixed with four pesticides (fenobucarb, procymidone, flutolanil, and tolclofosmethyl) at different temperatures, day lengths, and soil water contents. We compared plants' uptake and translocation abilities of the pesticides as affected by growth conditions. The root concentration factor (RCF) of pesticides tended to increase with rising temperature; however, but the influence of temperature on the transpiration stream concentration factor (TSCF) differed for each pesticide. The RCFs and TSCFs of pesticides were high for short days. The soil water content had little or no effect on the uptake and translocation of pesticides. These results showed that it is necessary to consider growth conditions, especially the temperature and day length in plant uptake models for these pesticides.

No abstract

The relationships between plant growth stage and pesticide-uptake ability were investigated via cultivation of Brassica rapa L. var. perviridis in soil to which was added four pesticides of relatively high log K OW : fenobucarb, procymidone, flutolanil, and tolclofos-methyl. The root concentrations of pesticides were low in very young seedlings with undeveloped root systems, highest in seedlings with developed root systems, and tended to decrease until the usual harvesting stage. Additionally, the shoot concentrations of tested pesticides showed the same trends as the roots. The pesticide-uptake abilities of roots were lowest in very young seedlings and then constant for seedlings until the harvesting stage. In contrast, the pesticide-translocation abilities from root to shoot were constant regardless of growth stage. The results indicated that changes in shoot concentrations with growth stage were affected by the development of the root system and pesticide-uptake ability of roots.