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Here we describe refined methods for the isolation and detection of a KDEL-tagged, plant-produced recombinant cholera toxin B subunit (CTB) that exhibits unique mucosal wound healing activity. The protein was transiently overexpressed in Nicotiana benthamiana, which generates some C-terminal KDEL truncated molecular species that are deficient in epithelial repair activity. With a new CHT chromatographical method described herein, these product-derived impurities were successfully separated from CTB with the intact KDEL sequence, as confirmed by mass spectrometry. In addition, an immunoassay capable of specifically detecting GM1 ganglioside-binding CTB with intact KDEL sequences was developed. Coupled together, these methods will aid in the quality control of KDEL-attached CTB produced in plant-based manufacturing systems towards a novel topical biotherapeutic for the treatment of acute and chronic mucosal inflammation.This study was designed to understand the dissipation of fipronil, cypermethrin, and tebuconazole in green onions and mustard greens grown in Central Vietnam. A field trial on ca. 400 m2 was implemented for two months with three different cropping regimes (natural plot-fully exposed to natural conditions; bed plot-fenced in by plastic sheets with an open roof; and protected plot-fully covered by plastic sheets) for each vegetable. A first-order kinetic model was successfully employed to interpret the dissipation data of each studied pesticide. A comparing the dissipation rate constants obtained under the same crop and growing regime between chemicals suggested that, among the chemicals examined, fipronil dissipated most readily. LW 6 The pesticides were also proven to be more persistent in crops grown in protected plots than those grown in natural plots. The half-lives for fipronil, cypermethrin, and tebuconazole fluctuated, occurring at 0.4-2.2 days, 2.0-6.0 days, and 0.9-3.3 days, respectively.The baseline sensitivity of Botrytis cinerea to fenpyrazamine was evaluated using 323 isolates collected in Japan prior to its launch. In this study, the isolates were classified as "sensitive" and "low-sensitive" according to their mycelial growth on 10 mg/L fenpyrazamine. However, their EC50 values for the germ-tube elongations from conidia were not significantly different between these two classes. In both a pot test and a field trial, diseases caused by the sensitive and low-sensitive isolates were effectively controlled by fenpyrazamine.Glutathione transferase (GST) is an important class of detoxification enzymes that are vital for defense against various xenobiotics and cellular oxidative stress. Previously, we had reported an unclassified glutathione transferase 2 in Bombyx mori (bmGSTu2) to be responsible for detoxifying diazinon. In this study, we aimed to identify the amino acid residues that constitute a hydrogen-bonding network important for GST activity. Site-directed mutagenesis of bmGSTu2 suggested that residues Asn102, Pro162, and Ser166 contribute to its catalytic activity.Orobanchaceae root parasitic weeds cause serious agricultural damage worldwide. Although numerous studies have been conducted to establish an effective control strategy for the growth and spread of root parasitic weeds, no practical method has been developed so far. Previously, metabolomic analyses were conducted on germinating seeds of a broomrape, Orobanche minor, to find novel targets for its selective control. Interestingly, planteose metabolism was identified as a possible target, and nojirimycin (NJ) selectively inhibited the germination of O. minor by intercepting planteose metabolism, although its precise mode of action was unclear. Here, transcriptome analysis by RNA-Seq was conducted to obtain molecular insight into the effects of NJ on germinating O. minor seeds. Differential gene expression analysis results suggest that NJ alters sugar metabolism and/or signaling, which is required to promote seed germination. This finding will contribute to understanding the effect of NJ and establishing a novel strategy for parasitic weed control.Five species of aquatic fungi and fungus-like organisms were used for toxicity assays with seven fungicides to determine the differences in species sensitivity. A microplate toxicity assay with adenosine triphosphate luminescence detection was used as an efficient and economical high-throughput assay. The obtained toxicity data were standardized based on the species sensitivity distribution method. Species sensitivity differed among the fungicides Rhizophydium brooksianum was most sensitive to hydroxyisoxazole, isoprothiolane, and ferimzone; Chytriomyces hyalinus was most sensitive to tricyclazole; Sporobolomyces roseus was most sensitive to ipconazole; Aphanomyces stellatus was most sensitive to orysastrobin and kasugamycin. Tetracladium setigerum was not the most sensitive species to any of the tested fungicides. The ranges of EC50s to fungal species were lower than to other aquatic organisms (primary producers, invertebrates, and vertebrates) for hydroxyisoxazole, kasugamycin, isoprothiolane, ipconazole, and ferimzone. These results suggest the usefulness of a battery of fungal species to assess the ecological effects of fungicides.The combination of herbicides with different modes of action has been adopted not only to improve weed control but also to increase the environmental sustainability of plant-protection products. In this study, we showed a synergistic effect of the auxin herbicide 2,4-D amine with the PSII-inhibiting herbicide metribuzin to control the global grass weed wild oat (Avena sterilis) population and investigated the underlying mechanisms. Pretreatment with 2,4-D amine did not change the foliar absorption of metribuzin but did increase metribuzin translocation to the roots and new leaves, although enhancement of the metribuzin metabolism was also observed. Considering that the expression level of the target site psbA gene is significantly higher in leaves than in roots, increased metribuzin translocation to new leaves is likely the major cause of the observed synergism, even though enhanced metribuzin metabolism may offset the metribuzin efficacy. This is the first report on the synergistic mechanism between 2,4-D amine and metribuzin in weed control.