Establishment of import tolerance for the insecticide thiacloprid in strawberry.

To promote exports, import tolerance (IT) of thiacloprid in strawberry was proposed using the Organization for Economic Cooperation and Development (OECD) maximum residue limit (MRL) calculator after conducting three different field trials. The pre-harvest interval of residual pattern and degradation dynamics of thiacloprid in strawberry were determined using ultra-performance liquid chromatography-tandem mass spectrometry. Samples were extracted with acetonitrile and a mixture of salts and dilution was performed for purification. A six-point matrix-matched calibration curve was constructed which provided excellent linearity with coefficient of determination (R2 ) of 0.9998 or more. Detection and quantification limits were 0.003 and 0.01 mg/kg, respectively. The method was validated in quintuplicate at three different concentrations, which resulted in acceptable recovery ranging from 80.86% to 101.71% with relative standard deviation of 6.50 or less among the three field sites. The developed method was applied to the field-treated sample harvested at different intervals. In the pre-harvest interval trial, the amount of thiacloprid residues ranged from 0.24 to 0.70 mg/kg in field site 1 (Nonsan), 0.16 to 0.50 mg/kg in field site 2 (Sunchang), and 0.36 to 0.50 mg/kg in field site 3 (Sacheon). By contrast, in the degradation trial, the observed residues were 0.03-0.81 mg/kg in field site 1 and 0.02-0.48 mg/kg in field site 2. Consequently, the IT of thiacloprid in strawberry using the OECD MRL calculator was proposed as 2 mg/kg, which is exactly the same as the MRL established by the Republic of Korea. In conclusion, the residue study proposes 2.0 mg/kg as the MRL of thiacloprid in strawberries.

Developmental toxicity of 3-phenoxybenzoic acid (3-PBA) and endosulfan sulfate derived from insecticidal active ingredients: Abnormal heart formation by 3-PBA in zebrafish embryos.

Pyrethroid and organochlorine insecticides are enormously used to control agricultural and indoor insect pests. The metabolites of pyrethroid and endosulfan were used to evaluate environmental toxicities using a representative animal model, zebrafish (Danio rerio) embryos in this study. The LC50 values in 3-phenoxy benzoic acid (3-PBA) and endosulfan sulfate (ES) were 1461 µg/L and 1459 µg/L, respectively. At the concentration of 2000 µg/L, spine curvature was observed in the ES-treated embryos. ES showed seizure-like events with an EC50 value of 354 µg/L. At the concentration of 1000 µg/L, the pericardial edema was observed in 3-PBA-treated embryos. The inhibition of heart development and the reduction of beating rates were observed in Tg(cmlc2:EGFP) embryos after the exposure to 3-PBA. Down-regulation of the vmhc gene coding ventricular myosin during heart development was significantly found in 3-PBA-treated embryos at 48 hpf, but recovered afterward. It indicates that ventricular malformation occurred at the initial stage of 3-PBA exposure. Considered together, both 3-PBA and ES need public concerns with periodic monitoring of these metabolites in households and agricultural areas to prevent humans and environmental organisms from their unexpected attacks.

Rice Senescence-Induced Receptor-Like Kinase (OsSRLK) Is Involved in Phytohormone-Mediated Chlorophyll Degradation.

Chlorophyll breakdown is a vital catabolic process of leaf senescence as it allows the recycling of nitrogen and other nutrients. In the present study, we isolated rice senescence-induced receptor-like kinase (OsSRLK), whose transcription was upregulated in senescing rice leaves. The detached leaves of ossrlk mutant (ossrlk) contained more green pigment than those of the wild type (WT) during dark-induced senescence (DIS). HPLC and immunoblot assay revealed that degradation of chlorophyll and photosystem II proteins was repressed in ossrlk during DIS. Furthermore, ultrastructural analysis revealed that ossrlk leaves maintained the chloroplast structure with intact grana stacks during dark incubation; however, the retained green color and preserved chloroplast structures of ossrlk did not enhance the photosynthetic competence during age-dependent senescence in autumn. In ossrlk, the panicles per plant was increased and the spikelets per panicle were reduced, resulting in similar grain productivity between WT and ossrlk. By transcriptome analysis using RNA sequencing, genes related to phytohormone, senescence, and chlorophyll biogenesis were significantly altered in ossrlk compared to those in WT during DIS. Collectively, our findings indicate that OsSRLK may degrade chlorophyll by participating in a phytohormone-mediated pathway.

Whole body dosimetry and risk assessment of agricultural operator exposure to the fungicide kresoxim-methyl in apple orchards.

This study examined dermal and inhalation exposure of agricultural operators to kresoxim-methyl during pesticide mixing/loading and speed sprayer application (10 replicates, each of 3000 L of spray suspension) in an apple orchard and performed risk assessment. For the whole body dosimetry (WBD) exposure protocol, outer clothing, inner clothing, gauze, and nitrile gloves were examined to measure dermal exposure. In contrast, an IOM (institute of occupational medicine) sampler with a glass fiber filter was used to measure inhalation exposure. Analytical method accuracy in the exposure matrices was evaluated by a field recovery study. The dermal and inhalation exposure amounts for mixing/loading were 9.7 mg [0.002% of the total mixed/loaded active ingredient (a.i.)] and 1.2 µg (1.7 × 10-6% of the total mixed/loaded a.i.), respectively. The body parts more exposed were the forearms (35.5%), chest & stomach (30.2%), and hands (17.9%). During application, the dermal and inhalation exposure amounts were 66.5 mg (0.009% of the total applied a.i) and 34.8 µg (4.6 × 10-5% of the total applied a.i.), respectively. The shins (18.5%) and chest & stomach (16.0%) were exposed to higher proportion of pesticide, followed by the thighs (15.8%) and back (14.7%). Comparing the exposure pattern as assessed by the WBD method in the present study with the patch method as in our previous study, the ADE (actual dermal exposure) as measured by the WBD method was 25 times less than that measured by the patch method. The daily exposure amounts of ADE and AIE (actual inhalation exposure) for mixing/loading were 711.8 µg/day and 4.3 µg/day, respectively, whereas the amounts of ADE and AIE for application were 1825.8 µg/day and 116.1 µg/day. In risk assessment of the mixing/loading and application scenarios, the AOEL (acceptable operator exposure level) of kresoxim-methyl was used as the reference dose to show that the RI (risk index) was much lower than 1, indicating that agricultural operators are at low risk of exposure to kresoxim-methyl.

Larvicidal and Nematicidal Activities of 3-Acylbarbituric Acid Analogues against Asian Tiger Mosquito, Aedes albopictus, and Pine Wood Nematode, Bursaphelenchus xylophilus.

Widespread concern for the occurrence of resistant strains, along with the avoidance of the use of highly toxic insecticides and their wide environmental dispersal, highlights the need for the development of new and safer pest control agents. Natural products provide inspiration for new chemical entities with biological activities, and their analogues are good lead compounds for the development of new pest control agents. For this purpose, we evaluated the larvicidal and nematicidal activities of 48 3-acylbarbituric acid analogues against the Asian tiger mosquito, Aedes albopictus and the pine wood nematode, Bursaphelenchus xylophilus, organisms of increasing global concern. Among the 48 3-acylbarbituric acid analogues, four compounds-10, 14d, 14g and 19b-showed >90% larvicidal activity against Ae. albopictus at 10 µg/mL concentration, and one (compound 10) showed the strongest larvicidal activity against Ae. albopictus, with a LC50 value of 0.22 µg/mL. Only compound 18 showed strong nematicidal activity against pine wood nematode. Most active compounds possessed similar physicochemical properties; thus, actives typically had ClogP values of around 1.40-1.50 and rel-PSA values of 16-17% and these similar cheminformatic characteristics reflect their similar structure. This study indicates that active 3-acylbarbituric acids analogues have potential as lead compounds for developing novel mosquito control agents.

Enantioselective effect of trifloxystrobin in early-stage zebrafish (Danio rerio) embryos: Cardiac abnormalities impacted by E,E-trifloxystrobin enantiomer.

Trifloxystrobin (TFS) is one of the extensively used strobilurin fungicides, which is composed of four enantiomers and its active form is E,E-TFS. In this study, we assess the acute toxicity of four enantiomers, E,E-, E,Z-, Z,E-, and Z,Z-TFS in zebrafish (Danio rerio) embryos. Among the four enantiomers, only E,E-TFS was found to be acutely toxic, with an estimated LC50 value of 0.68 mg/L. Treatment with E,E-TFS resulted in various phenotypic changes in the embryos, including pericardial and yolk-sac edema, spine curvature, and blood pooling. And it shortened the whole body length in the treated embryos by increasing the total intersegmental vessel numbers using a Tg(fli1a:EGFP) zebrafish line. Further study using Tg(cmlc2:EGFP) zebrafish line revealed that E,E-TFS treatment was associated with cardiac malformations, a failure of heart function, and a lowered heartbeat rate at the concentration of 0.25 mg/L. Also, the differential gene expression analysis identified significant down-regulation of vmhc and cacna1c genes encoding ventricular myosin heavy chain and calcium voltage-gated channel subunit alpha 1C, which are crucial for heart development. These results suggest the need for regular monitoring of E,E-TFS enantiomers after field application and further research into their potential chronic effects on environmental organisms.

Growth-inhibiting effects of Paeonia lactiflora root steam distillate constituents and structurally related compounds on human intestinal bacteria.

The growth-inhibiting activities of Paeonia lactiflora (Paeoniaceae) root steam distillate constituents and structurally related compounds against nine harmful intestinal bacteria and eight lactic acid-producing bacteria were compared with those of two antibiotics, amoxicillin and tetracycline. Thymol, α-terpinolene, (-)-perilla alcohol and (1R)-(-)-myrtenol exhibited high to extremely high levels of growth inhibition of all the harmful bacteria, whereas thymol and α-terpinolene (except for Lactobacillus casei ATCC 393) inhibited the growth of all the beneficial bacteria (MIC, both 0.08-0.62 mg mL(-1)). Tetracycline and amoxicillin exhibited extremely high level of growth inhibition of all the test bacteria (MIC, <0.00002-0.001 mg mL(-1)). 1,8-Cineole, geraniol, (-)-borneol, (1S,2S,5S)-(-)-myrtanol, nerol, (S)-(-)-ß-citronellol and (±)-lavandulol also exhibited inhibitory activity but with differing specificity and levels of activity. Structure-activity relationship indicates that structural characteristics, such as geometric isomerism, degrees of saturation, types of functional groups and types of carbon skeleton, appear to play a role in determining the growth-inhibiting activity of monoterpenoids. Global efforts to reduce the level of antibiotics justify further studies on naturally occurring P. lactiflora root-derived materials as potential preventive agents against various diseases caused by harmful intestinal bacteria such as clostridia.

Development of Simultaneous Analytical Method for Imidazolinone Herbicides from Livestock Products by UHPLC-MSMS.

A simultaneous analytical method, which used LC/MSMS for imidazolinone herbicides from livestock products (egg, milk, beef, pork, and chicken) for monitoring, was developed with a QuEChERS preparation. A weighed sample (5 g) in a 50 mL conical tube was added to 0.1 M potassium phosphate dibasic solution (5 mL) and shaken for 10 min. After shaking, 0.5 mL of 6 N HCl and 5 mL of acetonitrile were added, and this solution was shaken for 10 min. Additionally, QuEChERS extraction salts (original method, 4 g MgSO4, 1 g NaCl) were added to the sample in a 50 mL conical tube. The mixture was strongly shaken for 1 min and centrifuged at 3000× g for 10 min. The acetonitrile layer was purified with dSPE (150 mg MgSO4, 25 mg C18) and was centrifuged at 13,000× g for 5 min. The supernatant was filtered with a membrane filter (pore size: 0.2 µm) before analysis. The ME (%, matrix effect) range for almost all analytes was −6.56 to 7.11%. MLOD (method limit of detection) and MLOQ (method limit of quantitative) values were calculated by the S/N ratio. MLOQs were 0.01 mg/kg. The linear correlation coefficients (R2) were >0.99 with the range of 0.5~25 µg/kg for all of the imidazolinone herbicides. The recoveries (of imidazolinone herbicides) were in the range of 76.1~110.6% (0.01 mg/kg level), 89.2~97.1% (0.1 mg/kg level), and 94.4~104.4% (0.5 mg/kg level). These are within the validation criteria (to recover 70−120% with RSD <20%). The method demonstrated the simple, rapid, high throughput screening and quantitative analysis of imidazolinone herbicide residues for monitoring in livestock products.

Optimization of a Multi-Residue Analytical Method during Determination of Pesticides in Meat Products by GC-MS/MS.

In this study, a multi-residue analysis was developed for 32 compounds, including pesticides and metabolites, in five meat products using gas chromatography-tandem mass spectrometry (GC-MS/MS). The validation of the developed analytical method was also evaluated in accordance with Codex Alimentarius guidelines. Aminopropyl (NH2), C18, and florisil solid phase extraction (SPE) cartridges were used to evaluate and optimize the cleanup procedure of the tested samples prior to GC-MS/MS analysis. Based on the analytical performance, the C18 SPE cartridge was deemed to be the most suitable among the examined SPE cartridges. The optimized method demonstrated that 29 out of 32 tested compounds acquired good linearity (R2 ≥ 0.99), and 25 tested compounds displayed the method limit of quantification (MLOQ) ≤ 0.01 mg/kg. Out of the 32 tested compounds, only 21 compounds met the acceptable analytical criteria for the lard and tallow samples, compared to 27 compounds in the beef, pork, and chicken samples that falls within the acceptable standards for recovery (70-120%) and analytical precision (relative standard deviation RSD ≤ 20%). The average matrix effect was widely varied (20.1-64.8%) in the studied meat samples that were affected by either ion enhancement or suppression. In particular, in the lard sample, 13 compounds showed poor recovery and analytical precision due to ion suppression. Thus, the matrix effect (ME) was considered a critical factor during multi-residue pesticide analysis in different meat products. In conclusion, this developed analytical method can be used as a routine monitoring system for residual pesticide analysis in livestock products with acceptable analytical standards. Further meticulous analytical studies should be optimized and validated for multi-residue pesticide analysis in diversified meat products.

Ecofriendly shiitake authentication using bulk and amino acid-specific stable isotope models.

Accurate authentication of ecofriendly shiitake, such as organic and pesticide-free shiitake, is required to improve food safety and to increase reliability of the national agrofood certification system; however, this is a challenging task. Therefore, this study examined the feasibility of bulk and compound-specific isotope analyses to discriminate ecofriendly shiitake against conventional counterparts. Using the compound-specific isotope model, the classification accuracy was greater (100%) than that of the bulk isotope model (74.5%) for each original sample set. In the compound-specific model, a cutting score of -4.42 discriminated organic shiitake from pesticide-free shiitake and a cutting score of 4.87 discriminated organic shiitake from conventional shiitake. The isotope fractionation trend was less influenced by shiitake type and the amino acid synthetic pathway. Thus, the compound-specific isotope model of amino acids may be a good complementary authentication tool to overcome the limitations of bulk stable isotopes or a pesticide residue test.

Molecular evidence for suppression of swimming behavior and reproduction in the estuarine rotifer Brachionus koreanus in response to COVID-19 disinfectants.

The increased use of disinfectants due to the spread of the novel coronavirus infection (e.g. COVID-19) has caused burden in the environment but knowledge on its ecotoxicological impact on the estuary environment is limited. Here we report in vivo and molecular endpoints that we used to assess the effects of chloroxylenol (PCMX) and benzalkonium chloride (BAC), which are ingredients in liquid handwash, dish soap products, and sanitizers used by consumers and healthcare workers on the estuarine rotifer Brachionus koreanus. PCMX and BAC significantly affected the life table parameters of B. koreanus. These chemicals modulated the activities of antioxidant enzymes such as superoxide dismutase and catalase and increased reactive oxygen species even at low concentrations. Also, PCMX and BAC caused alterations in the swimming speed and rotation rate of B. koreanus. Furthermore, an RNA-seq-based ingenuity pathway analysis showed that PCMX affected several signaling pathways, allowing us to predict that a low concentration of PCMX will have deleterious effects on B. koreanus. The neurotoxic and mitochondrial dysfunction event scenario induced by PCMX reflects the underlying molecular mechanisms by which PCMX produces outcomes deleterious to aquatic organisms.

Antioxidant/anti-inflammatory activities and total phenolic content of extracts obtained from plants grown in Vietnam.

BACKGROUND: Medicinal plants have been used to treat various diseases since ancient times. Their specific activities, such as antioxidant, anti-inflammatory and anti-cancer, have been studied intensively. In particular, plants grown in Vietnam have attracted considerable attention among food chemists as ideal sources of natural medicinal chemicals. RESULTS: The methanol extracts from three edible Vietnamese-grown plants, Tram, Voi and Gac, tested with the DPPH assay showed antioxidant activities of 91.7 ± 0.4, 63.4 ± 0.7 and 3.7 ± 0.1% respectively. The malonaldehyde/gas chromatography assay also revealed strong antioxidant activity in Tram and Voi at a level of 25 µg mL(-1) (95.5 ± 0.3 and 78.5 ± 1.4% respectively). These results were confirmed by the thiobarbituric acid assay. The antioxidant activities correlated positively with the level of total phenolics in all plants. Tram exhibited dose response-related lipoxygenase-inhibitory activity, with values of 74.2 ± 3.1% at 5 µg mL(-1) , 62.0 ± 0% at 0.5 µg mL(-1) and 3.0 ± 1.5% at 0.05 µg mL(-1) . Conversely, Voi and Gac showed negative anti-lipoxygenase activity. CONCLUSION: The antioxidant/anti-inflammatory activities and total phenolic contents of the three edible plants grown in Vietnam revealed that they are good sources of supplements for human health.

Molecular mechanisms of developmental toxicities of azoxystrobin and pyraclostrobin toward zebrafish (Danio rerio) embryos: Visualization of abnormal development using two transgenic lines.

Azoxystrobin (AZ) and pyraclostrobin (PY) are strobilurin fungicides that inhibit fungal mitochondrial respiration. In this study, a representative model, zebrafish (Danio rerio), was used as a test species for acute and developmental toxicity. Survival and malformation rates were observed only PY-treated embryos, with an LC50 value of 77.75 ppb accompanied by a dramatic decrease in hatching rate, while AZ did not show great mortality. Morphological changes were observed in PY-treated embryos with the occurrence of pericadial edema at 25 ppb. A delay in growth was observed after treatment with pyraclostrobin at 50 ppb. Use of genetically engineered Tg(cmlc:EGFP) allowed fluorescence observation during heart development. PY interfered with normal heart development via upregulation of the nppa gene responsible for the expression of natriuretic peptides. Heart function was dramatically reduced as indicated by reduced heart rates. Increased expression of the nppa gene was also seen in AZ-treated embryos. The expression level of cyp24a1 was also up-regulated, while ugt1a1 and sult1st6 were down-regulated after treatment of zebrafish embryos with AZ or PY. Overall, strobilurin fungicides might inhibit normal heart formation and function within the range of concentrations tested.

Structural changes in mulberry (Morus Microphylla. Buckl) and chokeberry (Aronia melanocarpa) anthocyanins during simulated in vitro human digestion.

Mulberry and chokeberry are rich sources of anthocyanins. In this study, the effect of the anthocyanin composition on the anthocyanin profile changes during in vitro digestion (mimicking the physiological conditions) was investigated by UHPLC-(ESI)-qTOF and UHPLC-(ESI)-QqQ. The antioxidant activity before and after in vitro digestion was elucidated. Cyanidin-3-O-glucoside and cyanidin-3-O-galactoside were dominant in mulberry and chokeberry, respectively. Moreover, the loss of cyanidin-3-O-galactoside in the chokeberry extract after digestion was greater than that of cyanidin-3-O-glucoside in the mulberry extract. After digestion, phenolic acids including protocatechuic acid and various cyanidin conjugates were newly formed because of decomposition and changes in the cyanidin-glycosides. The phenolic acid and cyanidin conjugate levels varied depending on the cyanidin glycoside sources in the colonic fraction. Finally, antioxidant activity before and after digestion was higher in the chokeberry extract than in the mulberry extract. Moreover, this activity continuously decreased until intestinal digestion but increased in the colonic fraction.

Quantification of Morpholine in Peel and Pulp of Apples and Oranges by Gas Chromatography-Mass Spectrometry.

Morpholine salts of fatty acids have been used in wax coatings on the surfaces of fruit and vegetable commodities in China and the United States, etc. However, morpholine usage was prohibited in many other countries because of safety concerns. We optimized analytical methods to determine morpholine in the peel and pulp of fruits and vegetables by gas chromatography-mass spectrometry (GC-MS). This morpholine analysis method was applied to real samples of apples, citrus fruits, and vegetables from Korea, China, and the U.S. The method was validated using apple and citrus fruit peels and pulp. The method detection limit (MDL) was 1.3-3.3 µg/kg. The recovery rates of morpholine were 88.6-107.2% over a fortified level of 10-400 µg/kg. Intra-day and inter-day precisions were 1.4-9.4% and 1.5-2.8%, respectively. The morpholine concentrations were n.d. (not detected)-11.19 and n.d. (not detected)-12.82 µg/kg in apple and citrus peels, respectively. Morpholine was not detected in citrus or apple pulp samples or in vegetable samples.

Toxicity of dietary selenomethionine in juvenile steelhead trout, Oncorhynchus mykiss: tissue burden, growth performance, body composition, hematological parameters, and liver histopathology.

The steelhead trout (Oncorhynchus mykiss) is the species most at risk from selenium (Se) exposure in the San Francisco Bay Delta (SFBD). However, although steelhead trout are usually exposed to environmental Se in the juvenile stage, data to test their sensitivity to excess Se, especially its organic form, in the juvenile stage are scarce. Therefore, the objective of the current study was to assess the sensitivity of juvenile steelhead trout to ecologically relevant forms of Se using integrated sensitive endpoints. Fish (mean weight: 22.3 g) were fed one of five diets containing 1.1 (control), 8.8, 15.4, 30.8, and 61.6 µg Se/g diet dw (Se1.1, Se8.8, Se15.4, Se30.8, and Se61.6, respectively) in the form of selenomethionine for 4 weeks. After 4 weeks, Se significantly accumulated in a dose-dependent manner in all tissues at different rates. The growth rate and plasma cholesterol were significantly depressed in fish fed diets containing Se30.8 and above. Hematological parameters and mortality were significantly elevated in fish fed the Se61.6 diet. Marked histopathological alterations were observed in fish fed the Se8.8 diet (the lowest observed effect concentration, LOEC) and above. The current results suggest that the steelhead trout is more sensitive to excess Se than nonanadromous rainbow trout used in previous studies because of its lower LOEC despite the use of selenomethionine and the shorter experimental duration. Additionally, it should be noted that the current Se levels found in the SFBD are already a threat to the threatened population of steelhead trout on the central California coast.

Identification and characterization of potent CYP2B6 inhibitors in Woohwangcheongsimwon suspension, an herbal preparation used in the treatment and prevention of apoplexy in Korea and China.

Woohwangcheongsimwon is a traditional medicine for treating hypertension, arteriosclerosis, coma, and stroke in China and Korea. To assess potential interactions of herb and drug metabolism, commercially available Woohwangcheongsimwon suspensions were examined for their potential to inhibit the activity of nine human cytochrome P450 enzymes. The Woohwangcheongsimwon suspensions showed strong inhibition of CYP2B6 activity. To identify individual constituents with inhibitory activity, the suspension was partitioned using hexane, ethyl acetate, and dichloromethane, and each fraction was tested for its inhibitory effect on CYP2B6-catalyzed bupropion hydroxylation. The hexane fraction possessed inhibitory activity, and gas chromatography/mass spectrometry analysis identified borneol and isoborneol as major constituents of the hexane fraction. These two terpenoids moderately inhibited CYP2B6-catalyzed bupropion hydroxylase activity in a competitive manner, with K(i) values of 9.5 and 5.9 microM, respectively, as well as efavirenz 8-hydroxylase activity, with K(i) values of 22 and 26 microM, respectively. Additionally, reconstituted mixtures of borneol and isoborneol, at the same concentrations as in the Woohwangcheongsimwon suspension, had comparable potency in inhibiting bupropion hydroxylation. These in vitro data indicate that Woohwangcheongsimwon preparations contain constituents that can potently inhibit the activity of CYP2B6 and suggest that these preparations should be examined for potential pharmacokinetic drug interactions in vivo.

Dual role of SND1 facilitates efficient communication between abiotic stress signalling and normal growth in Arabidopsis.

Certain plant cells synthesize secondary cell walls besides primary cell walls. This biosynthesis is strictly controlled by an array of transcription factors. Here, we show that SND1, a regulator of cell-wall biosynthesis, regulates abscisic acid (ABA) biosynthesis to ensure optimal plant growth. In Arabidopsis, the lack of SND1 and its homolog NST1 leads to the deficiency of secondary cell walls, preventing snd1nst1 double mutant seedlings from growing upright. Compared to wild type seedlings, the snd1 knockout mutant seedlings accumulated less anthocyanin and exhibited low tolerance to salt stress. Compared to wild type seedlings, the snd1 knockout seedlings were more sensitive to salt stress. Although SND1 can bind to the promoter of Myb46, we observed that SND1 binds directly to the promoter of the ABI4 gene, thereby reducing ABA levels under normal growth conditions. Thus, plants adjust secondary cell wall thickening and growth via SND1. SND1 has a dual function: it activates the Myb46 pathway, fostering lignin biosynthesis to produce sufficient cell wall components for growth, while maintaining a low ABA concentration, as it inhibits growth. This dual function of SND1 may help plants modulate their growth efficiently.

Overexpression of Arabidopsis homogentisate phytyltransferase or tocopherol cyclase elevates vitamin E content by increasing gamma-tocopherol level in lettuce (Lactuca sativa L.).

Tocopherols, essential components of the human diet, are synthesized exclusively by photosynthetic organisms. To increase tocopherol content by increasing total flux to the tocopherol biosynthetic pathway, genes encoding Arabidopsis homogentisate phytyltransferase (HPT/V-TE2) and tocopherol cyclase (TC/VTE1) were constitutively overexpressed in lettuce (Lactuca sativa L.). Total tocopherol content of the transgenic plants overexpressing either of the genes was increased by more than 2-fold mainly due to an increase in gamma-tocopherol. However, chlorophyll content in the HPT/VTE2 and TC/VTE1 transgenic lines decreased by up to 20% and increased by up to 35%, respectively (P < 0.01). These results demonstrate that manipulation of the tocopherol biosynthetic pathway can increase or decrease chlorophyll content depending on the gene introduced.

Hapten syntheses and antibody generation for a new herbicide, metamifop.

To develop a competitive indirect enzyme-linked immunosorbent assay for metamifop, a new aryloxyphenoxypropionic acid herbicide, three structurally related haptens were synthesized. Hapten conjugates to keyhole limpet hemocyanin and bovine serum albumin were used as immunogens and plate-coating antigens, respectively. Various sets of polyclonal antibodies from rabbits and the coating antigens were screened for the assay in simple homologous and heterologous ELISA formats. A selected heterologous ELISA was optimized to show an average IC50 value as low as 20.1 ng/mL, detection ranges of 1.0-350 ng/mL, and a lowest detection limit of 0.1 ng/mL. The cross-reactivities of other aryloxyphenoxypropionic acid herbicides to the antibodies were less than 0.5% in the assays except fenoxaprop-P and fenoxaprop-P ethyl, having a diaryl ether group identical to that of metamifop. Molecular modeling studies revealed that the physicochemical properties of the diaryl ether group are the most important determinants of sensitivity and selectivity. The results strongly indicate that the selected set of ELISA is a highly sensitive and convenient tool for detecting metamifop.