The present study was conducted to monitor the level of triflumizole residues in fruits (apple and pear) and vegetable (cucumber) samples in order to assess risk posed by the presence of such residues to the consumer. Triflumizole was applied at a recommended dose rate to apple and pear pulps and to a cucumber sample. The samples were collected at harvesting time following several treatments (three and/or four treatments). Triflumizole was extracted with methanol and re-extracted into dichloromethane. The presence of triflumizole was determined by HPLC with UV detection at 238 nm following the cleanup of the extract by open preparative chromatographic column with Florisil. The versatility of this method was evidenced by its excellent linearity (> 0.999) in the concentration range between 0.2 and 4.0 mg/kg. The mean recoveries evaluated from the untreated samples spiked at two different fortification levels. 0.1 and 0.4 mg/kg, and ranged from 87.5${\pm}$0.0 to 93.3${\pm}$2.6 for the tested fruits and vegetable, respectively, and the repeatability (as relative standard deviation) from three repetitive determinations of recoveries were no larger than 6%. The calculated limit of detection was 0.02 mg/kg and the minimum detectable level of 4 ng for triflumizole was easily detected. When triflumizole was sprayed onto the apple trees three times at 50-40-30 and 40-30-21 days prior to harvesting and four times onto the pear trees at 40-30-21-14 days prior to harvesting, the mean residual amounts of 0.05 and 0.06 mg/kg for apples and pears, respectively, were not detected in all of the treatments. When the cucumber sample was fumigated four times at 7, 5, 3 and 1 day prior to harvesting, the mean residual amount was not detectable. Triflumizole can be used safely when sprayed (wettable powder, 30% active ingredient) and fumigated (10%) 4 times at 14 and 1 day prior to harvesting to protect the fruits and vegetable, respectively.
Root-knot disease caused by Meloidogyne incognita is major soil pathogen and cause severe economic damages to vegetable crops. In this study, we selected rhizobacteria for biocontrol of the root-knot nematode, M. incognita, and identified; performed bioassay of the bacterial extracts in cucumber seedlings. The crude extracts of strains CT16 and JC05 out of 180 strains inhibited egg hatching and increased juvenile mortality in vitro assay; based on 16S rRNA sequences analysis, the two strains were identified as Bacillus sp. CT16, and Neobacillus sp. JC05. After extracting the bacterial supernatants by using various organic solvents, n-butanol and n-hexane extracts of strain CT16 and n-butanol extract of strain JC05 showed inhibitory activity of egg hatching depending on concentrations. Subsequently, n-butanol extracts of two strains significantly suppressed formation of egg masses in cucumber seedling. Therefore, these results indicated that strains CT16 and JC05 could be used as potential biocontrol agents against M. incognita.
Pepper mild mosaic virus(PMMoV) and Cucumber mosaic virus (CMV) are important pathogens in various vegetable crops worldwide. We have found that hot water extract of Phellinus linteus mycelium strongly inhibit PMMoV and CMV infection. Based on these results, the inhibitor named as 'PLM-WE1' formulated from extract of Phellinus linteus mycelium was tested for its inhibitory effects on PMMoV and CMV infection to each local lesion host plant (Nicotiana glutinosa: PMMoV, Chenopodium amaranticolor: CMV). Pretreatment effect of PLM-WE1 against infections of each virus (PMMoV and CMV) to local host plant was measured to be 99.2% to PMMoV and 80.3% to CMV, and its permeability effect was measured to be 45.0% to PMMoV and 41.9% to CMV. Duration of inhibitory activity of PLM-WE1 against PMMoV infection on N. glutinosa was maintained for 3 days at 75% inhibition level and CMV infection on C. amaranticolor maintained for 3 days at 62% inhibition level. Inhibitory effects on systemic host plants of PLM-WE1 were measured to be 75~85% to PMMoV and 75% to CMV. Under electron microscope, PMMoV particles were not denatured or aggregated by mixing PLM-WE1. It is suggested that the mode of action of PLM-WE1 differ from that of inactivation due to the aggregation of viruses. The methanol extract of P. linteus mycelium was sequentially partitioned with haxane, ethyl acetate, BuOH and $H_2O$. The $H_2O$ fraction was showed high activity than the other fractions. The active compound was isolated with a partial acid hydrolysis, fractional precipitation with ethanol. The inhibitory effect of the precipitate isolated from 70% ethanol fraction was 99.1% to PMMoV and 88.0% to CMV. The structure of isolated compound was determined by $^1H$-NMR and $^{13}C$-NMR. This compound was identified as a polysaccharide consisting alpha or beta-glucan.
Pepper mild mosaic virus (PMMoV) and cucumber mosaic virus (CMV) are important pathogens in various vegetable crops worldwide. We have found that methanol extracts of Quercus dentate (Oaimyo Oak) gal/nut strongly inhibit PMMoV and CMV infection. Based on this result, the inhibitor named as "KN0912" formulated from the extract of Q. dentate gallnut was tested for its inhibitory effects on PMMoV or CMV infection to each local lesion host plant (Nicotiana glutinosa; PMMoV, Chenopodium amaranticolor; CMV). Pre-treatment effect of KN0912 against infections of each virus to local host plant was measured to be $75.1{\pm}0.5{\sim}97.5{\pm}1.5%$ to PMMoV and $70.6{\pm}2.2{\sim}99.0{\pm}1.0%$ to CMV in 1~10mg/ml conc. and the absorption effect of the antiviral composition of KN0912 to the inside of tobacco leaves tissue, was inhibited by 55.7% to PMMoV and 63.8% to CMV. The persistence of KN0912 treatment was maintained until after the 3 days high inhibitory effect by 98% to PMMoV and by 95.1% to CMV. Inhibitory effects on systemic host plants of KN0912 were measured to be 80~90% to PMMoV and 60~75% to CMV. From the change of morphological characteristics of PMMoV particles under EM, we are tentatively suggested that one mode of action of KN0912 is inactivation due to the destruction of virus particles.
The coat protein (CP) gene of Apple mosaic virus (ApMV), a member of the genus Ilarvirus, was selected for the design of virus-specific primers for amplification and molecular detection of the virus in cultivated apple. A combined assay of reverse transcription and polymerase chain reaction (RT-PCR) was performed with a single pair of ApMV-specific primers and crude nucleic acid extracts from virus-infected apple for rapid detection of the virus. The PCR product was verified by restriction mapping analysis and by sequence determination. The lowest concentration of template viral RNA required for detection was 100 fg. This indicates that the RT-PCR for detection of the virus is a 10$^3$times more sensitive, reproducible and time-saving method than the enzyme-linked immunosorbent assay. The specificity of the primers was verified using other unrelated viral RNAs. No PCR product was observed when Cucumber mosaic virus (Cucumovirus) or a crude extract of healthy apple was used as a template in RT-PCR with the same primers. The PCR product (669 bp) of the CP gene of the virus was cloned into the plasmid vector and result-ant recombinant (pAPCP1) was selected for molecule of apple transformation to breed virus-resistant transgenic apple plants as the next step. This method can be useful for early stage screening of in vitro plantlet and genetic resources of resistant cultivar of apple plants.
We selected eight resistant cultivars including 'Heukryongsamcheok', 'Heukgeumsolsamcheok' and 'Gangryeoksamcheok' showing high suppressive effect against cucumber downy mildew (CDM) through plastic film house and field trials in spring and autumn season in 2015. Of them, these three varieties, 'Heukryongsamcheok', 'Heukgeumsolsamcheok' and 'Gangryeoksamcheok' were used to evaluated suppressive effect against CDM by comparing disease severity (area under disease development progress curve, AUDPC) with those of three susceptible varieties under plastic film house condition. AUDPC of three resistant cultivars was in the range of 10.9 to 23.6, meanwhile those of three susceptible cultivars was in the range of 286.7 to 290.3. Consequently, we confirmed that cultivation of the selected resistant cultivars can reduce CDM in vivo. When disease severity (diseased leaf area) was investigated on a resistant cultivar ('Heukryongsamcheok') sprayed without any organic materials and susceptible cultivars sprayed single-, two times- and three times with Bordeaux mixture, sulfur-loess mixture and $Kocide^R$ (copper hydroxide), respectively, it was the lowest in the resistant cultivar plot compared to susceptible cultivar plots sprayed with any organic materials. In addition, we evaluated control effect of the selected resistant cultivar by comparing that of organic materials including lime sulfur mixture, Bordeaux mixture and garlic extract. As a result, Bordeaux mixture showed the highest control effect against CDM. AUDPC of Bordeaux mixture, resistant cultivar, lime sulfur, garlic extract and untreated control was 3.9, 10.6, 95.6, 24.9, and 258.7, respectively. Based on the above-mentioned results, we think that the resistant cultivars and Bordeaux mixture can be effectively used to control CDM as one of control measures under the farmhouse condition.
Yun, Young Beom;Byeon, Ri Na;Jang, Se Ji;Hyun, Kyu Hwan;Shin, Dong Young;Kim, Sang Su;Kim, Do Ik;Kwon, Oh Do;Kuk, Yong In
Weed & Turfgrass Science
/
v.2
no.3
/
pp.242-247
/
2013
Currently, methods for controlling weeds in organically produced crops have not been as effective as conventional methods. This research was carried out to determine the herbicidal effects of leaf, stem, fruit, root extracts of Trichosanthes kirilowii. The extraction methods used were water, boiling water and ethanol. The characteristics of potential herbicidal components among extraction methods were investigated by using the following solvent fractions: hexane, chloroform, ethyl acetate, butanol, and water. Generally, water extracts provided the best on inhibition of germination rate, plant height, and root length in cucumber and barley. Specifically, extractions made from fruit parts of T. kirilowii provided the greatest inhibition effect on plant growth in cucumber and barely. Inhibition of germination rate, plant height, and root length in cucumber and barley in solvent fractions was the best in water fractions, but there were no significant differences among the other fractions. Digitaria siliaris and Solanum nigrum were controlled 80-100% by 5% extractions of water fraction. However, there were no herbicidal effects from foliar treatment in cucumber, barley, black nightshade, and henry crabgrass by 5% extractions of the water fraction. These results show that extractions of T. kirilowii can be used for controlling some weeds in organically produced crops.
Journal of the Korean Society of Food Science and Nutrition
/
v.12
no.4
/
pp.342-349
/
1983
This study was undertaken to ascertain food and nutritional evaluating data on the processing of fermented sea cucumber (Stichopus japonicus) entrails. In the experiment, the crude proteolytic enzyme from the entrails tissue of raw and fermented sea cucumber during the days of ripening was extracted. The optimal activity condition for the crude enzyme and the compositional changes of amino acid of the protein and free amino acid in the raw and fermented sample were also investigated. 1. Less than three kinds of proteolytic enzymes that each enzyme has optimal activity condition at pH 3.1 $50^{\circ}C$(A-enzyme), pH 5.7 $50^{\circ}C$(B-enzyme) and pH 7.7 $45^{\circ}C$(C-enzyme), respectively were believed to be exist in the entrails tissue of sea cucumber. 2. A-enzyme and C-enzyme were strongly inhibited with the increase of the salt concentration, and B-enzyme was activated at the 1% salt concentration and was inhibited above the 5% salt concentration. 3. The result of the effect of several salt ions on the proteolytic activity showed that A-enzyme was slightly inhibited in the presence of all salt ions added, B-enzyme was activated in the presence of the all salt ions except $Cu^{2+}$ and C-enzyme was activated in the presence of $Ca^{2+}$ and $Mn^{2+}$, and inhibited by $Cu^{2+}$, $Co^{2+}$ and $Mg^{2+}$. 4. When the effects of the ripening days on the proteolytic activity of the crude enzymes were analysed, the activity of the A-enzyme was slightly weakened with the lapse of the fermentation days, whereas the B-enzyme was not influenced by the fermentation days. 5. In the analysis of amino acid composition of the protein of the samples, the 8 days fermented sea cucumber entrails showed the diminution of all kinds of amino acid. Apparently diminished amino acids were arginine, alanine, glutamic acid, glycine, serine, valine, threonine and lysine etc., and methionine, histidine and isoleucine were slightly decreased. 6. In the analysis of free amino acid composition of the 8 days fermented sample, glutamic acid, aspartic acid, leucine and lysine were rich, while histidine, methionine, proline and tyrosine were poor. The most of free amino acids were increased during the fermentation procedure and especially in lysine, histidine, threonine, glutamic acid, methionine, valine and leucine.
Park Sung-Min;Lee Jun-Seuk;Park Chi-Duck;Lee Jung-Hun;Jung Hyuck-Jun;Yu Tae-Shick
KSBB Journal
/
v.21
no.1
s.96
/
pp.42-48
/
2006
Bacillus subtilis KMU-13 was isolated from the Lillehammer forest soils at Norway and shown a strong antifungal activity on cucumber scab, Cladosporium cucumerinum KACC 40576. B. subtilis KMU-13 produced a maximum level of antifungal substance under incubation aerobically at $30^{\circ}C$, 180 rpm for 48 hours in LB broth containing 0.5% maltose and 0.5% bactopeptone and initial pH adjusted to 6.0. Butanol extract of cultured broth was confirmed inhibitory zone by plate assay and Rf 0.64 value substance by thin layer chromatography (TLC) represented high antifungal activity against C. cucumerinum KACC 40576 and also shown fungal growth inhibitory activity against Botytis cinerea KACC 40573, C. gloeosporioides KACC 40804, D. byoniae KACC 40669, F. oxysporum KACC 40037, F. oxysporum KACC 40052, F. oxysporum f. sp. radicis-lycopersici KACC 40537, F. oxysporum KACC 40902, M. cannonballus KACC 40940, P. cambivora KACC 40160, R. soiani AG-1 KACC 40101, R. solani AG-4 KACC 40142, and S. scleotiorum KACC by agar diffusion method.
Seo, Tae-Jin;Yang, Soo-Jeong;Lee, Bong-Choon;Kim, Kang-Min;Lee, Kui-Jae;Ju, Ho-Jong
Korean Journal of Organic Agriculture
/
v.24
no.3
/
pp.571-582
/
2016
Bacterial fruit blotch (BFB) is one of most important diseases in Cucurbitaceae due to infection of Acidovorax citrulli, causing huge economic losses damage worldwide. This seedborn disease spread rapidly at period of high temperature and humidity. The eco-friendly farming is getting popular. So far there was no effective agent to control BFB in eco-friendly farming. Therefore, effect of the material based on chinese nut-gall extract with antibacterial activity against BFB to was tested against A. citrulli. Different hosts showed various symptoms of BFB. Liquid formulation among exhibited most effective anti-bacterial activity on A. citrulli. Pot experiment in greenhouse showed the potential as an control agent of BFB in cucurbits. The treatment of material based on chinese nut-gall extract showed the positive effect on survival of the watermelon seedling and on the length of the cucumber seedling treated with A. citrulli. We cautiously conclude that the material based on chinese nut-gall extract used in this study may be good agents against major diseases of cucurbits in the future even though it is require to be tested with more study on field test.
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