• The Korean Journal of Pesticide Science
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• v.20 no.1
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• pp.56-65
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• 2016

Pine wood nematode, Bursaphlenchus xylophilus is a fatal damaging pest, infecting pinewood trees and is one of the serious forest pest in Korea. Trunk injection is one of the most effective method to protect the live pine trees from nematode infection. The aim of this research was to reveal the efficacy of the nematicidal materials from 46 Kyrgyzstan, 88 Vietnam plant materials and 21 active materials (${\alpha}$-terpinene, ${\alpha}$-pinene, ${\beta}$-thujaplicin, cinnamaldehyde, eugenol, emodin, geraniol, limonone, methyl palmitate, matrine, myrcene, methyl gallate, nicotine, quassin, resveratrol, rotenone, thymol, thujaplicin, terthiophene, tuberstemonine, ${\gamma}$-terpinene) isolated from plants for using as trunk injection agents. Eclipta prostrata extract was highly effective against pine wood nematode in plant extracts and terthiophene has the best nematicidal activity from active materials isolated from plants. Foliar spray of terthiophene on Japanese black pine (Pinus thunbergii) tree reduced proliferation of inoculated pine wood nematode at 22 days after treatment, however no efficacy was found over 22 days. Eclipta prostrata extract and terthiophene can be used as nematicide for further nematicidal efficacy test against any other plant parasitic nematodes and trunk injection efficacy against pine wood nematode.

• The Korean Journal of Pesticide Science
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• v.19 no.2
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• pp.141-150
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• 2015

Thirty-two actinomycetes isolates from Korean forest soil were screened for their nematicidal and reproduction suppression activity against pine wood nematode (PWN) which is widely spread in Korea. Culture filterates of 21 isolates showed more than 90% mortality at 2-fold concentration. Among them, AM210, SG16, YD116 and YD315 were more effective than others on reproduction of PWN. The YD116 isolate was identified as Streptomyces atratus by morphological and 16S rDNA analyses. Hydrazine hydrate, similar to hydrazidomycin which has cytotoxicity among substances from S. atratus against PWN, was tested for its nematicidal activity. Ten ppm of the hydrate showed 60.8% mortality. Additional studies are needed for practical use of the S. atratus YD116 isolate.

• Korean journal of applied entomology
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• v.59 no.4
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• pp.361-369
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• 2020

This study aimed to investigate the differences in the effects of nematicidal materials (Burkholderia rinojensis, fluazaindolizine, fluopyram, fosthiazate, hydrogen peroxide, and imicyafos) on eggs and larvae of the sugar beet cyst nematode, Heterodera schachtii and clover cyst nematode, H. trifolii, which have recently become problematic in highland Chinese cabbage cultivation areas in Korea. Comparison of the hatching rates according to the test materials showed differences between treatments. In the treatment with fluopyram, fosthiazate, and imicyafos, a very high hatching inhibitory effect of 99% was observed in both nematodes, and there was no difference in the efficacy of treatments between nematode species except for that with hydrogen peroxide. Regarding the nematocidal activity of treatments against hatched larvae, only the fluazaindolizine treatment, despite showing a high hatching inhibitory effect, showed a low nematicidal effect of less than 30% against both nematodes, whereas a mortality rate of more than 90% was observed in the other treatments. There was no difference between nematode species in the reactions of the 2nd instar juveniles to all nematicidal test materials after 24 and 48 h of treatment. Although there are limitations to the laboratory experiment, the reaction to the nematicidal materials of the two cyst nematodes that damage the same host was similar, and simultaneous control is suggested in the areas where the two nematodes occur together.

• Research in Plant Disease
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• v.22 no.4
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• pp.227-235
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• 2016

Plant disease caused by root-knot nematode is a major problem in crop production. Using of chemical pesticides, one of the most efficient methods to control nematodes, have raised issues in toxicity to humans and animals and environmental pollution. In this study, to select actinomycete strains that have potential to serve as a microbial agent for control of nematodes, we investigated nematicidal activity of culture broth from 670 Streptomyces isolates. A culture filtrate of KRA15-528 isolate that was identified as S. flavogriseus on the basis of 16S rRNA sequence analysis, showed strong nematicidal activity against second stage of juveniles of Meloidogyne incognita and inhibited egg hatching; exposure to 10% of culture filtrate resulted in 71% juvenile mortality at 48 hours afters treatment and suppressed egg hatching by 54% at 9 days after treatment. When the KRA15-528 culture filtrate was partitioned with ethyl acetate and butanol, ethyl acetate layer exclusively showed strong activity; 91%, 53%, 30% of mortality at 1,000, 500, $250{\mu}g/ml$, respectively. Additionally, the culture filtrate suppressed gall formation on cucumber plant by M. incognita with no phytotoxicity. These results suggest that S. flavogriseus KRA15-528 has potential to serve as a microbial nematicide for the control of root-knot nematode disease.

• Research in Plant Disease
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• v.20 no.1
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• pp.37-49
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• 2014

A total of 2,714 methanol extracts of 1,677 plant species in 178 families were screened for nemastatic activity against Rhabditis sp. by 96-well microplate bioassay. The plant extracts with a concentration of $5,000{\mu}g/ml$ were mixed with aqueous nematode solution containing about 20 Rhabditis sp. and their activity was examined daily for 7 days. Out of 2,714 plant extracts examined in this test, 2,362 (87.0%) showed no negative influence on the nematode activity, while 187 (6.9%) inhibited nematode activity about 50%, 95 (3.5%) inhibited nematode activity over 90%, and 70 (2.6%) rather enhanced nematode activity. Among those showing over 50% nemastatic activity, 25 extracts were randomly selected for further screening with Rhabditis sp. and with juveniles of Meloidogyne incognita. The screening revealed that 11 extracts (44%) were consistently nemastatic. The 282 extracts with nemastatic activity could be of use in future studies on nematicidal plants.

• The Korean Journal of Pesticide Science
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• v.14 no.2
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• pp.138-147
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• 2010

Nematicidal activity against pine wood nematode, Bursaphelenchus xylophilus, was tested in the pot and field by the treatment of microbial formulation after media optimization. The optimized media composition was glycerol 10 g, soybean meal 10 g, NaCl 1 g, $CaCO_3$ 2 g, $K_2HPO_4$ 0.125 g per liter and microbial complex formulation was made with liquid and powder type. Most effective antibiotics against symbiotic microorganism with nematode, kanamycin, was added to the formulation. The control effects against pine wood nematodes were checked by pot test and field test. In the result of treatment by trunk injection, five times treatment was more effective than one time and the treatment with the formulation of concentrated culture supernatant was the most effective in the nematicidal activity showing below 10% mortality in pine tree.

• Journal of Microbiology and Biotechnology
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• v.24 no.11
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• pp.1510-1515
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• 2014

Lysobacter capsici YS1215 isolated from soil previously showed nematicidal potential for biological control of the root-knot nematode. In this study, lactic acid, a nematicidal compound, was isolated from culture filtrate of YS1215, and its ovicidal activity was investigated. Purification and identification of lactic acid were performed by a series of column chromatographies and identified by $^1H$ and $^{13}C$ NMR spectra and GC-MS analysis. Our results showed that bacterial culture filtrate containing lactic acid significantly inhibited egg hatching. The lowest egg hatch rate (5.9%) was found at a high concentration ($25 {\mu}l/ml$) of lactic acid at 5 days after incubation, followed by 20 (15.2%), 15 (23.7%), 10 (29.8%), and $5(36.4%){\mu}l/ml$, while egg hatching in the control (sterile distilled water) was 44.5%. This is the first report of lactic acid as an ovicidal compound, and it may be considered as an alternative of chemical pesticide against root-knot nematodes.

• Korean Journal of Soil Science and Fertilizer
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• v.49 no.4
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• pp.401-408
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• 2016

The present study described the isolation of 2-methylbutyric acid (2-MBA) produced from Bacillus pumilus L1, to subsequently investigate its nematicidal activity for the control of the root-knot nematode. The results showed that 2-MBA could be purified by chromatographic techniques and was identified using nuclear magnetic resonance and liquid chromatography-mass spectrometry. Crude extract and partially purified compounds had a significant effect on the inhibition of egg hatchability and second-stage juvenile (J2) mortality. A dose-dependent effect of 2-MBA was observed for J2 mortality and egg hatchability. Egg hatchability was 69.2%, 59.9%, 32.7%, and 0.0% at 125, 250, 500, and $1000{\mu}g\;mL^{-1}$ of 2-MBA after 4 d of incubation, respectively. Meanwhile, J2 mortality was in the range of 24.4%-100.0% after 2 d of incubation, depending on the concentrations of 2-MBA used. A pot experiment also demonstrated that treatment of B. pumilus L1 culture caused a significant reduction in the number of galls, egg masses, and J2 population than that of the tap water (TW) control. However, as the B. pumilus L1 culture concentration was decreased, the efficacy of nematode control by treatment of B. pumilus L1 culture was reduced compared to that of TW. B. pumilus L1 inoculation at different concentrations also promoted cucumber plant growth. Therefore, our study demonstrated the potential of 2-MBA from B. pumilus L1 as a biocontrol agent against the root-knot nematode and a plant growth promoter for cucumber plants.

• Parasites, Hosts and Diseases
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• v.60 no.5
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• pp.345-352
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• 2022

Chitinase AO-801 is a hydrolase secreted by Arthrobotrys oligospora during nematode feeding, while its role remained elusive. This study analyzed the molecular characteristics of recombinant chitinase of Arthrobotrys oligospora (reAO-801). AO-801 belongs to the typical glycoside hydrolase 18 family with conserved chitinase sequence and tertiary structure of (α/β)₈ triose-phosphate isomerase (TIM) barrel. The molecular weight of reAO-801 was 42 kDa. reAO-801 effectively degraded colloidal and powdered chitin, egg lysate, and stage I larval lysate of Caenorhabditis elegans. The activity of reAO-801 reached its peak at 40℃ and pH values between 4-7. Enzyme activity was inhibited by Zn2⁺, Ca2⁺, and Fe3⁺, whereas Mg2⁺ and K⁺ potentiated its activity. In addition, urea, sodium dodecyl sulfate, and 2-mercaptoethanol significantly inhibited enzyme activity. reAO-801 showed complete nematicidal activity against C. elegans stage I larvae. reAO-801 broke down the C. elegans egg shells, causing them to die or die prematurely by hatching the eggs. It also invoked degradation of Haemonchus contortus eggs, resulting in apparent changes in the morphological structure. This study demonstrated the cytotoxic effect of reAO-801, which laid the foundation for further dissecting the mechanism of nematode infestation by A. oligospora.

• Journal of Microbiology and Biotechnology
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• v.18 no.5
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• pp.852-858
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• 2008

An extracellular protease (Mc1) was isolated from the nematode-trapping fungus Monacrosporium cystosporium by gel filtration, anion-exchange, and hydrophobic interaction chromatographies. This protease had a molecular mass of approximately 38 kDa and displayed an optimal activity at pH 7-9 and $56^{\circ}C$ (over 30 min). Its proteolytic activity was highly sensitive to the serine protease inhibitor PMSF (phenylmethylsulfonylfluoride, 0.1 mM), indicating that it belonged to the serine-type peptidase group. The Michaelis constant ($K_m$) and $V_max$ for substrate N-Suc-Ala-Ala-Pro-Phe-pNA were $1.67{\times}10^{-4}\;M$ and 0.6071 $OD_{410}$ per 30 s, respectively. This protease could degrade a broad range of substrates including casein, gelatin, BSA (bovine serum albumin), and nematode cuticle. Moreover, the enzyme could immobilize the free-living nematode Panagrellus redivivus and the pine wood nematode Bursaphelenchus xylophilus, suggesting that it might playa role in infection against nematodes. The encoding gene of Mc1 was composed of one intron and two exons, coding for a polypeptide of 405 amino acid residues. The deduced amino acid sequence of Mcl showed 61.4-91.9% identity to serine proteases from other nematode-trapping fungi. Our results identified that Mcl possessed biochemical properties including optimal reaction condition and substrate preference that are different from previously identified serine proteases.