• Korean journal of applied entomology
• /
• v.58 no.2
• /
• pp.101-109
• /
• 2019

An entomopathogenic bacterium, Xenorhabdus ehlersii KSY, is symbiotic to a nematode, Steinernema longicaudum, and exhibits high entomopathogenic virulence against lepidopteran insects. This study showed that the bacterial pathogenicity is induced by its inhibitory activity against eicosanoid biosynthesis of target insects, resulting in immunosuppression. To be applied for insect pest control, the bacteria should be infected to insect hemocoel. To deliver X. ehlersii to inset hemocoel, Bacillus thuringiensis (Bt) was mixed with the bacteria to breakdown the physical barrier (= midgut epithelium) from midgut lumen to hemocoel. The bacterial mixture significantly enhanced insecticidal activity of Bt only against larvae of Plutella xylostella and Maruca vitrata. For formulation, X. ehlersii cells were freeze-dried and mixed with sporulated Bt cells. The formulated bacterial mixture was applied to semi-field cultivating cabbage crop infested by P. xylostella. The bacterial mixture treatment showed over 95% control efficacy, while Bt alone gave 80% control efficacy. These results suggest that X. ehlersii can be applied to develop a novel insect control agent.

• The Korean Journal of Soil Zoology
• /
• v.5 no.2
• /
• pp.97-100
• /
• 2000

We report here an improved collecting method of the infective juveniles multiplied in the host insect with the entomopathogenic nematodes, Steinernema carpocapsae Weiser. The specific characteristics of this method involved the opening of the host insect hemocoel after the population of their infective juveniles reached at the maximum (6 days at 25$\^{C}$ after nematode treatment to nonimmunized host insects) to facilitate the escape of the multiplied nematodes. It also used 'Baermann funnel'method to select the infective juveniles effectively. This improved 'Baermann funnel'method was compared with a traditional collecting method, which was characterized with a combination of untreated host insects and 'White trap'collecting method, in both yield and pathogenicity of the collected infective juveniles to the fifth instar larvae of beet armyworm, Spodoptera exigua (H bner). More than 95% of the nematode populations collected by the two methods represented the morphological infective juveniles. To prove the nematodes to be infective juveniles functionally, pathogenicity and infective activity were compared in the nematodes collected by the two methods. They were not different in both pathogenicities and infective activities which were measured by the numbers of nematodes penetrated into the hemocoel of the insect hosts after exposure for the specific times to the same dote of infective juveniles. Significant difference between two collecting methods was found in the total yields of the infective juveniles per host insect About 50,000 infective juveniles per infected fifth instar larva of S. exigua after 6 day incubation at 25$\^{C}$ were collected only for 2 days by the improved 'Baermann funnel'method, while about 20,000 infective juveniles per host were collected for 10 days by the classical 'White trap'method.

• The Korean Journal of Pesticide Science
• /
• v.2 no.1
• /
• pp.18-23
• /
• 1998

In order to obtain leading compound for the development of new pesticide through the organic synthesis of natural products, the synthesis of ricinine, an active compound of Ricinus communis, was established and biological activities of synthetic compounds against insects were examined. The synthetic scheme of ricinine was composed of four steps by the spontaneous condensation of the cyanoacetyl chloride. A modified synthetic process was also estabilshed to enhance the synthetic yield by simple cyclization of ethoxymethylene malononitrile. In the bioassay results of synthetic ricinine and intermediates on four insects, the mortality of ricinine on brown planthopper (BPH, Nilaparvata lugens) and pea weevil(PW, Bruchus rufimanus) was 80% and 75% at the concentration of 1,000 ${\mu}g/ml$ respectively. Chloronorricinine and chlororicinic acid having chloride group in molecular structure gave 60% mortality on two-spotted mite (TSSM, Tetranychus urticae) at the concentration of 500 ${\mu}g/ml$. The mortality of compounds on house mosquito (HM, Culex pipens pallens) was meager at 10 ${\mu}g/ml$ level.

• Microbiology and Biotechnology Letters
• /
• v.19 no.3
• /
• pp.308-312
• /
• 1991

The delta-endotoxin from B. thuringiensis subsp. damstadienszs 73E10-2 was resistant to high concentration of salt (4M NaBr), organic solvents (50% acetone), denaturants (4 M urea), and neutral detergents (10% triton X-100). In contrast, the toxin was inactivated by treating with charged detergents as well as guanidine hydrochloride or carbon tetra-chloride. The delta-endotoxin is not a sulfhydryl activated toxin, but modification of the lysine side chains eliminated toxicity against mosquito larvae.

• Korean journal of applied entomology
• /
• v.30 no.4
• /
• pp.227-232
• /
• 1991

The pathogenicity of entomopathogenic nematodes Sfeinernema earpocapsae and Heferorhabditis baeferiophora was evaluated against forest insect pests, alder leaf beetle Agelastiea eoerulea, pellucid zygaenid Pryeria siniea, and box-tree pyralid Glyphodes perspeetalis. Alder leaf beetle larvae were exposed to S. earpocapsae at concentration of 0, 25, 50, and 100 nematodes and to H. baeferiophora at concentration of 0, 10, 20, and 40 nematodes per larva on alder leaves. Mortalities of 1st instar lavae were 85.4 $\pm$ 4.1-100%, 2nd instar larvae 80.0 $\pm$ 5.8-100%, and 3rd instar larvae 65.0 $\pm$ 10.8-100% in S. earpocapsae and those of 1st instar larvae were 82.5 $\pm$ 6.9-100%, 2nd instar larvae 77.5 $\pm$ 4.7-100%, and 3rd instar larvae 55.0 $\pm$ 13.5-100% in H. baeferiophora treatment. When pellucid zygaenid larvae were exposed to S. earpocapsae at concentration of 0, 5, 10, 20, 40, and 80 nematodes and to H. baeteriophora at concentration of 0, 2, 5, 10,20, and 40 nematodes per larva, mortalities were 98.9 $\pm$ 1.1-100% in S. earpocapsae and 26.7 $\pm$ 5.1-74.5 $\pm$ 6.2% in H. baeferiophora. The mortalities of box-tree pyralid larvae were 97.8 $\pm$ 1.5-100% in S. earpocapsae treated with concentration of 0,20,40, and 80 nematodes per larva and those were 92.0 $\pm$ 6.2-98.9 $\pm$ 1.1 % in H. baeferiophora treated with con'||'&'||'not;centration of 0, 10, 20, and 40 nematodes per larva.

• Korean journal of applied entomology
• /
• v.31 no.4
• /
• pp.452-460
• /
• 1992

To establish the economical and reliable routine bioassay system for developing new insecticidal compounds, effects of leaf-dipping time, application methods, insect species and their developmental stages on susceptibilities of insects to insecticides were studied. The stable insecticidal activity appeared at the dipping time for 30-60 seconds in leaf-dipping method, and the most effective application methods were leaf-dipping method for apterous green peach aphid adults, and third instars of diamond-back moth and tobacco cutworm, whereas seedling+insect spray method for adults or third instars of brown planthoppers. For two-spotted spider mite, leaf-dipping or intact plant spray method was favorable. In the bioassay for chitin synthesis inhibitors, the inoculation of third instars of brown planthopper, diamond-back moth, tobacco cutworm and green peach aphid, and larvae of two-spotted spider mite to the young host plants treated by spray method were adequate bioassay methods.

• Korean journal of applied entomology
• /
• v.55 no.2
• /
• pp.81-89
• /
• 2016

Double-stranded RNA (dsRNA) has been applied to control insect pests by its suppressive activity against specific target genes. Integrin is a heterodimer (${\alpha}$ and ${\beta}$) transmembrane protein and plays a critical role in cell-to-cell or cell-to-extracellular matrix interactions in eukaryotes. Suppression of ${\beta}$ subunit integrin gene expression by its specific dsRNA (= dsINT) induces significant mortality against target insects. Furthermore, a recombinant bacterium expressing dsINT is potent to kill target insects. However, it is necessary to develop a formulation technique of the dsRNA-expressing bacteria to apply the bacterial insecticide against field populations. This study formulated the recombinant bacteria by freeze-drying and tested its control efficacy against target insects. The formulation maintained significant insecticidal activity against last instar larvae of Spodoptera exigua. While a commercial Bacillus thuringiensis (Bt) insecticide exhibited only about 60% insecticidal activity against S. exigua last instar, an addition of the dsINT-expressing bacterial formulation significantly enhanced the Bt insecticidal activity. The dsINT-expressing bacterial formulation exhibited relative selectivity to target insects depending on sequence similarity. These results indicate that a freeze-dried form of dsRNA-expressing bacteria keeps its insecticidal activity.

• Microbiology and Biotechnology Letters
• /
• v.19 no.3
• /
• pp.303-307
• /
• 1991

The hemolyic polypeptide in delta-endotoxin from Bacillus thun'ngiensis subsp. darmstadiensis 73ElO-2 was purified by Sephadex G-IOO gel filtration and DEAE-cellulose ion exchange column chromatography. The purity of hemolysin was confirmed by ouchterlony test and SDS-PAGE. The molecular weight of the purified hemolysin was approximately 64 KDa by SDS-PAGE. The purified hemolysin has not mosquitocidal activity against larvae of Aedes agypti, but hemolytic activity on red blood cells of rat. There is no serological relationship between delta-endotoxin from B. thuringiensis subsp. israelensis and the purified hemolysin from the . strain 73ElO-2.

• Journal of Applied Biological Chemistry
• /
• v.55 no.2
• /
• pp.123-127
• /
• 2012

In order to develop eco-friendly biopesticide, an entomopathogenic bacterium Photorhabdus temperata M1021 has been lyophilized via freeze-drying along with protective agents such as skim milk, starch, sodium alginate, glucose and sodium glutamate to protect cells from lysis. Freeze-drying powder of P. temperata M1021 containing 7% skim milk (w/v) showed highest survival rate of 63% among all the protective agents used in trials. Furthermore, the freeze-dried microbial powder showed 75% of survival rate after stored at $4^{\circ}C$ for 4 weeks at air contact conditions. Injection toxicity of the freeze-dried sample was tested against larvae of Galleria mellonella. A dose of $2.0{\times}10^1$ cells of P. temperata M1021 killed 100% of the G. mellonella larvae within 4 days after injection. Moreover, $2.0{\times}10^0$ cells caused 50% mortality within the 4 days after injection. Freeze-dried P. temperata M1021 strains exhibited effective insecticidal activity and could be a better candidate for being used as a biopesticide.

• Microbiology and Biotechnology Letters
• /
• v.18 no.3
• /
• pp.301-304
• /
• 1990

In the mosquitocidal delta-endotoxins from Bacillus thuringiensis subsp, isruelensis and B. thuringiensis subsp. darmstudiensis 73E10-2, were contained an immunologically homologous protein. The homologous protein was confirmed from Ouchterlony test, irnmuno-electrophoresis, and enzyme linked immunoassay by polyclonal antibodies against the delta-endotoxins of both strains.