• Journal of Microbiology and Biotechnology
• /
• v.19 no.3
• /
• pp.314-322
• /
• 2009

Four different bacterial colonies were isolated from an old stock of an entomopathogenic nematode, Steinernema monticolum. They all showed entomopathogenicity to final instar larvae of beet armyworm, Spodoptera exigua, by hemocoelic injection. However, they varied in colony form, susceptibility to antibiotics, and postmortem change of the infected host insects. Biolog microbial identification and 16S rDNA sequence analyses indicate that these are four different species classified into different bacterial genera. Owing to high entomopathogenicity and a cadaver color of infected insect host, Serratia sp. was selected as a main symbiotic bacterial species and analyzed for its pathogenicity. Although no virulence of Serratia sp. was detected at oral administration, the bacteria gave significant synergistic pathogenicity to fifth instar S. exigua when it was treated along with a spore-forming entomopathogenic bacterium, Bacillus thuringiensis. The synergistic effect was explained by an immunosuppressive effect of Serratia sp. by its high cytotoxic effect on hemocytes of S. exigua, because Serratia sp. caused septicemia of S. exigua when the bacterial cells were injected into S. exigua hemocoel. The cytotoxic factor(s) was present in the culture medium because the sterilized culture broth possessed high potency in the cytotoxicity, which was specific to granular cells and plasmatocytes, two main immune-associated hemocytes in insects.

• Korean journal of applied entomology
• /
• v.38 no.3
• /
• pp.255-260
• /
• 1999

Pathogenicity and multiplication of entomopathogenic nematode, Steinernema carpocapsae Weiser, ere analyzed in two insect hosts, Spodoptera exiga (Hubner) and Sp. litura(Fabricius). The estimated ${LC}^{50}s$(lethal concentration of the infective juveniles to kill 50% of the host insect population) were not different between tow insect species or among their developmental stages on the filter paper assay, thought the actual numbers of the infected nematodes were varied among them. The significant variation, however occurred in nematode multiplication between tow inset hosts. Temperature also gave sosignificatn effect on nematode multiplication rate that it took 6 days after infection at $25^{\circ}C$, but did 12 days at $20^{\circ}C$ to show the maximal nematode population peak($\approx$500,000 infective juveniles(IJ) in a 5th instar larva of Sp. litura and $\approx$-100,000 IJ in a 5th instar larva of Sp. exigua).

• Korean journal of applied entomology
• /
• v.51 no.3
• /
• pp.255-263
• /
• 2012

High frequency sounds disrupt physiological processes, such as feeding behavior, development and immune responses of Spodoptera exigua. We analyzed high frequency sounds with respect to biochemical changes in S. exigua. High frequency sound (5,000 Hz, 95 dB) suppressed protein synthesis and secretion of midgut epithelium. It also significantly inhibited a digestive enzyme activity of phospholipase $A_2$. The gene expression of three different heat shock proteins and apolipophorin III was altered, particularly in midgut tissue in response to high frequency sound treatments. High frequency sound treatments significantly increased sugar and lipid levels in hemolymph plasma. These results suggest that high frequency sounds are a physiological stress that induces biochemical changes in S. exigua.

• Proceedings of the Korea Society of Environmental Biology Conference
• /
• 2002.11a
• /
• pp.123-126
• /
• 2002

Pollutants that are persistent, bioaccurnulative, and toxic have been linked to numerous adverse effects in human and animals, PBTs include heavy metals, polychlorinated biphenyls (PCBs), dioxins, polycyclic aromatic compounds (PACs) in addition to pesticides. This study focuses on toxic effects of the PBTs except pesticides on insects. Eight PBTs were selected from subgroups: three heavy metals (Pb, Hg, and Cd), two PCB mixtures (Aroclor mixtures 1 and 2), 2,3,7,8-tetrachlorodibenzo-p-dioxin, two monophenols (4-octylphenol and 4-nonylphenol), and tetrabutyltin, Beet armyworm, Spodoptera exigua, was used as test target insect species. Three physiological markers (metamorphosis, immune reaction, and follicle patency) were assessed in each exposure to different doses of the PCBs. Heat-shock proteins as molecular markers were also analyzed in response to the PCBs. All tested PBTs were toxic to metamorphosis from larvae to pupae when they were applied with diet. Two PCB mixtures were the most toxic compounds in this assay by giving significant toxicity at 0.005 ppm, while others had from 10 to 1000 ppm. Dioxin (0.1 ppb), tetrabutyltin (0.1 ppb), Pb (10 ppb), and Hg (0,01 ppb) were potent to inhibit immune reactions analyzed by inducing phenoloxidase activity and blocked phospholipase $A_2$ enzyme, Tetrabutyltin and dioxin significantly induced follicle cell patency, but their effects were lower than that of endogenous juvenile hormone, Dioxin, Pb, Hg, and Cd could induce the expression of heat shock proteins that were detected by immunoblotting against human HSP70 monoclonal antibody. HSP78 and HSP80 were upregulated in response to the PBTs. This expression was detected from the fat body and epidermis at as fast as 4h after injection. All these results clearly suggest that PBTs give significant ecotoxicity to insects that are valuable organisms in our environment.

• Korean journal of applied entomology
• /
• v.39 no.3
• /
• pp.149-152
• /
• 2000

Cryopreservation of infective juveniles of entomopathogenic nematode, Steinernema carpocapsae Weiser, was conducted at $-190^{\circ}C$ liquid nitrogen and its, efficacy was analysed on nematode survival and pathogenicity with glycerol pretreatments and storage periods. Infective juveniles were pre-treated before being frozen by incubating the nematodes in 22% glycerol for each of 6, 12, and 24 h, followed by 70% methanol at $0^{\circ}C$ for 10 minutes. Just after glycerol and methanol incubations, subsamp1es of the nematodes were resuspended in 0.85% saline and maintained during 24h for viability determination. Different glycerol incubation periods significantly affected the nematode susceptibility to methanol infiltration. Six hour incubation in glycerol resulted in much less nematode survival than did 12 h or 24 h incubation. About 70% of the infective juveniles frozen at $-190^{\circ}C$ for 5 months, preincubat-ed in glycerol at least for 12h, were able to survive after being resuspended in 30°C saline. They did not also show any change in their pathogenicity during cryopreservation. These results suggest an improved technique for long-term storage of the entomopathogenic nematodes.

• Korean journal of applied entomology
• /
• v.30 no.3
• /
• pp.212-218
• /
• 1991

Pathogenicity of Spodoptera exigua nuclear polyhedrosis virus (SeMNPV) against the host insect and 8 species of lepidopterous insects and cross infection of baculoviruses to third instar of S. exigua larvae were studied to determine as a biocontrol agent for S. exigua. The median lethal concentrations($LC_{50}$)of the SeMNPV to egg mass was $2.855\times10^5$ PIBs/ml and higher than that to the larvae of S. exigua. Mortality of the SeMNPV in third ins tar larvae was more increased than that in first and fifth instar of S. exigua larvae by 1.16 and 4.11 times, respectively. The median lethal times($LT_{50}$) to $1.56\times10^6$ PIBs/ml was in the range of 4.25 to 5.04 days. Infectivity of the SeMNPV against eight species of lepidopterous insects was showed only in the host insect, S. exigua. Autographa cali/ornica MNPV, Mamestra barassicae MNPV, and Trichoplusia ni MNPV were cross-infected to third instar of S. exigua larvae among ten of baculoviruses tested.

• Korean journal of applied entomology
• /
• v.37 no.2
• /
• pp.163-170
• /
• 1998

Larval morphological characters such as body color and stripe pattern were analyzed to get morphological genetic markers of beet armyworm, Spodoptera exigua (Hiibner). Body color was varied from light green to dark brown with diets. Stripe characters were classified by the presence of dorsal and lateral lines: three stripes with both lines, one stripe with dorsal line only, and zero stripe. Proportions of each stripe character increased with successive selections for its own character. Three stripe was dominant to one stripe when they were crossed. The estimated heritability in narrow sense (h2) of the stripe pattern was 0.50k0.42. About two fold more females than males were produced in zero stripe line. Stripe pattern was not significantly changed by different diets except in welsh onion which had lower proportion of three stripe individuals than that of the expected. Larval stripe pattern was also correlated with larval and pupal developmental rates and cold hardiness but not with insecticide susceptibility.

• Journal of Microbiology and Biotechnology
• /
• v.31 no.4
• /
• pp.529-539
• /
• 2021

The beet armyworm, Spodoptera exigua, is a serious insect pest infesting various vegetable crops. Two infectious insect viruses, baculovirus and iflavirus, are known to induce epizootics in S. exigua populations. Indeed, some laboratory colonies have appeared to be covertly infected by these viruses. Diagnostic PCR tests detected two different viruses: Spodoptera exigua multiple nucleopolyhedrosis virus (SeMNPV) and iflaviruses (SeIfV1 and SeIfV2). Viral extract from dead larvae of S. exigua could infect Sf9 cells and produce occlusion bodies (OBs). Feeding OBs to asymptomatic larvae of S. exigua caused significant viral disease. Interestingly, both SeIfV1 and SeIfV2 increased their titers at late larval stages. Sterilization of laid eggs with 1% sodium hypochloride significantly reduced SeMNPV titers and increased larval survival rate. Double-stranded RNA (dsRNA) specific to SeIfV1 or SeIfV2 significantly reduced viral titers and increased larval survival rate. To continuously feed dsRNA, a recombinant Escherichia coli HT115 expressing SeIfV1-dsRNA was constructed with an L4440 expression vector. Adding this recombinant E. coli to the artificial diet significantly reduced the SeIfV1 titer and increased larval survival. These results indicate that laboratory colony collapse of S. exigua is induced by multiple viral infections. In addition, either suppression of SeMNPV or SeIfV infection significantly increased larval survival, suggesting a cooperative pathogenicity between baculovirus and iflavirus against S. exigua.

• Entomological Research
• /
• v.48 no.5
• /
• pp.390-399
• /
• 2018

In order to precisely assess gene expression levels, the suitable internal reference genes must be served to quantify real-time reverse transcription polymerase chain reaction (RT-qPCR) data. For armyworm, Mythimna separata, which reference genes are suitable for assessing the level of transcriptional expression of target genes have yet to be explored. In this study, eight common reference genes, including ${\beta}$-actin (${\beta}$-ACT), 18 s ribosomal (18S), 28S ribosomal (28S), glyceraldehyde-3-phosphate (GAPDH), elongation fator-alpha ($EF1{\alpha}$), TATA box binding protein (TBP), ribosomal protein L7 (RPL7), and alpha-tubulin (${\alpha}$-TUB) that in different developmental stages, tissues and insecticide treatments of M. separata were evaluated. To further explore whether these genes were suitable to serve as endogenous controls, three software-based approaches (geNorm, BestKeeper, and NormFinder), the delta Ct method, and one web-based comprehensive tool (RefFinder) were employed to analyze and rank the tested genes. The optimal number of reference genes was determined using the geNorm program, and the suitability of particular reference genes was empirically validated according to normalized HSP70, and MsepCYP321A10 gene expression data. We found that the most suitable reference genes for the different experimental conditions. For developmental stages, 28S/RPL7 were the optimal reference genes, both $RPL7/EF1{\alpha}$ were suitable for experiments of different tissues, whereas for insecticide treatments, $28S/{\alpha}-TUB$ were suitable for normalizations of expression data. In addition, $28S/{\alpha}-TUB$ were the suitable reference genes because they have the most stable expression among different developmental stages, tissues and insecticide treatments. Our work is the first report on reference gene selection in M. separata, and might serve as a precedent for future gene expression studies.

• Korean journal of applied entomology
• /
• v.62 no.4
• /
• pp.347-354
• /
• 2023

Mythimna loreyi (Duponchel, 1827) (Lepidoptera: Noctuidae) is a pest that damages agricultural plants, such as rice, wheat, and maize. We sequenced the entire 15,314-bp mitochondrial genome of this species. It has a typical set of genes (13 protein-coding genes, two ribosomal RNA genes, and 22 transfer RNA genes) as well as one major non-coding A+T-rich region. Using concatenated sequences of 13 protein-coding genes and two rRNAs (13,376 bp, including gaps), phylogenetic analysis demonstrated that the sister relationship between M. loreyi and M. separata had the highest nodal support. The monophyly of each family (Noctuidae, Euteliidae, Nolidae, Erebidae, and Notodontidae) of the superfamily Noctuoidea was supported by the highest nodal support.