• Korean journal of applied entomology
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• v.52 no.1
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• pp.35-43
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• 2013

All tested Korean populations of the diamondback moth, Plutella xylostella, are known to be resistant especially against pyrethroid insecticides by mutation in its molecular target, para-sodium channel. Moreover, P. xylostella is able to develop resistance against most commercial insecticides. This study was performed to develop an efficient control technique against P. xylostella by a combined treatment of an endoparasitoid wasp, Cotesia plutellae, and a microbial insecticide, Bacillus thuringiensis. To investigate any parasitism preference of C. plutellae against susceptible and resistant P. xylostella, five different populations of P. xylostella were compared in insecticide susceptibilities and parasitism by C. plutellae. These five P. xylostella populations showed a significant variation against three commercial insecticides including pyrethroid, organophosphate, neonicotinoid, and insect growth regulator. However, there were no significant differences among five P. xylostella populations in their parasitic rates by C. plutellae. Moreover, parasitized larvae of P. xylostella showed significantly higher susceptibility to B. thuringiensis. As an immunosuppressive agent, viral ankyrin genes (vankyrins) encoded in C. plutellae were transiently expressed in nonparasitized larvae. Expression of vankyrins significantly enhanced the efficacy of B. thuringiensis against the third instar larvae of P. xylostella. Thus an immunosuppression induced by C. plutellae enhanced the insecticidal efficacy of B. thuringiensis. These results suggest that a combined treatment of C. plutellae and B. thuringiensis may effectively control the insecticide-resistant populations of P. xylostella.

• Korean journal of applied entomology
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• v.46 no.1 s.145
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• pp.57-62
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• 2007

Reproductive incompatibility is an Important factor to select a specific biologlcal control agent for successful augmentation of the corresponding endogenous population. An endoparasitoid, Cotesia plutellae (Kurdjumov), is an effective control agent to diamondback moth, Plutella xylostella (L.) and has been known to be classified into two groups in terms of reproductive incompatibility. This study analyzed an Korean population of C. plutellae in terms of morphological characters and mitochondrial DNA marker, which did not match with either of two reproductive incompatibility groups. These results suggest that a Korean population of C. plutellae can be involved in a novel reproductive group. For any augmentation program of C. plutellae in Korea, reproductive incompatibility should be seriously considered to select a particular exotic population.

• Korean journal of applied entomology
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• v.43 no.3 s.136
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• pp.225-231
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• 2004

An endoparasitoid wasp, Cotesia plutellae, has been used for a biological control agent against the diamondback moth, Plutellae xylostella. It has a symbiotic polydnavirus in their reproductive tract, which is required for its successful parasitization. Here, we measured a specific replication time of the polydnavirus during female development of C. plutellae. We, also, analyzed the reproductive potentials of female C. plutellae under mating or different host conditions. At $25^{\circ}C$, pupal C. plutellae began to develop adult tissues such as compound eyes and wings since day 2. At day 5, all adult tissues including antennae were developed and were ready to emerge. With polyclonal antibody raised against C. plutellae polydnavirus, an immunobloting could confirm virus replication at day 4 during pupal stage. Virus particles could be visualized by transmission electron microscope in the oviduct lumen of day 5 pupae. After adult eclosion, venom gland and ovarian calyx increased in size, though ovarioles did not. Mated females layed large number of eggs (over $60\%$) at first 4 days during their mean longevity of ca. 8 days at $25^{\circ}C$. Unmated females showed less active ovipositional behavior, where all the eggs developed into males. C. Plutellae parasitized both P. xylostella and fall webworm, Hyphantria cunea. However, C. Plutellae developed faster and showed higher successful paarasitization in P. xylostella than in H. cunea.

• Korean journal of applied entomology
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• v.43 no.3 s.136
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• pp.217-223
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• 2004

Polydnavirus is a symbiotic virus of some endoparasitic wasps and plays crucial roles in inhibiting immune responses and retarding development of the parasitized hosts. Cotesia plutellae bracovirus (CpBV) is a polydnavirus suggesting a major causative to change developmental physiology of the parasitized host. Here, we investigated whether CpBV can interrupt development of non-natural host. Beet armyworm, Spodoptera exigua, is used as a non-permissible host for parasitization of C. plutellae. Extract from the calyx region of C. plutellae contained CpBV, which was confirmed by immunoblotting with a polyclonal antibody raised against CpBV. One female equivalent of CpBV extract was injected into hemocoel of late 4th instar larvae of S. exigua. The injected larvae showed delayed larval period, decrease of body weight gain, and inability of pupal metamorphosis. These inhibitory effect of the CpBV extract was rescued by injection along with CpBV antibody, though the antibody itself did not give any effect on development of the larvae. This result clearly shows that CpBV can interrupt developmental physiology of a non-natural host for its symbiotic wasp.

• Korean journal of applied entomology
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• v.45 no.3 s.144
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• pp.375-380
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• 2006

A solitary endoparasitoid, Cotesia plutellae, parasitizes diamondback moth, Plutellae xylostella. It has been suggested that its antennae are a major sensory organ to search host and recognize host developmental and other physiological condition during parasitization. This research was performed to understand the parasitic behavior of C. plutellae by analyzing sensory types and their numbers on the antennae using scanning electron microscope. There was no significant difference in antennal length in both male and female C. plutellae, in which both sexes had 16 flagellomeres. Three different types of sensilla (trichoid, seta, and placodea sensilla) were located mostly on flagella and analyzed in their density on the different antennomeres. Trichoid sensillum was the major sensory type and showed about 87% density among all sensilla. Both trichoid and placodea types of sensilla exhibited even numbers on all flagellomeres with some decrease at terminal segments. In contrast, seta form of sensilla showed drastic increase in its density at distal part after 9th flagellomere. When distal half of flagellomeres were cut off, the C. plutellae could not parasitize host larvae. Even when only four distal flagellomeres were removed, the parasitism showed only 30%. These results indicate that C. plutellae antennae are required for parasitism and suggest that seta form of sensilla may play significant roles in recognizing host for parasitization.

• Korean journal of applied entomology
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• v.45 no.1 s.142
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• pp.15-24
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• 2006

Inhibitor kB (IkB)-like gene has been found in the genome of Cotesia plutellae bracovirus (CpBV), which is the obligatory symbiont of an endoparsitoid wasp, C. plutellae. The open reading frame of CpBV-IkB was 417 bp and encoded 138 amino acids. Four ankyrin repeat domains were found in CpBV-IkB, which shared high homology with other known polydnavirus IkBs. Considering a presumptive cellular IkB based on Drosophila Cactus, CpBV-IkB exhibited a truncated structure with deletion of signal-receiving domains, which suggested its irreversible inhibitory role in NFkB signal transduction pathway of the parasitized host in response to the wasp parasitization. CpBV-IkB was expressed only in the parasitized diamondback moth, Plutella flostella. Its expression was estimated by quantitative RT-PCR during parasitization period, showing a constitutive expression pattern from the first day of parasitization. An indirect functional analysis of CpBV-IkB was conducted and suggested a hypothesis of host antivirus inhibition.

• The Korean Journal of Pesticide Science
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• v.19 no.3
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• pp.279-286
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• 2015

Cotesia plutellae bracovirus (CpBV) is a polydnavirus symbiotic to C. plutellae parasitizing young larvae of the diamondback moth, Plutella xylostella. Several CpBV genes play important roles in suppressing immune responses of the parasitized larvae. This study tested a hypothesis that the CpBV genes inducing host immunosuppression could be applied to develop a potent recombinant baculovirus. Based on a previous study, a recombinant baculovirus expressing CpBV-ELP1 (AcMNPV-ELP1) was selected and multiplied using larvae of the beet armyworm, Spodoptera exigua. The recombinant viruses were produced in a yield of $5{\times}10^{10}$ polyhedral inclusion body (PIB)/larva. The cultured AcMNPV-ELP1 exhibited a much higher pathogenicity against S. exigua larvae. However, its insecticidal activity was varied among larval instars of S. exigua, in which first and late instars were high susceptible. Spray of the recombinant baculovirus ($5{\times}10^6PIB/mL$) exhibited higher control efficacy (${\approx}$ 88%) against S. exigua larvae infesting cabbage than a chemical insecticide, tebufenozide, at 7 days after treatment. These results indicate that AcMNPV-ELP1 mass-cultured using host insect system is highly pathogenic and can be applied to develop a novel microbial control agent.

• The Korean Journal of Pesticide Science
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• v.12 no.3
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• pp.283-290
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• 2008

Baculoviruses have been used to control some serious lepidopteran pests. However, their narrow target insect spectrum and slow efficacy are main limitations to be used in various applications. This study introduces a technique to overcome these limitations by inhibiting insect immune defence to enhance the viral pathogenicity. Polydnaviruses are an insect DNA virus group and symbiotic to some ichneumonid and braconid endoparasitoids. Cotesia plutellae bracovirus (CpBV) is a braconid polydnavirus and encodes several immunosuppressive genes. We selected seven CpBV genes and recombined them to wild type Autographa California multiple nucleopolyhedrovirus (AcNPV). A bioassay of these seven recombinants indicated that most recombinants had similar or superior efficacy to wild type AcNPV against beet armyworm, Spodoptera exigua, and diamondback moth, Plutella xylostella. Recombinant AcNPV with CpBV-ELP was the most potent in terms of lethal time by shortening more than 2 days compared to wild type AcNPV. This recombinant was further proved in its dose-dependent pathogenicity and its efficacy by spray application on S. exigua infesting cabbage cultivated in pots. We discussed the efficacy of CpBV-ELP recombinant AcNPV in terms of suppressing antiviral activity of target insects.

• Korean journal of applied entomology
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• v.56 no.1
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• pp.19-28
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• 2017

Polydnaviruses (PDVs) are a group of double-stranded DNA viruses symbiotic to some endoparasitoid wasps. Cotesia plutellae bracovirus (CpBV) is a PDV symbiotic to an endoparasitoid wasp, C. plutellae, parasitizing young larvae of Plutella xylostella. An early expressed gene, CpBV-ELP1, plays an important role in the parasitism by suppressing host cellular immunity by its cytotoxic activity against hemocytes. This study aimed to test its oral toxicity against insect pest by expressing it in a recombinant tobacco plant. A recombinant CpBV-ELP1 protein was produced using a baculovirus expression system and secreted to cell culture medium. The cell cultured media were used to purify CpBV-ELP1 by a sequential array of purification steps: ammonium sulfate fractionation, size exclusion chromatography, and ion exchange chromatography. Purified rCpBV-ELP1 exhibited a significant cytotoxicity against Spodoptera exigua hemocytes. CpBV-ELP1 was highly toxic to the fifth instar larvae of S. exigua by injection to hemocoel. It also showed a significant oral toxicity to fifth instar larvae of S. exigua by a leaf-dipping assay. CpBV-ELP1 was cloned into pBI121 vector under CaMV 35S promoter with opaline synthase terminator. Resulting recombinant vector (pBI121-ELP1) was used to transform Agrobacterium tumefaciens LBA4404. The recombinant bacteria were then used to induce callus of a tobacco (Nicotiana tabacum Xanthi) leaves and subsequent generation (T1) plants were selected. T1 generation tobacco plants expressing CpBV-ELP1 gave significant insecticidal activities against S. exigua larvae. These results suggest that CpBV-ELP1 gene can be used to control insect pests by constructing transgenic crops.