The behavioral responses of Plutella xylostella (Lepidoptera: Plutellidae) to four high power light emmitting diodes (HPLEDs) were tested at various illuminance intensity and light exposure time using a HPLED-equipped Y-maze chamber. Preference tests were conducted using the high power light emmitting diodes (HPLEDs) in the dark room at 27±0.5℃ and 60±0.5% relative humidity. Evaluated specific wavelengths were blue, green, yellow and red HPLEDs. The illuminance intensity was tested at 20, 40, 60, 80, and 100 lx and fixed at 30 min. As a result, the attraction rate of the green HPLED at 60 lx was the best effective (98.3%) to P. xylostella, followed by red HPLED of 60 lx (86.5%), yellow HPLED of 60 lx (83.6%), and blue HPLED of 40 lx (72.0%), respectively. Depending on the change of light exposure time (5, 10, 15, 20, and 25 min), green HPLED showed the potential attraction under the 15 min. These results may be used as information for developing an eco-friendly insect pest control system. Further research needs to be performed to evaluate the behavioral responses to single and multiple HPLED sources in the field.

This study was conducted to investigate the insecticidal capacity of recombinant baculoviruses to Plutella xylostella and Spodoptera exigua larvae. For recombinant viruses, Bacillus thuringiensis cry1-5 crystal protein gene was introduced into the genome by fusion of polyhedrin-cry1-5 under the control of polyhedrin gene promoter. Recombinant AcPolh5-3006BiKTT and AcPolh5-3006 AvTox2 based on BiKTT and AvTox2, respectively, were constructed under the control of early promoter from Cotesia plutellae bracovirus. Mortality of S. exigua larvae was significantly higher when they fed on cabbage coated with ApEGFP (wild type) over 5.0×106 PIBs/ml. For AcPolh5-3006BiKTT and AcPolh5-3006AvTox2, mortality of P. xylostella and S. exigua larvae was significantly higher when they fed on cabbage coated with recombinant baculoviruses over 5.0×106 PIBs/ml and 1.0×104 PIBs/ml, respectively. The value of LD50 was lower in the treatments with AcPolh5-3006BiKTT (P. xylostella:1.2×106, S. exigua:1.3×104) or AcPolh5-3006AvTox2 (P. xylostella:2.3×106, S. exigua:1.4×104) than the treatments with ApEGFP (P. xylostella: not estimated, S. exigua:5.0×105). Survival time (ST50) of P. sylostella larvae was much shorter at AcPolh5-3006BiKTT (29.6h) than AcPolh5-3006AvTox2 (46.2h) while that of S. exigua larvae was much shorter at AcPolh5-3006AvTox2 (95.1h) than AcPolh5-3006BiKTT (101.9h) or ApEGFp (130.7h). The two recombinant baculoviruses were more effective in S. exigua larvae but slower speed of action.

Parasitization by an endoparasitoid wasp, Cotesia plutellae, extends a larval period of Plutella xylostella and inhibits a larva-to-pupa metamorphosis. To determine antimetamorphic parasitic factor(s) in this host-parasitoid interaction, an effect of its symbiotic polydnavirus, Cotesia plutellae bracovirus (CpBV), was investigated by injecting purified virus particles to nonparasitized larvae of P. xylostella. Larvae injected with CpBV exhibited antimetamophosis in a viral dose-dependent manner. Also, the susceptibility to the viral injection was increased at young larval stages. Parasitized or virus-injected larvae shwed significant decrease in cell size of prothoracic gland and reduction in expression of ecdysone receptor (EcR) gene. However, they increased and maintained expression of insulin receptor (InR) gene. Twenty four CpBVsegments were individually injected to nonparasitized larvae. Only two segments (S22 and S27) had significant antimetamorphic effect. Subsequent RNA interference using double stranded RNA (dsRNA) was performed in each of encoded genes in each segment. Protein tyrosine phosphatase, ELP, and three hypothetical genes were determined to be antimetamorphic factors.

To evaluate potential attractive activity of high power light emitting diodes (HPLEDs) against Plutella xylostella (Lepidoptera: Plutellidae), the specific wavelengths, illuminance intensities (20, 40, 60, 80, 100 lux), and light exposure times were investigated at 5 minute intervals. The evaluated specific wavelengths were blue (450±10 nm), green (520±5 nm), yellow (590±5 nm), red (660±10 nm), and white (450-630 nm). Based on the highest attraction rate, the HPLEDs treated to 60 lux intensity against P. xylostella were significantly more attractive than other illuminance intensities when light exposure time was 10 min. Attraction rate under optimal conditions showed that the green HPLED had the highest attraction rate (98.3%), followed by red HPLED (89.3%), blue HPLED (86.7%), yellow HPLED (76.7%), and white HPLED (70.0%), respectively. These results indicated that phototatic effect of the green HPLED against P. xylostella showed the greatest attraction at 60 lux intensity and 10 min light exposure time. Further research needs to be performed to evaluate the phototatic behavioral responses to single and multiple HPLED sources in the field.

From the methanol crude extracts of the tree of heaven (Ailanthus altissima) leaves, the antifeedant substance was isolated and bioassayed with different concentrations against diamondback moth (Plutella xylostella) larvae. The antifeeding activity was evaluated by measuring the feeding area during 24 hr after inoculation. Methanol extracts showing antifeeding activity at 5000 ppm was subsequently fractionated into hexane, chloroform, ethyl acetate and water layer. Third larvae of diamondback moth was tested to each fraction layer. Chloroform layer shows the highest antifeeding activity and the layer was purified by silica gel open column chromatography. The C22 and C23 fractions showed higher antifeeding ratio with 96 and 86%, respectively, and then these two fractions were re-isolated by ODS open column chromatography. As a result, both fractions in methanol 40% (v/v) showed antifeeding ratio over 90%. The C221 fraction showed insecticidal activity in all fraction, however, C231 fraction was showed the antifeeding activity only in C2311 fraction. The C2311 fraction judging to have antifeeding activity was re-isolated and purified by HPLC and recycling, and finally obtained the bioactive substances (C23111) with antifeeding ratio with 88%. The structure of bioactive materials isolated was confirmed by LC-mass and 1H-NMR(500 MHz), 13C-NMR(100 MHz).

To find an alternative for synthetic pesticides, methanol extract from plant samples were tested for their insecticidal activity against insect. The extract of Asiasarum sieboldii had strongly insecticidal activity against Plutella xylostella. Roots of A. sieboldii were extracted with methanol, and the concentrated extract was partitioned with n-hexane, ethylacetate, n-buthanol and H2O. The highest activity was shown in the hexane fraction. Activity-guided fractionation led to the isolation of two amides from hexane fration through the repeated silica gel column chromatographic separations. From the interpretation of spectropic data including NMR, MS, IR, the chemical structures of compounds were determined as dodeca-2E,4E,8Z,10Z-tetraenoic acid isobutylamide and dodeca-2E,4E,8Z, 10E-tetraenoic acid isobutylamide. These compounds showed insecticidal activity on P. xylostella by 96.7% at 100ppm. The liquid formulation controlled on cabbage effectively. The extract and compounds from A. sieboldii showed insecticidal activity against Nilaparvata lugens. As a naturally occurring pesticide, A. sieboldii could be useful as a new botanic insecticide.

The methanol extracts and essential oils from 9 medicinal plants, Ulmus davidiana var. japonica, Cryptomeria japonica, Hedera rhombea, Prunus mume, Taxus cuspidatal, Paulownia coreana, Kalopanax pictus, Paris verticillata, and Ixeris dentata were tested for their insecticidal activity against larvae of P. xylostella by topical application method. And the methanol extracts and essential oils obtained from Paris verticillata and Ixeris dentata were subjected to a screening test for their antifeeding activities, pupation and adult emergence of P. xylostella by a leaf-dipping method at a concentration of LC50 value. The methanol extract from all parts of P. verticillata and the essential oil from all parts of I. dentata exhibited potent activity with LC50 of 6.34 and 6.53(g/L) 5days after treatment. And the essential oil from all parts of P. verticillata and I. dentata have shown strong antifeeding activity compared to those of methanol extracts against larvae of P. xylostella. Also, the methanol extract and essential oil of P. verticillata and I. dentata gained 37.5%, 5.0% of pupation and 22.5%, 2.5% of adult emergence, respectively.

In this study, we compared global proteome profiles and the expression pattern of defense-related genes in Chinese cabbage when infested by Myzus persicae and Plutella xylostella. Four-week-old Chinese cabbage was exposed to each insect for 24 h, and then proteins and total RNA were extracted from leaves. To elucidate the herbivore-induced differentially expressed proteins in Chinese cabbage, proteins were separated by two-dimensional gel electrophoresis, and visualized by staining with Coomassie G250. Approximately 1600 protein spots were separated and 249 protein spots showed reproducible changes in expression. Among them, nine proteins whose expressions were markedly up-regulated in M. persicae-infested group were identified using matrixassisted laser desorption/ionization time of flight mass spectrometry. The identified herbivore-responsive proteins (ribulose-1,5-bisphosphate carboxylase/ oxygenase, ATP synthase CF1, putative mismatch binding protein Mus3, and integrase core domain-containing protein) were involved in regulation of photosynthesis, carbohydrate metabolism and DNA repair. The expression levels of chitinase, b-1,3-glucanse, peroxidase, PR1, and PR4 in herbivore-infested Chinese cabbage were analyzed by reverse transcription-polymerase chain reaction. The results clarify the response of Chinese cabbage to two herbivore attack at the protein level.

Parasitization by Cotesia plutellae inhibits pupal metamorphosis of diamondback moth, Plutella xylostella. Two questions are raised : (1) which parasitic factor(s) is responsible for the antimetamorphosis and (2) how the parasitized larvae are altered in endocrine signals. This study addressed both questions. When C. plutellae bracovirus (CpBV), a parasitic factor of the wasp, alone was injected to nonparasitized P. xylostella larvae, it significantly inhibited pupal metamorphosis in a dosedependent manner. Corpora allata (CA) and prothoracic gland (PTG) were compared in both nonparasitized and parasitized P. xylostella. In both groups, size and shape of CA were not different. However, PTG was detected on prothoracic tracheal trunk in nonparasitized larvae, but not detected in parasitized. CpBV injection to nonparasitized larvae inhibited the growth of PTG. Transcriptional factor, broad complex, was partially cloned and expressed in nonparasitized P. xylotella. In parasitized or CpBV-injected larvae, broad complex gene was not expressed during late larval stage.

An endoparasitoid wasp, Cotesia plutellae parasitized young larval of diamondback moth, Plutella xylostella. Parasitized larva exhibit significant immunosuppression and fail to metamorphose to pupal stage. Especially, during last instar of parasitized P. xylostella, massive nutrients divert from host to wasp development. HTIF (host translation inhibitory factor) encoded in C. Plutella bracovirus (CpBV) play a crucial role in suppressing host usage of amino acids. However, its inhibitory activity is selective by discriminating mRNAs based on their 5’UTR secondary structures. Our RT-PCR and proteomic analysis indicated that arginine kinase mRNA was inhibited by HTIF, but imaginal disc growth factor was not. Arginine kinase and IDGF were persistently expressed in parasitized P. .xylotella with the gradual decrease at the late parasitisation period. Expression of arginine kinase and IDGF were also tissue specific in the gut/epidermis and haemocyte but not in fat bodies. Subsequent analysis of these gene functions by RNA interference explained the benefit of parasitoid for the mRNA discrimination by HTIF.

This study was performed to investigate the antifeeding activity of Perilla frutescens extracts against diamondback moth, Plutella xylostella larvae and to confirm the electrophysiological responses of two sensilla (LST=lateral styloconic sensillum, MST=medial styloconic sensillum) in maxilla galea, a chemoreceptor. Crude extract of P. frutescens in methanol was showed antifeeding activity approximately 70%, and subsequently separated into four fractions - n-hexane(H), chloroform(C), ethylacetate(E), and water(W). Antifeeding activity was only showed in n-Hexane fraction around 99%. H1, H2, H3, H4, H5 were isolated from n-Hexane fraction using an open column chromatography, and the bioassay showed strongest antifeeding activity in H1 fraction. Using an oscilloscope, electrophysiological responses of two sensilla showed more seven times activity in MST. H11, H12, H13, H14 were separated from H1 fraction and antifeeding activity was showed highest in H11 fraction. H11 fraction was examined electrophysiological responses at doses of 100, 10, and 1 ppm, and MST of P. xylostella responded at a dose of 100 ppm. H11 fraction was separated using HPLC and identified using GC/MS and NMR. Finally, the structure of active compound proved to be farnesene with molecular weight of 204, and a formula of C15H24

The purpose of this work was to study the effects of electron beam irradiation on development and reproduction and evaluate the DNA damage in Plutella xylostella. Adults and pupae of P. xylostella were irradiated with 30, 50 and 100 Gy electron beam. Hatchability and fecundity of adults declined as increased irradiation doses. When pupae were irradiated with 100 Gy, fecundity of emerged adults significantly decreased and no eggs hatched. However, the adults longevity and emergence of pupae did not change. Assessment of DNA damage in cells obtained from adults and pupae of P. xylostella was carried out using single-cell electrophoresis (comet assay). Electron beam-radiated adults and pupae showed that tail length and percentage of DNA damage at all the doses were significantly larger than the control batch. Our results suggest that electron beam induces sterility through the DNA damage and this technique could contribute to analytical identification of an effective disinfestation and quarantine treatment.

Four major agricultural insect pests, Bemisia tabaci, Myzus persicae, Plutella xylostella and Tetranychus urticae, were irradiated with 30, 50, 70, or 100 Gy electron-beam. Longevity, egg hatching, emergence, and fecundity of the test insects were measured. Hatchability of B. tabaci, P. xylostella and T. urticae declined with increasing irradiation doses, and all B. tabaci and T. urticae eggs were dead at 100 Gy. When eggs of B. tabaci, P. xylostella and T. urticae were irradiated, hatch was inhibited. B. tabaci adults grown from 70 Gy irradiated eggs did not lay eggs. Fecundity of P. xylostella from the 100 Gy irradiated eggs decreased. When B. tabaci, P. xylostella, M. persicae and T. urticae nymphs/larvae were irradiated, the results were similar as those of the hatched eggs. When P. xylostella pupae were irradiated with 100 Gy, fecundity of emerged adults decreased and no eggs hatched. When B. tabaci, P. xylostella, M. persicae and T. urticae adults were irradiated with 70 and 100 Gy, fecundity decreased and egg hatch of B. tabaci was inhibited. However, the longevity of adults did not change and electron-beam irradiation of all developmental stages had no effect on the longevity of adults.

The diamondback moth, Plutella xylostella, is one of the most important pests of cole crops in the world and is the first insect to evolve resistance to Bt toxins in open-field populations. To search for useful molecular markers for Bt reistance monitoring, the PCR-based restriction fragment length polymorphism (RFLP) profiles of three aminopeptidase N (PxAPN1, PxAPN2 and PxAPN4) were determined for 15 representative regional field populations of P. xylostella. Most regional samples had similar RFLP patterns, whereas PxAPN1 from four regions and PxAPN4 from two regions showed different banding patterns after restriction enzyme treatment, but no differences were found in PxAPN2 among populations. The DNA sequence analysis revealed that a point mutation at the restriction site was responsible for the polymorphism of PxAPN1 but no mutations were observed in PxAPN4. Comparing amino acid sequences of PxAPNs from regional populations with reference PxAPNs (GenBank accession no. AAB70755) revealed that four regional populations possessed a point mutation in the Cry1A binding site of PxAPN1 and five regional populations possessed a deletion of eight amino acids in PxAPN4. These RFLP patterns were consistently observed in Southern regions of Korea, including Kyungsangnam-Do and Jeju-Do. The functional association of these RFLP with Bt resistance is currently under investigation

Diamondback moth (DBM, Plutella xylostella L.), the most destructive pest of cruciferous crops, is well-known as typical subtropical insect pest. A number of biological agents such as diseases, parasites and predators can affect populations of DBM in the fields negatively. In previous reports, we suggested Cotesia glomerata, Diadegma semiclusum, and Microplitis plutellae as promising natural enemies to DBM control at highland areas, but these species are larval parasitoids. In 2004~2009, we searched highland fields cultivating various cruciferous crops for PUPAL parasitoids which can supplement the unsatisfactory parasitism in the augmentative release of larval parasitoids. We obtained adults of hymenopteran parasitoid from DBM pupae in early July at Hoengseong region (asl 540 m), and then identified as Diadromus sp., although being a critical species so far. This parasitoid showed high rate of parasitism, about 13.2%, in mid October at the same region. Development period from oviposition to emergence of Diadromus sp. ranged from 14 to 18 days under 23℃ condition. Adult longevity, > three weeks, was longer when it was provided with 10% sugar solution as food than with water only or without food. Male adults mated as soon as emerging from parasitized DBM pupae, and laid eggs into DBM pupae for a week. Parasitism by Diadromus sp. was highest on 7th day after emergence. In contrast, lethality of DBM pupae which were not parasitized by wasps showed 60% on average for seven days. Parasitized DBM pupae could be stored at 10℃ for two weeks. The simultaneous augmentative release of larval parasitoids and pupal parasitoids can be an important component to integrated DBM management program in the future.

This study was conducted to investigate the insecticidal capacity of several recombinant baculoviruses to P. xylostella and S. exigua larvae. NeuroBactrus was constructed as follows: the cry1-5 of Bacillus thuringiensis 2385-1 was inserted into Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) genome by fusion of polyhedrin-cry1-5-polyhedrin under the control of polyhedrin gene promoter, and insect-specific neurotoxin from the scorpion Androctonus australis (AaIT) under the control of early promoter from Cotesia plutellae bracovirus was introduced by fusion of orf603 partial fragment in the opposite direction of polyhedrin gene, respectively. Other recombinant baculoviruses derived from the NeuroBactrus - NBt-DelA (deleted AaIT), NBt-Del5 (deleted cry1-5), and NBt-DelA5 (deleted AaIT and cry 1-5) - were manufactured in serial passages in vitro. The data were analyzed by SPSS. The value of LC50 was lower when P. xylostella larvae fed on cabbage coated with NeuroBactrus (4068.4) than when it fed on cabbage coated with AcMNPV (4.5x106). Survival time (ST50) of P. xylostella larvae (2.54days) was shorter when it fed on cabbage coated with NeuroBactrus than when it fed on cabbage coated with other recombinant baculoviruses (7.54days, 7.68days, and 8.26days) and AcMNPV (9.67days). S. exigua larvae presented the same results.

The life table statistics of diamondback moth on the transgenic Chinese cabbage (line: SKCP) with myrosinase gene was compared with that on the non transgenic Chinese cabbage (line: SC) at 25±1℃. Adult life span and number of progeny of P. xylostella on the SC and SKCP lines were similar to each other. The sex ratio, developmental period, intrinsic rate of increase and finite rate of increase of P. xylostella on SC and SKCP were not significantly different in both treatments (sex rati t=-1.60; df=220; P=0.1108, developmental period: t=-0.55; df=220; P=0.5803, intrinsic rate of increase: t=-0.11; df=45; P=0.9172). However, the finite rate of increase and net reproduction were significantly different in between SC and SKCP lines (finite rate of increase: t=2.26; df=45; P=0.0287, net reproduction: t=2.08; df=45; P=0.0442). This work was supported financially by Biogreen21 project of Rural Development Administration (No. 20070301-034-010)

The diamondback moth, Plutella xylostella is a one of the most important pests of various cruciferous crops and has a geographically wide ranging habitat. The heavy dependence on chemical pesticides has created severe pesticide resistance problems. In recent years, Bacillus thuringiensis product have been widely used for P. xylostella control bus genetic resistance in populations to some B. thuringiensis strains, compounded by cross-resistance to several different B. thuringiensis toxins, has also been identified. Such recent resistance problems serve to emphasize the urgent need for alternative control agents and their use within an integrated pest management approach. Baculoviruses have been used as agents for the biological control of certain insect pest species. the granuloviruses (GVs), based on the structure of the occluded virus and the occlusion body (OB). Several reports have showed P. xylostella granulovirus (PxGV) as a promise control agent for P. xylostella. However, it is very difficult to study GV because its OB, granule, has very small size and could be observed exactly under the electron microscopy (EM). This study was performed to develop rapid quantification method for granule of PxGV. After the exact quantification of granule with latex beads using EM, the universal extraction method of viral DNA was established for consistent experiment. The number of granules was calculated by the quantification of PCR products for granuline gene using spectrophotometer and densitometer. This novel calculation method for granule would be useful to study GV.

Bacillus thuringiensis (Bt) is effective to control the diamondback moth, Plutella xylostella. However, its relative slow and unstable control efficacy limits its wide use by farmers. To facilitate pathogenic rate of Bt, a bacterial mixture technique has been developed in this study. Two entomopathogenic bacteria, Xenorhabdus nematophila (Xn) and Photorhabdus temperata temperata (Ptt), possess high immunosuppressive activity against several lepidopteran insects. The mixture treatments using Bt + Xn or Bt + Ptt significantly enhanced Bt pathogenicity in median lethal concentration and time. Though live Xn and Ptt bacterial cells gave significant effect on the pathogenicity, their 48 h culture broth after removing the bacterialcells still possessed the synergistic effect on the Bt pathogenicity. The larvae fed with the bacterial culture broth suffered significant immunosuppression in response bacterial to infection

A polydnavirus, Cotesia plutellae bracovirus (CpBV), possesses segmented genome located on chromosome(s) of an endoparasitoid wasp, C. plutellae. An episomal viral segment (CpBV-S3) consists of 11,017 bp encoding two putative open reading frames (ORFs). ORF301 shows amino acid sequence homologies (28~50%) with RNase T2s of various organisms. It also contains BEN domain in C-terminal region. ORF302 is a hypothetical gene, which is also found in other bracoviruses. Both genes were expressed in larvae of Plutella xylostella parasitized by C. plutellae. ORF301 and ORF302 were transiently expressed in hemocyte, fat body, gut, and epidermis of P. xylostella. To analyze effects of these genes on the parasitism, the segment of CpBV-S3 was injected to non parasitized larvae of P. xylostella, in which the two genes were expressed at least for four days post-injection. The P. xylostella larvae injected with CpBV-S3 exhibited significant immunosuppression, such as reduction in total hemocyte population, suppression of immune associated genes including cecropin, pro-phenoloxidase (PO) and serpin1, and impairment in nodule formation behavior of hemocytes in response to bacterial challenge. Each gene expression in the treated larvae was inhibited by co-injecting respective double strand RNA (dsRNA) specific to each ORF. Injection of dsRNA of ORF301 could rescue the immunosuppression by the viral segmenttreated larvae, but not by ORF302 specific dsRNA. The larval injected with CpBV-S3 exhibited an enhanced susceptibility to baculovirus infection. These results indicate that ORF301 of CpBV-S3, which containing BEN domain, suppresses both cellular and humoral immune responses in P. xylostella.