주변 국가인 태국, 캄보디아, 베트남, 라오스 등에서 벼멸구(Nilaparvata lugens)와 흰등멸구(Sogatella furcifera)를 채집하던 중, 벼멸구와 형태가 아주 유사한 이삭멸구(N. muiri)와 벼멸구붙이(N. bakeri), 그리고 흰등 멸구와 형태가 아주 유사한 흰등멸구붙이(S. kolophon), 피멸구(S. vibix) 그리고 애멸구(Laodelphax striatellus)가 동시에 채집이 되는 등 형태적 차이점이 거의 없어 전문가도 쉽게 구분하지 못하는 문제가 있음이 확인되었다. 따라서 형태가 유사한 상기 멸구류의 종 동정을 확실히 할 수 있는 PCR용 프라이머의 개발을 위해 벼멸구 및 흰등멸구의 미토콘드리아 내 COI 영역을 특이적으로 검출할 수 있는 프라이머 세트를 제작하고 종 동정 효과를 확인하였다.

매년 국내로 비래해 오는 해충인 벼멸구는 그 기원이 중국 또는 중국 남부일 것으로 예상해왔으나, 이에 대한 유전학적 근거는 Mun et al. (1999)에 의해 제시된 세 가지 COI haplotype 비교가 유일하다. Mun et al. (1999)은 국내에 서 확인된 두 가지 haplotype 유형이 인도차이나반도 이남의 균일한 한 가지 haplotype 집단 유형과 중국에서 확인 된 또 다른 haplotype 집단 유형임을 근거로 국내 벼멸구의 기원을 중국으로 특정한 바 있다. 본 연구는 국내 및 동남아시아 5개국(부탄, 미얀마, 캄보디아, 라오스 및 태국)으로부터 직간접적으로 확보한 개체들을 대상으로 GBS (genotyping by sequencing) 및 NGS 기법을 통해 PCA를 포함한 다양한 집단유전학적 분석을 수행하였다. 그 결과 인도차이나반도의 벼멸구 집단은 크게 북부와 남부로 나뉘며, 국내 개체들은 북부에 비해 남부(캄보디 아, 태국)에 더 가깝다는 사실을 확인하였다. 따라서 벼멸구의 국내 비래는 중국으로부터의 기원 이전에 장마전 선이 형성될 무렵부터 인도차이나반도 남쪽의 고온다습한 서풍이 남남서풍으로 바뀌면서 중국 내륙을 거쳐 국내로 비래하는 경로를 따르는 것으로 보인다. 하지만 태안의 개체 중에는 인도차이나반도 집단들의 외군으로 확인되는 개체가 있었고, 이는 인도차이나반도 외의 샘플링되지 않은 다른 지역에서도 벼멸구가 국내로 비래할 수 있다는 가능성을 제시하였다. 따라서 국내로 유입되는 벼멸구의 유전적 기원을 확인하기 위해서는 인도차이 나반도 남쪽 지역에서 시작한 동아시아 여름 몬순의 바람이 한국으로 도착하는 경로에 위치한 다른 지역에서의 추가적인 샘플링 및 지속적인 관심과 추적이 필요할 것이다.

Nilaparvata lugens, brown planthoppers, is one of important pests on rice. Korean N. lugens is migrated from China and causing severe damages on rice in early September in Korea. For identifying biotypes of these N. lugens based on complete mitochondrial genomes, we completed mitochondrial genome of N. lugens captured in Hadong-gun, Gyoungsang-nam province in Korea. The circular mitogenome of N. lugens is 17,610 bp including 13 protein-coding genes, two ribosomal RNA genes, 22 transfer RNAs, and a single large non-coding region of 2,424 bp. The base composition was AT-biased (89.5%). 112 single nucleotide polymorphisms (SNPs) and 59 insertions and deletions are identified by comparing with Chinese N. lugens. Based on phylogenetic trees together with those of N. lugens captured in China, no clear phylogenetic relationship along with five biotypes, requiring more researches to achieve biotype identification including detailed analysis of sequence variations on mitochondrial genomes and whole genome analysis of N. lugens in near future.

Development, survival, and reproduction of brown planthopper (BPH) Nilaparvata lugens Stål (Hemiptera: Delphacidae), were studied in laboratory at 25±2°C, 65±5% RH and a 16L : 8D hours photoperiodism on five rice cultivars of: Dongjin 1ho, Chungchungbyeo, Jangseongbyeo, Chinnongbyeo and Jungmo 1045. BPH nymphs successfully survived on all rice cultivars, although survival rate was lowest on Jangseongbyeo (36.0%). Developmental time of immature stages ranged from 11.7±0.59 d on Jungmo 1045 to 12.8±0.59 d on Chinnongbyeo. Reproductive period and female longevity were longest on Dongjin 1ho, Chinnongbyeo and Jungmo 1045 while highest fecundity of N. lugens being observed on these three rice cultivars. Highest and lowest net reproductive rates were calculated on rice cultivars, Jungmo 1045 and Jangseongbyeo, respectively. Mean generation time was the longest on rice cultivar Dongjin 1ho. Respective descending order of intrinsic rates of population increase were on Jungmo 1045, Chinnongbyeo, Dongjin 1ho, Chungchungbyeo and Jangseongbyeo. These population parameters showed that N. lugens can successfully survive and reproduce on Chinnongbyeo and Jungmo 1045.

관주처리에 의한 벼멸구 흡즙 섭식 독성, 잎 침지처리에 의한 접촉독성과 식물체내 침투에 의한 섭식독성, 미량국소처리에 의한 표피 접촉 독성 등, 처리 방법에 따른 카보퓨란의 벼멸구에 대한 살충 작용 특성을 분석하였다. 카보퓨란 관주처리는 시간에 따른 벼멸구의 살충력 변화가 2 가지 패턴으로 나타났다. 상대적으로 높은 농도(8~30 ppm)의 처리에서는 급격히 사충율이 올라가다가 시간이 지남에 따라 증가율이 낮아지는 로그곡선의 형태와 낮은 농도(1~4 ppm)의 처리에서는 사충율 증가 패턴이 S 자 모양의 곡선 형태를 보였다. 관주처리 시 높은 농도에서는 처리 효과가 즉시 나타나 하루 만에 치사율이 40% 이상 도달하였으나 상대적으로 낮은 1~2 ppm의 농도에서는 처리 5일 후에야 80~100%에 달하는 사충율을 얻을 수 있었다. 침지처리는 사충율이 처리 1일 후 약효가 급격히 증가하고 이 후 증가율이 낮아졌다. 국소처리도 침지처리와 비슷한 경 향을 보이나 침지처리와는 달리 처리 후 3일 이후에는 사충율이 거의 증가하지 않았다. 약효 최적 조사 시간은 관주처리에서 처리 후 2일 (42시 간)이었으며 침지처리와 국소처리에서는 처리 후 1~2일이 약효 조사 적기였다.

Both Nilaparvata lugens Stål and Laodelphax striatellus Fallén has been known as the major rice pests which were irregularly migrated into Korea by wind current from China. We re-arranged and compared the resistance profiles to detect the existence convergent adaptation by using the previously published dataset (N. lugens at 2014, 17:711-716 and L. Striatellus at 2016, 19:247-252 in J. Asia Pac. Entomol., respectively). Both species commonly exhibited the high level of resistance to imidacloprids. Especially, the immigrant populations of L. Striatellus exhibited higher resistance level to indigenous populations, suggesting that they would settle down in Korea with insecticide resistance traits being predisposed. To detect the convergent adaptation to insecticide, pearson correlation analysis were employed by comparing mean dosage of LD50 and resistance ratio to nine insecticides. Strikingly, high and similar dosage responses were observed between the two species with high correlation coefficients of 0.928 (df=7, P <0.001) and 0.950 (df=7, P <0.001), respectively, in the comparison of LD50 vs. resistance ratio. This finding indicates that convergent adaption has occurred in both species through consistent selection by insecticides with similar usage patterns.

Nilaparvata lugens Stål is one of the important migratory pests of rice paddy fields in Korea. Resistance levels to nine insecticides were monitored in 12 local strains and correlation analysis was conducted to determine cross-resistance relationships among the tested insecticides. The local strains revealed 1.3- to 28.0-, 1.6- to 6.0-, 2.8- to 237.0-, 0.6- to 0.9-, and 0.7- to 1.3-fold resistance to carbamates, organophosphates, neonicotinoids, fipronil and etofenprox, respectively. Organophosphates revealed moderate correlations with benzofuranyl methylcarbamate (r = 0.566 - 0.614, p > 0.01). Three neonicotinoids were not correlated each other, but imidacloprid and clothianidin were moderately correlated with several benzofuranyl methylcarbamate and organophosphate insecticides (r = 0.590 - 0.705, p > 0.05), indicating that unknown common factors (such as detoxification enzymes) might contribute to resistance to both insecticides. Fipronil and etofenprox exhibited low levels of resistance and cross-resistance with other insecticides, suggesting their potential as an effective insecticide for field application. Resistance level monitoring and correlation analysis would be valuable for the selection of appropriate insecticides to control insecticide-resistant N. lugenes, a typical migratory pest in Korea.

Brown planthopper, Nilaparvata lugens is giving enormous damage to rice production. In this work, virulence of four Beauveria bassiana isolates against brown planthoppers was investigated by applying fungal granules on the water of pots, and we further examined the growth of hypha on the rice plants from the water surcae to explain the insecticidal mode of action. We used Beauveria bassiana (Bb) ERL 836, 1170, 1575 and 1578 isolates, which produced ~2×108 conidia/g of millet grain in a solid culture. Rice seeding was grown in breeding boxes at 28±2℃ for 5 days. Mycotized millet grains were treated on the water in a box at 1 g/box and the rice seeding was infested with 18~25 brown planthopper adults per box. A chemical pesticide and water treatment served as controls. Among the treatments, Bb ERL 836-treated plants had the lowest damage, rather than the other fungal treatments in laboratory assays. Hyphal growth on the stem of rice plants was observed in Petridish conditions under a fluorescent microscopy. This work suggests a possible control of brown planthoppers using entomopathogenic fungi.

We developed and characterized five polymorphic microsatellites of Nilaparvata lugens from hybridization method using biotin enrichment strategy and two polymorphic microsatellites from Next Generation Sequencing. Also 11 microsatellites that developed by Sun et al. (2011) are employed to carry out genetic analysis of N. lugens in Southeast Asia. The number of alleles per locus ranged from 2 to 12 with an average of 4.63 alleles per locus. The mean observed heterozygosity of the eleven populations ranged from 0.031 to 0.938 and the expected hetetrozygosity ranged from 0.031 to 0.881. Signifiant genetic differentiation was detected among the three N. lugens populations as the FST ranged from 0.028 (Cheong Do and Ha Long) to 0.161 (CH and BN). The results of microsatellite marker suggested that found N. lugens migrated to Korea at least two times in different period or once. Genetic distance of N. lugens between Korea and Hi Pong were mostly closed and genetic distance of Ha Long and HCM were relatively closed. In this study, development of microsatellites should facilitate the study of future population genetics of N. lugens, and eventually elucidate the route of N. lugens migration to Korea. Thus, combining satisfactory microsatellite markers and intensive surveillance methods in paddy field could be easy to understand to the N. lugens migration mechanism.

The resistance levels to carbamate (CB) and organophosphate (OP) insecticides were determined by topical application in 14 field strains of Nilaparvata lugens. The resistance levels to CB and OP were 1.3~47.5-fold and 1.4~14.4-fold higher than a susceptible strain, respectively. A quantitative sequencing (QS) protocol was established to determine the allele frequencies of four acetylcholinesterase point mutations putatively associated with CB and OP resistance. The allele frequencies of the four mutations (G119A, F/Y330S, F331H and I332L) in field strains ranged from ca. 0.0~51.7%, 0.0~88.9%, 2.5~47.7%, 5.1~56.0% and 6.7~57.3%, respectively. The F331H and I332L were tightly linked each other, suggesting these mutations may occur simultaneously. In the correlation analysis, G119A was not well correlated with actual resistance levels (r2= < 0.232), whereas F331H and I332L showed a better correlation with the resistance levels of benzofuranyl methylcarbamates (r2= 0.595). This finding indicates that F331H and I332L mutation frequencies may be used as molecular markers for detecting carbamate resistance in N. lugens. A QS protocol detecting the F331H and I332L mutation frequencies would be employed as a supportive tool for rapid monitoring of CB insecticide resistance levels in N. lugens

The resistance levels against carbamates (CB) and organophosphates (OP) were determined through bioassay and quantitative sequencing (QS) methods in 16 field populations of Nilaparvata lugens. The resistance levels to CB and OP by bioassay were 1.3~47.5-fold and 1.4~14.4-fold higher than a susceptible strain, respectively. The QS protocol was established to determine the allele frequencies of eight point mutations on acetylcholinesterase putatively associated CB and OP resistance. The allele frequencies of four mutations in local populations (G119A, F/S330Y, F331H and I332L) ranged from ca. 0.0~51.7%, 1.0~44.3%, 8.5~57.3% and 7.12~56.6%, respectively. The average prediction limits were –9.6±5.1~7.7±2.9%. The F330Y, F331H and I332L were tightly linked each other, suggesting these mutations may occur simultaneously. In the correlation analysis, G119A was not well correlated with both insecticides (r2= less 0.25), whereas F/S330Y, F331H and I332L showed better correlation with the resistance levels of carbamate (r2=0.590) than organophosphate (r2=0.235). This finding indicates that F/S330Y, F331H and I332L mutation frequencies are suitable for detecting carbamate resistance in N. lugens. QS will be applicable for the rapid monitoring of resistance levels to CB insecticides in N. lugens.

A carbofuran-resistant strain (CAS) showed ca. 41.1- and 15.1-fold resistance compared to a susceptible strain (SUS) and a non-selected field strain (FM), respectively. Enhanced activities of carboxylesterase and P450 were found as ca. 3- and 1.6-fold higher in CAS strain, suggesting these enzymes play a minor role in carbofuran resistance. Interestingly, the insensitivity of acetylcholinesterase (AChE) to carbofuran was revealed to be ca. 5.5- and 3.7-fold higher in CAS strain compared to, indicating that AChE insensitivity mechanism is associated with carbofuran resistance. In the western blot analysis, two kinds of AChEs were found and type-1 AChE (Nlace1) was identified as the major AChE in N. lugens. The open reading frame of Nlace1 is composed of 2,106 bp (ca. 78 Kd) and revealed 52.5% and 24.3% identity compared with Nephotettix cincticeps and Drosophila melanogaster, respectively. In the screening of point mutations, four amino acid substitutions (G119A, F/S330Y, F331I and H332L) were identified in the CAS strain that likely contribute to the AChE insensitivity. The allele frequencies of these mutations increased in the survived populations following the selection by LC50 of carbofuran, confirming that they are in fact associated with reduced sensitivity to carbofuran in N. lugens. These point mutation can be useful for the monitoring of resistance levels in conjunction with QS methods.

Role of flavonoids in plant against herbivores is well known. Many studies showed that flavonoids are deterrent or have anti-feeding effects to herbivores. In this study phenolic compounds were analyzed as defense materials to brown planthopper (BPH) feeding in four rice varieties using HPLC. Two of them, Chunambyeo and Nackdongbyeo, are BPH-susceptible, and the other two, Chungmo-1006 and Hanambyeo, are BPH-resistant. Changes in flavonoids were investigated by the function of damage period by BPH, Twelve flavonoids were characterized in the four varieties damaged by BPH. Among those flavonoids, apigenin and qurecetin were significantly higher in Chungmo-1006 damaged by BPH. Thus, resistance of Chungmo-1006 to BPH may be attributed to the increase of the two flavonoids contents.

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.