일천궁과 토천궁에서 발생하는 총채벌레류의 분자동정

정충렬,정대희,박홍우,김현준,전권석,윤정범 한국약용작물학회 2019 한국약용작물학회지 Vol.27 No.1

Background: Cnidium officinale Makino and Ligusticum chuanxiong Hort. are important medicinal crops in Korea. However, there is insufficient information on the identification of thrips, which attack these plants. Until now, one species of thrips has been recorded as a main pest. Methods and Results: To identify the thrips emerging in C. officinale Makino and L. chuanxiong Hort., these plants were independently cultivated in two local areas. Thirty individuals of each plant species were selected randomly and surveyed for the presence of thrips. After confirming the existence of thrips, 100 thrips individuals were collected from each crop using the beating method. To identify thrips species, we performed PCR-restriction fragment length polymorphism (RFLP)-based analysis using ITS2 primer sets. Six thrips species were identified: western flower thrips (Frankliniella occidentalis), flower thrips (F. intonsa), onion thrips (Thrips tabaci), chrysanthemum thrips (T. nigropilosus), chilli thrips (Scirtothrips dorsalis), and grass thrips (Anaphothrips obscurus). The proportion of these species differed between the host plant species. Conclusions: Six thrips species were major pests of two medicinal crops. Integrated pest management is required to control these thrips species, and will enhance the yield and quality of C. officinale and L. chuanxiong.

시설재배 국화에서 총채벌레의 종 동정 및 보독 바이러스 동시 검출을 위한 다중 진단법 적용

윤주연(Ju-Yeon Yoon),윤정범(Jung-Beom Yoon),서미혜(Mi-Hye Seo),최승국(Seung-Kook Choi),조인숙(In-Sook Cho),정봉남(Bong-Nam Chung),양창열(Chang Yeol Yang),Venkata Subba Reddy Gangireddygari 한국식물병리학회 2020 식물병연구 Vol.26 No.4

이번 연구는 국화에서 문제되는 총채벌레의 종 동정 및 보독 바이러스인 토마토반점위조바이러스(Tomato spotted wilt virus, TSWV)를 동시에 확인할 수 있는 진단방법을 개발하였다. 이는 총채벌레 1마리에서 추출한 핵산에 꽃노랑총채벌레및 대만총채벌레의 ITS2 부분에 특이적인 프라이머와 TSWV 외피단백질(N) 유전자 특이적인 프라이머를 동시에 넣어 reverse tran scription‒polymerase chain reaction을 수행하여 DNA를증폭시키는 방법으로 전기영동하여 각각 287, 367, 777 bp의DNA 단편의 크기를 비교함으로써 총채벌레의 종 동정 및 총채벌레의 TSWV 보독 여부를 동시에 확인할 수 있다. 충청남도 태안 및 경상남도 창원의 국화 시설하우스에서 총채벌레를 포집하여 총채벌레 우점종과 총채벌레의 TSWV 보독율을 조사한결과, 태안의 국화 시설하우스에서는 꽃노랑총채벌레가 83.7% 로 우점하고 있으며 채집된 총채벌레 중 72.9%가 TSWV를 보독하고 있었으며, 창원에서는 꽃노랑총채벌레가 92.2%를 차지하고 있으며 84.0%의 총채벌레에서 TSWV가 진단되었다. 이러한결과는 Frankliniella occidentalis가 우점종이며 온실의 국화 식물에서 TSWV의 전반에 중요한 역할을 한다는 것을 확인해준다. 이번 연구는 국화 시설하우스에서 총채벌레를 통한 TSWV 의 시설하우스내 유입시기 및 확산 경로 등 바이러스의 역학연구를 위한 간편진단법으로 활용 가능함을 예시해준다. We have developed a simultaneous diagnostic method that can identify both the species of thrips and tomato spotted wilt virus (TSWV) that are problematic in chrysanthemum plants. This is a method of amplifying DNA by performing reverse transcription-polymerase chain reaction by simultaneously adding primers specific to TSWV coat protein (N) gene and primers specific to the internal transcribed spacer 2 region of Frankliniella occidentalis and F. intonsa using total nucleic acid extracted from one thrips. The sizes of DNA fragments for TSWV, F. occidentalis, and F. intonsa were 777, 287, and 367 bp, respectively. These results showed species identification of thrips and whether thrips carrying TSWV can be simultaneously confirmed. Further usefulness of the simultaneous diagnostic method was made from greenhouse survey at chrysanthemum greenhouses in Taean (Chungcheongnam-do) and Changwon (Gyeongsangnam-do) to investigate the identification of thrips species and the rate of thrips carrying TSWV. Of thrips collected from the greenhouses, 83.7% thrips was F. occidentalis and 72.9% F. occidentalis carried TSWV in Taean. Similarly, the diagnostic method showed that 92.2% thrips was F. occidentalis and 84.0% F. occidentalis carried TSWV in Changwon. These results confirm that F. occidentalis is a dominant thrips species and the thrips species plays a crucial role in the transmission of TSWV in chrysanthemum plants in the greenhouses. Taken together, this study showed a simple diagnostic method for thrips identification and epidemiological studies of the timing and spread of TSWV through thrips in chrysanthemum greenhouses in South Korea.

Estimating optimum rate of chrysanthemum flower model traps to manage Frankliniella intonsa in strawberry greenhouse

Bishwo P. Mainali,Un Taek Lim 한국응용곤충학회 2008 한국응용곤충학회 학술대회논문집 Vol.2008 No.10

Chrysanthemum flower model trap (CFMT) developed to attract the flower thrips was evaluated for its optimal trap density that is required to manage the population of flower thrips in a strawberry greenhouse. CFMTs were installed in different rates per unit area (10m×5m), i.e., control (no trap), low (5 traps), medium (10 traps), and high (20 traps), in three different strawberry greenhouses at Namhoo, Andong. From each plot, 10 flowers were sampled every week from 16 May 2008 to 6 June 2008. Five traps were also randomly collected from each plot every week to record the number of flower thrips trapped. The seasonal number of both female and male thrips collected in the flowers was significantly lower in the plots with high density of traps compared to the control. In the plot with medium level of traps, less seasonal number of male F. intonsa were found in the strawberry flowers compared to the plot with low level of traps or control. When the numbers of thrips in the flowers were plotted against the numbers of thrips trapped in the CFMTs, significant correlation was found in female thrips only. However, there was no significant difference in seasonal number of male and female thrips trapped in the CFMTs randomly collected from the plots among the treatments. In conclusion CFMT used in medium or high density in the strawberry greenhouse reduced infestation of F. intonsa in the flowers and can be a good addition to integrated pest management tactics against anthophilous thrips.

강원도 아스파라거스 파총채벌레 방제를 위한 종합적 방제방안

전신재 ( Jeon,S. J. ),김삼규 ( S. K. Kim ) 강원대학교 농업생명과학연구원(구 농업과학연구소) 2019 강원 농업생명환경연구 Vol.31 No.2

아스파라거스를 가해하는 총채벌레류 해충을 방제하기 위한 방법 중 가장 효과적은 방법은 살충제를 처리하는 것이나 아스파라거스의 경우 매일 수확하기 때문에 수확기에는 살충제 처리가 용이하지 않다. 아스파라거스에 발생하는 해충을 방제하기 위해 적용가능한 살충제는 총 114종이 등록되어 있으며 그 중 총채벌레 방제용 살충제는 60종이 등록되어 있다. 봄 수확기에 살충제를 처리할 경우에는 등록된 약제 살포 3-5일 경과 후 잔류허용기준 또는 정량한계 미만으로 검출되므로 잔류특성이 검증된 약제 살포 후 수확 한 아스파라거스를 저온저장고에 3일 이상 저장 후 출고하면 아스파라거스 농약잔류 안전성에 문제가 되지 않는다. 끈끈이트랩을 이용한 총채벌레 방제시 노란색과 파란색 끈끈이트랩 중 노란색 끈끈이 트랩의 파총채벌레 유인효과가 뛰어났다. 이산화탄소를 이용한 총채벌레 방제의 경우 60% 농도, 20°C, 24시간 또는 40%, 24°C, 24시간 처리 조건일 때 파총채벌레 성충과 유충에 대한 높은 살충효과를 나타냈다. 아스파라거스를 저장하는 2-4°C의 경우에는 고농도의 이산화탄소를 72시간 처리에도 유충이 살아남아 방제효과가 높지 않았다. 곤충병원성 곰팡이인 Beauveria bassiana와 Lecanicillium attenuatum은 토양에서 자연적으로 발생하는 곰팡이로 약제 살포가 어려운 봄 수확기 이전 토양에 존재하는 총채벌레 번데기를 방제하여 초기 총채벌레 밀도를 감소시키는데 큰 도움을 준다. Thrips are among the most important pests of asparagus. The most effective method to control this pest is to insecticides spraying. Currently, a total of 114 pesticides have been registered for the control of pests of asparagus, 60 of which are registered for thrips control. However, crops are harvested every day, applying insecticides to control thrips is not ideal. Moreover, a recent study has shown that depending on the type of registered insecticide used, less than the residual or quantitative limit could be detected at 3-5 days after spraying. Therefore, if the asparagus is harvested after the application of insecticides, it should be stored in a cold chamber for 3 or more days before being moved to market. The use of sticky traps to control thrips is effective not only for monitoring them but also for reducing their numbers without applying insecticides. Yellow and blue sticky traps were usually used, but the yellow traps have been shown to attract more thrips than the blue ones. A recent study showed the high insecticidal effects of carbon dioxide on adult and larval thrips, at 60% concentration (20°C for 24 hours) or 40% concentration (24°C for 24 hours). Soap was also shown to have a high insecticidal effect after thrips-infested asparagus was immersed in soap water (diluted 1,000 times) for 15 minutes. Entomopathogenic fungi, such as Beauveria bassiana and Lecanicillum attenuatum, which occur naturally in soil, also help to reduce the initial thrips population by attacking the pupae in the soil in the spring harvest period.

상이한 지역별 노지 고추재배지의 총채벌레 연중 발생 및 토마토반점위조바이러스 발병

Abdisa Eticha,권지윤,진가현,김용균 한국응용곤충학회 2024 한국응용곤충학회지 Vol.63 No.1

Thrips infest hot peppers (Capsicum annuum) cultivating in open fields and give serious economic damages. This study reports their yearly occurrence from transplanting to harvest at three different places in Andong, an intense hot pepper-cultivating area. Two main occurrence peaks were detected in June and September. Two dominant thrips were the flower flowers, Frankliniella occidentalis and F. intonsa, which comprised of over 87% of the total occurring thrips. Other thrips did not follow the occurrence peaks of the two dominant species. Tomato spotted wilt virus (TSWV) was detected from the dominant two species, in which higher viruliferous rate was recorded in F. intonsa. Although the hot peppers were the resistant varieties against TSWV, some of them cultivating in the fields exhibited a characteristic disease symptom infected by the virus over the growing seasons. TSWV was isolated from the viruliferous thrips and assessed in NSs sequences encoded in S RNA segment of the virus. Compared to the known resistance breaking (RB) strains, the TSWV isolated from the viruliferous thrips in Andong did not show any RB mutations. 노지에서 재배되는 고추(Capsicum annuum)에 총채벌레가 발생하여 심각한 경제적 피해를 주고 있다. 본 연구는 고추 주산지인 안동을 중 심으로 상이한 세 지역의 재배지에서 고추 정식에서 수확 시기까지 총채벌레의 연중 발생을 보고한다. 총채벌레의 최대 발생기는 6월과 9월을 중 심으로 나타났다. 이 가운데 두 주요 총채벌레는 꽃노랑총채벌레(Frankliniella occidentalis)와 대만총채벌레(F. intonsa)였으며, 전체 총채벌레 발 생의 87% 이상을 차지하였다. 기타 총채벌레는 이상의 두 우점종의 발생 패턴과는 상이하였다. 토마토반점위조바이러스(Tomato spotted wilt virus: TSWV)가 이들 우점종 개체들에서 검출되었으며, 바이러스 보독율은 대만총체벌레에서 높았다. 이들 지역에서 재배되는 고추가 TSWV 에 내병성 품종들이지만, 일부 고추 개체들에서는 이 바이러스 감염에 따른 전형적인 병징을 보였다. 이에 보독충에서 TSWV를 분리하여 이 바 이러스 S RNA 게놈에 존재하는 비구조단백질 일종을 암호화하는 NSs 유전자의 염기서열을 분석하였다. 기존에 알려진 저항성붕괴(resistance breaking: RB) 계통의 서열에 비해 안동에서 발생한 보독충에서 분리된 TSWV의 서열과는 상이하여, 이 지역에서는 RB 돌연변이체가 존재하 지 않는 것으로 판명되었다.

Screening of lethal genes for feeding RNA interference against Frankliniella occidentalis (western flower thrips) using leaf disc-mediated dsRNA delivery

Seung Hee Han,Ju Hyeon Kim,Kyungmun Kim,Si Hyeock Lee 한국응용곤충학회 2019 한국응용곤충학회 학술대회논문집 Vol.2019 No.04

Frankliniella occidentalis is a major pest in agriculture. Following overuse of insecticides, high resistance has developed due to its high reproduction rate and short generation time. To control the resistant strains of the thrips, the ingestion RNAi- based control was established. A total of 67 genes were selected, and their double-stranded RNAs (dsRNA) were delivered to thrips via the leaf disc-feeding method. Among the genes screened, the dsRNA of Toll-like receptor 6 (TLR6) and coatomer protein subunit epsilon (COPE) resulted in the highest mortality (3.8- and 2.8-fold faster LT50 compared to control, respectively) when ingested by thrips. The dsRNA-fed thrips showed 53% and 83% reduced transcription levels of TLR6 and COPE, respectively. This result demonstrates that the observed mortality of thrips following dsRNA ingestion was due to RNAi, and this lethal genes can be employed as a practical tool to control thrips in the field.

Pantoea Bacteria Isolated from Three Thrips ( Frankliniella occidentalis , Frankliniella intonsa , and Thrips tabaci

Jin Gahyeon,Kim Yonggyun 한국미생물·생명공학회 2023 Journal of microbiology and biotechnology Vol.33 No.6

Gut symbionts play crucial roles in host development by producing nutrients and defending against pathogens. Phloem-feeding insects in particular lack essential nutrients in their diets, and thus, gut symbionts are required for their development. Gram-negative Pantoea spp. are known to be symbiotic to the western flower thrips (Frankliniella occidentalis). However, their bacterial characteristics have not been thoroughly investigated. In this study, we isolated three different bacteria (BFoK1, BFiK1, and BTtK1) from F. occidentalis, F. intonsa, and T. tabaci. The bacterial isolates of all three species contained Pantoea spp. Their 16S rRNA sequences indicated that BFoK1 and BTtK1 were similar to P. agglomerans, while BFiK1 was similar to P. dispersa. These predictions were supported by the biochemical characteristics assessed by fatty acid composition and organic carbon utilization. In the bacterial morphological analysis, BFoK1 and BTtK1 were distinct from BFiK1. All these bacteria were relatively resistant to tetracycline compared to ampicillin and kanamycin, in which BFoK1 and BTtK1 were different from BFiK1. Feeding ampicillin (100,000 ppm) reduced the bacterial density in thrips and retarded the development of F. occidentalis. The addition of BFoK1 bacteria, however, rescued the retarded development. These findings indicate that Pantoea bacteria are symbionts to different species of thrips.

Cuticular hydrocarbon pattern as a chemotaxonomy marker to assess six species of thrips

Chen Ting,Li Qunchen,Qi Guojun,Gao Yan,Zhao Chenyin,Lu Lihua 한국응용곤충학회 2020 Journal of Asia-Pacific Entomology Vol.23 No.4

Thrips constitute several families of slender insects with fringed wings and unique asymmetrical mouthparts. They have become globally important pests, infesting a variety of agriculturally important crops. Species of thrips are difficult to identify due to their small size and similarities in morphology. Recently, in addition to morphology, both molecular and non-molecular taxonomic tools have been used to identify species differences. Insect cuticular hydrocarbons have been widely used in chemotaxonomy. In this study, a Thermal Separation Probe was used to identify the cuticular hydrocarbons of Frankliniella occidentalis, Frankliniella intonsa, Thrips palmi, Thrips hawaiiensis, Haplothrips chinensis and Gynaikothrips ficorum. We analyzed the hydrocarbon com position of adults in all 6 species, and in the larvae of F. occidentalis, T. hawaiiensis and T. palmi. The results showed that the composition of cuticular hydrocarbons differed between species. All 6 species of adults and 3 species of larvae were easily distinguishable by quantitative analysis of hydrocarbon profiles. These results provide a possible method for the identification of thrips.

Fungal library for developing biopesticides to control thrips

Dongwei Li,Sihyeon Kim,Jong Cheol Kim,So Eun Park,Mi Rong Lee,Sehyeon Baek,Tae Young Shin,Jae Su Kim 한국응용곤충학회 2018 한국응용곤충학회 학술대회논문집 Vol.2018 No.10

Thrips is one of the serious insect pests in many economic crops, in which general managements rely mainly on chemical insecticides. However, the overuse of chemicals led to the development of insect resistance and environmental residual issues. Therefore, thrips management requires the consideration of alternative strategies which are less harmful to the environment and working on different target points. In this work, we isolated entomopathogenic fungi from soil, and identified them with morphological and molecular biology methods, followed by a preliminary virulence assay against Tenebrio molitor larvae. Selected fungal isolates were adjusted to 1×107 conidia/ml for an indoor virulence assay against thrips adults, and highly virulent isolates were added to a thrip-pathogenic fungal library. Biological characteristics of efficacious isolates were investigated comparing to a previously commercialized fungal isolate. This entomopathogenic fungal library could be used as a valuable resource for developing effective strains to control thrips in agricultural fields.

Building of entomopathogenic fungal library to manage melon thrips, Thrips palmi Karny

Dongwei Li,Sihyeon Kim,Se Jin Lee,Mi Rong Lee,Jong Cheol Kim,So Eun Park,Tae Young Shin,Jae Su Kim 한국응용곤충학회 2019 한국응용곤충학회 학술대회논문집 Vol.2019 No.04

Melon thrips, Thrips palmi Karny (Thysanoptera: Thripidae) is a worldwide polyphagous pest. The management of this pest have mainly relied on chemical agents. However, the overuse is harmful to the environment and results in insects resistance. Therefore, alternative eco-friendly control methods that have different mode of action, such as biological control, are necessary to overcome the current issue. In this study, we isolated entomopathogenic fungi from Korean soil, and characterized them via morphological and molecular techniques and pathogenicity assay against Tenebrio molitor larvae. The isolated fungi were screened for virulence against T. palmi under laboratory conditions and the results were used to establish a thrips-pathogenic fungal library. The highly virulent isolates were selected and further characterized for optimum culture conditions and application as biopesticide in the field.