풀무치에 대하여 살충활성을 보유한 곤충병원성 진균의 생리활성 평가

이미롱,김종철,이세진,김시현,이석주,박소은,이왕휴,김재수,Lee, Mi Rong,Kim, Jong Cheol,Lee, Se Jin,Kim, Sihyeon,Lee, Seok Ju,Park, So Eun,Lee, Wang Hyu,Kim, Jae Su 한국응용곤충학회 2017 한국응용곤충학회지 Vol.56 No.3

풀무치 (Locusta migratoria) (Orthoptrea: Acridiade)는 전 세계적으로 작물 생산에 심각한 문제를 야기하는 돌발 해충이다. 그러나 우리나라의 경우 풀무치를 방제하기 위한 방제제 및 적용에 대한 연구는 미흡한 실정이다. 본 연구에서는 풀무치에 병원성을 갖는 풀무치병원성 진균 라이브러리를 구축하였으며, 풀무치 방제에 이용 가능한 생물학적 방제제로서의 가능성을 평가 하였다. 먼저 갈색거저리 유충-baiting 시스템을 이용하여 다양한 지역에서 채집된 토양에서 곤충병원성 진균을 발굴 하였다. 풀무치 병원성 검정을 진행하기 위하여 국립 농업 과학원에서 풀무치를 분양 받았으며, 고체 배양된 곤충병원성 진균을 곤충 사육 상자에 처리하여 (2 g/box), 풀무치 약충 (3-4령충)에 대한 곤충병원성 진균의 병원성을 평가 하였다. 그 결과 곤충병원성 진균 처리 3-7일차에 풀무치의 머리, 복부, 다리 표면에서 진균이 증식하는 mycosis를 확인 할 수 있었다. 특히, Metarhizium anisopliae, M. lepidiotae, Clonostachys rogersoniana에서 높은 병원성이 나타나는 것이 확인 되었다. 확보된 34개의 풀무치병원성균주의 특성을 파악하기 위하여 열안정성 및 포자생산성을 확인 한 결과, Paecilomyces, Purpureocillium 균주가 다른 균주에 비해 열에 대한 높은 안정성안 나타나는 것을 확인 하였으며, 대부분의 균주에서 $1{\times}10^8conidia/gram$ 이상의 포자수를 생산 하는 것을 확인 하였다. 또한 온실 조건에서 비교적 병원성이 높았던 M. anisopliae 고체 배양된 균주를 토양에 처리하여 병원성을 확인한 결과, 85.7%의 높은 방제효과를 확인 할 수 있었다. 본 실험을 통하여 풀무치가 이동하면서 토양에 정착된 곤충병원성 진균에 접촉되어 치사 될 수 있을 것으로 판단되며, 효과적인 풀무치 방제가 가능 할 것이라고 판단된다. Locusts, Locusta migratoria (Orthoptera: Acrididae) are periodical unpredictable agricultural pests worldwide and cause serious damage to crop production; however, little consideration has been given to the management of this pest. Herein, we constructed a locust-pathogenic fungal library and confirmed that some fungi could be used as resources for locust management. First, the entomopathogenic fungi were collected from sampled soils using a Tenebrio molitor-based baiting system. For the locust assay, a locust colony was obtained from the National Institute of Agricultural Science and Technology. A total of 34 entomopathogenic fungal granules, which were produced by solid cultures, were placed in the plastic insect-rearing boxes (2 g/box) and nymphs of locust were contained in the box. In 3-7 days, mycosis was observed on the membranous cuticles of the head, abdomen, and legs of locusts. In particular, Metarhizium anisopliae, M. lepidiotae, and Clonostachys rogersoniana exhibited high virulence against the locust. Given that the 34 isolates could be used in field applications, their conidial production and stability (thermotolerance) were further characterized. In the thermotolerance assay, Paecilomyces and Purpureocillium isolates had higher thermotolerance than the other isolates. Most of the fungal isolates produced ca. >$1{\times}10^8conidia/g$ on millet grain medium. In a greenhouse trial, the granular application of M. anisopliae isolate on the soil surface resulted in 85.7% control efficacy. This work suggests that entomopathogenic fungi in a granular form can be effectively used to control the migratory locust.

Long-term storage stability of Beauveria bassiana ERL836 granules as fungal biopesticide

김종철,이미롱,김시현,이세진,박소은,백세현,Leila Gasmi,신태영,김재수 한국응용곤충학회 2019 Journal of Asia-Pacific Entomology Vol.22 No.2

Beauveria bassiana (Balsamo) Vuillemin (Hypocreales: Cordycipitaceae) ERL836 has been commercialized under the name ChongchaeSak to control an agricultural insect pest, the western flower thrips Frankliniella occidentalis Pergande (Thysanoptera: Thripidae), in the Republic of Korea. As soon as it was launched in 2017, it became a popular product and has received a positive response. However, study of the storage stability of the fungus ERL836 has yet to be investigated. To determine the optimum conditions for long-term storage, we assessed conidial viability and insecticidal activity of B. bassiana ERL836 according to storage temperature and culture substrate. Viability of B. bassiana ERL836 conidia from mycotized grains (millet and rice) stored at low (4 °C) and moderate (25 and 30 °C) temperatures was maintained at > 85% for 24 and 18 months, respectively, along with insecticidal activity. In contrast, the samples stored at 37 °C showed low germination rate (about 80% germination rate for only 5 months). This result suggests that low and moderate temperatures (4 to 30 °C) conserve B. bassiana ERL836 viability and virulence.

Tenebrio molitor-mediated entomopathogenic fungal library construction for pest management

김종철,이미롱,김시현,이세진,박소은,나유신,이관석,신태영,김재수 한국응용곤충학회 2018 Journal of Asia-Pacific Entomology Vol.21 No.1

Entomopathogenic fungi are soil-dwelling microorganisms that infect host insects, and some have been used asbiological control agents. Construction of an entomopathogenic fungal library provides a strong platform for thedevelopment of highly effective biopesticides, but the traditional method using antibiotic media has low isolationefficiency. Herein, to increase the efficiency of isolation, a Tenebrio molitor pathogenicity-based fungalcollection method was used for the construction of a library. The isolation efficiency using the T. molitor larvaebasedpathogenicity assay was 55.4%, which was significantly higher than the method of antibiotic medium withdodine (6.0%). We named it the Jeonbuk (Chonbuk) National University Entomopathogenic Fungi (JEF) library. It consisted of 279 isolates belonging to 12 genera and 29 species. Main features of the species-representative 15isolates were characterized in terms of morphology, virulence, conidiogenesis, thermotolerance and productionof biologically active materials such as virulence-related enzymes. Some of the species-representative isolatesshowed higher virulence against T. molitor, Riptortus pedestris and Plutella xylostella in laboratory conditions. Theproduction of enzymes such as chitinase, Pr1 protease and lipase all related to pathogenesis in LB medium washigher than in the SDB and PDB media. In the thermotolerance assay, a Purpureocillium lilacinum isolate showedhigher thermotolerance than other isolates. This work reports an efficient library method using T. molitor forbuilding a library for development of fungal biopesticides for pest management.

Solid cultures of thrips-pathogenic fungi Isaria javanica strains for enhanced conidial productivity and thermotolerance

박소은,김종철,이세진,이미롱,김시현,Dongwei Li,백세현,한지희,김정준,구경봉,신태영,김재수 한국응용곤충학회 2018 Journal of Asia-Pacific Entomology Vol.21 No.4

Entomopathogenic fungi have great potential to control agricultural and horticultural insect pests, however optimizing conidial production systems to demonstrate high productivity and stability still needs additional efforts for successful field application and industrialization. Although many virulent entomopathogenic fungal isolates have been viewed as potential candidates in a laboratory environment, very few of the isolates are being used in practice for application in agricultural fields as commercial products. I. javanicus is an entomopathogenic fungus that is parasitic to various diverse coleopteran and lepidopteran insects and thought good candidate as biopesticdes. In this work, the basic characteristics of two entomopathogenic fungi, I. javanica FG340 and Pf04, were investigated in morphological examinations, genetic identification, and virulence against Thrips palmi, and then the feasibility of various grains substrates for conidial production was assessed, particularly focusing on conidial productivity and thermotolerance. Isaria javanica FG340 and Pf04 conidia were solid-cultured on 12 grains for 14 days in a Petri dish. Of the tested Italian millet, perilla seed, millet and barley-based cultures showed high conidial production. The four-grain media yielded > 1 × 10 9 conidia/g of I. javanica FG340 and Pf04. Pf04 strain had enhanced thermotolerance up to 45 °C when cultured on Italian millet. In application, it was easy to make a conidial suspension using the cultured grains, and several surfactants were tested to release the conidia. This work suggests several possible inexpensive grain substrates by which to promote conidial production combined with enhanced stability against exposure to high temperature.

식물 및 미생물 유래 유기농자재 살충효과: 단감해충 감꼭지나방, 톱다리개미허리노린재

김종철,유정선,송민호,이미롱,김시현,이세진,김재수,Kim, Jong Cheol,Yu, Jeong Seon,Song, Min Ho,Lee, Mi Rong,Kim, Sihyeon,Lee, Se Jin,Kim, Jae Su 한국응용곤충학회 2016 한국응용곤충학회지 Vol.55 No.4

단감 해충의 효과적인 방제를 위하여, 현재까지 주로 화학 살충제가 사용되어 왔으나, 최근 화학 살충제의 과다 사용은 해충 저항성과 농약 잔류 문제라는 심각한 문제를 야기하고 있다. 본 연구에서는 친환경 유기농자재 (Eco-Friendly Organic Pesticides; EFOP)의 주요 단감 해충인 감꼭지나방과 톱다리개미허리노린재에 대한 살충 활성을 검토하였다. 현재 유통되고 있는 친환경 유기농자재 11 종을 실험실 조건에서 대상 해충에 분무 처리하였다. 대조 약제는 화학 살충제인 buprofezin+dinotefuran (20+15) 수화제를 사용하였다. 감꼭지나방에 대한 유기농자재의 살충 효과를 평가하는 과정에서, 감꼭지나방 유충의 개체수 부족 문제로 인하여 배추좀나방 유충을 1차 실험충으로 사용하였으며, 세 종류의 친환경 유기농자재를 사전 선발하였다. 선발된 친환경 유기농자재는 목초액 (EFOP-1), 회화나무, 양명아주, 멀구슬나무의 혼합추출물 (EFOP-2), Bacillus thuringiensis subsp. aizawai NT0423 (EFOP-11)이었으며, 선발된 유기농자재의 감꼭지나방 유충에 대한 살충력 실험 결과, EFOP-2의 처리구에서 생충율이 27.7% (5 일차), 13.3% (7 일차), 6.7% (10 일차)로 가장 높은 살충 효과를 보였다. 톱다리개미허리노린재에 대한 살충 효과 평가에서는 회화나무, 양명아주, 멀구슬나무의 혼합추출물 (EFOP-2, EFOP-9)과 고삼, 데리스의 혼합추출물 (EFOP-10)이 높은 살충 효과를 보였다. 특히 EFOP-2의 처리구에서 생충율이 20.0% (5 일차), 16.7% (10 일차)로 가장 높은 살충 효과를 보였다. 결론적으로, 친환경 유기농자재인 EFOP-2를 이용하여 단감의 주요 해충인 감꼭지나방과 톱다리개미허리노린재를 포함한 다양한 노린재 해충에 대한 방제가 가능할 것으로 판단된다. Chemical pesticides have been used to control persimmon pests, however the overuse of the pesticides caused insect resistance, followed by failure in pest management and residual problems. Herein we investigate the potential of eco-friendly organic pesticides (EFOP) on the control persimmon pests, Stathmopoda masinissa (persimmon fruit moth) and Riptortus pedestris (bean bug). Ten commercially available plant-derived organic pesticides and one microbial pesticide were sprayed on the target insects in laboratory conditions. The chemical pesticide, buprofezin+dinotefuran wettable powder served as a positive control. In the first bioassay against persimmon fruit moth, alternatively Plutella xylostella larvae were used due to the lack of persimmon fruit moth population from fields, and three organic pesticides showed high control efficacy, such as pyroligneous liquor (EFOP-1), the mixture of Chinese scholar tree extract, goosefoot and subtripinnata extracts (EFOP-2) and Bacillus thuringiensis subsp. aizawai NT0423 (EFOP-11). When the three selected organic pesticides were treated on the persimmon fruit moths, the EFOP-2 treatment showed the highest control efficacy: 27.7% (5 days), 13.3% (7 days) and 6.7% (10 days) of survival rates. In the bioassay against bean bugs, the mixture of Chinese scholar tree, goosefoot and subtripinnata extracts (EFOP-2 and EFOP-9) and the extracts of sophora and derris (EFOP-10) showed high control efficacy, particularly the highest in the treatment of EFOP-2: 20.0% (5 days) and 16.7% (10 days) of survival rates. These results suggest that the mixture of Chinese scholar tree, goosefoot and subtripinnata extracts (EFOP-2) has high and multiple potential in the management of the persimmon pests.

Management of pine wilt disease vectoring Monochamus alternatus adults using spray and soil application of Metarhizium anisopliae JEF isolates

김종철,Se Jin Lee,김시현,이미롱,Sehyeon Baek,박소은,김준헌,Tae Young Shin,Jae Su Kim 한국응용곤충학회 2020 Journal of Asia-Pacific Entomology Vol.23 No.1

Chemical control is widely used to control the Japanese pine sawyer beetle, Monochamus alternatus, but strong chemical regulations require an environmentally sound management strategy. In this work, we investigated the use of entomopathogenic fungi and their application as a means of practical pest management. Thirty-two diverse species of fungal isolates were assayed against adult pine sawyer beetles using a contact method under laboratory conditions, and four isolates showed over 70% virulence consequently. These isolates, two each of Beauveria bassiana and Metarhizium anisopliae were sprayed on the adult beetles at 1 × 10 7 conidia/ml in plastic containers, respectively. The M. anisopliae-treated adult beetles showed 67% mortality. M. anisopliae isolates JEF-197 and JEF-279 demonstrated dosage-dependent insecticidal activity. Following the laboratory experiments, semi-field trials were conducted in young pine trees under high (RH 94%) and low (RH 35%) humidity conditions. In the high humidity conditions, most of the adult beetles stayed on the top of the branches. When the two M. anisopliae isolates were sprayed on the beetles, they showed ca. 50–70% insecticidal activity 11 days after application. In contrast, in low humidity conditions, the adult beetles tried to move off the branches and onto the soil. When the beetles reached the JEF-197 and JEF-279-treated soil, we measured > 90% insecticidal activity. This work suggests that M. anisopliae was the most virulent entomopathogenic fungus against adult Japanese pine sawyer beetles, and this forest insect could be ecologically controlled by the spray and soil application of the M. anisopliae isolates.