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Kafe 바로가기주관연구기관 | 서울대학교 Seoul National University |
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연구책임자 | 제연호 |
참여연구자 | 최재영 , 유금 , 푸젠리 |
보고서유형 | 최종보고서 |
발행국가 | 대한민국 |
언어 | 한국어 |
발행년월 | 2012-02 |
과제시작연도 | 2011 |
주관부처 | 농촌진흥청 |
과제관리전문기관 | 농촌진흥청 Rural Development Administration |
등록번호 | TRKO201400000770 |
과제고유번호 | 1395021125 |
DB 구축일자 | 2014-05-10 |
□ 과제명 : 해충 방제용 미생물 살충제의 현장 활용증진 기술개발
▶ 연구목적 :
○ 해충방제용 미생물살충제의 적절한 현장처리 모델 및 광범위 적용기술 구축
○곤충병원 미생물을 이용한 친환경 해충방제용 미생물살충제의 개발
○난방제 해충의 방제를 위한 새로운 고효율의 우수 농업 미생물살충제 자원 개발
▶ 주요연구내용 :
○ 1세부과제명 : 유통 세균살충제의 평가 및 신규 바이러스살충제의 제작기술 개발
- Bt제 5품목 살충효과 (배추좀나방, 파밤나방, 담배거세미나방) 검토 완료
- Bt제의
□ 과제명 : 해충 방제용 미생물 살충제의 현장 활용증진 기술개발
▶ 연구목적 :
○ 해충방제용 미생물살충제의 적절한 현장처리 모델 및 광범위 적용기술 구축
○곤충병원 미생물을 이용한 친환경 해충방제용 미생물살충제의 개발
○난방제 해충의 방제를 위한 새로운 고효율의 우수 농업 미생물살충제 자원 개발
▶ 주요연구내용 :
○ 1세부과제명 : 유통 세균살충제의 평가 및 신규 바이러스살충제의 제작기술 개발
- Bt제 5품목 살충효과 (배추좀나방, 파밤나방, 담배거세미나방) 검토 완료
- Bt제의 상온 경시안정성 평가완료
- 유통 Bt제의 문제점 파악 (나방제 해충에 대한 약효 미흡)
- 신규 바이러스 자원 SpliNPV, SpliGV 분리 및 특성규명 완료
- SpliNPV 유래 살충성 chitinase의 혼용 가능성 평가 완료
○ 1협동과제명 : 유통 곰팡이살충제의 평가 및 신규 곰팡이살충제의 제작기술 개발
- 곰팡이살충제 3품목중 확보 가능한 1개 제품 ‘보타니가드’의 유효성분 및 실내에서의 살충효과 (온실가루이) 검토 완료
- 곰팡이살충제의 보관 기간 및 온도에 따른 활성 변호 조사 완료
- 유통 곰팡이 살충제의 문제점 파악 (판매자, 사용자의 기피 및 제품에 대한 불신)
- 우리나라 전국 토양에서 신규한 곤충병원성 곰팡이 자원의 분리 및 특성규명 완료
- 난방제 해충에 대한 신규 곰팡이 살충제의 병원력 검정 및 선발 곰팡이 자원의 산업화를 위한 배양기술 및 제제화 기술 구축
○ 2협동과제명 : 유통 미생물살충제의 농약절감 모델 개발 및 신규 세균성 미생물 살충제의 제작 기술 개발
- Bt제와 화학농약의 교호 살포를 통한 농약 절감 모델 개발
- 진도 지역에서 미생물 살충제인 Bt제와 화학농약과의 교호살포를 통한 농약 사용 절감모델을 개발하였음.
- 신규 살충성 세균 (Bt) 자원 확보
- 담배거세미나방에 높은 병원성을 보이는 Bt. kurstaki KB099를 분리 확보하고 특성 규명을 하였음.
- Tannnic acid와 Bt. kurstaki KB100 균주와의 혼합처리를 통한 살충효과 향상.
○ 3협동과제명 : 과채류 해충방제를 위한 미생물살충제의 현장적용 연구
- 신규 병원성미생물의 가루이에 대한 살충력 검정
- 유통 미생물살충제(B. bassiana)의 대한 살충력 검정
- 미생물살충제(B. bassiana)에 대한 농자재의 효과 억제 시험
- 토마토 해충방제용 유통 미생물살충제의 약해평가
- 토마토 해충에 대한 화학농약과 미생물살충제 간의 혼용 및 로테이션 효과검증
○ 4협동과제명 : 엽채류 해충방제를 위한 미생물 살충제의 현장적용연구
- 등록된 미생물살충제 6종의 배추좀나방, 배추흰나비, 담배거세미나방, 도둑나방에 대한 살충효과 검정
- 화학살균제과 미생물살충제의 상호작용 평가
- 배추좀나방에 대한 B.t.제의 살충효과 추이 분석 및 B.t.제 살포간격 결정
- 배추좀나방에 효과적인 식물추출물(고삼추출물) 및 친환경농자재 선발
- 배추에서 B.t제와 선발된 친환경 농자재의 교호살포 효과 표장 검정
○ 5협동과제명 : 미생물살충제의 현장적용 및 효율증진 연구
- 유통 미생물 살충제 중 주요 해충에 대한 약효 평가 완료 (제품 또는 충에 따라 효과 상이)
- 곰팡이 살충제 보타니가드와 혼합 사용 가능 농약 (살균제 : 에타복삼, 코퍼하이드록사이드; 살충제 : 아바멕틴, 티아메톡삼, 디노테퓨란, 델타메트린 등; 식물추출물 : 새로이, 도열이, 파라핀153, 나비콘에프,응박사, 올캐치 등)
- 곰팡이 살충제와 친환경농자재 또는 화학농약 적정처리간격 구명 (활화산, 카메레온 7일간격, 은하수 4일간격 살포 필요)
Commercially available six Bacillus thuringiensis (Bt) products were bio-assayed against Plutella xylostella (diamondback moth), Spodoptera exigua (beet armyworm) and Spodoptera litura (Tobacco cutworm) in laboratory conditions. All the products had >80% of control efficacy 48 h post application. Bt
Commercially available six Bacillus thuringiensis (Bt) products were bio-assayed against Plutella xylostella (diamondback moth), Spodoptera exigua (beet armyworm) and Spodoptera litura (Tobacco cutworm) in laboratory conditions. All the products had >80% of control efficacy 48 h post application. Bt products, SC, TB-WP, TB-SO had excellent control efficacy aginst beet armyworms and SC and TB-WP against tobacco cutworms, but most Bt products had significantly lower control performance against older stages of larva.
Alternative strategies are necessary to control such pests, where Bt products are not practically available. All the Bt products were stable in maintaining their spore viability and control efficacy. No significant phytotoxicity was not observed. Tobacco cutworm is a noctuid moth that is considered an agricultural pest. The larvae feed on a wide range of plants and have been recorded on plants from 40 plant families (mostly dicotyledons). It is a major pest of many crops. To better understand Spodoptera litura granulovirus (SpliGV), the nucleotide sequence of the SpliGV DNA genome was determined and analyzed. The genome of the SpliGV was completely sequenced. The nucleotide sequence of the SpliGV genome was 124,121 bp long with 61.2% A+T content and contained 133 putative open reading frames (ORFs) of 150 or more nucleotides. The 133 putative ORFs covered 86.3% of the genome. Among these, 31 ORFs were conserved in most completely sequenced baculovirus genomes, 38 were granulovirus (GV)-specific, and 64 were present in some nucleopolyhedroviruses (NPVs) and/or GVs. We proved that 9 of the ORFs were SpliGV specific. The genome of SpliGV is 124,121 bp in size. One hundred thirty-three ORFs that putatively encode proteins of 50 or more amino acid residues with minimal overlap were determined. No chitinase or cathepsin genes, which are involved in the liquefaction of the infected host, were found in the SpliGV genome, explaining why SpliGVinfected insects do not degrade in a typical manner. The DNA photolyase gene was first found in the genus Granulovirus. When phylogenic relationships were analyzed, the SpliGV was most closely related to Trichoplusia ni granulovirus (TnGV) and Xestia c-nigrum granulovirus (XecnGV), which belong to the Type I-granuloviruses (Type I-GV).
The commercial mycoinsecticide 'BotaniGard' (BOT) was investigated its pathogenicity and storage stability. BOT was high pathogenicity against white fly, Trialeurodes vaporariorum in laboratory condition and stored at various condition (5, 25, 37°C and room temperature) retain viability of >87% for 6 months except 37°C condition.
Entomopathogenic fungal species have been investigated for their potential as the biological control agents due to their role of natural enemies for insects. To address the requirements of a potential in Korea based biocontrol effort using entomopathogenic fungi, we investigated the occurrence of various entomopathogenic fungi in 1080 soil samples representing from various areas and conditions in Korea. Entomopathogenic fungi were isolated from soils using selective medium containing dodine and antibiotics. After morphological selection and identification, the isolated fungi were identified finally using the internal transcribed spacer (ITS) region sequences of the nuclear ribosomal analysis.
As a result, entomopathogenic fungi were found to occur in 31.67% (342 isolates) of the soil samples studied. The most abundant species were Beauveria spp. (125 isolates) and Metarhizium spp. (82 isolates). The occurrence of entomopathogenic fungi were predominantly recovered more often from mountain and riparian area soils like as natural habitats than agricultural habitats. We isolated various pathogenic fungi to develop of mycoinsecticide against Tetranychus urticae, Myzus persicae, Spodoptera exigue using 342 isolate fungi from Korean soils or cadaver and then evaluated their virulence and thermotolerance to select best of one. Finally, we studied mass production and formulation of selected fungus Paceilomyces lilacinus 4-5 to develop eco-friendly mycoinsecticide against Myzus persicae.
Environmental friendly control model of beet armyworm for the reduction of pesticide use was established. Three kinds of combinations treated with chemical pesticides and microbial pesticides which were indoxacarb WP-chlorfluazuron EC-B.t. var. kurstaki CAB141, indoxacarb WP-methoxyfenozide+spinosad SC-chlorfluazuron EC-B.t. var. aizawai CAB109 and indoxacarb WP-methoxyfenozide+spinosad SC-chlorfluazuron EC-B.t. var.
kurstaki CAB141 were showed the more 78% of average mortality. It was showed that control effects of three combinations were the average 10% higher than single treatment, and the damage rates was reduced over 43% that meaned 10% more than single-treated.
It is supposed that these combination treatments might be minimized the development of resistance according to repetitive same chemical spraying, and the environmental friendly control methods with processing by reducing the number of times of chemical pesticides, can be control beet armyworm used by alternating sprays of chemical and microbial pesticides. The characteristics of endotoxin protein of Bacillus thuringiensis subsp. kurstaki KB099 strain showed a high bioactivity against Spodoptera litura that is one of the serious agricultural insect pests, and six genes, Cry1Aa, Cry1Ab, Cry1Ac, Cry1C, Cry1D and Cry1I, were confirmed. For the strengthen of insecticidal activity of Bacillus thuringiensis against beet armyworm that is one of the serious agricultural insect pests, it was selected tannic acid among some of the protease inhibitor that is the best of inhibitor of proteolytic degradation. In a field experiment with beet armyworm, in case of B. thuringiensis treatment alone, once, twice and three times of control effects with cumulative processing were 61.8%, 80.4% and 47.3%, respectively. However, in case of combined treatments with B. thuringiensis and 40 mM of tannic acid, their control effects were highly increased as 83.9%, 89.4% and 66.8%, respectively.
All experiments were carried out for investigation of physiological and biological characteristics of Lecanicillium lecanii Btab01 strain and microbial agents(Beauveria bassiana) and their insectcidal activity against major pests of tomato. In vitro bioassays to Bemisia tabaci nymphs, mortality was not significantly different either conidia (93.9%) or blastospore (98.3%), mortality and LT50 was significantly different following concentrations, 105cfu/㎖ (5.8%, 16.1d), 106cfu/㎖ (33.8%, 7.3d), 107cfu/㎖ (77.3%, 5.1d) and 108cfu/㎖ (98.5%, 3.5d). Mortality according to different R.H. conditions was made by hours of moist-chamber treatment, was appeared 0h (0%), 24h (20.3%) and 168h (100%). In moist-chamber treatment, Btab01 added oil (98.6%) was shown more high mortality than Btab01 alone (79.9%). Not in moist-chamber treatment, only “o”was appeared 25.7% mortality. Activity of commercial microbial agent(BOT) against whitefly decreased low temperature and low humidity. BOT was 79% and 35% of insecticidal activity against aphid at 100% and 60% humidity. BOT didn't damaged tomato at recommended concentration but it showed harmful effect at high concentration and at mixed treatment with other pesticides. Some fungicides inhibited growth of B. bassiana on PDA and its virulence against whitefly. Rotated treatment technique of BOT and insecticide show high control value against whitefly and could release chemical pesticide in the future.
Efficacy of several Bacillus thurigiensis commercial products (B.t.product) were tested on Plutella xylostella, Mamestra brassicae, Spodoptera litura in laboratory. As a result, the mortality rates of P xylostella and M. brassicae were more than 80% and 60% in all of B.t product treatments, respectively. The mortality rates of S. litura were, however, very low. The insecticidal activity of B.t. products were different according to formulation, the wettable powers of B.t. were more effective than the suspension concentrates. The insecticidal property maintained high until nine days after B.t. products sprayed. Although the colony of B. thuringiensis was not formed on tryptic soy agar medium plates containing antibiotics such as oxytetracycline, streptomycine, validamycin-A, oxolinic acid, bioassay with P. xylostella showed that the antibiotics did not affect the insecticidal activities of B.t. products. This is regarded that the antibiotics did not affect crystal toxins. Matrine which is extracted from Sophora flavescens had a higher insecticidal property to P. xylostella than Rotenon, pyrethrin, matrine, neem oil, canola oil. After comparing insecticidal proporty of six eco-friendly matarial products to P. xylostella, D-products was selected as a component of a alternating spray system to control P. xylostella. The alternating spry system showed high control efficiency to P. xylostella in chinese cabbage fields.
Most commercial Bt products in Korean market showed high control efficacy against
larva of diamond back moth, but there was no concentration increase effect to promote
mortality. The Bt products had low virulence against 4th instar of tobacco cutworm.
Susceptibility of Spodoptera exigua was comparatively high to most Bt products compared
with S. litura, and the mixture of each two products had higher control efficacy to S.
exigua than single product treatment. After storage of a mycopesticide BotaniGard (BOT)
at room temperature and 4℃ for 4 months the product had no viability reduction and
can kill tobacco whitefly, western flower thrips and onion thrips. Twelve chemical
pesticides which are mainly using in cucumber did not influence on the spore
germination and mycelial growth of B. bassiana GHA. Among 11 fungicides using cucumber
disease control ethaboxam and copper hydroxide was not influence on the viability of B. bassiana
GHA. Fenarimol and tetraconazol was very harmful to the fungal isolate. Azoxystrobin, dimethomorph and propineb impeded mycelical growth and spore germination of BOT spores regardless of both pesticide doses and mixing time. Among 25 eco-friendly products tested most product did not influence on B. bassiana GHA except the mixture of Japanese apricot + ginkgo nut extracts and the wettable sulphur powder. For effective use of mycopesticide with other control agents we conducted the study of the safe treatment period between mycopesticide and pesticides or eco-friendly agricultural materials using pest or disease control. When dimethomorph and fenarimol was sprayed 7 days before BOT treatment, whitefly control efficacy by BOT was high. Triflumizol, propineb and tetraconazol required only 1 day interval to control whitefly with BOT. The mixture of Japanese apricot + ginkgo nut extracts and the wettable sulphur powder required the interval of 7 days and the mixture of Japanese honeysuckle + jeffer sonia + Korean pasque flower extracts required 4 days interval to get high control efficacy with BOT.
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