초록 ▼

- 주요인시목해충 표본수집 및 형태분류 (세부 1)
•각 연차별 전국의 주요 산림지역 8개소 이상 채집조사 실시
•인시목 산림곤충 1,139종 6,113점의 표본제작 및 분류 동정을 통한 연구소재 확보
•정확한 분류동정을 위해 채집된 표본의 외부형태 및 해부검경 실시
•DNA바코드 작성완료 911종에 대한 종정보 데이터시트 작성완료
- 산림해충 DNA바코드작성 (세부 2)
•미량 시료를 대상으로한 mt DNA 추출 및 유전자 분석실시
•인시목 산림곤충 911종의 인시목 곤충 COI DNA sta

- 주요인시목해충 표본수집 및 형태분류 (세부 1)
•각 연차별 전국의 주요 산림지역 8개소 이상 채집조사 실시
•인시목 산림곤충 1,139종 6,113점의 표본제작 및 분류 동정을 통한 연구소재 확보
•정확한 분류동정을 위해 채집된 표본의 외부형태 및 해부검경 실시
•DNA바코드 작성완료 911종에 대한 종정보 데이터시트 작성완료
- 산림해충 DNA바코드작성 (세부 2)
•미량 시료를 대상으로한 mt DNA 추출 및 유전자 분석실시
•인시목 산림곤충 911종의 인시목 곤충 COI DNA standard 648 bp 바코딩 추출 및 분석
•인시목 산림병해충 DNA바코딩 DB화 방안 수립 - 기 확립된 standard DNA를 분석하여 Species-specific COI gene (약 501∼618 bp)을 DB화에 적용
- 산림해충 DNA바코드 예찰현장적용시험(세부 3)
•9개도 유아등 포획 인시류 828종 7,351개체 확보
•전국 우점비율 순위에 따른 50종 및 각 도별 10종의 우점종을 선정
•DNA 바코드로 외형적 혼동이 되는 미국흰불나방 개체들이 동종임을 확인하여 지역별 발생예찰조사를 통한 방제적기 판단에 활용
- 산림해충 증거표본관리 및 보존(세부 4)
•자나방과 및 밤나방과 소장표본 346종 2,194개체 동정재검토 및 목록작성 실시
•DNA 증거표본의 관리 및 보존방안에 대한 매뉴얼을 작성
•보유표본 관리체계 및 보존방안 사례조사 (4식)
•그 외 국외 문헌 등의 검색을 통한 기주식물 정보확보

Abstract ▼

Ⅳ. Results of research development
1. Collecting and classification of lepidopteran pests (Section 1)
We divided into two teams of collecting of forestry lepidopteran pests to obtain the materials for this study intensively and conducted the collect for the adult of pests. Collecting survey wa

Ⅳ. Results of research development
1. Collecting and classification of lepidopteran pests (Section 1)
We divided into two teams of collecting of forestry lepidopteran pests to obtain the materials for this study intensively and conducted the collect for the adult of pests. Collecting survey was conducted monthly from April to October which insects begin to emerge mainly. In the first year(2012), we carried out collecting survey 12 times in Gyeonggi Namnyangju area, Gangwon Pyeongchang, Yangyang, Samcheok, Jeongseon area, Chungbuk Boeun area, Chungnam Gongju area, Daejeon Yuseong-gu area, Jeonnam Wan-do area, Gyeongbuk Ulleung and Gyeongnam Millyang area. And second year(2013), we carried out collecting survey 13 times in Gangwon Inje, Hongcheon, Hoengseong, Samcheok area, Chungnam Gongju, Daejeon Yuseong-gu area, Jeonbuk Muju, Jeonnam Jangheung area, Gyeongbuk Ulleung and Gyeongnam Tongyeong area. Last year(2014), we carried out collecting survey 8 times in Gyeongnam Hapcheon-gun area, Incheon Mt.Gyeyang, Gyeongbuk Pohang and Ulleung-do. For accurate identification and classification of insects were collected, we examined related organization's sample.
This year we used common insect collecting survey method such as a light trap. The subject of this study were considered moth characteristic, which usually set at night. Therefore, we selected point where the vegetation of the forest area is good and easy to collect . When we carry out collecting survey, we install a light at night then collect the lepidopteran insect by vial tube (diameter 1cm × length 5cm) or poison bottle which attracted to light. Also we used middle size vial tube(diameter 3.5cm × length 11cm) to protect the sample from anesthetic. The bucket traps were additionally installed besides the light trap to obtain various species inhabiting the investigation area.
In the first year of research, we collected the main taxonomic group Geometridae and secured 305 species 2,023 specimens. The second year of research, we collected the main taxonomic group Noctuidae and secured 443 species 2,040 specimens. Last year of research, we collected a microlepidoptera and some other taxonomic groups. So we collected 403 species 2,020 specimens. Lastly we collected 1,139 species 6,113 specimens. To carry out this research, we performed morphological character and genital structure for the correct classification and identification. Then we made labels and a list of the completed sample.
From this research, the 1,139 collected species samples are selected for DNA barcode extraction. The analysis was carried out with secured samples. As a result, we successed to extract 911 species. Then we established the diagnostic information for forecasting and related species information based on DNA barcode, morphological character, ecological character and appearance time. Species which contain both morphological classification and DNA barcode data were selected as a information DB. The informations of the species are considered to be used as a basis for future DB construction, as well as to serve as an information sheet for pest diagnosis.

2. Forest pest DNA barcode extension (Section 2)
In this study, a rapid identification of the target gene was achieved, which is characterized by using only a minute amount of tissue sample (about 10∼25 mg from hind leg), and a universal primer pair (5′primer; 5'-GGTCAACAAATCATAAAGATATTGG-3' and 3′primer; 5'-TAAACTTCAGGGTGACCAAAAAATCA-3') to preserve the dried insect specimens. The experiment is accomplished by three steps; first, COI amplification by PCR with the universal primers for DNA library template of the corresponding insect tissue, second, COI sequencing by 3' primer, and thirdly, COI identification using 5' primer as an indicator. This approach was able to obtain the barcodes of mitochondrial COI DNA (648 bp) from a total of 911 Lepidopteran forest pest species which have been morphologically identified previously. and to construct the DNA barcode DB including the species-specific 558 bp of COI domain.
This achievement may lead a followed massive insect bar code and DB construction, and contribute to its effective and convenient application for the rapid and accurate identification and diagnosis of the unknown pest species occurred in forest as a preliminary demonstrational application has been successful to the population of fall armyworm, Hyphanria cunea.

3. Field application of lepidopteran forest pests monitoring techniques using DNA barcodes (Section 3)
Using light traps, approximately 7,351 macrolepidopteran individuals belong to 490 species were collected with the help of Forest Environment Research Institute located in 9 provinces from 2012 to 2014. A database was constructed based on “family name”, “scientific name (Korean name)”, “collection site”, “collection month”, “collection date”, “number of individuals”. Considering rank of dominant rate, 50 dominant species on a national scale and 10 local dominant species in each Province were selected, and made a list of target species for barcoding. In result of analysis of dominance rank on a national scale, the first dominant species was Spodoptera depravata during the whole survey period, and there were few differences between dominant rates of other species. In the first year of survey, the most dominant species of in each Province was Athetis dissimilis in Gyeonggi, Ivela auripes in Gangwon, Geometra dieckmanni in Chungbuk, Agrotis tokionis in Chungnam, Ambulyx japonica in Jeonbuk, Nordstromia japonica and Glyphodes perspectalis in Jeonnam, Spodoptera depravata in Gyeongbuk, Amphipyra livida in Gyeongnam, Lithosia quadra in Jeju Province. In the second year of survey, the most dominant species of in each Province was Niphonyx segregata in Gyeonggi, Orthosia limbata in Gangwon, Kunugia undans in Chungbuk and Chungnam, Theretra japonica in Jeonbuk, Deilephila elpenor in Jeonnam, Spodoptera depravata in Gyeongbuk, Obeidia tigrata in Gyeongnam, Lithosia quadra in Jeju Province. In the third year of survey, the most dominant species of in each Province was Rhamnosa angulata in Gyeonggi, Ivela auripes in Gangwon, Meteima mediorufa in Chungbuk, Epatolmis caesarea in Chungnam, Theretra japonica in Jeonbuk, Theretra japonica, Palpita nigropunctalis, Marumba gaschkewitschii and Marumba sperchius in Jeonnam, Athetis gluteosa in Gyeongbuk, Acronicta rumicis in Gyeongnam, and Phalera assimilis in Jeju Province. There has been some confusion on identification of fall webworm, Hyphanria cunea in monitoring site due to morphological variation (especially wing spot) of fall webworm adult. The 35 individuals of fall webworm adult which seemded to be morphologically different from each other were identified by same species with applying DNA barcode technique. This result was applied to monitoring site of fall webworm and the optimum time of control was determined by local monitoring survey with pheromone trap.

4. The management and preservation of the voucher specimens of forest insect pests (section 4)
We reviewed and re-identified the lepidopteran specimens which were deposited in the Insect Collection of Korea National Arboretum (KNA) and we also collected related data on management system for preservation of the voucher specimens that the DNA were extracted.
The studies on management of determined specimens were based on the specimens in the Insect Collection of KNA. We involved the newly found species in the studies.
In 2012, the first year of this project, we revised and re-identified 430 specimens of 50 species belonging to family Geometridae in KNA. As results, we found that the misidentification for 32 specimens and we added 150 newly identified specimens of 30 species.
In 2013, the second year of this project, we continued to revise and re-identify 670 specimens of 30 species of Geometridae, as well as, taxonomic revision of the lower taxa of the family. As results, we confirmed that six specimens were misidentified and we newly identified 1,394 specimens of 249 species. In addition, we revised the scientific name and the Korean name of the subfamily Geometrinae based on the recent systematic references on this subfamily. We found that 8 genera of 10 species were already changed.
In 2014, we revised and re-identified 464 specimens of 51 species belong to Noctuidae. As results, we found the one specimen of one species was misidentified and we newly identified 220 specimens of 65 species with a revision of the scientific name and the Korean name.
We had surveyed the references to establish the management system and preservation of determined specimens from several foreign institutions (e.g. National Museum of Natural History, Naturalis Biodiversity Center, Natural History Museum, London, England).
Consequently, we suggested a manual on a treatment and efficient management of specimens, preservation of determined specimens that were used for molecular experiments and we also showed a plan to construct a database on diagnostic information for identification of lepidopteran insects.
We hope that the results of this study will be helpful for identification, diagnosis of insect pests, biodiversity studies, biological studies. In addition, the results can be used as fundamental data to identify for monitoring of forest insect pests. Also, the results on management of voucher specimens that are used for molecular experiments will be very helpful for further studies and they we be used to identify and classify the insect pests.

목차 Contents

• 표 지 ... 1
• 제 출 문 ... 2
• 보고서 요약서 ... 5
• 요 약 문 ... 6
• SUMMARY ... 13
• CONTENTS ... 22
• 목 차 ... 24
• 제 1 장 연구개발과제의 개요 ... 26
• 제 1 절 연구개발의 필요성 ... 26
• 제 2 장 국내외 기술개발 현황 ... 27
• 제 3 장 연구개발수행 내용 및 결과 ... 29
• 제 1 절 주요인시목해충 표본수집 및 형태분류 ... 29
• 1. 현지조사 및 표본채집 ... 29
• 2. 채집표본의 분류·동정 및 유관기관 표본조사 ... 33
• 3. DNA 작성용 표본선정 및 진단정보 작성 ... 41
• 4. 연구과제 수행 중 현장적용 사례 ... 42
• 제 2 절 산림해충DNA바코드작성 (세부 2) ... 46
• 1. 연구개발 내용 및 방법 ... 46
• 2. 연구개발 결과 ... 51
• 제 3 절 산림해충 DNA바코드 예찰현장적용시험 (세부 3) ... 57
• 1. 전국적인 예찰조사 자료 수집 ... 57
• 2. 인시목 수집시료의 표본 확보 및 목록 정리 ... 57
• 3. 인시목 DNA바코드 예찰현장적용 ... 78
• 제 4 절 산림해충 증거표본관리 및 보존 (세부 4) ... 81
• 1. 증거표본 관리체계 및 보존방안 수립 ... 81
• 2. 산림생물표본관내 인시목 표본에 대한 자료구축 ... 84
• 제 4 장 목표달성도 및 관련분야에의 기여도 ... 99
• 제 1절 목표달성도 ... 99
• 1. 주요인시목해충 표본수집 및 형태분류 (세부 1) ... 99
• 2. 산림해충DNA바코드작성 (세부 2) ... 100
• 3. 산림해충 DNA바코드 예찰현장적용시험 (세부 3) ... 101
• 4. 산림해충 증거표본관리 및 보존 (세부 4) ... 102
• 제 2절 관련분야에의 기여도 ... 103
• 제 5 장 연구개발결과의 활용계획 ... 104
• 제 6 장 연구개발과정에서 수집한 해외과학기술정보 ... 105
• 1. 전문 연구문헌의 수집 ... 105
• 2. 연구재료 확보 및 현장 진단을 위한 샘플 처리기법 수집 ... 107
• 제 7 장 참고문헌 ... 110
• 끝페이지 ... 120