보고서 정보
주관연구기관 |
국립농업과학원 National Institute of Agricultural Sciences |
보고서유형 | 최종보고서 |
발행국가 | 대한민국 |
언어 |
한국어
|
발행년월 | 2016-02 |
과제시작연도 |
2015 |
주관부처 |
농촌진흥청 Rural Development Administration(RDA) |
등록번호 |
TRKO201600003146 |
과제고유번호 |
1395041348 |
사업명 |
농업기초기반연구 |
DB 구축일자 |
2016-06-25
|
DOI |
https://doi.org/10.23000/TRKO201600003146 |
초록
Ⅳ. 연구개발결과
- 외래해충 미국선녀벌레의 집단 유전자 구조 분석을 위한 mtCO1, SSR 등 유전자 마커 영역 유전정보 확보(국내, 미국, 유럽) 및 이동 경로 추정
- 외래해충 미국선벌레 구기(mouthparts)의 미세구조 및 주변 감각기 분포 정보
- 미국선녀벌레에서 7 개의 EPG 섭식파형 분리 및 섭식행동 정량적 분석 기술 개발
- 외래해충 미국선녀벌레에 살충효과가 있는 식물추출물 선발 및 제형화를 통한 약효 검증(인삼밭에서 계피정유 이용시 80% 방제효과)
Abstract
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The citrus flatid planthopper, Metcalfa pruinosa Say (Hemiptera: Flatidae), is a very polyphagous native insect in North America and currently a serious pest in Europe and South Korea. In order to clarify there relationships among different populations of M. pruinosa from South Korea and foreign cou
The citrus flatid planthopper, Metcalfa pruinosa Say (Hemiptera: Flatidae), is a very polyphagous native insect in North America and currently a serious pest in Europe and South Korea. In order to clarify there relationships among different populations of M. pruinosa from South Korea and foreign countries, 251 COI sequences from 251 samples collected from six countries, South Korea, France, Italy, Spain, Slovenia, and USA were analyzed, together with 7 published COI sequences (Canada). A total of 19 haplotypes was detected from the 258 COI sequences, and three haplotypes, hap-1, 3, and 9, were detected in South Korea. The hap-1 was from most samples of seven populations of South Korea, together with samples from the USA, France, Italy, Slovenia, and Spain. Hap-3 was found in a few GG population samples and some from France, USA, and Slovenia, and the hap-9 was found in the GG population and USA. The MJ network revealed the population of M. pruinosa may have undergone specific genetic-level variation and this species has genetically spread in South Korea. In the BI tree, the 258 COI sequences were divided into three groups, and their phylogenetic analysis suggested that most of individuals of M. pruinosa in South Korea are closely related to populations in Italy, Spain, Slovenia, France, and USA. We investigated the population genetic structure and estimated migration route of M. pruinosa among thirty locations across South Korea using six microsatellite markers developed by Kim et al. (2015). The number of alleles per locus ranged from 6 to 36 with an average of 21.33 alleles per locus. The mean observed heterozygosity of the thirty population ranged from 0.03 to 1.000 and expected heterozygosity ranged from 0.404 to 0.850. A population assignment test using the first-generation migrant detection method suggested that nationwide dispersal of M. pruinosa may have occurred across South Korea. By using developed primer sets of microsatellite, population genetic studies could be better understand the migration routes and the population structure of M. pruinosa.
Survival rates of M. pruinosa during total nymphal stage in 25℃ temperature condition were two times higher in 14:10 (L:D) photoperiod than in 12:12 (L:D), but developmental duration of nymphs and longevity of adults were not significantly different in both photoperiod conditions. Time to reach cumulative 50% hatching rate of overwintered-eggs in field was investigated as from mid May to late May in South Korea. In comparison of host preference and suitability of M. pruinosa among maize, perilla, pumpkin, bell pepper, and sweet potato, perilla showed the highest, but bell pepper and sweet potato were the lowest. Types of chemosensitive sensilla in the labium tip and internal and external structure of stylets in M. pruinosa were investigated in this study. In the tip of labium, 20 and 26 chemosensitive sensilla were observed from 2nd ∼ 3rd nymphal stage and 4th ∼ 5th, respectively. To understand the feeding behaviour of M. pruinosa, stylet penetration behaviour of M. pruinosa was investigated with an electrical penetration graph (EPG) system. This study reports seven EPG waveforms related to M. pruinosa feeding behaviour: np (no stylet penetration), Mp1 (initiation of stylet penetration), Mp2 (stylet movement and salivation), Mp4 (phloem feeding), Mp4-H (honeydew excretion), Mp5 (xylem feeding) and Mp6 (unknown). To determine respective feeding behaviour related to the Mp4 and Mp5 waveforms, stylets were cut with a laser beam, and the location of the stylet tip within plant tissue was examined. We found plant sap was exuded from the severed stylets only when the Mp4 waveform was observed, suggesting phloem sap ingestion. The stylet tip was located in the xylem region, indicating xylem-feeding activity, when the Mp5 waveform was observed. The analysis of 24 different EPG parameters suggests that M. pruinosa stylets reached the vascular bundle of a plant within ca. 5 min and spend ca. 70% of the time feeding on xylem and phloem feeding. This is the first study that reports seven distinctive EPG waveforms with respect to the feeding behaviour of M. pruinosa which could help determine host specificity and host plant susceptibility.
The insecticidal activity of 120 plant essential oils and control efficacy of six experimental spray formulations (SF) containing 0.25, 0.5, 1, 2.5, 5, and 10% of the selected oils was examined against both nymph and adult of the citrus flatid planthopper, Metcalfa pruinosa using direct contact applications (leaf dipping and spray). Reponses varied according to dose (1,000 and 500 mg/L). When exposed at 1,000 mg/L for 24 h using leaf dipping assay, 19 essential oils showed strong mortality (100%) among 120 essential oils screened. At 500 mg/L, 100% mortality was observed in cinnamon technical, cinnamon green leaf, cinnamon #500, cassia tree, citronella java and pennyroyal followed by origanum, thyme white, grapefruit, savory, fennel sweet, aniseed and cinnamon bark showed considerable mortality (93.3-80%) against nymphs of M. pruinosa. The moderate mortality (73.3-60%) was found in thyme red, tagette, calamus, lemoneucalptus and geranium. Oils applied as SF-10% sprays provided 100 % mortality against adult M. pruinosa. One hundred mortalities were achieved in cinnamon technical at >SF-0.5 formulation, in cinnamon #500, cinnamon green leaf and penny royal at >SF-2.5. The toxicity of the cinnamon oil constituents and 21 cinnamaldehyde related compounds to the nymph of M. pruinosa was evaluated using a leaf dipping bioassay. Based on 24 h LC50 values, hydro-cinnamicacid (1.55 mg/cm3) was the most toxic compound, followed by geranic acid (1.59 mg/cm3). The LC50 of 11 compounds including cinnamaldehyde is between 1.60 and 4.94 mg/cm3. Low and no toxicity were observed with the other 15 compounds (5.24–13.47 mg/cm3) and 2 compounds, respectively. Also the toxicity of the cinnamon oil constituents and 21 cinnamaldehyde related compounds to the adult of M. pruinosa was evaluated using a direct spray method. The toxicity of eugenol (10.81 mg/L) was the most toxic compound against adults of M. pruinosa, followed by geranic acid (30.68 mg/L). The LC50 of 9 compounds including cinnamaldehyde, is between 59.16 and 96.70 mg/L. Low and no toxicity were observed with the other 15 compounds (105.44–255.76 mg/L) and 3 compounds, respectively.
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