DNA bacoding is a popular DNA diagnostic technique especially for specific and generic level identification during various quarantine activities. BOLD Systems, an internet accessible DNA barcode data portal, currently includes over 12,000 DNA barcodes for the fruit fly family Tephritidae, of which 8,940 barcodes are open to public. These data represent 655 tephritid species, majority of which are pest species. These figures are rapidly increasing as the tephritid barcoding research is increasing every year. Therefore, BOLD has established as the most important DNA barcode source for tephritid identification. It is, therefore, also important to understand the limitations and problems of the DNA barcoding analysis. We here discussed the following potential problems and their possible solutions: (1) misidentification of species listed in BOLD; (2) sibling species with identical or near-identical barcodes; (3) NUMT (nuclear mitochondrial DNA) or pseudogene; and (4) introgression by hybridization between closely related species.

금지급 과실파리 침입시 대응조치(박멸프로그램) 실행과정에서 박멸프로그램의 종료는 3세대기간 동안 미포획되는 상황을 기준으로 하고 있다. 본 연구에서는 카람볼라과실파리(Bactocera carambolae), 구아바과실파리(B. correcta), 남방고추과실파리(B. latifrons), 작은퀸즐랜드과실파리(B. neohumeralis), 일본귤과실파리(B. tsuneonis) 등 5종에 대한 기존 온도발육자료를 바탕으로 세대기간 추정에 필요한 적산온도 모형을 제시하였다. 각 종의 세대기간 추정에 필요한 발육 영점온도와 적산온도는 칼람볼라과실파리 11.8℃와 384.6 Degree days(DD), 구아바과실파리 11.6℃와 454.5 DD, 남방고추과실파리 10.4℃와 517.5 DD, 작은퀸즐랜드과실파리 9.2℃와 467.8 DD로 추정되었다. 일화성인 일본귤과실파리의 경우는 월동용의 우화모형(발육영점은 11.3℃, 625 DD)을 이용하는 방법을 제안하였다.

Monitoring is an important component in the detection, surveillance and eradication of invasive insect pests. In particular, the availability of useful attractants and effective traps is critical in establishing a new surveillance program for quarantine pests. A powerful, species-specific attractant is desirable when precision early detection is needed. However, such an ideal attractant is not often available when the introduction of a new surveillance program is urgent. In reality, the combination of available monitoring means is adopted, which in turn requires the subsequent development of optimum monitoring strategy with these available means. Various species of fruit flies, belonging to the genus Bactrocera in particular, are serious threat to Korea, and developing systematic monitoring systems is needed against the invasion of the fruit flies. Effective male attractants, such as methyl eugenol and cue lure, and traps are currently available for some species such as B. dorsalis, B. tryoni, and B. carambolae. However, such attractants are yet to be developed for many other fruit fly species. The strong attraction of the striped fruit fly, B. scutellata, being present in large numbers in Korean fields to cue lure can be a potential obstacle in monitoring other species of fruit flies attracted to the same compound. Various attractants based on protein hydrolysates are also useful in monitoring fruit flies since they are attractive to both males and females of fruit flies. However, it is desirable to develop these protein-based baits into more species-specific attractants. Monitoring techniques for some important quarantine fruit flies are further discussed.

외래 과실파리에 대한 모니터링은 검역에 필수적이다. 과실파리 유인물질을 왁스 재질로 제형화하여 비교적 장기간 모니터링 할 수 있는 기술이 금지급 과실파리 모니터링에 유효하였다. 본 연구는 이 왁스 제형의 유용성을 확장시키기 위해 금지급 과실파리가 자생하고 있는 태국 현지에 적용하였다. 포획된 개체들을 분자 동정한 결과 기존에 유인력이 입증된 오리엔탈과실파리(Bactrocera dorsalis)와 오이과실파리(Zeugodacus cucurbitae)는 물론이고 Zeugodacus caudata의 검역 과실파리가 유인된다는 것을 밝혔다.

우리나라는 1996년 WTO/SPS(동식물검역협정)에 따라 검역병해충제도를 정비하였으며, 해외병해충의 위험으로부 터 자국내 생물자원과 자연환경을 보호하기 위하여 다양한 노력을 하고 있다. 현재까지의 수입식물 검역과정에서 검출된 병해충을 분석한 결과 병해충 검출건수는 수입량이 늘어나면서 증가하는 추세이다(00년 6,233건 → 17년 12,749건). 하지만 식물의 수입 유형에 따라 곡류, 사료류 등 비재식용 식물의 경우 병해충 검출률은 07년 이후 감소하고 있으며(07년 9.8% → 17년 3.3%), 묘목류, 종자류 등 재식용 식물의 경우 11년 이후로 지속적으로 증가하고 있어(11년 7.9% → 17년 22.0%) 재식용 식물을 통한 해외 병해충의 유입 위험도가 가증되고 있는 상황이다. 따라서 우리는 검역본부에서 운영하고 있는 자체 전산시스템인 병해충정보시스템(PIS)을 이용하여 2000년 이후 식물검역과 정에서 발견된 병해충 검출동향과 수입 유형에 따른 검출동향을 비교분석하고, 2016년 대비 2017년 병해충 검출동향을 파악하였다.

감귤그린병(Citrus Huanglongbing Disease)은 전 세계적으로 감귤산업에 가장 큰 피해를 주는 병해로 국내 미분포종인 귤나무이(Diaphorina citri Kuwayama)가 매개충으로 알려져 있다. 바이러스에 감염된 나무는 5~12년내에 대부분 고사하며 열매는 크지 못하고 색이 들지 않는 증상을 보여 상품성을 저해시킨다. 현재 접목, 묘목, 매개충등의 유입으로 전 세계적으로 확산되고 있는 실정이다. 귤나무이의 국내 분포가능성이 꾸준히 제시되면서 유입 대비를 위한 예찰방법 및 동정법 개발이 필요한 실정이다. 제주도에는 총 33종이 분포하는 것으로 알려져 있으며(권, 1983; Cho & burckhardt, 2017) 2016년부터 지금까지의 조사로 11종의 나무이와 10종의 기주식물을 확인되었다. 귤나무가 속해있는 운향과를 기주로 하는 나무이는 발견되지 않았다.

지난 18년간(2000~2018년 3월) 검역현장에서 검출된 해충의 종류는 1,980종이다. 이중 딱정벌레목은 전체의 42.6% (곤충은 1,707종으로 86.2%)로 검역적으로 매우 중요하며, 여기에서 가는납작벌레과(Silvanidae)는 딱정벌레목 머리대 장상과에 포함되고, 이들의 검출 종은 21종, 횟수는 15,516회로 가장 많은 비율을 차지한다. 최근 18년간 검출률 결과 Ahasverus sdvana(쌀머리대장)로 6,404회로 가장 많이 검출되었으며, Slivanus didentatus(두니가는납작벌레)는 6,225회, Oryzaephilus surinamensis(머리대장가는납작벌레)는 1,123회, Oryzaephilus mercator(곡물가는납작벌레)는 415 회의 검출률을 보여준다. 가는납작벌레과는 딱정벌레목중 가장많은 검출률을 보이는 과로 검역현장에서 매우 중요한 과에 속한다. 본 연구는 검역과정에서 검출되는 가는납작벌레과 곤충 중 검역현장에서 최초로 발견된 Monanus 속의 한 종을 발견하여 보고 한다.

Craspedoxantha genus has been distributed in worldwide as 7 species in Afrotropical and 2 species in Oriental Regions. It is generally known that C. marginalis is preferred Asteraceae including Vernonia spp. for host plant. However, C. marginalis was first found in cut-flower of Phaenocoma prolifera, which was imported from South Africa, under the plant quarantine inspection in Korea. Therefore, we first report that P. prolifera as a new host plant for C. marginalis and provide information of the morphological characteristics and DNA barcoding sequences on male and female for identification.

The Lucid technology as an interactive fully illustrated identification tool, has developed and assisted in the diagnosis of agricultural and quarantine pests and invasive organisms over the past years. In this study, lepidopterans specifically relating to quarantine pests were selected for a lucid key development based on the border inspection data from 1996 to 2016 in South Korea. They belong to four families: Sphingidae, Noctuidae, Pyralidae and Crambidae, and also categorize into the three groups: regulated, potential regulated and non-quarantine pests. The key includes diagnostic characters from head, thorax, fore- and hind-wings for each species examined.

In the 21st century, the risk of the exotic pest being invaded is increasing due to the rise of trading activities. Oriental fruit fly, Bactrocera dorsalis (Diptera: Tephritidae), has been considered to very destructive invader as it is highly intrusive and has a wide host range. In this study, 2,299 samples were collected from 88 locations in 12 countries by quarantine and investigation. Among them, 608 individuals were used for COI DNA barcoding analysis based on Neighbor Joining method with P-distance model. Population genetics analysis was conducted for 510 individuals selected from 47 locations of 12 Southeast Asian countries using 15 microsatellite markers. COI DNA barcoding results showed that B. dorsalis did not support any clade as a geographical isolation but almost indicated mixed populations of each country. Population genetics analysis showed similar pattern of genetic structure between neighboring countries across borders. Most domestic quarantine groups were more similar in genetic structure to Taiwan, China and Thailand in the order of appearance.

The nematode Bursaphelenchus xylophilus and its vector insect Monochamus alternatus are major quarantine pests of timber products in Korea. Due to the phaseout of methyl bromide as a fumigant for plant quarantine and pre-shipment treatments, the need for an alternative fumigant is essential. Based on preliminary studies on the efficacy of ethanedinitrile (C2N2) to these two pests, three quarantine trials on logs covered with PVC-tarpaulins were conducted at three dosages and three temperatures potential for inhalation exposure was assessed by monitoring atmospheric levels of C2N2 in relation to the threshold limit value (TLV, 10 ppm, v/v). Concentration × time products (Ct) of 398.6, 547.2 and 595.9 g h m-3 were obtained for each quarantine trial. A 100% mortality of B. xylophilus and M. alternatus larvae at 23±4°C and 10±4°C occurred with a load factor of pine logs of 46% and at 3±1°C with a load factor of 30%. During all fumigant procedures, atmospheric levels of C2N2 20m downwind from the PVC tarpaulin were below the TLV. During aeration, levels 10 m and 5 m downwind were below the TLV after a maximum of 0.4 h and 1 h, respectively. For the purpose of an emergent quarantine or phytosanitary treatment, specific doses of C2N2 at the temperatures that occurred during fumigation could control B. xylophilus and M. alternatus larvae without significant inhalation risk to workers.

Plant quarantine is preventing the introduction and adaption of exotic pests in homeland, and then eventually promoting and improving domestic agriculture and forestry. Therefore, we conducted a survey of intercepted pests on hand-carried plants from all over the world to Korea. Totally 7,291 of hand-carried plants had been inspected by quarantine inspectors at 2017. Gimpo, Daegu, and Incheon International Airport showed more inspection cases than other areas by November 10, 2017, meanwhile, Gimhae and Incheon International Airport and Incheon Harbor showed more intercepting pest rate than other areas. In Animal and Plant Quarantine Agency, inspector found 490 cases of intercepted pests on 32 countries including China with 116 imported goods.

과실파리류는 국내 60종의 검역 해충 가운데 42종을 차지하고 있다. 이들 대부분은 자연계 산물인 methyl eugenol (ME) 또는 raspberry ketone에 특이적으로 유인된다. 이들 유인물질을 파라핀 왁스를 이용하여 제형화하였고, 이를 대만에서 야외 유인 효과를 검증하였다. 이들 왁스 제형은 모두 델타트랩에 설치하고 8월 말경에 모니터링 한 결과 ME 왁스 제형은 오리엔탈과실파리(Bactrocera dorsalis)를 매일 60-80 마리 포획하 였다. 반면에 Cuelure 왁스제형은 오이과실파리(Bactrocera cucurbitae)와 B. tau를 유인하였다. 이러한 결과는 본 왁스제형이 이들 검역 해충에 대 한 국내 모니터링에 응용될 수 있다는 것을 제시하고 있다.

The spotted wing drosophila (SWD), Drosophila suzukii, is classified exotic quarantine pest in Australia and EU, soSWD-free must be supported by evidence of surveys and phytosanitary measures in exported host agricultural commoditiesin Korea. From a quarantine control point of view, SWD is importantly considered as model insect pest for exotic fruitflies (Bactrocera dorsalis) in Korea as well because of similarity in ecological cycles. In evaluations of ethyl formate(EF) only and combined cold treatment to kill eggs and larvae of SWD, the combined EF fumigation applied at LCt50%(50% killed lethal concentration X time) and cold treatement (5℃) for > 5 days showed the promise to new conceptfor eradicating quarantine pest and these could be helpful to pre-develop exotic fruit fly management in Korea.

The bulb mite (BM), Rhizoglyphus echinopus, is hard to control in the harvested onion and garlic. Methyl bromide (MeBr) fumigation is the only option to eradicate BM at present. However, MeBr causes phytotoxic damage (index 3) to onion and garlic and highly toxic to fumigators. To replace MeBr, we evaluated ethyl formate (EF) and phosphine gas (PH3) to BM at 5℃ which is current practical fumigation temperature. The >140 g·h m-3 of EF for 24 h could completely control BM and this CT (concentration × time) product was slightly (index 1) phytotoxic to onion and garlic. Fumigation of 2 g PH3 required >120 h exposure to kill 100% of BM without any pyhtotoxic damage. Further researches are being planned to shorten the exposure time with synergistic effect between EF and PH3.

Rapid growth in trade of agricultural products has increased the risk of introduction of exotic pests into new area. In order to fulfill quarantine obligations, several treatments are in practice: fumigation with methyl bromide (MB), application of extreme temperatures, controlled atmospheres, and combinations of them. Although MB was most effective and widely applied for quarantine treatment, its use was banned by the Montreal Protocol. Ionizing irradiation phytosanitary treatments – gamma radiation, electron beam, and X-ray are being used as an alternative to MB currently. For applying ionizing irradiation as a quarantine treatment, there is a research protocol which should be followed, International Standards for Phytosanitary Measure 18 [ISPM #18] and Phytosanitary Treatment Regulation of Korea (Notice 2014-35). We have evaluated the effect of gamma radiation, electron beam, and X-ray on H. armigera as quarantine treatments. In this presentation, ISPM #18 and Notice 2014-35 will be talked with the example of H. armigera.

Tomato ringspot virus (ToRSV)는 Group IV(+) sense ssRNA viruses, Nepovirus속으로 분류되며 식물병원성을 가진 국내 미보고 관리급 검역바이러스이다. 본 연구에서는 우리나라 검역현장에서 ToRSV를 검사하기 위한 reverse transctiption(RT)-polymerase chain reaction(PCR)과 nested PCR 진단시스템을 개발하였다. ToRSV 특이적으로 진단할 수 있는 RT-PCR은 프라이머 조합9 (F120/R20, 549 bp)와 조합31(F60/R80, 741 bp)]이며, 각각의 nested PCR 결과 439와 363 bp를 증폭할 수 있다. 한편, 본 연구에서 개발한 유전자변형-양성대조구는 실험실 오염으로 인한 거짓양성반응을 확인할 수 있다. 본 연구에서 개발한 ToRSV 검역진단시스템은 향후 식물검역에 기여할 것으로 기대된다.

With the increasing trend of global trades and protection of agro-ecosystem in importing and exporting countries, quarantine and pre-shipment(QPS) fumigation for perishable commodities are critical in terms of quality maintenance of fumigated commodities as well as eradicating quarantine pest. There are limited use of methyl bromide(MB) fumigation on fruits and vegetables due to not only worker safety issue at low temperature condition but phytotoxic damage to valuable commodities. We are here presented that commercial development of methyl bromide (MB) alternatives such as ethyl formate and phosphine gas formulated with carbon dioxide, that are increasing consumer and worker safety and avoiding phytotoxic damage in routine MB fumigation.

Pelargonium zonate spot virus (PZSV)는 group IV (+) ssRNA viruses, Bromoviridae에 속하는 식물 병원체로, 일반적으로 토마토, 국화, 아티초크 및 제라늄에 감염된다. 본 연구는 검역 현장에서 PZSV를 신속하고 특이적으로 진단 할 수 있는 PCR module을 개발하는 것을 목적으로 하였다. PZSV를 검출하기 위한 RT-PCR 프라이머 선발 결과, 각각 513 및 320 bp를 증폭하는2개 조합을 선발하으며, 더욱 높은 검출감도로 검출할 수 있을 뿐아니라 RT-PCR을 검증할 수 있는 nested PCR 프라이머 조합을 개발하였다. 또한, 제한효소 Xho I 부위를 삽입한 유전자변형-양성대조구 플라스미드를 설계하여, PCR module에서 대조구로부터 오염을 검증할 수 있도록 개발하였다. 본 연구에서 개발한 PCR module은 토마토, 국화, 아티초크 및 제라늄 등에서 PZSV를 간편, 신속 및 특이적으로 검출하여, 지속적으로 식물검역에 활용할 수 있을 것으로 기대된다.

Phytophagous insects detect volatile compounds produced by host and non-host plants, using species-specific sets of olfactory receptor neurons (ORNs). To investigate the relationship between the range of host plants and the profile of ORNs, single cell recordings were carried out to identify ORNs and corresponding active compounds in female Uraba lugens (Lepidoptera: Nolidae), an oligophagous eucalypt feeder. Based on the response profiles to 39 plant volatile compounds, 13 classes of sensilla containing 40 classes of ORNs were identified in female U. lugens. More than 95% (163 out of 171) of these sensilla contained 16 classes of ORNs with narrow response spectra, and 62.6% (107 out of 171) 18 classes of ORNs with broad response spectra. Among the specialized ORNs, seven classes of ORNs exhibited high specificity to 1,8-cineole, (±)-citronellal, myrcene, (±)-linalool and (E)-β-caryophyllene, major volatiles produced by eucalypts, while nine other classes of ORNs showed highly specialized responses to green leaf volatiles, germacrene D, (E)-β)-farnesene and geranyl acetate that are not produced by most eucalypts. We hypothesize that female U. lugens can recognize their host plants by detecting key host volatile compounds, using a set of ORNs tuned to host volatiles, and discriminate them from non-host plants using another set of ORNs specialized for non-host volatiles. The ORNs with broad response spectra may enhance the discrimination between host and non-host plants by adding moderately selective sensitivity. Based on our finding, it is suggested that phytophagous insects use the combinational input from both host-specific and non-host specific ORNs for locating their host plants, and the electrophysiological characterization of ORN profiles can be used for predicting the range of host plants in phytophagous insects.