[논문]시설재배지 고추를 가해하는 총채벌레류와 TSWV 유전자 서열 변이

김철영; 최두열; 강정훈; 아흐메드샤비르; 길의준; 권기면; 이관석; 김용균

doi:10.5656/ksae.2021.11.0.037

시설재배지 고추를 가해하는 총채벌레류와 TSWV 유전자 서열 변이
Thrips Infesting Hot Pepper Cultured in Greenhouses and Variation in Gene Sequences Encoded in TSWV 원문보기

한국응용곤충학회지 = Korean journal of applied entomology, v.60 no.4, 2021년, pp.387 - 401

김철영 (안동대학교 식물의학과) , 최두열 (안동대학교 식물의학과) , 강정훈 (안동대학교 식물의학과) , 아흐메드샤비르 (안동대학교 식물의학과) , 길의준 (안동대학교 식물의학과) , 권기면 ((주) 생물이용연구소) , 이관석 (국립농업과학원 직물보호과) , 김용균 (안동대학교 식물의학과)

초록
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시설재배지를 대상으로 고추 정식 이후 황색 끈끈이트랩으로 총채벌레 발생을 모니터링하였다. 아울러 토마토반점위조바이러스(Tomato spotted wilt virus: TSWV)가 유발하는 고추 칼라병을 유관으로 조사하였다. 고추 정식 직후(3월 말) 낮은 밀도로 대만총채벌레(Frankliniella intonsa)가 트랩에 포획되었으며 4월 중순부터는 꽃노랑총채벌레(Frankliniella occidentalis)도 발견되었다. 이후 5월부터는 두 종이 전체 총채벌레의 98% 이상을 차지하였고, 이 가운데 대만총채벌레가 꽃노랑총채벌레보다 다소 많은 발생 밀도를 보였다. 전체 총채벌레의 발생 피크를 보면 5월 중순에 낮은 피크를 기점으로 6-7월에 발생 최성기를 보였다. 이후 총채벌레의 발생은 급격하게 감소하였다. 포획된 꽃노랑총채벌레의 암수 비율이 일정하지 않았는데 이는 이 곤충의 특이적 단성생식 가능성으로 이에 대한 실험적 증거를 제공하였다. 지역간 꽃노랑총채벌레의 유전적 거리를 COI 서열로 비교한 결과 원거리에서 채집한 꽃노랑총채벌레 집단과는 차이를 보였지만 안동지역 내에서 발생한 꽃노랑총채벌레는 COI 서열에서 높은 유사성을 보였다. 이들 주요 두 종의 총채벌레가 전파할 것으로 추정되는 고추 칼라병이 일부 시설재배지를 중심으로 발견되었으며 항혈청 및 분자진단을 통해 확인되었다. 더불어 감염 고추에서 채집된 꽃노랑총채벌레에서도 분자진단을 통해 TSWV를 검출하였다. 감염 TSWV의 게놈 구조를 비교하기 위해 기능성 단백질을 갖는 NS_S, N, NS_M의 유전자 서열을 분석하였다. 서로 다른 지역별 이들 유전자는 다수의 점돌연변이가 존재하였고 이들 가운데는 아미노산 서열 차이를 초래하는 오류 돌연변이를 포함하였다. 추출된 TSWV를 비보독충 꽃노랑총채벌레에 섭식 처리한 유충과 성충 모두에서 감염으로 일어났으나, 유충에게서만 바이러스 증식이 일어나는 것을 확인하였다.

Abstract ▼ AI-Helper

Thrips infesting hot peppers were monitored in greenhouses using yellow sticky traps. In addition, the hot peppers infected with tomato spotted wilt virus (TSWV) were observed during the monitoring period. The flower thrips (Frankliniella intonsa) were initially trapped at a low density just after transplanting seedlings of hot peppers at late March. The western flower thrips (Frankliniella occidentalis) were trapped after mid April. These two thrips represented more than 98% of the total thrips attracted to the traps after May, in which F. intonsa showed higher occurrence frequency than F. occidentalis. The total number of thrips had two peaks at mid May with a small and short-term peak and at June-July with a large and long-term peak. The trapped thrips exhibited inconsistent sex ratios, suggesting a seasonal parthenogenesis. Different geographical populations were varied in cytochrome oxidase I sequences, in which local populations in Andong shared a high sequence similarity. TSWV-infected hot peppers, which might be mediated by these two thrips species, were observed and confirmed by an immunoassay kit and a molecular diagnosis using RT-PCR. In addition, the TSWV was detected in F. occidentalis collected from the infected hot peppers. Three open reading frames (NSS, N, and NSM) of the isolated TSWV genomes were sequenced and showed multiple point mutations containing missense mutations among geographical variants. When the isolated TSWV was fed to nonvirulent thrips of F. occidentalis, the virus was detected in both larvae and adults. However, the viral replication occurred in larvae, but not in adults.

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표/그림 (9)

그림 Fig. 1. Morphological characters determining two Frankliniella species at adult stage. (A) F. occidentalis female. ① Dorsal view of whole body ② Flagellar segments (I-VIII) of an antenna. Brackets indicate dark black stigma areas at IV and V segments ③ Head showing ocular seta (OS) and postocular seta (POS) ④ Four pairs of setae on prothorax. (B) F. occidentalis male. ① Dorsal view of whole body ② Flagellar segments (I-VIII) of an antenna. Brackets indicate dark black stigma areas at IV and V segments ③ Abdominal tip showing yellow testis (TE) ④ Head showing setae ⑤ Four pairs of setae on prothorax. (C) F. intonsa female. ① Dorsal view of whole body ② Flagella segments of an antenna. Brackets indicate lightly black stigma areas at IV and V segments. ③ Head showing a seta ④ Four pairs of setae on prothorax. (D) F. intonsa male. ① Dorsal view of whole body ② Flagella segments of an antenna. Brackets indicate lightly black stigma areas at IV and V segments ③ Head showing a seta. ④ Four pairs of setae on prothorax.
그림 Fig. 2. Seasonal occurrence peaks of thrips in greenhouses cultivating hot pepper (C. annuum) in three Andong places (AD1-AD3). Monitoring used yellow sticky traps. In each place, three traps were installed and monitored every week from March 31 to August 3. Upper and lower panels show the occurrences of F. occidentalis (WFT) and total thrips, respectively. Insets indicate the thrips monitoring data at early season.
그림 Fig. 3. Relative occurrence frequencies of F. occidentalis (Fo) and F. intonsa (Fi) in greenhouses cultivating hot pepper (C. annuum) in three Andong places (AD1-AD3). Others indicate thrips not determined into these two species. In each place, three traps were installed and monitored every week from March 31 to July 27. (A) Comparison of total numbers of thrips occurrence in three places. (B) Relative frequencies of different thrips species in AD3.
그림 Fig. 4. Parthenogenesis of F. occidentalis. (A) Unequal sex ratios of F. occidentalis and F. intonsa in greenhouses cultivating hot pepper (C. annuum) in a Andong place (AD3). In each place, three traps were installed and monitored every week from March 31 to July 27. (B) Progeny analysis in different mating types. Each mating type used 10 pairs with a kidney for oviposition. Progeny number represented the number of the hatched larvae from the kidney.
그림 Fig. 5. Sequence variation in cytochrome oxidase I (COI) of the western flower thrips (WFT), F. occidentalis. Three Andong local populations ('Hahoe 1', 'Hahoe 2', and 'Poongsan') were compared with Goesan and laboratory ('Lab') populations. (A) Sequence alignment. Point mutations are denoted in a black-colored shade. (B) A phylogeny analysis of the WFT populations with other known haplotypes ('H1-H38'). Phylogeny analysis was performed by ClustalW alignment using MEGA6.06. Bootstrap values at the branches were obtained with 1,000 repetitions. Two separate monophyletic lineages are called 'WFTL' and 'WFTG'.
그림 Fig. 6. Detection of TSWV in hot pepper (C. annuum) and its infesting thrips, F. occidentalis. (A) Suspected plant host cultivated in greenhouse in Andong and its collection at May 15. (B) A positive test result of the plant sample against TSWV using an immunoassay kit. (C) RT-PCR diagnosis against thrips. 'P' represents the plant RNA sample. 'N' represents a negative control without template. Tests represent five insect samples. Each sample used three thrips to extract RNA.
그림 Fig. 7. Replication of TSWV in F. occidentalis. First (L1) and second (L2) instar larvae, and adults were fed for 3 h with kidney soaked into viral extract from the hot pepper infected with TSWV. After the viral treatment, the thrips were reared on fresh kidney and randomly selected for RNA extraction using RT-qPCR to quantify the virion numbers. Different color backgrounds indicate different developmental stages: blue for L1, yellow for L2, green for prepupa, violet for pupa, and red for adult. Each treatment was replicated three times. Each replication used xx individuals for RNA extraction. Different letters above standard deviation bars indicate significant difference among means at Type I error = 0.05 (LSD test).
그림 Fig. 8. Amino acid sequence analysis of three TSWV genes of different viral isolates collected from different localities cultivating hot pepper (C. annuum). (A) Sequence variation in N genes of four isolates: Hahoe, Yongsang, Goesan, and Jeonju. These were compared with known sequences uploaded in GenBank. Out-groups include N genes of GNRNV (Groundnut necrosis virus), TYRV (Tomato yellow ring virus), and WSMV (Wartermelon silver mottle orthotospovirus). (B) Sequence variation in NSS genes of four isolates: Hahoe, Yongsang, Goesan, and Jeonju. These were compared with known sequences uploaded in GenBank. Out-groups include NSS genes of GRNV (Groundnut necrosis virus), TYRV (Tomato yellow ring virus), and WSMV (Wartermelon silver mottle orthotospovirus). (C) Sequence variation in NSM genes of four isolates: Hahoe, Yongsang, Goesan, and Jeonju. These were compared with known sequences uploaded in GenBank. Out-groups include EP genes of GNRV (Groundnut necrosis virus), TYRV (Tomato yellow ring virus), and WSMV (Wartermelon silver mottle orthotospovirus).
표 Table 1. Polymorphism of three genes of TSWV among different local population

참고문헌 (43)

Almasi, A., Csillery, G., Csomor, Z., Nemes, K., Palkovics, L., Salanki, K., Tobias, I., 2015. Phylogenetic analysis of tomato spotted wilt virus (TSWV) NSs protein demonstrates the isolated emergence of resistance-breaking strains in pepper. Virus Genes 50, 71-78.

상세보기
Culbreath, A.K., Todd, J.W., Brown, S.L., 2003. Epidemiology and management of tomato spotted wilt in peanut. Annu. Rev. Phytopathol. 41, 53-75.

상세보기
German, T.L., Ullman, D.E., Moyer, J.W., 1992. Tospoviruses: diagnosis, molecular biology, phylogeny, and vector relationships. Annu. Rev. Phytopathol. 30, 315-348.

상세보기
Groves, R.L., Walgenbach, J.F., Moyer, J.W., Kennedy, G.G., 2001. Overwintering of Frankliniella fusca (Thysanoptera: Thripidae) on winter annual weeds infected with Tomato spotted wilt virus and patterns of virus movement between susceptible weed hosts. Phytopathology 91, 891-899.

상세보기
Han, M.J., Kim, I.S., Ahn, S.B., Lee, M.L., Hong, K.J., Lee, G.H., Ku, D.S., 1998. Distribution and host plants of recently introduced western flower thrips, Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae) in Korea. RDA J. Crop Prot. 40, 83-88.
Hunter, W.B., Ullman, D.E., 1989. Analysis of mouthpart movements during feeding of Frankliniella occidentalis (Pergande) and F. schultzei trybom (Thysanoptera: Thripidae). Intl. J. Insect Morph. Embryol. 18, 161-171.
Jahn, M., Paran, I., Hoffmann, K., Radwanski, E.R., Livingstone, K.D., Grube, R.C., Aftergoot, E., Lapidot, M., Moyer, J.W., 2000. Genetic mapping of the Tsw locus for resistance to the Tospovirus Tomato spotted wilt virus in Capsicum spp. and its relationship to the Sw-5 gene for resistance to the same pathogen in tomato. Mol. Plant-Microbe Interact. 13, 673-682.

상세보기
Katayama, H. 1997. Effect of temperature on development and oviposition of western flower thrips Frankliniella occidentalis (Pergande). Jpn. J. Appl. Entomol. Zool. 41, 225-231.
Katayama, H., Ikeda, F. 1995. Overwintering of western flower thrips Frankliniella occidentalis Pergande in the western region of Shizuoka Prefecture. B. Shizuoka Agric. Exp. Stn. 40, 63-73.
Kim, C., 2000. Review of disease incidence of major crops in 2000. Korean J. Pestic. Sci. 5, 1-11.
Kim, J.H., Choi, G.S., Kim, J.S., Choi, J.K., 2004. Characterization of tomato spotted wilt virus from Paprika in Korea. Plant Pathol. J. 20, 297-301
Kim, T.Y., Jang, C., Kang, H.W., Choi, J.H., Lee, H.W., Lee, J.W., Lee, D.H., Yang, S.K., Lee, S.Y., Min, C.G., Lee, D.W., 2021. Comparison of pest occurrence and viral disease incidence rate with reduced the application of pesticides in red pepper field. Korean J. Pestic. Sci. 25, 1-10.
Kindt, F., Joosten, N.N., Peters, D., Tjallingii, W.F., 2003. Characterisation of the feeding behaviour of western flower thrips in terms of electrical penetration graph (EPG) waveforms. J. Insect Physiol. 49, 183-191.

상세보기
Ko, S.J., Kang, B.R., Choi, D.S., Kim, D.I., Lee, G.S., Kim, C.S., Choi, H.S., 2013. Pattern of the occurrence of tomato spotted wilt virus in Jeonnam province. Res. Plant Dis. 19, 273-280.
Korea Crop Protection Association (KCPA), 2020. Agrochemicals user's guide book, Korea Crop Protection Association, https://www.koreacpa.org/ko/use-book. (Accessed at Aug. 18. 2021).
Korean Statistical Information Service (KOSIS), 2020. Area of cultivation of outdoor vegetables. https://kosis.kr/statHtml/statHtml.do?orgId101&tblIdDT_1ET0013&vw_cdMT_ZTITLE&list_idK1_15&seqNo&lang_modeko&languagekor&obj_var_id&itm_id&conn_pathMT_ZTITLE. (Accessed Aug. 18. 2021).
Lee, G.S., Lee, J.H., Kang, S.H., Woo, K.S., 2001. Thrips species (Thysanoptera: Thripidae) in winter season and their vernal activities on Jeju island, Korea. J. Asia Pac. Entomol. 4, 115-122.

원문보기 상세보기
Lee, S., Lee, J., Kim, S., Choi, H., Park, J., Lee, J., Lee, K., Moon, J., 2004. The incidence and distribution of viral diseases in pepper by cultivation types. Res. Plant Dis. 10, 231-240.
Lim, U.T., Kim, E., Mainali, B.P., 2013. Flower model traps reduced thrips infestations on a pepper crop in field. J. Asia Pac. Entomol. 16, 143-145.

원문보기 상세보기
Moon, H.C., Cho, I.K., Im, J.R., Goh, B.R., Kim, D.H., Hwang, C.Y., 2006. Seasonal occurrence and damage by thrips on open red pepper in Jeonbuk Province. Korean J. Appl. Entomol. 45, 9-13.
Okazaki, S., Okuda, M., Komi, K., Yoshimatsu, H., Iwanami, T., 2007. Overwintering viruliferous Frankliniella occidentalis (Thysanoptera: Thripidae) as an infection source of tomato spotted wilt virus in green pepper fields. Plant Dis. 91, 842-846.

상세보기
Park, H.H., Kim, K.H., Park, C.G., Choi, B.R., Kim, J.J., Lee, S.W., Lee, S.G., 2009. Damage analysis and control threshold of Frankliniella occidentalis Pergande (Thysanoptera: Thripidae) on greenhouse eggplant and sweet pepper. Korean J. Appl. Entomol. 48, 229-236.
Park, J.D., Kim, D.I., Kim, S.G., 2002. Seasonal occurrence and damaged aspects of Frankliniella occidentalis Pergande (Thysanoptera: Thripidae) by cultural environments and varieties of chrysanthemum. Korean J. Appl. Entomol. 41, 177-181.
Park, Y., Kim, K., Kim, Y., 2014. Rapid cold hardening of Thrips palmi (Thysanoptera: Thripidae). Environ. Entomol. 43, 1076-1083.

상세보기
Reitz, S.R., Gao, Y., Kirk, W.D.J., Hoddle, M.S., Leiss, K.A., Funderburk, J.E., 2020. Invasion biology, ecology, and management of western flower thrips. Annu. Rev. Entomol. 65, 17-37.

상세보기
Roggero, P., Masenga, V., Tavella, L. 2002. Field isolates of tomato spotted wilt virus overcoming resistance in pepper and their spread to other hosts in Italy. Plant Dis. 86, 950-954.

상세보기
Rotenberg, D., Jacobson, A.L., Schneweis, D.J., Whitfield, A.E., 2015. Thrips transmission of tospoviruses. Curr. Opin. Virol. 15, 80-89.

상세보기
Rugman-Jones, P.F., Hoddle, M.S., Stouthamer, R. 2010. Nuclearmitochondrial barcoding exposes the global pest western flower thrips (Thyasnoptera: Thripidae) as two sympatric cryptic species in its native California. Mol. Entomol. 103, 877-886.
Rural Development Administration (RDA), 2020. Agricultural technology guide, Pepper. Jinhan M&B Inc. Seoul. Korea.
SAS Institute, Inc., 1989. SAS/STAT User's Guide. SAS Institute, Inc., Cary, NC.
Seo, J., Yi, Y., Kim, B., Hwang, J.M., Choi, S.W., 2011. Disease occurrence on red-pepper plants surveyed in Northern Kyungbuk province, 2007-2008. Res. Plant Dis. 17, 205-210.
Seo, M.H., Lee, S.C., Yang, C.Y., Yoon, J.B., Park, J., 2018. Monitoring occurrence status of thrips populations on field-cultivated pepper at major cultivated region in west coast, Korea. Korean J. Environ. Biol. 36, 544-549.
Shalileh, S., Ogada, P.A., Moualeu, D.P., Poehling, H.M., 2016. Manipulation of Frankliniella occidentalis (Thysanoptera: Thripidae) by tomato spotted wilt virus (Tospovirus) via the host plant nutrients to enhance its transmission and spread. Environ. Entomol. 45, 1235-1242.

상세보기
ShuQi, H., Ying, L., Lei, Q., ZhiHua, L., Chao, X., Lu, Y., FuRong, G., 2017. The influence of elevated CO 2 concentration on the fitness traits of Frankliniella occidentalis and Frankliniella intonsa (Thysanoptera: Thripidae). Environ Entomol. 46, 722-728.

상세보기
Skendzic, S., Zovko, M., Zivkovic, I.P., Lesic, V., Lemic, D., 2021. The impact of climate change on agricultural insect pests. Insects 12, 440.

상세보기
Stafford, C.A., Walker, G.P., Ullman, D.E., 2011. Infection with a plant virus modifies vector feeding behavior. Proc. Natl. Acad. Sci. USA 108, 9350-9355.

상세보기
Tentchev, D., Verdin, E., Marchal, C., Jacquet, M., Aguilar, J.M., Moury, B., 2011. Evolution and structure of tomato spotted wilt virus populations: evidence of extensive reassortment and insights into emergence processes. J. Gen. Virol. 92, 961-973.

상세보기
Ullah, M.S., Lim, U.T., 2015. Life history characteristics of Frankliniella occidentalis and Frankliniella intonsa (Thysanoptera: Thripidae) in constant and fluctuating temperatures. J. Econ. Entomol. 108, 1000-1009.

상세보기
Webster, C.G., Reitz, S.R., Perry, K.L., Adkins, S., 2011. A natural M RNA reassortant arising from two species of plant- and insect-infecting bunyaviruses and comparison of its sequence and biological properties to parental species. Virology 413, 216-225.

상세보기
Woo, K.S., Paik, W.H., 1971. Studies on the thrips (Thysanoptera) unrecorded in Korea (I). Korean J. Plant Prot. 10, 69-73.
Yoon, J.Y., Her, N.H., Cho, I.S., Chung, B.N., Choi, S.K., 2021. First report of a resistance-breaking strain of tomato spotted wilt orthotospovirus infecting Capsicum annuum carrying the Tsw resistance gene in South Korea. Plant Dis. doi: 10.1094/PDIS-09-20-1952-PDN.

상세보기
Yoon, J.Y., Yoon, J.B., Seo, M.H., Choi, S.K., Cho, I.S., Chung, B.N., Yang C.Y., Gangireddygari, V.S.R., 2020. Application of multiplex RT-PCR for simultaneous identification of tomato spotted wilt virus and thrips species in an individual thrips on chrysanthemum. Res. Plant Dis. 26, 264-271.
Zhang, Z.J., Wu, Q.J., Li, X.F., Zhang, Y.J., Xu, B.Y., Zhu, G.R., 2007. Life history of western flower thrips, Frankliniella occidentalis (Thysan., Thripae), on five different vegetable leaves. J. Appl. Entomol. 131, 347-354.

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저장형식	Text(ASCII format) Excel format RefWorks Direct Export RIS format (for Reference Manager, ProCite, EndNote), Scholar's Aids, Mendeley
메일정보	받는사람 (필수) @ 보내는사람 (선택) @ 제목 내용 KISTI 검색결과 이메일 서비스
안내	총 건의 자료가 검색되었습니다. 다운받으실 자료의 인덱스를 입력하세요. (1-10,000) 검색결과의 순서대로 최대 10,000건 까지 다운로드가 가능합니다. 데이타가 많을 경우 속도가 느려질 수 있습니다.(최대 2~3분 소요) 다운로드 파일은 UTF-8 형태로 저장됩니다. 파일의 내용이 제대로 보이지 않을실 때는 웹브라우저 상단의 보기 -> 인코딩 -> 자동선택 여부를 확인하십시오. ~ Text(ASCII format) Excel format

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