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1 농약과학회지 (Korean J. Pestic. Sci.) Vol. 19, No. 2, pp (2015) Open Access Online ISSN Print ISSN ORIGINAL ARTICLES / CONTROL 살선충제 Imicyafos 입제의 2 종뿌리혹선충에대한방제효과 김형환 * 정영학 1 김동환 하태기 2 윤정범 박정규 1 추호렬 1 국립원예특작과학원원예특작환경과, 1 경상대학교응용생명과학부, 농업생명과학연구원, 2 ( 주 ) 경농중앙연구소 Effects of Imicyafos GR against Two Species of the Root-knot Nematodes (Meloidogyne incognita and Meloidogyne hapla) Hyeong Hwan Kim*, Young Hak Jung 1, Dong Hwan Kim, Tae Ki Ha 2, Jung Beom Yoon, Chung Gyoo Park 1 and Ho Yul Choo 1 Horticultural & Herbal Crop Environment Division, National Institute of Horticultural & Herbal Science, RDA, Wanju , Republic of Korea 1 Division of Applied Life Science (BK21+)/Institute of Agriculture and Life Sciences, Gyeongsang National University, Jinju, Gyeongnam , Republic of Korea 2 Central Research Institude Kyung Nong Corporation, Gyeongju, Gyeongbuk , Republic of Koreea (Received on November 7, Revised on April 20, Accepted on June 1, 2015) Abstract Efficacy of novel nematicide, imicyafos GR was evaluated against two species of the root-knot nematodes in pot and greenhouse conditions. When tested in pots, the population of Meloidogyne incognita and M. hapla was reduced sixty days after treatment, with mortality rate of 91.5% and 90.6%, respectively. Suppression effect of imicyafos GR on root galling in tomato was tested. The number of root galls caused by M. incognita and M. hapla was reduced 60 days after nematode inoculation, with the efficacy of 94.2% and 95.1%, respectively. Under greenhouse conditions planted with watermelon, melon, cucumber, and tomato, the efficacy of imicyafos GR on M. incognita persisted up to 60 days after treatment, showing 90% of control efficacy. Moreover, the number of root galling was more reduced than fosthiazate treatment, with the potential as a control agent. Key words Meloidogyne incognita, Meloidogyne hapla, imicyafos GR, control efficacy 서 론 현재까지전세계적으로약 500여종의식물기생성선충이보고되어있는데그중, 뿌리혹선충 (Meloidogyne spp.) 은 78종이분포하는것으로기록되어있다 (Jepson, 1987). 국내에서시설원예재배지에서식하고있는뿌리혹선충으로 M. arenaria ( 땅콩뿌리혹선충 ), M. cruciani, M. hapla ( 당근뿌리혹선충 ), M. hispanica, M. incognita ( 고구마뿌리혹선충 ), M. javanica ( 자바니카뿌리혹선충 ) 등 6종이알려져있는데이중에서 M. arenaria, M. hapla, M. incognita, M. javanica 등 4종이농업상중요뿌리혹선충이다 (Choi and Choi, *Corresponding author hhkim8753@korea.kr 1982; Cho et al., 2000; Kim et al., 2001). 뿌리혹선충은국내에재배되고있는과채류, 엽채류및화훼류등원예작물전반에걸쳐연작장해의중요한요인으로알려져있으며, 뿌리혹선충 2령충이작물에기생되면구침으로작물의조직을찔러영양분을섭취하면서뿌리에혹을만들어양분과수분의이동을저해하고, 탈취하므로지상부의생육이불량해진다. 뿌리혹선충은양분과수분의흡즙에의한직접적인피해와더불어선충의침입에의해생긴뿌리혹주변세포조직을와해시켜토양에서식하는각종병원균이침입하여이차적으로토양전염성병을일으키기도한다 (Choo et al., 1990; Park et al., 1995). 뿌리혹선충이작물의유묘기나정식초기부터발생밀도가많으면조기에고사되며, 반촉성재배에서정식초기에토양중선충의밀도가낮더라도작물의생육후기에이르면선충의밀도가 101
2 102 김형환 정영학 김동환 하태기 윤정범 박정규 추호렬 급격히증가하여뿌리전체에혹이형성되어생산량을감소시킨다 (Kim et al., 2001). 우리나라는공정육묘장의증가로대부분의원예작물재배농가에서는유묘를자가육묘하기보다는구입하여사용하고있어육묘기에뿌리혹선충에의한감염은극히드물어뿌리혹선충과접촉하게되는것은정식과더불어시작되기때문에방제는주로정식전 후에행해지고있다. 뿌리혹선충의방제방법에는답전윤환, 객토, 태양열소독, 약제방제, 저항성품종, 저항성윤작 (Park et al., 1995; Chon, 1996; Kim and Choi, 2001), 휴경, 재배시기조절, 길항식물과유치작물재배, 잔재물소각, 담수, 기생된작물잔재물제거, 접목, 유기물토양개선과생물적방제, 작물경작개선 (Bridge, 1996) 등과같이다양한방법들이알려져있다. 그러나국내원예작물재배에서는동일한작물을연작하는경우가많으며, 작기와작기사이가짧고, 뿌리혹선충에감수성인작물의재배가많고, 윤작시경제성이문제가되어다양한뿌리혹선충의방제법이있음에도불구하고최근까지방제효과의불확실성과번거로움으로인해살선충제를이용한방제법을선호하고있는실정이다. 현재국내에서는뿌리혹선충의방제용으로카두사포스 카보설판입제, 에토프로포스입제등 14종류의약제가수박, 참외, 오이, 토마토등 20종의원예작물에등록되어있다. 그중에서카두사포스캡슐현탁제, 포스티아제이트액제, 메탐소듐액제등 3종류를제외한 11종류의약제가정식전에처리하는입제이다 (KCPA, 2014). 최근작물의생육중에도뿌리혹선충의방제가가능한살선충캡슐현탁제, 유제등이등록되고있으나, 여전히작물을정식하기전에토양혼화처리되는입제형태의제품이대부분이다. 뿌리혹선충을방제하기위해매년반복적으로같은토양에살선충제입제처리가잦아지면서살선충제저항성이발생하는경우가많아져선충의방제효과가떨어지고있어농작물의피해가증가하고있기때문에새로운살선충제의개발이시급한상황이다. 따라서국내시설원예작물중에서뿌리혹선충에피해가많은몇몇작물을대상으로최근에개발된살선충제인 imicyafos 입제의방제효과를실증시험하였다. 재료및방법 뿌리혹선충실내포트검정에이용된고구마뿌리혹선충 (Meloidogyne incognita) 과당근뿌리혹선충 (M. hapla) 은경농중앙연구소의유리온실에서토마토유묘를이용하여증식하였다. 증식은직경 16.0 cm 높이 13.5 cm 플라스틱포트에 45~50일된토마토 ( 선명, 농우 ) 유묘를정식하여 40~45일동안뿌리혹이형성되도록관리하였다. 그리고뿌리혹이형성된뿌리를 1cm 길이로잘라서새로운토마토유묘를심을포트에상토 ( 바로크, 서울바이오 ) 와함께섞은후토마토유묘를정식한다음누대증식하였다. 포트검정에사용된뿌리혹선충은누대증식중이던고구마뿌리혹선충과당근뿌리혹선충의유충을사용하였다. 포트방제효과검정실내포트에서 imicyafos 입제 ( 네마킥, 5%) 의고구마뿌리혹선충과당근뿌리혹선충에대한처리별방제효과에대한시험은직경 16.0 cm 높이 13.5 cm 플라스틱포트에 45~50 일된토마토 ( 선명, 농우 ) 유묘를상토 ( 바로크, 서울바이오 ) 가담겨있는포트에심었다. 그리고처리방법에관계없이정식과함께각각의증식된뿌리혹선충의유충을분리하여포트당 1,000마리를접종하였다. 뿌리혹선충의알분리는누대증식중이던토마토뿌리를수거하여깨끗이씻어서뿌리속의알을분리하기위하여개량된 sodium hypochloride 방법 (Baker et al., 1985) 을사용하였다. 분리방법은깨끗이씻은뿌리를 1cm 간격으로잘라서 200 ml의 1% NaOCl 용액이들어있는믹서기에넣고고속으로 1분간회전시켰다 (Kim and Lee, 2008). 그리고믹서기내의뿌리찌꺼기, 알은 75 µm와 28 µm 체를통과시키고 28 µm 체에거른알이부화할때까지상온 (25 ± 2 o C) 에서보관하였다. 건전한유충을시험에활용하기위하여일별부화율을현미경하에서관찰하면서부화율이최적의정점에달하는날에 1차체에거른후 2차깔때기법으로거른후획득한유충을시험에이용하였다. 실험은처리별로뿌리혹선충종에따라한개의포트를 1반복으로하여 10반복으로조사하였다. 조사는뿌리혹선충의생태에비추어약제처리후 30일과 60일째포트내의상토중 100 g을개량깔때기법 (Kaya and Stock, 1997) 으로선충의밀도를조사하고, 60일째토마토에형성된뿌리혹의난낭수를조사 (Taylor and Sasser, 1978) 하여방제효과를처리간비교, 분석하였다. 포장방제효과시험 Imicyafos 입제 ( 네마킥, 5%) 약제의고구마뿌리혹선충대한방제효과시험은시설재배수박, 참외, 멜론, 오이와토마토재배지에서수행되었으며, 다음과같이각작물별로약제를처리하여효과를조사하였다. 작물별시험은시설수박 ( 삼복꿀수박 ) 은경남함안에서 2012년 4월 1일, 시설참외 ( 금관 ) 는경북예천에서 2012년 7월 2일, 시설멜론 ( 뷰티 ) 은경북경주에서 2012년 6월 1일, 시설오이 ( 조은백다다기 ) 는경북경주에서 2012년 9월 3일, 시설토마토 ( 선명 ) 는경북경주에서 2012년 6월 8일에수행하였다. 각작물별로 imicyafos 5% 입제를 10a 당 6 kg 처리한후로터리작업을하고, 작물을정식한다음물을관주하였다. 대조약제로는 fosthiazate 5% 입제를사용하였고, 10a 당 6
3 살선충제 Imicyafos 입제의 2 종뿌리혹선충에대한방제효과 103 kg을동일한날짜에처리하였다. 토양중고구마뿌리혹선충의발생밀도는 imicyafos 입제와대조약제인 fosthiazate 입제를처리하기전토양을채취하여사전밀도를조사하였고, 약제처리후 30일째와 60일째동일한방법으로토양을채취하여선충의생충수를조사하였다. 토양내고구마뿌리혹선충의살아있는밀도조사는작물별로다소차이는있지만, 단동및연동구분없이 330 m 2 를 1개구역으로정하고임의로 5곳을지정한다음지점당토양 100 g을채취하여 500 g을취하여골고루섞고다시이중에서 100 g의토양을개량깔때기법 (Kaya and Stock, 1997) 을이용하여뿌리혹선충의유충을추출하여생충수를조사하였다. 조사는 330 m 2 1개구역을 1반복으로 3반복으로토양내고구마뿌리혹선충의밀도를조사하였다. 또한살선충제처리후 60일째는작물체의주위를야삽으로조심스럽게파고, 작물체를뽑은다음흐르는수돗물에뿌리에묻어있던흙을제거하고, 난낭수를조사하였다. 난낭수조사는 Phloxin B 용액에 20분간염색하여붉게염색된난낭의수를계수하였고 (Taylor and Sasser. 1978), 10주를 1반복으로 30주를조사하였다. 통계분석 Imicyafos 입제의방제효과를작물별로구분하여뿌리혹선충에대한방제효과를 Tukey test로처리평균간차이로분산분석하였다 (SAS Institute, 2008). 결과및고찰 포트시험방제효과토마토포트시험에서 imicyafos 입제처리후 30일째와 60일째생충률은각각고구마뿌리혹선충이 3.4%, 12.5%, 당근뿌리혹선충이 4.9%, 12.2% 로 fosthiazate 입제와유의차는없었으나방제가는유의적차이가있었다 (Table 1). 특 히 60일후난낭수에서 fosthiazate 입제처리보다 imicyafos 입제처리가고구마뿌리혹선충과당근뿌리혹선충의난낭형성을더많이억제하였다. Fosthiazate는미국의 Georgia에서땅콩뿌리혹선충방제 (Minton et al., 1993), Florida에서담배뿌리혹선충방제 (Rich et al., 1994) 에사용되어그효과가인정된약제로국내뿐만아니라외국에서도널리사용되고있다. 본연구에서이용한 imicyafos 입제는 fosthiazate 입제보다뿌리혹선충에대한방제효과가더뛰어나효과적으로뿌리혹선충을방제할수있을것으로기대된다. 포장시험방제효과시설재배수박, 참외, 멜론, 오이, 토마토에서 imicyafos 입제를처리하여방제효과를조사한결과고구마뿌리혹선충에대하여 imicyafos 입제는처리후 30일째와 60일째까지 90% 이상높은방제가를나타내었다 (Table 2). 멜론을제외한모든처리구에서 imicyafos 입제처리후 30일째생충률과방제가는대조약제인 fosthiazate 입제처리구와유의적차이가나지않았다. 하지만처리후 60일째방제가에서 imicyafos 입제처리가 fosthiazate 입제처리보다방제가가더높았는데, 이는 imicyafos 입제처리가상대적으로약효지속성이더높기때문인것으로생각되어진다. 난낭수조사에서도모든작물에서약효지속성이더뛰어난 imicyafos 입제처리에서방제효과가더뛰어났다. Imicyafos는유기인계침투성살선충제로침투한약제가신경자극계로이동하여 Acethylcholin 수용체에부착하여 Acethylcholinesterase (AChE) 의작용을억제함으로써신경전달을저해하여뿌리혹선충을죽인다. Imicyafos와 fosthiazate 는유기인계 (organophosphorus) 살선충제이고, 침투이행성약제로대표적인비훈증약제이다 (Wada et al., 2011). 두약제모두뿌리썩이선충 (Pratylenchus penetrans) 에방제효과 Table 1. effect of nematicides, Imicyafos GR and Fosthiazate GR against the Meloidogyne incognita and M. hapla on tomato in pot Nematode species M. incognita M. hapla Treatment Survival rate a 30 DAT 60 DAT efficacy b Survival rate efficacy No. of egg mass/plant efficacy c Imicyafos GR 63.4b d 95.2a 612.5b 91.5a 669.0c 94.2a Fosthiazate GR 66.4b 91.0b 617.8b 87.8b 630.3b 80.7b 71.3a a a - Imicyafos GR 64.9b 93.7a 612.2b 90.6a 610.3c 95.1a Fosthiazate GR 67.7a 90.0b 625.0b 80.8b 647.3b 76.8b 67.5a a a - a ; Survival rate of juveniles/100 g soil. b ; efficacy; (Survival ratio of M. incognita juveniles in control-survival ratio of M. incognita juveniles in treatment)/survival ration of M. incognita juveniles in control 100. c ; efficacy; (No. of egg mass in control-no. of egg mass in treatment)/no. of egg mass in control 100. d ; Means within a column followed by different letters are significantly different (Tukey's HSD test P < 0.001).
4 104 김형환 정영학 김동환 하태기 윤정범 박정규 추호렬 Barker, K. R., D. P. Schmitt and J. L. Imbriani (1985) Nematode population dynamics with emphasis on determining damage potential to crops. In: An advanced treatise on Meloidogyne, Vol, ed. by Barker, K. R., C. C. Carter and J. N. Sasser. pp North Carolina State University, Raleigh, North Carolina, USA. Bridge, J. (1996) Nematode management in sustainable and subsistence agriculture. Annu. Rev. Phytopathol. 34: Cho, M. R., S. Y. Na and M. S. Yiem (2000) Biological control of Meloidogyne arenaria by Pasteuria penetrans. J. Asia- Table 2. effect of nematicides, Imicyafos GR and Fosthiazate GR against the Meloidogyne incognita on 5 crops in greenhouse 30 DAT 60 DAT Predensity a Survival Survival No. of egg Crop Treatment rate b efficacy c rate efficacy mass/plant efficacy d Watermelon Oriental melon Melon Cucumber Tomato Imicyafos GR 148.3a e 664.3b 96.9a 11.7b 92.6a 6.3c 96.1a Fosthiazate GR 137.7a 619.0b 85.2a 28.8b 81.9b 32.7b 79.3b 123.0a 137.1a a a - Imicyafos GR 676.0a 668.2b 95.2a 21.5b 90.8a 6.3c 96.1a Fosthiazate GR 661.3a 617.8b 89.5a 53.4b 77.9b 33.0b 79.6b 670.7a 180.0a a a - Imicyafos GR 678.0a 669.4b 95.4a 17.2b 93.9a 4.7c 97.0a Fosthiazate GR 665.3a 621.9b 89.3b 38.7b 86.2a 36.0b 77.2b 669.7a 205.5a a a - Imicyafos GR 675.3a 612.2b 94.4a 25.5b 92.8a 4.7c 96.4a Fosthiazate GR 683.3a 621.7b 89.7a 72.5b 78.4b 48.3b 61.3b 681.7a 214.8a a a - Imicyafos GR 671.7a 666.2b 96.4a 11.2b 95.0a 6.0c 96.3a Fosthiazate GR 663.0a 622.6b 86.0a 54.9b 75.0b 45.0b 72.4b 673.3a 173.5a a a - a ;Pre-density of juveniles/100 g soil. b ; Survival rate of juveniles/100 g soil. c ; efficacy; (Survival ratio of M. incognita juveniles in control-survival ratio of M. incognita juveniles in treatment)/survival ration of M. incognita juveniles in control 100. d ; efficacy; (No. of egg mass in control-no. of egg mass in treatment)/no. of egg mass in control 100. e ; Means within a column in each crop followed by different letters are significantly different (Tukey's HSD test P<0.001). 가우수하며, 토양내자유생활선충 (free-living nematode), 비표적미생물 (non-target organism) 및박테리아섭식선충 (bacterial-feeding nematode) 에는영향이적다고하였다 (Sturz and Kimpinskim, 1999; Wada et al., 2011). 그러나 imicyafos가토양내선충개체군의독성범위에있어서는 fosthiazate보다영향이더적고, 작기내내지속적으로뿌리썩이선충과같은식물기생성선충의밀도를억제할수있었다 (Wada et al., 2011). 또한약제의처리후자유생활을하는토양선충의개체군에대한영향이 imicyafos는토양표면에한정적이며일시적인반면 fosthiazate는보다넓다고하였다 (Kimpinski et al., 2005). 또다른 imicyafos 약제의특징으로 cadusafos 등다른살선충제와달리약제에접촉된선충을다시분리해정상적인기주에접종하여도뿌리혹을형성하지못하게되는데, 이러한특성은본실험에서 imicyafos 입제처리가 fosthiazate 입제처리보다난낭형성이억제된원인이라할수있다. 따라서 imicyafos 입제는뿌리혹선충뿐만아니라뿌리썩이선충에대하여방제효과가뛰어나며, 특히난낭형성을억제하고장기간지효성이있어재배기간이긴작물에도효과적일것으로생각되어진다. 본연구에서는 imicyafos 입제에따른뿌리혹선충방제효과시험만이루어졌으나, imicyafos 액제에대한방제효과연구결과가더해진다면토양혼화처 리외에정식후에도뿌리혹선충을효과적으로방제할수있는방법도개발할수있을것으로기대되므로금후 imicyafos 액제를이용한뿌리혹선충방제효과시험이필요하다고생각된다. 감사의글 본성과물은농촌진흥청연구사업 (PJ ) 의지원에의해이루어진것입니다. Literature Cited
5 살선충제 Imicyafos 입제의 2 종뿌리혹선충에대한방제효과 105 Pacific Entomol. 3(2): Choi, D. R. and Y. E. Choi (1982) Survey on plant parasitic nematodes in cropping by controlled horticulture. Korean J. Plant Protec. 21(1):8-14. Choo, H. Y., S. M. Lee, J. B. Kim and Y. D. Park (1990) Relationship of root-knot nematode, Meloidogyne incognita to pathogenesis of Rhizoctonia solani on cucumber, pepper, and tomato. Korean J. Plant Pathol. 6(3): Jepson, S. B. (1987) Identification of root-knot nematodes (Meloidogyne species). CAB International, Wallingford, Oxon, United Kingdom. 265 pp. Kaya, H. K. and S. P. Stock (1997) Techniques in insect nematology. In: lacey L. A. (ed.), Manual of Techniques in insect pathology. Academic Press, New York. pp Kim, D. G. and S. K. Choi (2001) Effects of incoporation method of nematicides on reproduction of Meloidogyne arenaria. Korean J. Appl. Entomol. 40(1): Kim, D. G., D. R. Choi and S. B. Lee (2001) Effects of control methods on yields oriental melon in fields infested with Meloidogyne arenaria. Res. Plant Dis. 7(1): Kim, D. G. and J. H. Lee (2008) Economic threshold of Meloidogyne incognita for greenhouse grown cucumber in Korea. Res. Plant Dis. 14: Kim, D. G., Y. K. Lee and B. Y. Park (2001) Root-knot nematode species distributing in greenhouses and their simple identification scheme. Res. Plant Dis. 7(1): Kimpinski, J., R. A. martin and A. V. Sturz (2005) Nematicides increase grain yields in spring wheat cultivars and suppress plant-parasitic and bacterial-feeding nematodes. J. Nematol. 37: Korea Crop Protection Association (KCPA) (2014) Agrochemicals use guide Minton, N. A., T. B. Brenneman, K. Bondari and G. W. harrison (1993) Activity of fosthiazate against Meloidogyne arenaria, Frankliniella spp. and Sclerotium rolfsii in peanut. Peanut Science 20: Park, S. D., S. D. Park, T. Y. Kwon, H. S. Jun and B. S. Choi (1995) The occurrance and severity of damage by root-knot nematode Meloidogyne incognita in controllled fruit vegetable field. RDA. J. Agri. Sci. 37(1): Rich, J. R., R. A. Dunn, W. D. Thomas, J. W. Breman and R. S. Tervola (1994) Evaluation of fosthiazate for management of Meloidogyne javanica in Florida flue-cured tobacco. J. Nematol. 26(4): SAS Institute (2008) Sas/STAT user s guide: Statistics, version 9.3 Institute Cary, N.C., U.S.A. Sturz, A. V. and J. Kimpinski (1999) Effects of fosthiazate and aldicarb on populations of plant-growth-promoting bacteria, root-lesion nematodes and bacteria-feeding nematodes in the root zone of potatoes. Plant Pathol. 48: Taylor, A. L. and J. N. Sasser (1978) Biology, identification and control of root-knot nematodes (Meloidogyne species). North Carolina State Univ. Raleigh. 111 pp. Wada S., K. Tovota and A. Takada (2011) Effects of the nematicide imicyafos on soil nematode community structure and damage to radish caused by Pratylenchus penetrans. J. Nematol. 43(1):1-6. 살선충제 Imicyafos 입제의 2 종뿌리혹선충에대한방제효과 김형환 * 정영학 1 김동환 하태기 2 윤정범 박정규 1 추호렬 1 국립원예특작과학원원예특작환경과, 1 경상대학교응용생명과학부, 농업생명과학연구원, 2 ( 주 ) 경농중앙연구소 요약살선충제인 imicyafos 입제처리에따른방제효과를알아보기위하여고구마뿌리혹선충과당근뿌리혹선충을대상으로포트시험과포장시험을수행하였다. 포트시험결과 imicyafos 입제처리 60 일후생충수와난낭수조사에서고구마뿌리혹선충은 91.5%, 94.2%, 당근뿌리혹선충은 90.6%, 95.1% 로각각높은방제효과를나타내었다. 시설재배수박, 참외, 멜론, 오이, 토마토에서 imicyafos 입제를처리하여방제효과를조사한결과고구마뿌리혹선충에대하여 imicyafos 입제는처리후 30 일째와 60 일째까지 90% 이상의방제가를나타내었으며, 약효의지속성도뛰어났다. 특히 imicyafos 입제를처리가 fosthiazate 입제처리보다난낭수형성을더억제하는것으로나타났다. 따라서살선충제인 imicyafos 입제는고구마뿌리혹선충이나당근뿌리혹선충방제에실용성이있을것으로판단된다. 색인어 고구마뿌리혹선충, 당근뿌리혹선충, Imicyafos 입제, 방제효과
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