臺灣博碩士論文加值系統

小菜蛾為本省十字花科蔬菜之重要害蟲，其對本省早期的殺蟲劑已產生高抗藥性，造成小菜蛾防治不易，為降低殺蟲劑使用對環境衝擊及對人體影響，發展優質防治藥劑以降低小菜蛾抗藥性是重要課題。
本研究首先探討各種供試藥劑對小菜蛾之藥效及抗藥性，並選用不同化學藥劑混合複方或化學與微生物藥劑混合複方對小菜蛾做生物檢定，以檢視其藥效及抗藥性。藥效之檢定方法以劑量－反應之生物檢定分析法進行檢驗，其結果以對數－機率統計分析其LC50及LC90，以LC50及LC90為基礎之相對抗性比(Resistance Ratio)進行抗藥性分析比較。而下列混合藥劑之共同毒效係數分析法(Co-toxicity coefficent)主要進行下列四類混合複方：(1)雙變動比例之化學製劑混合複方；(2)單變動比例之化學製劑混合複方；(3)雙變動比例之微生物製劑與化學藥劑複方；(4)單變動比例之微生物製劑與化學藥劑複方，每個劑型中分別進行生物檢定測試後，可得到LC50及LC90之數據，並進行數據分析計算出毒效指數(toxicity index)，再進行其共同毒效係數(Co-toxicity coefficient)之分析。
結果顯示：本省小菜蛾對傳統之化學藥劑之抗藥性以阿巴汀(Abametin)抗藥性比值高達5000倍，新型藥劑克凡派(Chlorfenapyr)則無抗藥性。混合藥劑之複方中，化學藥劑間之混合對小菜蛾之共同毒效倍數分析，以雙變動比例混合之賽滅寧+因得克(Cypermethrin+Indoxacarb)最具協力效果，達8倍以上。單變動比例混合，以芬化利+賜諾殺(Fenvalerate+Spinosad)之效果最佳，協力作用為2倍，且其LC50及LC90均在10ppm以下，微生物藥劑與化學藥劑混合複方之雙變動比例混合，以蘇力菌(Bta)+ 賜諾殺(Spinosad)之協力作用高達8倍最佳，而單變動比例混合，阿巴汀(Abametin)+蘇力菌(Bta)之協力效果可達2倍，其LC50及LC90均在5ppm以下。
研究顯示微生物製劑與化學藥劑之適當比例混合，具有開發防治小菜蛾之潛力，主要因藥劑對小菜蛾之作用機制不同，田間產生抗藥性之機率相對減緩。

The diamondback moth(DBM), plutella xylostella (L.) is one of key insect pest of crucifers in Taiwan. DBM has became one of most difficult control insects because of depdloping resistance to chemical insecticides used extensively against it in the past 50 yr in Taiwan. In order to decrease the insecticides pollution to environment and to reduce the side effect for human health, how to develop the reasonable and efficacy control agents to against diamondback moth became the urgent issue now.
The objectives of this study were to investigate the susceptibility of DBM to different chemical insecticides and new registered insecticide in Taiwan and to monitor potential resistance evolution after field introduction. The other objective were to develop the chemical insecticide mixtures and microbial–chemical insecticide mixtures to search the best and most potent uses of toxicants to control DBM. To analyze the joint toxicity of two kinds of mixture, co-toxicity coefficient analysis method were used. The tested mixture divided into four group.
(1) Changes in the proportion of dual-mixed compound of insecticides.
(2) Changes in proportion of single insecticide in the insecticide
mixtures.
(3) Changes in the proportion of dual-mixed compound of insecticides and microbial agent.
(4) Changes in proportion of single insecticide in the insecticide
and microbial agent mixtures.
The LC50 and LC90 Value were calculated from each bioassay test. Based on LC50 and LC90 , the toxicity index and co-toxicity coefficient of each mixture was analyzed .
The results showed that the Kaoshung field population of DBM possessed 5000 fold tolerance to conventional insecticide, Abamectin , otherwise, the new registered insecticides, Chlorfenapyr have no resistance to DBM. Mixtures of insecticide, (in change in the proportion of dual-mixed groups), Cypermethrin + Indoxacarb showed 8 fold increase in toxicity of insecticide over sole treatment. Mixtures of insecticide in change in the proportion of single-mixed groups, Fenvalate + Spinosad showed 2 fold increase in toxicity over sole treatment. It’s LC50 and LC90 value were within 10 ppm. Mixture of insecticides and Bacillus thuringiensis aizawa (Bta) in changes of the proportion of dual-mixed groups, Bta + Spinosad exhibited 8 times more toxic than each sloe treatment.
Mixtures of insecticides and Bta in change of the proportion of single-mixed groups, Abamectin + Bta indicated 2 folds synergistic effect over each sole treatment. It’s LC50 and LC90 values were under 5ppm.
It is apparent that Bta and insecticides mixtures in appropriated ratio combinations were significantly more toxic than insecticide alone. These insecticide and Bta mixtures seem to be a potential new formulation for controlling DBM in the future. These new formulations posses different toxicity mechanism to DBM and seem to induce resistance to DBM slowly in field use.

中文摘要 I
Abstract III
誌謝 V
目錄 VI
表目錄 IX
圖目錄 XI
圖目錄 XI
第一章 前言 1
第二章 文獻回顧 4
2.1 小菜蛾之防治 4
2.2 抗藥性 5
2.3 蘇力菌 9
2.4 複方藥劑 15
第三章 材料與方法 17
3.1 化學藥劑推薦使用濃度與蘇力菌相容性測定 17
3.2 蘇力菌對小菜蛾之生物檢定 20
3.3 不同來源品系的小菜蛾對蘇力菌之感受性之檢測 21
3.4 化學藥劑對小菜蛾之毒效測定 23
3.5 混合藥劑對小菜蛾之毒效檢定 25
3.5.1 化學藥劑複方對小菜蛾之藥效測定 25
3.5.2 微生物製劑與化學藥劑複方對小菜蛾之藥效測定 25
3.5.3 結果數據分析 26
第四章 結果 30
4.1 化學藥劑與微生物製劑相容性測定 30
4.2 蘇力菌對耐、感性小菜蛾篩選 31
4.3 蘇力菌對小菜蛾之生物檢定 34
4.4 化學藥劑對小菜蛾之毒效測定 38
4.5 混合藥劑對小菜蛾之毒效檢定 44
4.5.1 化學藥劑複方對小菜蛾之藥效測定 44
4.5.2 微生物製劑與化學藥劑複方對小菜蛾之藥效測定 54
第五章 討論 59
5.1 化學藥劑與微生物製劑相容性測定 59
5.2 蘇力菌對耐、感性小菜蛾篩選 59
5.3 蘇力菌對小菜蛾之生物檢定 60
5.4 化學藥劑對小菜蛾之生物檢定 60
5.5 混合藥劑對小菜蛾之生物檢定 62
5.5.1 化學藥劑複方對小菜蛾之藥效測定 62
5.5.2 微生物製劑與化學藥劑複方對小菜蛾之藥效測定 65
第六章 結論 68
第七章 參考文獻 70
第八章 附錄 78
表目錄
Table 1 不同地區小菜蛾對大寶天機之毒效測定32
Table 2 不同地區小菜蛾對大寶天機之抗性比值33
Table 3 菜蛾實驗結果之半致死濃度與政府推薦濃度之比較35
Table 4 不同地區小菜蛾對蘇力菌β外毒素之抗性比值37
Table 5 各殺蟲劑對美國敏感品系之小菜蛾毒效測定結果39
Table 6 各殺蟲劑對高雄抗藥品系之小菜蛾毒效測定結果40
Table 7 高雄抗藥品系與美國敏感品系之抗藥性比值(LC50)42
Table 8 高雄抗藥品系與美國敏感品系之抗藥性比值(LC90)43
Table 9 化學藥劑混合複方對美國敏感品系之共同毒效係數46
Table 10 化學藥劑混合複方對高雄抗藥品系之共同毒效係數47
Table 11 測試藥劑與政府推薦濃度之比較LC50 and LC90 49
Table 12 化學藥劑複方共同毒效係數與政府推薦濃度之比較51
Table 13 化學藥劑複方對高雄品系小菜蛾之共同毒效係數53
Table 14 化學藥劑與微生物製劑複方對高雄抗藥品系小菜蛾之共同毒效係數55
Table 15 化學藥劑與微生物製劑複方對美國敏感品系小菜蛾之共同毒效係數57
Table 16 化學藥劑與微生物製劑複方對高雄抗藥品系小菜蛾之共同毒效係數58
圖目錄
圖 一、防治甘藍小菜蛾化學藥劑稀釋至推薦施用濃度後之酸鹼值。30
圖 二、小菜蛾餵食β-exotoxin 72 小時後的結果。比例尺長1 cm。36

1.周桃美、高靜華、鄭允，「十字花科蔬菜三種鱗翅目害蟲抗藥性之探討」，中華農業研究，第33捲，第3期，第331-336頁(1984)。
2.高靜華、鄭允，「小菜蛾寄生蜂之量產技術及田間應用研究」，農作物害蟲與害螨生物防治研討會，第133-145頁(2002)。
3.廖華盛，「實用農藥」，得力興業股份有限公司研發部，台中，第622-813頁(2005)。
4.鄭允、高靜華，小菜蛾之化學防治與抗藥性，中華昆蟲特刊第四號，第一屆璊蜱學研討會，第15-27頁(1989)
5.Ayalew, G., Ethiopian A., “Comparison of yield loss on cabbage from Diamondback moth, Plutella xylostella L. (Lepidotera: Plutellidae) using two insecticides,” Crop Protection Vol. 25, No. 9, pp.915-919(2005).
6.Bues, R., Bouvier, J., Boudinhon, C., “Insecticide resistance and mechanisms of resistance to selected strains of Helicoverpa armigera (Lepidoptera: Noctuidae) in the south of France,” Crop Protection, Vol. 24, No. 9, pp. 814-820(2005).
7.Bynum, E. D., Plapp, F. W., Archer, T. L., “Comparison of banks grass mite and twospotted spider mite (Acari: Tetranychidae): Responses to insecticides alone and in synergistic combinations,” Journal of Economic Entomology, Vol .90, No. 5, pp.1125-1130(1997).
8.Chandler, L. D., “Compatibility of insect growth regulators and bacterial insecticides for management of Spodoptera spp. Proceedings Beltwide Cotton Conferences,” San Antonio, TX, USA, January 4-7, Vol. 2, pp.1043-1045(1995).
9.Chandler, L. D., “Compatibility of insect growth regulators Beltwide Cotton Conferences,” San Antonio, TX, USA, January 4-7, Vol. 2, pp.1043-1045(1995).
10.Cleveland, C. B., Mayer, M. A., and Cryer, S. A., “An ecological risk assessment for spinosad use on cotton,” Pest Management Science, Vol. 58, No. 1, pp.70-84(2001).
11.Curtis, C. F., “Theoretical models of the use of insecticide mixtures for the management of resistance,” Bull. Entomol, Vol. 75, No. 9, pp.259-265(1985).
12.Damo, M. C., and Solsoloy, A. D., “Efficacy of Bacillus thuringiensis-chemical insecticide mixtures for cotton bollworm, Helicoverpa armigera (Hubn.) control,” Philippine-Entomologis,. Vol. 11, No. 1, pp.57-64(1997).
13.Devi, K. U., Reddy, N. N. R., Sridevi, D., Sridevi. V., and Mohan, C. M., “Esterase-mediated tolerance to a formulation of the organophosphate insecticide monocrotophos in the entomopathogenic fugus.Beauveria bassiana (balsamo) Vuill: a promising biopesticide,” Pest Management Science, Vol. 60, No. 4, pp.408-412(2004).
14.Dobrowolski, M., and Popowska, N. E., “Possibility of application of Beauveria bassiana (Bals.) Vuill. spores to control the pine bark bug (Aradus cinnamomeus Panz., Hemiptera-Heteroptera),” Prace- Instytutu- Badawczego- Lesnictwa, Vol. 866, No. 872, pp.95-102(1999).
15.Fadare. T. A., and Amusa, N. A., “Comparative efficacy of microbial and chemical insecticides on four major lepidopterous pests of cotton and their (insect) natural enemies,” African Journal of Biotechnology, Vol. 2, No. 11, pp.425-428(2003).
16.Huang, H. S., Hu, N. T., Yao, Y. E., Wu, C. Y., Chiang , S. W., Sun, C. N., “Molecular cloning and heterologous expression of a glutathione S-transferase involved in inssecticde resistance from the Diamondbackmoth, Plutella xylostella,” Insect Biochemistry and Molecular Biology , Vol.28, No.9, pp.651-658(1998).
17.Irigaray, F. J. S.D., Marco, M. V., and Ignacio, P. M., “The entomopathogenic fungus Beauveria bassiana and its compatibility with trifumuron:effect on the twospotted spicler mite Tetranychus urticae,” Biological Control, Vol.26, No. 2, pp.168-173(2003).
18.Li, A., Yang, Y., Wu, S., Wu, S., Li, C., and Wu, Y., “Investigation of resistance mechanisms to fipronil in diamondback moth (Lepidoptera:Plutellidae),” Journal of economic entomology , Vol. 99, No. 3, pp.914-919(2006).
19.Li, K., Xin, B., Torello, W. A., “Effect of organic fertilizers derived dissolved organic matter on pesticide sorption and leaching,” Environmental Pollution, Vol. 134, No. 2, pp.187-194(2005).
20.Mohan, M., and Gujar, G. T., ”Local variation in susceptibility of the diamondbackmoth, Plutella xylostella L. (Lepidoptera) to insecticides and role of detoxification enzymes,” Crop Protection, Vol. 22, No. 3, pp.495-504 (2003).
21.Mohan, M., and Gujar, G. T., “Toxicity of Bacillus thuringiensis strains and commercial formulation to th diamondback moth, Plutella xylostella(L.),” Crop Protection, Vol. 20, No. 4, pp.311-316(2002).
22.McCann, F. S., Annis, G. D., Shapiro, R. D., Piotrowslri, W., Lahm, G. P., Long, J. K., Lee, K. C., Hugher, M. M., Myers, B. J., Griswold, S. M., Reeves, B. M., March, R. W., Sharpe, P. L., Lowder, P., Barette, W. E., and Wing, K. D., ”The discovery of indoxacarb: oxadiazines as a new class of pyrazoline-type insecticides,” Pest Management Science, Vol. 57, No. 2, pp.153-164(2001).
23.Mourao, S. A., Vilela, E. F., Zanuncio, J. C., Zanuncio, L., and Tuelher, E.S., “Selectivity insecticides and fugicides to the entomopathogenic fungus,” Beauveria bassiana. Ento Soc. of Brasil. Jan/ Mar.Vol. 32, No. 1, pp.103-106(2003).
24.Nakata, M., Fukushima, A., Ohkawa, H., “A monoclonal antibody-based ELISA for the analysis of the insecticide flucythrinate in environmental and crop samples,” Pest management science, Vol. 57, No. 3, pp.269-277(2001).
25.Olmert, I., and Kenneth, R. G., “Sensitivity of the Entomopathogenic Fungi, Beauveria bassiana, Verticillum lecanii and Verticillium sp. to fungicides and insecticides,” Environ. Entomo, Vol. 3, No. 1, pp.33-38(1974).
26.Prabhaker, N., Toscano, N. C, Henneberry, T. J., “Evaluation of insecticide rotations and mixtures as resistance management strategies for bemisia argentifolii (Homoptera: Aleyrodidae),” Journal of Economic Entomology, Vol. 91, No. 4, pp. 820-826(1998).
27.Purwar, J. P., and Sachan, G. C., “Synergisitic effect of entomogenous fungi on some insecticides against Bihar hairy caterpillar Spilarctia oblique (Lepidoptera: Arctiidae),” Microbiological research, Vol. 161, No.1, pp.38-42(2006).
28.Quintela, E. D., and Mccoy, C. W., “Pathogenicity enhancenent of Metarhizium anisopliae and Beauveria bassiana to first instar of Diaprepev abbreviator (Coleoptera: Curculionidae) with sublethal doses of imidacloprid,” Environmental Entomology, Vol. 26, No. 5, pp. 1173-1182 (1997).
29.Ruwandi, A., and Cedric, G., “Resistance of Brassica, especially B. juncea (L.) Czern, genotypes to the diamondback moth, Plutella xylostella (L.),” Crop Protection, Vol. 17, No. 1, pp. 85-94(1998).
30.Saenz, D. C., Irigaray, F. J., Marco, M.V., and Perez, M. I., “The entomopathogenic fungus Beauveria bassiana and its compatibility with triflumuron: effects on the twospotted spider Tetranychus urticate,” Biological Control, Vol.26, No. 2, pp.168-173(2003).
31.Sapunaru, T., and Patras, A.,”Experimental results concerning the efficiency of new stocks of Beauveria bassiana Vuill in controlling Colorado beetle (Leptinotarsa decemlineata Say),” Moldova, Vol. 29, No. 1-2, pp.161-166(1996).
32.Sarfraz, M., and Keddie, B. A., “Conserving the efficacy of insecticides against Plutella xylostella (L.) (Lep., Plutellidae),” Journal of Applied Entomology,Vol.129, No.3, pp.149-157(2005).
33.Sayyed, A. H., Omar, D., and Wright, D. J., “Genetics of spinosad resistance in a multi-resistant field-selected population of Plutella xylostella,” Pest Management Science, Vol. 60, No. 8, pp.827-932(2004).
34.Shelton, A. M., Sances, F. V., Hawley, J., Tang, J. D.; Boune, M.; Jungers, D., Collins, H. L., Farias, J., “Assessment of insecticide resistance after the outbreak of diamondback moth (Lepidoptera: Plutellidae) in California in 1997,” Journal of Economic Entomology, Vol. 93, No. 3, pp. 931-936(2000).
35.Sonoda, S., and Tsumuki, H., “Studies on glutathione S-transferase gene invloed in chlorxuazuron resistance of the diamondback moth, Plutella xyloststella L. (Lepidoptera: Yponomeutidae),” Pesticide biochemistry and physiology, Vol. 82, No. 1, pp.94-101(2005).
36.Silva, L., Silva, R. E. F., and Heineck, M. A., “In vitro evaluation of effect of different insecticides on the sporulation of the fugus Nomuraea rileyi (Farlow) Samson,” Anais- da- Sociedade - Entomologica- do- Brasil, Vol. 22, No. 1, pp.99-103.(1993).
37.Simon, J.Y., and Mccord, E. J., “Lack of cross resistance to indoxacarb in insecticide-resistant Spodoptera frugiperda (Lepidoptera: Noctuidae ) and Plutella xylostella (Lepidoptera: Yponomeutidae),” Pest Management Science, Vol. 63, No. 1, pp. 63-67( 2007).
38.Sun, J.D., Wu, Lin, G.Y., A., Zeng, M. S., Wang, Q. S., Xu, D. Y., “Investigation and demonstration of the integrated control of the tea weevil by a mixture of pesticides and microbes,” Tech Bulletin, Vol. 60, No. 3, pp.32-34(1993).
39.Sun, Y. P., and Johnson, E. R., “Analysis of joint action of insecticides against house flies,” Journal of Economic Entomology, Vol. 53, No. 5, pp.887-892(1960).
40.Tanja, H. S., David, M. T., Andrew, J. T., Denholm, A. L. D., Duce, I. R., Martin, S. W., “Toxicological, electrophysiological, and molecular characterisation of knockdown resistance to pyrethroid insecticides in the diamondback moth, Plutella xylostella (L.),” Pesticide Biochemistry and Physiology, Vol. 59, No. 3, pp. 169-182(1998).
41.Wiwat, C., Thaithanun, S., Pantuwatana, S., Bhumiratana, A., “Toxicity of Chitinase-Producing Bacillus thringiensis ssp. Kurstaki HD-4 (G) toward Plutella xylostella,” Journal of Invertebrate Pathology, Vol.76, No. 4, pp.270-277(2000).
42.Wendt, R. L., Pirzadeh, P., Woin, P., “Effects of metsulfuron methyl and cypermethrin exposure on freshwater model ecosystems,” Aquatic Toxicology, Vol. 63, No. 3, pp. 243-256 (2003).
43.Zemaity, T. E., Refai., M. S. S., and Refai, E. L. S. A., “Joint action of fenvalerate mixtures against the cotton leafworm, Spodoptera littoralis (Boisd),“ Annals of Agricultura, Vol. 32, No. 3, pp.1741-1749(1987).
44.Zhao, J.Z., Collins, H.L., Li, Y. X., Mau, R. F. I., Thompson, G. D., Mhertlein, J.T., Andaloro, R. B., Shelton, A. M., “Monitoring of diamondback moth (Lepidoptera: Plutellidae) resistance to spinsad, indoxacarb, and emamectin benzoate,” Journal of Economic Entomology, Vol. 99, No. 1, pp. 176-181(2006).
45.Zhou, P., Lu, Y., Liu, B., Gan, J. J., “Dynamics of fipronil residue in vegetable-field ecosystem,” Chemosphere, Vol. 57, No. 11, pp. 1691-1696 (2004).
46.Zhao, J. Z., Collins, H. L., Li, Y. X., Mau, R. F. L.; Thompson, G. D.; Hertlein, M.,Andaloro, J. T.; Boykin, R.; Shelton, A. M., “Monitoring of diamondback moth (Lepidoptera: Plutellidae) resistance to spinosad, indoxacarb, and emamectin benzoate,” Journal of Economic Entomology, Vol. 99, No. 1, (2006).