| Project/Area Number |
14206005
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
植物保護
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| Research Institution | University of Tsukuba |
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Principal Investigator |
KONO Yoshiaki University of Tsukuba, Graduate School of Life and Environmental Sciences, Professor, 大学院・生命環境科学研究科, 教授 (10225386)
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| Co-Investigator(Kenkyū-buntansha) |
TOMITA Takashi National Institute of Infectious Diseases, Department of Medical Entomology, Head of Laboratory, 昆虫医科学部, 室長 (20180169)
SHIOTSUKI Takahiro University of Tsukuba, Institute of Agrobiological Sciences, Developmental Biology Department, Head of Laboratory, 発生分化研究グループ, チーム長
KOBAYASHI Jun Yamaguchi University, Faculty of Agriculture, Professor, 農学部, 教授 (70242930)
本田 洋 筑波大学, 農林学系, 助教授 (90126160)
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| Project Period (FY) |
2002 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥49,010,000 (Direct Cost: ¥37,700,000、Indirect Cost: ¥11,310,000)
Fiscal Year 2004: ¥12,350,000 (Direct Cost: ¥9,500,000、Indirect Cost: ¥2,850,000)
Fiscal Year 2003: ¥15,990,000 (Direct Cost: ¥12,300,000、Indirect Cost: ¥3,690,000)
Fiscal Year 2002: ¥20,670,000 (Direct Cost: ¥15,900,000、Indirect Cost: ¥4,770,000)
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| Keywords | acetylcholinesterase / insecticide resistance / insensitivity to insecticide / amino acid substitution / gene expression / purification of cnzyme / purification of enzyme / organophosphate / 酸素精製 / コガタアカイエカ / 分子機構 / アブラムシ / ハダニ / 殺虫剤感受性 / cDNA |
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| Research Abstract |
Molecular mechanism of acetylcholinesterase (AChE) insensitivity to organophosphates and carbamates was studied, and the insensitivity was elucidated to be caused by amino acid substitutions in the active cite of the enzyme. In cyclorrhapha dipterans, only one gene of AChE exists in their genome, and several amino acids were replaced in the insensitive AChE. In the orthologous AChE of other insects, however, no amino acid substitution has been found in relation to the insensitivity. After the discovery of the second AChE (Ace paralogous AChE, AP-AChE) gene in an aphid and a mosquito species in 2002, we cloned AP-AChE cDNA from several species of resistant insects and mites ; Myzus persicae, Aphis gossypii, Nephotettix cincticeps, Tetranychus urticae and Tetranychus kanzawai, and found species specific amino acid substitutions causing the insensitivity at the important position in the active site of the enzyme. When 6 recombinant AChEs having respective amino acid substitutions were expressed in baculovirus-insect cultured cell system, AChEs produced showed specific characteristics in the reduction of sensitivity to insecticides.
Purification of AChE and gene expression of two AChE were also studied in the mosquitoes.
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