0.2%苦皮藤素乳油抗药性初步研究
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摘要
0.2%苦皮藤素乳油是一种新型的植物性杀虫剂。为了明确0.2%苦皮藤素乳油商品化后抗药性的发展情况，为指导该乳油的正确、合理使用提供依据。作者以室内饲养的敏感品系小菜蛾为抗性选育虫种，对0.2%苦皮藤素乳油的抗性形成规律、交互抗性和抗性机理进行了初步研究，获得如下结果：
经过20代的抗性选育，抗性增长21.57倍。和常见的化学杀虫剂及微生物杀虫剂（Bt和阿维菌素）相比，其抗性发展速度是最慢的，且呈明显的“S"型。前期(F0～F6)慢、中期(F6～F17)快、后期(F17～F20)基本稳定。敏感品系(F0)和抗性品系不同代(F5、F10、F15和F20)的各项生物学指标（4龄虫重、蛹重、雌雄比、产卵量和孵化率）无明显差异，表明这种植物性杀虫剂连续使用后不会引起种群退化。
选育的抗性品系对10种常用杀虫剂的敏感度测定的结果表明，对杀虫双、杀螟丹和叶蝉散有明显交互抗性（抗性倍数分别为4.63、4.11和3.71），对溴氰菊酯、氯菊酯、氯氰菊酯和Bt有明显负交互抗性(抗性倍数分别为0.22、0.01、0.26和0.66)。以高抗杀螟丹、杀虫双和溴氰菊酯小菜蛾品系为试虫，测试了它们对0.2%苦皮藤素乳油的敏感度。结果表明，这三品系对该乳油分别有1.06、1.76和3.61倍的交互抗性。
用聚丙烯酰胺凝胶电泳比较了敏感品系和抗性品系的酯酶同功酶的差异。结果表明，E4～5是敏感品系的特异性酯酶同功酶，E10是抗性品系的特异性酯酶同功酶，推测E10可能是苦皮藤素的解毒酶。从电泳图谱推断该抗性不是由于基因扩增而产生，E10可能是随着选育代数的增加，敏感品系的基因结构发生了改变或者基因的表达产物发生了变异的结果，使编码E4～5的基因突变为编码E10的基因，或者该基因的表达产物E4～5变为E10,从电泳的图谱的比较可以推测E10可能是与抗药性有关的同功酶，编码E10的基因可能是抗性基因。磷酸酯酶和羧酸酯酶活性测定的结果表明，敏感品系和抗性品系之间无明显差异，说明抗性的产生与这两种酶的活性无关。
0.2% Celangulin EC is a new kind of botanical insecticide. It is essential to study the resistance of 0.2% Celangulin EC for sake of guiding to be used in the field. The preliminary studies to Celangulin resistance on the trend of resistance development, cross-resistance and mechanism of resistance were carried. The selective strain is the susceptible strain of diamondback moth. The main results are as follows:
After 20 generations selection, the resistance of moth increased by 21.57 folds. The speed of resistance was slower than chemical and microbial insecticides, such as Bt and avermectin. The time course of resistance development appeared to be "S" shape. The resistance development was slow in the earlier stage(F0～F6), faster in the mid-stage(F6～F17),and stable in the later stage(F17～F20). There was no obvious difference between the susceptible(F0) and the selective strain(F5, F10, F15 and F20) in biological characteristics, which shows 0.2% Celangulin EC can't result in degeneration of the strain.
The sensitivities of the Celangulin-resistance strain to 10 insecticides were tested. The results showed the resistant strain has positive cross-resistance to dimehypo, cartap and isoprocarb(4.63, 4.11 and 3.71 folds), and negative cross-resistance to deltamethrin, cypermethrin, permecthrin and Bt(0.22, 0.26, 0.01 and 0.66 folds). The sensitivities of cartap, dimehypo and deltamethrin-resistance strains to 0.2% Celangulin EC showed the strains has 1.06, 1.76 and 3.61 folds resistance to 0.2% Celangulin EC, respectively.
PAGE demonstrated that the resistance of 0.2% Celangulin EC was related to special esterases. Maybe enzymes 4～5 are special esterases of susceptible strain and enzyme 10 is special esterase of the resistance strain. Enzyme 10 is an important detoxicative enzyme. Maybe the gene amplification is not a reason of resistance development, but the genes of coding enzymes 4～5 altered expression or changed structure, so that enzyme 10 was expressed. The gene of coding enzyme 10 concerns resistance, probably. The result of study on activities of phosphatase and carboxylesterase showed the resistance wasn't related to phosphatase and carboxylesterase.
经过20代的抗性选育，抗性增长21.57倍。和常见的化学杀虫剂及微生物杀虫剂（Bt和阿维菌素）相比，其抗性发展速度是最慢的，且呈明显的“S"型。前期(F0～F6)慢、中期(F6～F17)快、后期(F17～F20)基本稳定。敏感品系(F0)和抗性品系不同代(F5、F10、F15和F20)的各项生物学指标（4龄虫重、蛹重、雌雄比、产卵量和孵化率）无明显差异，表明这种植物性杀虫剂连续使用后不会引起种群退化。
选育的抗性品系对10种常用杀虫剂的敏感度测定的结果表明，对杀虫双、杀螟丹和叶蝉散有明显交互抗性（抗性倍数分别为4.63、4.11和3.71），对溴氰菊酯、氯菊酯、氯氰菊酯和Bt有明显负交互抗性(抗性倍数分别为0.22、0.01、0.26和0.66)。以高抗杀螟丹、杀虫双和溴氰菊酯小菜蛾品系为试虫，测试了它们对0.2%苦皮藤素乳油的敏感度。结果表明，这三品系对该乳油分别有1.06、1.76和3.61倍的交互抗性。
用聚丙烯酰胺凝胶电泳比较了敏感品系和抗性品系的酯酶同功酶的差异。结果表明，E4～5是敏感品系的特异性酯酶同功酶，E10是抗性品系的特异性酯酶同功酶，推测E10可能是苦皮藤素的解毒酶。从电泳图谱推断该抗性不是由于基因扩增而产生，E10可能是随着选育代数的增加，敏感品系的基因结构发生了改变或者基因的表达产物发生了变异的结果，使编码E4～5的基因突变为编码E10的基因，或者该基因的表达产物E4～5变为E10,从电泳的图谱的比较可以推测E10可能是与抗药性有关的同功酶，编码E10的基因可能是抗性基因。磷酸酯酶和羧酸酯酶活性测定的结果表明，敏感品系和抗性品系之间无明显差异，说明抗性的产生与这两种酶的活性无关。
0.2% Celangulin EC is a new kind of botanical insecticide. It is essential to study the resistance of 0.2% Celangulin EC for sake of guiding to be used in the field. The preliminary studies to Celangulin resistance on the trend of resistance development, cross-resistance and mechanism of resistance were carried. The selective strain is the susceptible strain of diamondback moth. The main results are as follows:
After 20 generations selection, the resistance of moth increased by 21.57 folds. The speed of resistance was slower than chemical and microbial insecticides, such as Bt and avermectin. The time course of resistance development appeared to be "S" shape. The resistance development was slow in the earlier stage(F0～F6), faster in the mid-stage(F6～F17),and stable in the later stage(F17～F20). There was no obvious difference between the susceptible(F0) and the selective strain(F5, F10, F15 and F20) in biological characteristics, which shows 0.2% Celangulin EC can't result in degeneration of the strain.
The sensitivities of the Celangulin-resistance strain to 10 insecticides were tested. The results showed the resistant strain has positive cross-resistance to dimehypo, cartap and isoprocarb(4.63, 4.11 and 3.71 folds), and negative cross-resistance to deltamethrin, cypermethrin, permecthrin and Bt(0.22, 0.26, 0.01 and 0.66 folds). The sensitivities of cartap, dimehypo and deltamethrin-resistance strains to 0.2% Celangulin EC showed the strains has 1.06, 1.76 and 3.61 folds resistance to 0.2% Celangulin EC, respectively.
PAGE demonstrated that the resistance of 0.2% Celangulin EC was related to special esterases. Maybe enzymes 4～5 are special esterases of susceptible strain and enzyme 10 is special esterase of the resistance strain. Enzyme 10 is an important detoxicative enzyme. Maybe the gene amplification is not a reason of resistance development, but the genes of coding enzymes 4～5 altered expression or changed structure, so that enzyme 10 was expressed. The gene of coding enzyme 10 concerns resistance, probably. The result of study on activities of phosphatase and carboxylesterase showed the resistance wasn't related to phosphatase and carboxylesterase.
引文
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