新烟碱类杀虫剂胁迫下中、意蜂行为和生理响应的比较研究

发布时间：2018-01-13 22:37

本文关键词：新烟碱类杀虫剂胁迫下中、意蜂行为和生理响应的比较研究　出处：《福建农林大学》2017年硕士论文　论文类型：学位论文

【摘要】：新烟碱类杀虫剂是目前全球范围内应用最广泛的一类农药,它们的大量使用对害虫有很好的防治作用,但对不属于靶标类的授粉昆虫也产生了一定危害。我国是农业大国,蜜蜂作为最主要的授粉昆虫在农业经济发展中起着至关重要的作用。本文通过研究新烟碱类杀虫剂(吡虫啉和噻虫胺)对我国饲养的两个主要蜂种——中华蜜蜂(中蜂,Apis cerana cerana)和意大利蜜蜂(意蜂,Apis mellifera L.)的行为、免疫基因表达量和主要解毒酶活性的影响,分别评估了这两种新烟碱类杀虫剂对中、意蜂健康的危害。通过比较分析,进一步探究中、意蜂对新烟碱类杀虫剂免疫机制和解毒机制的差异性。首先通过测定吡虫啉和噻虫胺对中、意蜂24h内口试LD50,评估新烟碱类杀虫剂对蜜蜂的急性致死毒性。吡虫啉对意蜂24 h内口试LD50为8.6 ng/只、中蜂1.9ng/只,噻虫胺对意蜂24h内口试LD50为2.0ng/只,中蜂0.6ng/只。中蜂相比于意蜂对新烟碱类杀虫剂更敏感。通过蔗糖反应行为实验和联想性学习行为实验,分析杀虫剂对蜜蜂伸吻学习记忆能力的影响。实验结果表明,新烟碱类杀虫剂对中蜂学习记忆能力的影响更大。学习实验中,经过吡虫啉处理的意蜂从第四次训练开始,对照组与处理组伸吻反应率差异逐渐缩小,但经过处理的中蜂没有观察到明显的恢复趋势。通过测定蜜蜂体内抗菌肽等免疫基因的表达量,经统计分析得出,新烟碱类杀虫剂能够引起蜜蜂相关免疫基因表达量的上调,但上调趋势有所差异。Eater基因在中蜂经过吡虫啉经过处理后2 h,相对表达量达到峰值,而意蜂在经过处理后24 h达到峰值。Defensin族基因在经过处理后的中蜂体内表达量受到明显抑制,但在意蜂体内表达量明显上调。通过新烟碱类杀虫剂对两种蜜蜂解毒酶比活力的检测,然后对蜜蜂不同组织部位解毒酶的比活力进行统计分析,结果显示,意蜂和中蜂解毒酶的比活力没有明显差异,但新烟碱类杀虫剂能够引起蜜蜂体内多酚氧化酶、羧酸酯酶、谷胱甘肽-s-转移酶三种解毒酶的比活力升高,同时抑制蜜蜂脑部乙酰胆碱酯酶的比活力,说明解毒酶也参与了蜜蜂对新烟碱类杀虫剂的防御机制且新烟碱类杀虫剂作为神经毒剂参与了蜜蜂脑部信号的传导。
[Abstract]:New nicotinic insecticides are the most widely used pesticides in the world at present. Their extensive use has a good effect on pest control. But it is harmful to pollination insects that do not belong to target category. China is a big agricultural country. Bee, as the most important pollinating insect, plays an important role in the development of agricultural economy. In this paper, new nicotinic insecticides (imidacloprid and thiamethylamine) were studied. The two main species of bees raised in China, the Chinese bee (Chinese bee), are introduced in this paper. The behavior of Apis cerana cerana and Apis mellifera L., the effect of immune gene expression and main detoxification enzyme activity. The health hazards of these two new nicotinic insecticides were evaluated respectively. The difference of the immune and detoxification mechanisms of Italian bee to new nicotinic insecticides. First, LD50 was determined by measuring imidacloprid and thiaclomide pairs and oral test of Italian bee within 24 hours. To evaluate the acute lethal toxicity of new nicotinic insecticides to honeybees, the oral LD50 of imidacloprid to Italian bees was 8.6ng / mouse, and that of Chinese wasp was 1.9ng. within 24 hours. The oral test LD50 of thiamethylamine was 2.0ng / mouse in 24 hours. The wasp was more sensitive to the new nicotinic insecticides than the Italian wasp. The experiment of sucrose reaction and associative learning behavior was carried out. The effects of insecticides on the ability of learning and memory of honeybee were analyzed. The results showed that the effect of new nicotine insecticides on the ability of learning and memory was greater. From 4th times of training, the difference of the rate of spray-kiss reaction between the control group and the treatment group was reduced gradually after the treatment of imidacloprid. However, no obvious recovery trend was observed in the treated Chinese bee. The expression of antimicrobial peptides and other immune genes in the honeybee was determined by statistical analysis. New nicotinic insecticides could induce the up-regulation of honeybee related immune genes, but the upregulation trend was different. The relative expression of Eater gene reached the peak at 2 h after treated with imidacloprid. However, the expression of the defensin gene reached the peak at 24 h after treatment, and the expression of the gene in the treated wasp was significantly inhibited. The specific activity of detoxification enzyme of two kinds of bees was detected by the new nicotinic insecticides, and then the specific activity of detoxification enzyme in different tissues of honeybee was statistically analyzed. There was no significant difference in the specific activity of detoxifying enzymes between Italian bee and Chinese bee, but the specific activity of polyphenol oxidase, carboxylic acid esterase and glutathione-stransferase were increased by new nicotine insecticides. At the same time, the specific activity of acetylcholinesterase in bee brain was inhibited, which indicated that detoxification enzyme was also involved in the defense mechanism of honeybee against new nicotinic insecticides and that neotinic insecticides were involved in the transmission of brain signals as nerve agents.
【学位授予单位】：福建农林大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S482.3;S895.9

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