候梦赏,程雪芬,邱园妹,朱雅楠,赵必安,李志国,苏松坤,2020,吡虫啉胁迫对意大利蜜蜂哺育蜂免疫解毒相关基因表达及酶活力的影响[J].环境昆虫学报,(6):1415-1423
吡虫啉胁迫对意大利蜜蜂哺育蜂免疫解毒相关基因表达及酶活力的影响
Effects of imidacloprid on immune detoxification-related gene expression and immune detoxification enzymes activity in nurse bees of Apis mellifera ligustica
  
DOI:
中文关键词:  意大利蜜蜂哺育蜂  吡虫啉  免疫  解毒酶  存活率  酶活力
英文关键词:nurse bees of Apis mellifera ligustica  imidacloprid  immunity detoxifying enzyme  survival rate  enzyme activity
基金项目:国家现代农业产业技术体系(蜜蜂)项目(CARS-44-KXJ4);国家自然科学基金项目(31702192)
作者单位
候梦赏,程雪芬,邱园妹,朱雅楠,赵必安,李志国,苏松坤 福建农林大学动物科学学院蜂学学院福州350002 
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中文摘要:
      蜜蜂作为世界上最重要的授粉性昆虫,在采集过程中易接触到杀虫剂,前人研究表明新烟碱类杀虫剂吡虫啉(imidaclorprid)影响意大利蜜蜂Apis mellifera ligustica(简称“意蜂”)的存活和舞蹈、采集等行为。本研究旨在探究亚致死剂量吡虫啉胁迫对意大利蜜蜂哺育蜂(8日龄成年工蜂)免疫解毒相关基因表达、免疫解毒酶系活力及存活率的影响。结果显示哺育蜂连续取食3 d和9 d含0.1 ng/μL吡虫啉的蔗糖液后,其存活率与对照组(饲喂含等量丙酮的蔗糖溶液)无显著差异;连续饲喂11 d含0.1 ng/μL吡虫啉的50%蔗糖溶液后,其存活率与对照组有显著差异。荧光定量PCR检测及双抗体一步夹心法酶联免疫吸附试验结果显示哺育蜂取食吡虫啉3 d后,蜜蜂体内免疫基因多酚氧化酶基因(PPOA3,GB43738),Abaecin类抗菌肽基因(ABA,GB18323),葡萄糖脱氢酶基因(GLD, GB43007)和解毒基因细胞色素P450基因(CYP450 6a2,GB49876),细胞色素B561基因(CYB561 2-like,GB40148),葡萄糖醛酸转移酶(UDP-glucuronosyltransferase,GB52179)的表达及蜂体内体内细胞色素P450酶(cytochrome P450,CYP450)含量均有上调趋势,超氧化物岐化酶(superoxide dismutase,SOD)和过氧化氢酶(catalase,CAT)均有显著下调趋势;哺育蜂取食吡虫啉9 d后,PPOA3,ABA,GLD,CYP450 6a2,CYB561 2-like,UDP-glucuronosyltransferase的表达及蜂体内体内细胞色素P450酶含量均有下调趋势,多酚氧化酶(polyphenol oxidase,PPO),超氧化物歧化酶和过氧化氢酶酶活力均有显著下调趋势。本研究在分子水平上提供了亚致死剂量吡虫啉是通过扰乱蜜蜂正常的免疫系统进而影响蜜蜂行为的证据,以期为维护蜜蜂健康提供一定的理论依据。
英文摘要:
      Honeybees, Apis mellifera ligustica are one of the most important pollinating insects in the world. However, nurse bees are easily accessed to pesticides during the collection process. Previous study documents have shown that neonicotinoid pesticides, imidaclorprid, affects the survival rate, dance and collection behavior of bees. The purpose of the experiment was to investigate the effects of sublethal doses imidacloprid on the expression of immune detoxification-related genes, the activity of immune detoxification enzymes, and the survival rate of nurse bees (8-day worker bees). The results showed that the nurse bees of 8-day were continuously fed with 50% sucrose solution containing 0.1 ng/μL imidacloprid for 3 d and 9 d showed no significant difference; after 11 days of continuous feeding with 50% sucrose solution containing 0.1 ng/μL imidacloprid, the survival rate was significantly different from the control group (fed with sucrose solution containing the same amount of acetone). The results of real-time quantitative PCR and double-antibody one-step sandwich enzyme-linked immunosorbent assays showed that nurse bees of 8-day took sucrose solution containing 0.1 ng/μL imidacloprid for 3 d. The polyphenol oxidase gene (PPOA3, GB43738), Abaecin antibacterial peptide gene (ABA, GB18323), glucose dehydrogenase gene (GLD, GB43007) and detoxification gene cytochrome P450 gene (CYP450 6a2, GB49876), cytochrome B561 gene (CYB561 2-like, GB40148), the expression of UDP-glucuronosyltransferase (GB52179) and the content of cytochrome P450 (CYP450) in the body of the bees are all up-regulated. Superoxide dismutase (SOD) and Catalase (CAT) had a significant downward trend; nurse bees of Apis mellifera ligustica fed with sucrose solution containing 0.1 ng/μL imidacloprid for 9 d, then PPOA3, ABA, GLD, CYP450 6a2, CYB561 2-like, the expression of UDP-glucuronosyltransferase and the content of cytochrome P450 enzymes in bees were down-regulated. Polyphenol oxidase (polyphenol oxidase, PPO), superoxide dismutase and catalase enzymes activities had a significant downward trend. This study provides evidence at the molecular level that sublethal doses of imidacloprid can affect bees’ behavior by disrupting the bees' normal immune system, in order to provide some theoretical basis for the maintenance of bee health.
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