亚致死剂量呋虫胺对意大利蜜蜂采集蜂免疫解毒相关基因表达和酶活性的影响

Effects of sublethal doses of dinotefuran on genes expression and enzymes activities related to immune and detoxification in foragers of Apis mellifera ligustica

为确定呋虫胺对意大利蜜蜂Apis mellifera ligustica采集蜂的亚致死浓度,探究亚致死剂量呋虫胺对意大利蜜蜂采集蜂免疫解毒相关基因的表达及酶活性的影响,本研究采用饲喂法测定呋虫胺对意大利蜜蜂采集蜂的毒力曲线。应用qRT-PCR技术研究LC_(20)和LC_(50)浓度呋虫胺对蜜蜂Abaecin,Apidaecins,Defensin-1,Hymenoptaecin,AChE-2,PPO,GST及Cyp 450 mRNA相对表达量的影响。测定蜜蜂体内多酚氧化酶、乙酰胆碱酯酶、超氧化物歧化酶、谷胱甘肽S-转移酶和过氧化氢酶活性的变化。结果表明,呋虫胺胁迫意大利蜜蜂采集蜂的LC_(20)和LC_(50)为0.458 mg·L^(-1)和0.988 mg·L^(-1)。LC_(20)组中PPO、Apidaecins、Defensin-1和Hymenoptaecin基因表达量与对照组相比有显著(P<0.05)上调趋势,LC_(50)组中AChE-2、PPO、GST、Cyp450、Apidaecins和Defensin-1表达量有显著上调趋势,Abaecin和Hymenoptaecin基因表达量有显著下调趋势,且LC_(50)组中的abaecin和Hymenoptaecin表达量均显著低于LC_(20)组,AChE-2、PPO、GST、Cyp 450和Apidaecins基因表达量显著高于LC_(20)组。LC_(20)组与对照组相比,AChE、CAT和SOD活性均显著下降;LC_(50)组与对照组相比,AChE、PPO、CAT和SOD活性均显著下降。呋虫胺抑制Abaecin和Hymenoptaecin基因表达,诱导Apidaecins、Defensin-1、AChE-2、PPO、GST和Cyp 450基因的表达,影响正常的免疫系统。AChE、PPO、CAT和SOD活性的下降预示着呋虫胺对蜜蜂解毒系统的损害,本试验为探究呋虫胺对意大利蜜蜂生理和代谢的影响提供理论依据。

To determine sublethal doses of dinotefuran to Apis mellifera ligustica and explore the effects of sublethal doses of dinotefuran on the expression of immune detoxification genes and enzyme activities on Apis mellifera ligustica,in this study,the feeding trials was used to determine the virulence curve of dinotefuran to Apis mellifera ligustica.The effects of LC_(20) and LC_(50) dinotefuran on the relative expressions of Abaecin,Apidaecins,Defensin-1,Hymenoptaecin,AChE-2,PPO,GST and Cyp450 mRNA were investigated by qRT-PCR.Determination of changes in the activities of PPO,AChE,SOD,GST,CAT in Apis mellifera ligustica.The LC_(20) and LC_(50) of dinotefuran on Apis mellifera ligustica determined by feeding trials were 0.458 mg·L^(-1) and 0.988 mg·L^(-1).Compared with the control group,the expression levels of PPO,Apidaecins,Defensin-1,Hymenoptaecin genes were significantly(P<0.05)up-regulated in the LC_(20) group.Apidaecins,In LC_(50) group,the expression of AChE-2,PPO,GST,Cyp450,Apidaecins,Defensin-1 showed a significant up-regulation trend,and the expression of Hymenoptaecin and Abaecin showed a significant down-regulation trend.In additon,the expression levels of Abaecin and Hymenoptaecin in the LC_(50) group were significantly lower than those in the LC_(20) group.The expression levels of AChE-2,PPO,GST,Cyp450,Apidaecins in the LC_(50) group were significantly higher than those in the LC_(20) group.Compared with the control group,the activities of AChE,CAT and SOD of LC_(20) group decreased significantly.Compared with the control group,the activities of AChE,PPO,CAT and SOD of LC_(50) group decreased significantly.In conclusion,dinotefuran inhibits gene expression of Abaecin and Hymenoptaecin,and has an inducing effect on Apidaecins,Defensin-1,AChE-2,PPO,GST,Cyp450 affects the normal immune system of Apis mellifera ligustica.The decline of AChE,PPO,CAT and SOD enzyme activities indicates the damage of dinotefuran to the detoxification system of Apis mellifera ligustica.Our stduy provides a further basis for further research on the effects of sublethal dose dinotefuran on the metabolism and physiology of Apis mellifera ligustica.