摘要
为探究亚致死剂量吡虫啉胁迫对红光熊蜂工蜂的存活率、解毒酶活力与解毒基因表达量的影响,采用cDNA末端快速扩增技术(RACE)克隆红光熊蜂工蜂(21日龄)解毒基因,检测其表达量变化与抗氧化酶的活性,并通过饲喂外源褪黑素以提高熊蜂的抗药性.结果表明:随着胁迫时间的延长,吡虫啉处理组(IMI组)存活率显著下降,但始终低于吡虫啉+褪黑素处理组(IMI+MT组);IMI组4种抗氧化酶的活性均随吡虫啉胁迫时间的延长呈先上升后下降的趋势,始终低于IMI+MT组;IMI组8种解毒基因的表达量除AChE,DDTase和GLD基因外,其余表达量均随吡虫啉胁迫时间的延长呈先激活后抑制的状态,且明显高于对照组(CK组)而低于IMI+MT组;IMI组DDTase基因的表达量与CK组差异不明显,但始终低于IMI+MT组,说明吡虫啉对DDTase的作用不明显,而褪黑素诱导DDTase表达效果较显著(P<0.05);IMI组AChE和GLD基因的表达量均低于CK组,随吡虫啉胁迫时间的延长而呈下降趋势,但IMI+MT组二者的表达量始终高于IMI组.短期亚致死剂量吡虫啉胁迫对红光熊蜂影响不显著,长期胁迫则影响其存活率,红光熊蜂对吡虫啉的代谢解毒是一个多酶系、多基因协同参与的理化过程,褪黑素可提高红光熊蜂的抗药性.
To explore the sublethal dose of imidacloprid stress on the effect of Bombus ignitus workers,detoxification enzyme activity and detoxification gene expression.The detoxification genes were cloned by the RACE(rapid amplification of cDNA ends)technique,the expression levels and antioxidant enzyme activity of them in the 2l day-old of Bombus ignitus after treatment were detected,melatonin was also fed to improve the resistance of bumblebees.The results showed that,With the increasing duration of the stress period,The significant decrease in survival in the imidacloprid processing group(IMI group),But always lower than imidacloprid+melatonin processing group(IMI+MT group);Expression of the eight detoxification genes in the IMI group,in addition to AChE,DDTase and GLD,the rest of the expression levels increased with the extension of imidacloprid stress,and significantly higher than CK group and lower than IMI+MT group;The expression of DDTase in the IMI group was not significantly different from that in the CK group,but were consistently lower than those in the IMI+MT group,suggesting that the effect of imidacloprid on DDTase is insignificant.However,melatonin induced DDTase expression significantly(P<0.05);The expression level of both AChE and GLD in the IMI group was lower than that in the CK group,detrend with the extension of imidacloprid stress.However,the expression level in the IMI+MT group was always higher than that in the IMI group.In conclusion,short-term sublethal dose of imidacloprid stress has no significant effect on Bombus ignitus,while long-term stress affects their survival rate,and the metabolic detoxification of pyridacloprid is a physical and chemical process involving multiple enzyme lines and multiple genes,and melatonin can improve the resistance of Bombus ignitus.
作者
王丹丹
王星
张晓婉
刘丹梅
WANG Dan-dan;WANG Xing;ZHANG Xiao-wan;LIU Dan-mei(Institute of Advanced Characteristic Agriculture Studies,Liaodong University,Dandong 118003,China)
出处
《东北师大学报（自然科学版）》
CAS
北大核心
2024年第1期125-132,共8页
Journal of Northeast Normal University(Natural Science Edition)
基金
国家自然科学基金资助项目(31972363,31872048)
辽宁省教育厅科研项目(LJKZ1124)
辽宁省科学事业公益研究项目(20202681)。
关键词
红光熊蜂
吡虫啉
抗氧化酶
解毒酶
克隆
基因表达
Bombus ignitus
imidacloprid
antioxidase
detoxification enzyme
cloning
gene expression
