Honey Bee Research and Extension Laboratory, Department of Entomology and Nematology, University of Florida, Natural Area Drive, Gainesville, FL 32611, USA.
J Insect Physiol. 2012 Aug;58(8):1042-9. doi: 10.1016/j.jinsphys.2012.03.015. Epub 2012 Apr 9.
Honey bee (Apis mellifera) larvae reared in vitro were exposed to one of nine pesticides and/or were challenged with the parasitic mite, Varroa destructor. Total RNA was extracted from individual larvae and first strand cDNAs were generated. Gene-expression changes in larvae were measured using quantitative PCR (qPCR) targeting transcripts for pathogens and genes involved in physiological processes, bee health, immunity, and/or xenobiotic detoxification. Transcript levels for Peptidoglycan Recognition Protein (PGRPSC), a pathogen recognition gene, increased in larvae exposed to Varroa mites (P<0.001) and were not changed in pesticide treated larvae. As expected, Varroa-parasitized brood had higher transcripts of Deformed Wing Virus than did control larvae (P<0.001). Varroa parasitism, arguably coupled with virus infection, resulted in significantly higher transcript abundances for the antimicrobial peptides abaecin, hymenoptaecin, and defensin1. Transcript levels for Prophenoloxidase-activating enzyme (PPOact), an immune end product, were elevated in larvae treated with myclobutanil and chlorothalonil (both are fungicides) (P<0.001). Transcript levels for Hexameric storage protein (Hsp70) were significantly upregulated in imidacloprid, fluvalinate, coumaphos, myclobutanil, and amitraz treated larvae. Definitive impacts of pesticides and Varroa parasitism on honey bee larval gene expression were demonstrated. Interactions between larval treatments and gene expression for the targeted genes are discussed.
体外饲养的蜜蜂(Apis mellifera)幼虫暴露于九种农药中的一种或受到寄生螨(Varroa destructor)的侵袭。从单个幼虫中提取总 RNA,并生成第一链 cDNA。使用针对病原体和参与生理过程、蜜蜂健康、免疫和/或外来化合物解毒的基因的定量 PCR(qPCR)测量幼虫中的基因表达变化。肽聚糖识别蛋白(PGRPSC)的转录物水平增加,这是一种病原体识别基因,在暴露于壁虱的幼虫中(P<0.001),而在农药处理的幼虫中没有变化。正如预期的那样,感染变形翅病毒的寄生蜂幼虫的转录物水平高于对照幼虫(P<0.001)。壁虱寄生,加上病毒感染,导致抗菌肽 abaecin、 hymenoptaecin 和 defensin1 的转录物丰度显著升高。免疫终产物原酚氧化酶激活酶(PPOact)的转录物水平在被 myclobutanil 和 chlorothalonil(均为杀菌剂)处理的幼虫中升高(P<0.001)。六聚体储存蛋白(Hsp70)的转录物水平在 imidacloprid、fluvalinate、coumaphos、myclobutanil 和 amitraz 处理的幼虫中显著上调。证明了农药和壁虱寄生对蜜蜂幼虫基因表达的明确影响。讨论了针对靶基因的幼虫处理和基因表达之间的相互作用。