Kakani Evdoxia G, Sagri Efthymia, Omirou Michalis, Ioannides Ioannis M, Mathiopoulos Kostas D
Department of Biochemistry and Biotechnology, University of Thessaly, Greece.
Pest Manag Sci. 2014 May;70(5):743-50. doi: 10.1002/ps.3564. Epub 2013 Jul 3.
The olive fruit fly, Bactrocera oleae (Rossi) (Diptera: Tephritidae), is the most important pest of olives. Its control is based mostly on organophosphate (OP) insecticides, a practice that has led to resistance development. OP resistance in B. oleae has been associated with three mutations in the acetylcholinesterase (AChE), the product of ace gene. The current study presents new diagnostic tests for the detection of the ace mutations and aims at monitoring the frequency of the Δ3Q mutation, which appears associated with resistance at higher OP doses in natural olive fly populations.
An allele-specific polymerase chain reaction (PCR), a PCR-RFLP (restriction fragment length polymorphism) and a Taq-Man test were developed for the Δ3Q mutation detection and a new duplex quantitative PCR assay was designed for the G488S and I214V mutations. Moreover, the frequency of Δ3Q mutation was examined in ten populations of eight countries around the Mediterranean basin. The highest frequencies (10%) were found in Greece and Italy, whereas a gradual decrease of Δ3Q frequency towards the western Mediterranean was noted.
Robust tests for insecticide resistance mutations at their incipient levels are essential tools to monitor the increase and geographical spread of such mutations. Three different tests were developed for AChE-Δ3Q that indicated its association with OP applications across the Mediterranean.
地中海实蝇(Bactrocera oleae (Rossi),双翅目:实蝇科)是橄榄最重要的害虫。其防治主要基于有机磷(OP)杀虫剂,这种做法已导致抗性产生。地中海实蝇对OP的抗性与乙酰胆碱酯酶(AChE,ace基因的产物)的三个突变有关。本研究提出了用于检测ace突变的新诊断测试,并旨在监测Δ3Q突变的频率,该突变在天然橄榄蝇种群中似乎与较高OP剂量下的抗性相关。
开发了一种等位基因特异性聚合酶链反应(PCR)、一种PCR-限制性片段长度多态性(RFLP)和一种Taq-Man测试用于检测Δ3Q突变,并设计了一种新的双重定量PCR测定法用于检测G488S和I214V突变。此外,在地中海盆地周边八个国家的十个种群中检测了Δ3Q突变的频率。在希腊和意大利发现了最高频率(10%),而注意到向西地中海方向Δ3Q频率逐渐降低。
针对初期水平的杀虫剂抗性突变的可靠测试是监测此类突变增加和地理扩散的重要工具。针对AChE-Δ3Q开发了三种不同的测试,表明其与地中海地区OP的使用有关。