通过 RNAi 鉴定出与瓦螨生存或繁殖有关的重要基因。

Genes important for survival or reproduction in Varroa destructor identified by RNAi.

机构信息

Department of Entomology, Michigan State University, East Lansing, MI, USA.

Ecology, Evolutionary Biology and Behavior, Michigan State University, East Lansing, MI, USA.

出版信息

Insect Sci. 2019 Feb;26(1):68-75. doi: 10.1111/1744-7917.12513. Epub 2017 Dec 1.

DOI:10.1111/1744-7917.12513

PMID:28748595

Abstract

The Varroa mite, (Varroa destructor), is the worst threat to honey bee health worldwide. To explore the possibility of using RNA interference to control this pest, we determined the effects of knocking down various genes on Varroa mite survival and reproduction. Double-stranded RNA (dsRNA) of six candidate genes (Da, Pros26S, RpL8, RpL11, RpP0 and RpS13) were synthesized and each injected into Varroa mites, then mite survival and reproduction were assessed. Injection of dsRNA for Da (Daughterless) and Pros26S (Gene for proteasome 26S subunit adenosine triphosphatase) caused a significant reduction in mite survival, with 3.57% ± 1.94% and 30.03% ± 11.43% mites surviving at 72 h post-injection (hpi), respectively. Control mites injected with green fluorescent protein (GFP)-dsRNA showed survival rates of 81.95% ± 5.03% and 82.36 ± 2.81%, respectively. Injections of dsRNA for four other genes (RpL8, RpL11, RpP0 and RpS13) did not affect survival significantly, enabling us to assess their effect on Varroa mite reproduction. The number of female offspring per mite was significantly reduced for mites injected with dsRNA of each of these four genes compared to their GFP-dsRNA controls. Knockdown of the target genes was verified by real-time polymerase chain reaction for two genes important for reproduction (RpL8, RpL11) and one gene important for survival (Pros26S). In conclusion, through RNA interference, we have discovered two genes important for mite survival and four genes important for mite reproduction. These genes could be explored as possible targets for the control of Varroa destructor in the future.

摘要

瓦螨（Varroa destructor）是全球范围内对蜜蜂健康最严重的威胁。为了探索利用 RNA 干扰控制这种害虫的可能性，我们确定了敲除各种基因对瓦螨生存和繁殖的影响。合成了 6 个候选基因（Da、Pros26S、RpL8、RpL11、RpP0 和 RpS13）的双链 RNA（dsRNA），并将每种 dsRNA 注入瓦螨体内，然后评估螨的生存和繁殖情况。注射 Da（无女儿）和 Pros26S（蛋白酶体 26S 亚基三磷酸腺苷酶基因）的 dsRNA 导致螨的存活率显著降低，分别在注射后 72 小时（hpi）有 3.57%±1.94%和 30.03%±11.43%的螨存活。用绿色荧光蛋白（GFP）-dsRNA 注射的对照螨的存活率分别为 81.95%±5.03%和 82.36%±2.81%。注射另外 4 个基因（RpL8、RpL11、RpP0 和 RpS13）的 dsRNA 对生存没有显著影响，使我们能够评估它们对瓦螨繁殖的影响。与 GFP-dsRNA 对照相比，每个基因的 dsRNA 注射都导致每只螨虫产生的雌性后代数量显著减少。对于两个重要的繁殖基因（RpL8、RpL11）和一个重要的生存基因（Pros26S），通过实时聚合酶链反应验证了靶基因的敲低。总之，通过 RNA 干扰，我们发现了两个对螨生存重要的基因和四个对螨繁殖重要的基因。这些基因可以作为未来控制瓦螨的可能靶点进行探索。