Mookhploy Wannapha, Krongdang Sasiprapa, Chantawannakul Panuwan
Bee Protection Laboratory, Department of Biology, Faculty of Science, Chiang Mai University, Chiang Mai 50200, Thailand.
Graduate School, Chiang Mai University, Chiang Mai 50200, Thailand.
Insects. 2021 Jan 19;12(1):82. doi: 10.3390/insects12010082.
Honeybees are globally threatened by several pathogens, especially deformed wing virus (DWV), as the presence of DWV in western honeybees is indicative of colony loss. The high mortality rate is further exacerbated by the lack of effective treatment, and therefore understanding the immune and apoptosis responses could pave an avenue for the treatment method. In this study, DWV was directly injected into the white-eyed pupae stage of western honeybees (). The DWV loads and selected gene responses were monitored using the real-time PCR technique. The results showed that honeybee pupae that were injected with the highest concentration of viral loads showed a significantly higher mortality rate than the control groups. Deformed wings could be observed in newly emerged adult bees when the infected bees harbored high levels of viral loads. However, the numbers of viral loads in both normal and crippled wing groups were not significantly different. DWV-injected honeybee pupae with 10 and 10 copy numbers per bee groups showed similar viral loads after 48 h until newly emerged adult bees. Levels of gene expression including immune genes (, and ) and apoptosis genes ( and ) were analyzed after DWV infection. The expressions of immune and apoptosis genes were significantly different in infected bees compared to those of the control groups. In the pupae stage, the immune genes were activated by injecting DWV ( and ) or ( and ), a positive control. On the contrary, the expression of apoptosis-related genes (, , , and genes) was suppressed at 96 h post-infection. In DWV-infected newly emerged adult bees, , and genes were upregulated. However, these genes were not significantly different between the normal and crippled wing bees. Our results suggested that DWV could activate the humoral immunity in honeybees and that honeybee hosts may be able to protect themselves from the virus infection through immune responses. Apoptosis gene expressions were upregulated in newly emerged adult bees by the virus, however, they were downregulated during the initial phase of viral infection.
蜜蜂在全球范围内受到多种病原体的威胁,尤其是变形翅病毒(DWV),因为西方蜜蜂体内存在DWV表明蜂群会损失。由于缺乏有效的治疗方法,高死亡率进一步加剧,因此了解免疫和凋亡反应可为治疗方法开辟一条道路。在本研究中,将DWV直接注射到西方蜜蜂的白眼蛹阶段()。使用实时PCR技术监测DWV载量和选定基因的反应。结果表明,注射最高浓度病毒载量的蜜蜂蛹的死亡率明显高于对照组。当受感染的蜜蜂携带高水平的病毒载量时,在新出现的成年蜜蜂中可以观察到变形的翅膀。然而,正常翅膀组和残缺翅膀组的病毒载量数量没有显著差异。每只蜜蜂注射10和10拷贝数的DWV蜜蜂蛹在48小时后直到新出现的成年蜜蜂都显示出相似的病毒载量。在DWV感染后,分析了包括免疫基因(、和)和凋亡基因(和)在内的基因表达水平。与对照组相比,感染蜜蜂中免疫和凋亡基因的表达有显著差异。在蛹阶段,通过注射DWV(和)或(和)(阳性对照)激活免疫基因。相反,在感染后96小时,凋亡相关基因(、、、和基因)的表达受到抑制。在DWV感染的新出现的成年蜜蜂中,、和基因上调。然而,这些基因在正常翅膀和残缺翅膀的蜜蜂之间没有显著差异。我们的结果表明,DWV可以激活蜜蜂的体液免疫,并且蜜蜂宿主可能能够通过免疫反应保护自己免受病毒感染。病毒在新出现的成年蜜蜂中上调了凋亡基因的表达,然而,在病毒感染的初始阶段它们被下调。