Department of Agricultural Technology, Faculty of Science and Technology, Thammasat University, Rangsit Centre, Khlong Nueng, Klong Luang, Pathum Thani, Thailand.
School of Crop Production Technology, Institute of Agricultural Technology, Suranaree University of Technology, Suranaree, Muang, Nakhon Ratchasima, Thailand.
Curr Microbiol. 2022 Apr 30;79(6):173. doi: 10.1007/s00284-022-02822-8.
Wolbachia infections affect the reproductive system and various biological traits of the host insect. There is a high frequency of Wolbachia infection in the leafhopper Yamatotettix flavovittatus Matsumura. To investigate the potential roles of Wolbachia in the host, it is important to generate a non-Wolbachia-infected line. The efficacy of antibiotics in eliminating Wolbachia from Y. flavovittatus remains unknown. This leafhopper harbors the mutualistic bacterium Candidatus Sulcia muelleri, which has an important function in the biological traits. The presence of Ca. S. muelleri raises a major concern regarding the use of antibiotics. We selectively eliminated Wolbachia, considering the influence of antibiotics on leafhopper survival and Ca. S. muelleri prevalence. The effect of artificial diets containing different doses of tetracycline and rifampicin on survival was optimized; high dose (0.5 mg/ml) of antibiotics induces a high mortality. A concentration of 0.2 mg/ml was chosen for the subsequent experiments. Antibiotic treatments significantly reduced the Wolbachia infection, and the Wolbachia density in the treated leafhoppers sharply declined. Wolbachia recurred in tetracycline-treated offspring, regardless of antibiotic exposure. However, Wolbachia is unable to be transmitted and restored in rifampicin-treated offspring. The dose and treatment duration had no significant effect on the infection and density of Ca. S. muelleri in the antibiotic-treated offspring. In conclusion, Wolbachia in Y. flavovittatus was stably eliminated using rifampicin, and the Wolbachia-free line was generated at least two generations after treatment. This report provides additional experimental procedures for removing Wolbachia from insects, particularly in host species with the coexistence of Ca. S. muelleri.
沃尔巴克氏体感染会影响宿主昆虫的生殖系统和各种生物学特性。在叶蝉 Yamatotettix flavovittatus Matsumura 中，沃尔巴克氏体的感染频率很高。为了研究沃尔巴克氏体在宿主中的潜在作用，生成非感染沃尔巴克氏体的品系非常重要。抗生素消除 Y. flavovittatus 中的沃尔巴克氏体的效果尚不清楚。该叶蝉携带共生菌 Candidatus Sulcia muelleri，其在生物学特性中具有重要功能。Ca. S. muelleri 的存在使得抗生素的使用成为一个主要关注点。我们考虑了抗生素对叶蝉生存和 Ca. S. muelleri 流行率的影响，选择性地消除了沃尔巴克氏体。优化了含有不同剂量四环素和利福平的人工饲料对生存的影响；高剂量（0.5mg/ml）的抗生素会导致高死亡率。选择 0.2mg/ml 的浓度用于后续实验。抗生素处理显著降低了沃尔巴克氏体的感染，处理叶蝉中的沃尔巴克氏体密度急剧下降。沃尔巴克氏体在四环素处理的后代中再次出现，无论是否接触抗生素。然而，沃尔巴克氏体无法在利福平处理的后代中传播和恢复。抗生素处理对 Ca. S. muelleri 的感染和密度没有显著影响。总之，使用利福平稳定地消除了 Y. flavovittatus 中的沃尔巴克氏体，并在处理后至少两代产生了无沃尔巴克氏体的品系。本报告提供了从昆虫中去除沃尔巴克氏体的额外实验程序，特别是在共存 Ca. S. muelleri 的宿主物种中。
