Localization of endosymbionts Portiera and Wolbachia in Bemisia tabaci based on improved fluorescence in situ hybridization

Lu Zhenhua,Yan Xiaohui,Ma Yajie,Shan Yongpan,Song Xianpeng,Liu Qingmei,Wang Dan,Hu Hongyan,Ma Yan

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Cotton Science ›› 2022, Vol. 34 ›› Issue (1) : 23-32. DOI: 10.11963/cs20210027
RESEARCH REPORTS

Localization of endosymbionts Portiera and Wolbachia in Bemisia tabaci based on improved fluorescence in situ hybridization

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Abstract

[Objective] Whitefly, Bemisia tabaci is one of the most invasive insect pest, and its endosymbiont Wolbachia plays an important role in population differentiation and speciation of this pest insect. The aim of this study is to identify the localization and distribution of the primary endosymbiont Portiera and the secondary endosymbiont Wolbachia, by using the fluorescence in situ hybridization (FISH). [Method] Whitefly samples were subjected to whole-mount FISH using two specific fluorescent probes to detect Portiera and Wolbachia in the B. tabaci at different developmental stages. The abdominal tissue samples of adult B. tabaci dissected in phosphate buffer saline (PBS) were stained using an optimized FISH protocol to recognize the localization of the endosymbionts. [Result] When whitefly samples were subjected to whole-mount FISH, hybridization signals were observed at different developmental stages of the B. tabaci. Portiera and Wolbachia were localized inside of the bacteriocytes in eggs and nymphae. However, the localization of these two bacteria in adults of B. tabaci was obscured. The staining results of the dissected samples showed that Portiera and Wolbachia were located inside of the bacteriocyte and ovary of B. tabaci. [Conclusion] An optimized fluorescence staining method of insect tissue was established in this study. The method can be used for the observation of the localization of these two endosymbionts in the bacteriocyte and ovary of B. tabaci, and can provide references for the observation of fluorescence staining of other insects and tissue.

Keywords

Bemisia tabaci / endosymbiont / Portiera / Wolbachia / localization / fluorescence in situ hybridization / method improvement

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Lu Zhenhua , Yan Xiaohui , Ma Yajie , Shan Yongpan , Song Xianpeng , Liu Qingmei , Wang Dan , Hu Hongyan , Ma Yan. Localization of endosymbionts Portiera and Wolbachia in Bemisia tabaci based on improved fluorescence in situ hybridization[J]. Cotton Science, 2022, 34(1): 23-32. https://doi.org/10.11963/cs20210027

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