A single-pair method to screen Rickettsia-infected and uninfected whitefly Bemisia tabaci populations
A single-pair method to screen Rickettsia-infected and uninfected whitefly Bemisia tabaci populations
复制标题
单对法筛查感染和未感染立克次体的粉虱烟粉虱群体
DOI：
10.1016/j.mimet.2019.105797
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发表时间：
2020-01-01
影响因子：
2.2
通讯作者：
Qiu, Bao-Li
中科院分区：
文献类型：
--
作者：
Liu, Yuan;Fan, Ze-Yun;Qiu, Bao-Li
细菌内共生菌如立克次体和沃尔巴克氏菌在其寄主如粉虱、蚜虫、木虱和粉虱的发育和行为中起着重要的作用。越来越多的研究强调了建立缺乏或含有某些内共生体的实验昆虫种群的重要性，因为它们是揭示单个共生体生物学作用的基础。本研究以烟粉虱立克次体为例，探索了一种“单对筛选”的方法，建立了烟粉虱立克次体感染种群和未感染种群，为进一步的实验利用奠定了基础。原宿主人群立克次体感染率较低(35%)。当新出现的烟粉虱成虫时，随机选择未交配的雄性和雌性，放入覆盖着健康植物叶片的叶笼中，以便产卵F1代。产卵6天后，取其亲本进行聚合酶链式反应。将亲本为立克次体阳性或阴性的F1代用于产生F2代，F3、F4和F5代也是如此；如果F1至F5代立克次体感染状况一致，则该群体可作为立克次体阳性或阴性品系进行进一步实验。此外，我们的系统发育分析表明，立克次体在不同世代的母体传播过程中具有很高的保真度。
Bacterial endosymbionts such as Rickettsia and Wolbachia play prominent roles in the development arid behaviour of their insect hosts, such as whiteflies, aphids, psyllids and mealybugs. Accumulating studies have emphasized the importance of establishing experimental insect populations that are either lacking or bearing certain species of endosymbionts, because they are the basis in which to reveal the biological role of individual symbionts. In this study, using Rickettsia as an example, we explored a "single-pair screening" method to establish Rickettsia infected and uninfected populations of whitefly Bemisia tabaci MEAM1 for further experimental use. The original host population had a relatively low infection rate of Rickettsia (< 35%). When B. tabaci adults newly emerged, unmated males and females were randomly selected, and released into a leaf cage that covered a healthy plant leaf in order to oviposit F1 generation eggs. Following 6 days of oviposition, the parents were recaptured and used for PCR detection. The F1 progeny, for which parents were either Rickettsia positive or negative, were used to produce the F2 generation, and similarly in turn for the F3, F4 and F5 generations respectively; if the infection status of Rickettsia was consistent in the F1 to F5 generations, then the populations can be used as Rickettsia positive or negative lines for further experiments. In addition, our phylogenetic analyses revealed that Rickettsia has high fidelity during the maternal transmission in different generations.