Aseptic rearing procedure for the stinkbug Plautia stali (Hemiptera: Pentatomidae) by sterilizing food-derived bacterial contaminants
Aseptic rearing procedure for the stinkbug Plautia stali (Hemiptera: Pentatomidae) by sterilizing food-derived bacterial contaminants
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DOI：
10.1007/s13355-017-0495-y
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发表时间：
2017-08-01
影响因子：
1.3
通讯作者：
Koga, Ryuichi
中科院分区：
文献类型：
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作者：
Nishide, Yudai;Onodera, Naoko T.;Koga, Ryuichi
臭虫Plautia Stali Scott是一种臭名昭著的农业害虫，其后中肠含有对其生长和生存至关重要的特定细菌，被强调为共生研究的实验模型。Stali的一些共生菌是可培养的，可以在环境中自由生活和获得，而且一些自由生活的环境细菌有可能与P.Stali建立共生关系。在这种情况下，预计这种环境细菌可能偶尔会污染和感染实验室中的实验昆虫，这可能会影响共生的功能分析。在这里，我们报告了这样的污染事件发生在实验室饲养条件下的P.Stali。从表面灭菌的鸡蛋中分离出共生菌的初生若虫，在灭菌的塑料容器中用高压灭菌水饲养时，大部分可能是由于脱离共生而死亡，但只有一小部分能成虫，成虫都感染了与泛托亚和肠杆菌结盟的伽马变形杆菌。从作为P.Stali食物的花生和大豆中检测到各种细菌，主要是芽孢杆菌，也包括泛细菌和肠杆菌。花生和大豆的高压灭菌消除了这些细菌，但对寄主的存活产生了负面影响，而花生和大豆的乙醇灭菌去除了泛细菌和肠杆菌，但不能去除芽孢杆菌，对寄主的存活率没有负面影响。在这些结果的基础上，我们建立了一套实用的Stali无菌培养程序，这将使我们能够可靠和严格地分析模式共生系统中宿主-共生体的相互作用。
The stinkbug Plautia stali Scott is a notorious agricultural pest whose posterior midgut hosts specific bacteria essential for its growth and survival, highlighted as an experimental model for symbiosis studies. Some symbiotic bacteria of P. stali are cultivable, found free-living in and acquired from the environment, and, furthermore, some free-living environmental bacteria are potentially capable of establishing symbiotic association with P. stali. In this context, it is expected that such environmental bacteria may occasionally contaminate and infect the experimental insects maintained in the laboratory, which could potentially affect the functional analyses of the symbiosis. Here we report that such contamination events do occur under a laboratory rearing conditions for P. stali. When symbiont-deprived newborn nymphs from surface-sterilized eggs were reared in sterilized plastic containers with autoclaved water, most of them died as nymphs presumably as a result of aposymbiosis, but only a small fraction could attain adulthood and the adult insects were all infected with gamma-proteobacteria allied to Pantoea and Enterobacter. A variety of bacteria, mainly Bacillus and also Pantoea and Enterobacter, were detected from peanuts and soybeans provided as food for P. stali. Autoclaving of peanuts and soybeans eradicated these bacteria but negatively affected the host survival, whereas ethanol sterilization of peanuts and soybeans removed Pantoea and Enterobacter, but not Bacillus, without negative effects on the host survival. On the basis of these results, we established a practical procedure for aseptic rearing of P. stali, which will enable reliable and strict analyses of host-symbiont interactions in the model symbiotic system.