The paratransgenic potential of Lactobacillus kunkeei in the honey bee Apis mellifera
The paratransgenic potential of Lactobacillus kunkeei in the honey bee Apis mellifera
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DOI：
10.3920/bm2014.0115
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发表时间：
2015-01-01
影响因子：
5.4
通讯作者：
Diep, D. B.
中科院分区：
文献类型：
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作者：
Rangberg, A.;Mathiesen, G.;Diep, D. B.
蜜蜂（Apis mellifera）是一种对人类社会具有重要价值的家养昆虫，在农业上是农作物的传粉者，在科学研究中是模式动物。然而，由于蜂群衰竭失调（CCD）现象，蜜蜂在世界范围内经历了大规模的死亡，导致欧洲和美国农作物歉收的前景令人担忧。CCD的原因是复杂的和有争议的，但几种蜜蜂病原体被认为是参与。副转基因是一种特洛伊木马策略，其中内源性微生物用于表达拮抗病原体发育的效应分子。为了在蜜蜂中使用，副转基因必须依赖于候选副转基因微生物必须满足以获得成功结果的一组标准：（1）候选微生物必须是可遗传修饰的以表达效应分子;（2）经修饰的生物体在重新引入时对蜜蜂健康没有不利影响;以及（3）它必须与其他非致病性蜜蜂相关微生物一起存活。乳酸菌（LAB）是脊椎动物和无脊椎动物中常见的肠道细菌，有些细菌在其宿主中具有天然有益的特性。在目前的工作中，我们的目标是找到一个潜在的副转基因的候选人在这个细菌组中使用的蜜蜂。在与蜜蜂肠道微生物群相关的分离的LAB中，我们发现在觅食季节，嗜果糖的昆氏乳杆菌是最主要的物种。四种遗传上不同的L.选择kunkeei进行进一步评估。我们证明了（1）L. kunkeei是可转化的;（2）转化细胞对蜜蜂的存活没有明显的不利影响;（3）转化细胞在重新引入后在蜜蜂的肠道环境中存活良好。研究表明，L. kunkeei符合副转殖的三个标准，可以作为进一步研究蜜蜂中这种策略的合适候选者。
The honey bee (Apis mellifera) is a domestic insect of high value to human societies, as a crop pollinator in agriculture and a model animal in scientific research. The honey bee, however, has experienced massive mortality worldwide due to the phenomenon Colony Collapse Disorder (CCD), resulting in alarming prospects for crop failure in Europe and the USA. The reasons for CCD are complex and much debated, but several honey bee pathogens are believed to be involved. Paratransgenesis is a Trojan horse strategy, where endogenous microorganisms are used to express effector molecules that antagonise pathogen development. For use in honey bees, paratransgenesis must rely on a set of criteria that the candidate paratransgenic microorganism must fulfil in order to obtain a successful outcome: (1) the candidate must be genetically modifiable to express effector molecules; (2) the modified organism should have no adverse effects on honey bee health upon reintroduction; and (3) it must survive together with other nonpathogenic bee-associated microorganisms. Lactic acid bacteria (LAB) are common gut bacteria in vertebrates and invertebrates, and some have naturally beneficial properties in their host. In the present work we aimed to find a potential paratransgenic candidate within this bacterial group for use in honey bees. Among isolated LAB associated with bee gut microbiota, we found the fructophilic Lactobacillus kunkeei to be the most predominant species during foraging seasons. Four genetically different strains of L. kunkeei were selected for further assessment. We demonstrated (1) that L. kunkeei is transformable; (2) that the transformed cells had no obvious adverse effect on honey bee survival; and (3) that transformed cells survived well in the gut environment of bees upon reintroduction. Our study demonstrates that L. kunkeei fulfils the three criteria for paratransgenesis and can be a suitable candidate for further research on this strategy in honey bees.