Salt concentration in substrate modulates the composition of bacterial and yeast microbiomes of Drosophila melanogaster

Ekaterina Y. Yakovleva, Irina Danilova, Irina Maximova, Alexander Shabaev, Anastasia Dmitrieva, Andrey Belov, Alexandra Klyukina, Ksenia Perfilieva, E. Bonch-Osmolovskaya, Alexander Markov
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Abstract

Aim: Microbiomes influence the physiology and behavior of multicellular organisms and contribute to their adaptation to changing environmental conditions. However, yeast and bacterial microbiota have usually been studied separately; therefore, the interaction between bacterial and yeast communities in the gut of Drosophila melanogaster (D. melanogaster ) is often overlooked. In this study, we investigate the correlation between bacterial and yeast communities in the gut of D. melanogaster . Methods: We studied the shifts in the joint microbiome of Drosophila melanogaster , encompassing both yeasts and bacteria, during adaptation to substrate with varying salt concentrations (0%, 2%, 4%, and 7%) using plating for both yeasts and bacteria and NGS-sequencing of variable 16S rRNA gene regions for bacteria. Results: The microbiome of flies and their substrates was gradually altered at moderate NaCl concentrations (2% and 4% compared with the 0% control) and completely transformed at high salt concentrations (7%). The relative abundance of Acetobacter , potentially beneficial to D. melanogaster , decreased as NaCl concentration increased, whereas the relative abundance of the more halotolerant lactobacilli first increased, peaking at 4% NaCl, and then declined dramatically at 7%. At this salinity level, potentially pathogenic bacteria of the genera Leuconostoc and Providencia were dominant. The yeast microbiome of D. melanogaster also undergoes significant changes with an increase in salt concentration in the substrate. The total yeast abundance undergoes nonlinear changes: it is lowest at 0% salt concentration and highest at 2%-4%. At a 7% concentration, the yeast abundance in flies and their substrate is lower than at 2%-4% but significantly higher than at 0%. Conclusions: The abundance and diversity of bacteria that are potentially beneficial to the flies decreased, while the proportion of potential pathogens, Leuconostoc and Providencia , increased with an increase in salt concentration in the substrate. In samples with a relatively high abundance and/or diversity of yeasts, the corresponding indicators for bacteria were often lowered, and vice versa . This may be due to the greater halotolerance of yeasts compared to bacteria and may also indicate antagonism between these groups of microorganisms.
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底物中的盐浓度调节黑腹果蝇细菌和酵母微生物群的组成
目的:微生物群影响多细胞生物的生理和行为,并有助于它们适应不断变化的环境条件。然而,酵母和细菌微生物群通常是分开研究的,因此黑腹果蝇肠道中细菌和酵母群落之间的相互作用常常被忽视。本研究调查了黑腹果蝇肠道中细菌和酵母菌群落之间的相关性。方法:我们研究了黑腹果蝇在适应不同盐浓度(0%、2%、4% 和 7%)的基质过程中,包括酵母菌和细菌在内的联合微生物组的变化,对酵母菌和细菌都进行了平板培养,对细菌进行了可变 16S rRNA 基因区域的 NGS 测序。结果与 0% 的对照组相比,中等 NaCl 浓度(2% 和 4%)下苍蝇及其底物的微生物组逐渐发生了变化,而在高盐浓度(7%)下则完全发生了转变。随着 NaCl 浓度的增加,可能对黑腹蝇虫有益的醋杆菌的相对丰度下降,而耐盐性更强的乳酸杆菌的相对丰度先是增加,在 NaCl 浓度为 4% 时达到峰值,然后在 7% 时急剧下降。在这一盐度水平下,白念珠菌属和普罗维登菌属的潜在致病菌占优势。随着基质中盐浓度的增加,黑腹酵母的酵母微生物组也发生了显著变化。酵母菌的总丰度发生了非线性变化:盐浓度为 0% 时酵母菌的丰度最低,盐浓度为 2%-4% 时酵母菌的丰度最高。在盐浓度为 7% 时,苍蝇及其底物中的酵母菌丰度低于 2%-4% 时的水平,但明显高于 0% 时的水平。结论随着基质中盐浓度的增加,对苍蝇可能有益的细菌的丰度和多样性降低了,而潜在病原体(白念珠菌和普罗维登菌)的比例增加了。在酵母菌丰度和/或多样性相对较高的样本中,相应的细菌指标通常会降低,反之亦然。这可能是因为与细菌相比,酵母菌具有更强的耐盐性,也可能表明这两类微生物之间存在拮抗作用。
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