Faculty of Life and Environmental Sciences, University of Tsukuba, Tsukuba, Ibaraki 305-8572, Japan.
J Biosci Bioeng. 2012 Jul;114(1):73-9. doi: 10.1016/j.jbiosc.2012.02.021. Epub 2012 May 8.
The effects of low-shear modeled microgravity (LSMMG) on a microbial community filtered through a 0.2-μm filter were investigated, and the potential application of LSMMG in the screening of microorganisms was evaluated. Pond water was passed through a 0.2-μm filter and the filtrate inoculated into two kinds of media (Schneider's insect medium, and ten-times-diluted Schneider's insect [0.1-Sch] medium). The cultures were incubated under LSMMG and normal-gravity and the microbial cell growth rates compared. Cell growth rates, final cell concentrations, and substrate consumption rates were higher in the LSMMG culture than in the normal-gravity culture. The microbial communities obtained under the various culture conditions were subjected to denaturing gradient gel electrophoresis (DGGE), revealing three different groups of microorganisms: (i) microorganisms whose growth rates were increased by LSMMG; (ii) microorganisms whose growth rates were suppressed or inhibited by LSMMG; and (iii) microorganisms whose growth rates were not affected by LSMMG. Sequence analysis of the microorganisms whose growth rates were increased by LSMMG showed that some had high similarity with unculturable microorganisms. When these microorganisms that displayed similarity with unculturable microorganisms were cultivated on agar plates, some of the DGGE bands present in the LSMMG culture were also present. We show that it is possible to isolate and cultivate uncultured microorganisms by using combinations of LSMMG, normal-gravity, and agar plate culturing techniques.
研究了低剪切模拟微重力 (LSMMG) 对通过 0.2μm 过滤器过滤的微生物群落的影响,并评估了 LSMMG 在微生物筛选中的潜在应用。池塘水通过 0.2μm 过滤器过滤,滤液接种到两种培养基( Schneider 昆虫培养基和十倍稀释的 Schneider 昆虫 [0.1-Sch] 培养基)中。在 LSMMG 和常重力下培养培养物,并比较微生物细胞生长率。LSMMG 培养物中的细胞生长率、最终细胞浓度和底物消耗率均高于常重力培养物。在不同培养条件下获得的微生物群落进行变性梯度凝胶电泳(DGGE)分析,揭示了三种不同的微生物群:(i)LSMMG 促进生长的微生物;(ii)LSMMG 抑制或抑制生长的微生物;和(iii)LSMMG 对生长速率没有影响的微生物。LSMMG 促进生长的微生物的序列分析表明,其中一些与不可培养的微生物具有高度相似性。当在琼脂平板上培养与不可培养微生物显示相似性的这些微生物时,LSMMG 培养物中存在的一些 DGGE 条带也存在。我们表明,通过使用 LSMMG、常重力和琼脂平板培养技术的组合,可以分离和培养不可培养的微生物。