Reitschuler Christoph, Lins Philipp, Wagner Andreas Otto, Illmer Paul
University of Innsbruck, Institute of Microbiology, Technikerstr. 25, A-6020 Innsbruck, Austria.
Anaerobe. 2014 Oct;29:73-9. doi: 10.1016/j.anaerobe.2013.10.002. Epub 2013 Oct 26.
PCR-DGGE, qPCR and sequencing highlighted a quite homogenous archaeal community prevailing in secondary calcite deposits, so-called moonmilk, within the cold alpine Hundalm cave in Tyrol (Austria). Furthermore, the depth profile of this moonmilk could prove that the Archaea are located in oxygen-rich near- and oxygen-depleted sub-surface layers. To gather these communities we therefore applied an aerobic and anaerobic cultivation approach in oligotrophic and methanotrophic media. The mixed moonmilk community was analyzed with a combination of molecular methods using qPCR, PCR-DGGE and sequencing. Anaerobic and aerobic cultures were additionally investigated with GC and HPLC analyses. It was possible to initially cultivate and enrich the supposed aerobic/microaerophilic and anaerobic archaeal fraction, representing the natural archaeal community. While the naturally less abundant near-surface Archaea are closely related to members of the Thaumarchaeota (Nitrosopumilus maritimus), the highly abundant anaerobic Archaea are more distantly related to members within the Euryarchaeota. It is possible that these cultivable moonmilk-born Archaea represent new ecotypes or are so far undescribed. Based on the sequencing results and the production of very low amounts of methane, a corresponding methanogenic community is thought to represent only a minor abundant archaeal fraction. On a physiological level the cultivated moonmilk community is cold-adapted and basically of oligotrophic and organotrophic character.
聚合酶链反应-变性梯度凝胶电泳（PCR-DGGE）、定量聚合酶链反应（qPCR）和测序结果表明，在奥地利蒂罗尔州寒冷的高山洪德尔姆洞穴内的次生方解石沉积物（即所谓的月奶石）中，存在一个相当均匀的古菌群落。此外，月奶石的深度剖面证明，古菌位于富氧的近地表层和贫氧的次表层。因此，为了收集这些群落，我们在贫营养和甲烷营养培养基中采用了好氧和厌氧培养方法。使用qPCR、PCR-DGGE和测序等分子方法组合对混合的月奶石群落进行了分析。此外，还通过气相色谱（GC）和高效液相色谱（HPLC）分析对厌氧和好氧培养物进行了研究。最初有可能培养和富集假定的好氧/微好氧和厌氧古菌部分，代表天然古菌群落。虽然自然状态下近地表数量较少的古菌与奇古菌门（嗜盐栖热孢菌）的成员密切相关，但数量众多的厌氧古菌与广古菌门中的成员关系较远。这些可培养的源自月奶石的古菌有可能代表新的生态型，或者迄今为止尚未被描述。根据测序结果以及极少量甲烷的产生情况，相应的产甲烷群落被认为仅代表少量丰富的古菌部分。在生理水平上，培养的月奶石群落适应寒冷，基本上具有贫营养和有机营养的特征。
