Archaeal diversity in a terrestrial acidic spring field revealed by a novel PCR primer targeting archaeal 16S rRNA genes
Archaeal diversity in a terrestrial acidic spring field revealed by a novel PCR primer targeting archaeal 16S rRNA genes
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DOI：
10.1111/j.1574-6968.2011.02267.x
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发表时间：
2011-06-01
影响因子：
2.1
通讯作者：
Yamagishi, Akihiko
中科院分区：
文献类型：
--
作者：
Kato, Shingo;Itoh, Takashi;Yamagishi, Akihiko
利用本研究设计的一种新的古菌特异性引物，通过PCR技术对日本箱根大和谷温泉区古菌16 S rRNA基因的系统发育多样性进行了研究。利用一个新设计的正向引物和一个已报道的正向引物与反向引物，从热水（78 ℃）和泥浆（28 ℃）样品中构建了古细菌16 SrRNA基因克隆文库。在热水样品库中发现的大多数细菌类型与培养的（超）嗜热菌有关。用两种不同引物扩增的文库，从热水样品中检测到的细菌类型及其检测频率相似。与此相反，在来自泥浆样品的文库中发现了与培养的微藻具有低相似性（< 95%）的微藻类型。其中，有些类型与热浆菌门的成员比较接近（80-93%），有些类型与泉古菌门和广古菌门的亲缘关系不明显，但与奇姆古菌门和科尔古菌门的亲缘关系较近。两个库的泥浆样品中的荧光素酶类型及其检出频率存在显著差异。我们的结果从基于PCR的分析，使用重新设计的引物表明，更多样化的，未培养的微藻存在于酸性环境中，在较低的温度比以前认识到的。
The phylogenetic diversity of archaeal 16S rRNA genes in a thermoacidic spring field of Ohwakudani, Hakone, Japan, was investigated by PCR-based analysis using a novel Archaea-specific primer designed in the present study. Clone libraries of archaeal 16S rRNA genes were constructed from hot water (78 degrees C) and mud (28 degrees C) samples by PCR using a newly designed forward primer and a previously reported forward primer with reverse primers. Most phylotypes found in the libraries from the hot water sample were related to cultured (hyper)thermophiles. The phylotypes and their detection frequencies from the hot water sample were similar for the libraries amplified with the two different primer sets. In contrast, phylotypes having a low similarity (< 95%) to cultured Archaea were found in the libraries from the mud sample. Some of the phylotypes were relatively close to members of Thermoplasmata (80-93% similarity) and the others were not clearly affiliated with Crenarchaeota and Euryarchaeota, but related to Thaumarchaeota and Korarchaeota. The phylotypes and their detection frequencies were significantly different between the two libraries of the mud sample. Our results from the PCR-based analysis using the redesigned primer suggest that more diverse, uncultured Archaea are present in acidic environments at a low temperature than previously recognized.