Discovery of a novel rumen methanogen in the anaerobic fungal culture and its distribution in the rumen as revealed by real-time PCR.
Discovery of a novel rumen methanogen in the anaerobic fungal culture and its distribution in the rumen as revealed by real-time PCR.
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厌氧真菌培养物中新型瘤胃产甲烷菌的发现及其在瘤胃中的分布（实时 PCR 揭示）
DOI：
10.1186/1471-2180-14-104
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发表时间：
2014-04-23
期刊：
影响因子：
4.2
通讯作者：
Zhu WY
中科院分区：
文献类型：
--
作者：
Jin W;Cheng YF;Mao SY;Zhu WY
背景属于瘤胃簇C（RCC）的新型古菌可能在瘤胃甲烷产生中发挥重要作用，受到越来越多的关注。然而，目前关于瘤胃中 RCC 的信息因未成功从瘤胃中分离出无菌纯 RCC 而受到限制。本研究通过分子和培养方法鉴定了厌氧真菌亚培养物中生长的RCC。结果通过16S rRNA基因克隆文库鉴定并证明了真菌亚培养物中存在的新RCC物种。有趣的是，新的 RCC 物种在所有次代培养转移中都在真菌培养物中存活下来，甚至在第 62 次次代培养中也是如此，与此相反，其他产甲烷菌在次代培养过程中消失了。进一步的研究表明，继代培养转移频率显着影响真菌继代培养中新 RCC 物种的相对丰度。 5天和7天的转移频率增加了RCC物种的相对丰度（P<0.05）。此外，定量实时 PCR 显示，高浓度日粮不会影响古细菌的丰度，但会在数量上减少瘤胃中新型 RCC 物种的丰度。此外，瘤胃液体部分中 RCC 物种的相对丰度在数值上高于瘤胃上皮和固体部分。最后，成功获得含有RCC物种的纯化真菌培养物。 PCR和测序分析表明，该新的RCC物种含有amcrA基因，该基因在产甲烷过程中发挥着至关重要的作用，因此可以被鉴定为产甲烷菌。结论在本研究中，一个新的RCC物种被鉴定为产甲烷菌，并且与厌氧真菌密切相关。这种与厌氧真菌共培养的新方法可能为培养和研究尚未鉴定的产甲烷菌提供一种可行的方法。
BackgroundThe novel archaea belonging to Rumen Cluster C (RCC), which may play an important role in methane production in the rumen have received increased attention. However, the present information on RCC in the rumen is limited by the unsuccessful isolation of axenic pure RCC from the rumen. In the present study, RCC grown in anaerobic fungal subcultures was identified by the molecular and culture methods.ResultsA novel RCC species existing in the fungal subcultures was identified and demonstrated by the 16S rRNA gene clone library. Interestingly, the novel RCC species survived in the fungal cultures over all the subculture transferring, even in the 62ndsubculture, in contrast to the other methanogens, which disappeared during subcultures. Further work showed that subculture transfer frequency significantly affected the relative abundance of the novel RCC species in the fungal subcultures. The five-day and seven-day transfer frequencies increased the relative abundance of the RCC species (P<0.05). In addition, quantitative real-time PCR revealed that high concentrate diets did not affect the abundance of archaea, but numerically reduced the abundance of the novel RCC species in the rumen. In addition, the relative abundance of the RCC species was numerically higher in the rumen liquid fraction than in the rumen epithelium and solid fractions. Finally, a purified fungal culture containing the RCC species was successfully obtained. PCR and sequencing analysis showed that the novel RCC species contained amcrA gene, which is known to play a crucial role in methanogenesis, and thus could be identified as a methanogen.ConclusionIn this study, a novel RCC species was identified as a methanogen and closely associated with anaerobic fungi. This novel approach by using co-culture with anaerobic fungi may provide a feasible way to culture and investigate not yet identified methanogens.
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影响因子：
4.4
作者：
Koike, S.;Handa, Y.;Kobayashi, Y.
通讯作者：
Kobayashi, Y.
影响因子：
3.5
作者：
BRYANT, MP;BURKEY, LA
通讯作者：
BURKEY, LA
影响因子：
4.4
作者：
King, Erin E.;Smith, Rachel P.;Wright, Andre-Denis G.
通讯作者：
Wright, Andre-Denis G.
DOI：
10.1099/ijs.0.033712-0
发表时间：
2012-08-01
影响因子：
2.8
作者：
Dridi, Bedis;Fardeau, Marie-Laure;Drancourt, Michel
通讯作者：
Drancourt, Michel
影响因子：
4.4
作者：
Paul, Kristina;Nonoh, James O.;Brune, Andreas
通讯作者：
Brune, Andreas