Identification and characterization of anaerobic gut fungi using molecular methodologies based on ribosomal ITS1 and 18S rRNA
Identification and characterization of anaerobic gut fungi using molecular methodologies based on ribosomal ITS1 and 18S rRNA
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DOI：
10.1099/00221287-146-2-393
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发表时间：
2000-02-01
期刊：
影响因子：
2.8
通讯作者：
Tuckwell, DS
中科院分区：
文献类型：
--
作者：
Brookman, JL;Mennim, G;Tuckwell, DS
肠道真菌是一类特殊的游动孢子型真菌，它们占据着瘤胃厌氧环境这一独特的生态位，它们表现出两种基本形式，对于多中心物种，它们的核在菌丝体中迁移；对于单中心物种，它们的核物质集中在游动孢子囊中，使用常规技术很难区分这些真菌的菌株。在这项研究中，基于DNA的方法被用来检验从不同地理位置和宿主动物收集的两个属的单中心肠道真菌之间的关系。对16个单中心分离物和4个多中心分离物的核糖体ITS1序列进行了PCR扩增和测序，并与已发表的序列进行了比对和系统发育分析，这些分析清楚地区分了这两个属，并反映了先前发表的生理结论：Neocallimastix spp.构成一个比相对分化的Piromyces属更接近的属。这些分析将两个模式种N，Frontalis和N，hurley ensis放在一起，但与最近文献中的建议相反，将它们与另一个商定的种N.patriciarum分开，研究了原位杂交和狭缝印迹法作为检测和区分单中心性肠道真菌的潜在方法。利用核糖体序列的DNA狭缝杂交分析能够区分肠道真菌属，因此在这些生物的生态学研究中具有相当大的潜力。
The gut fungi are an unusual group of zoosporic fungi occupying a unique ecological niche, the anaerobic environment of the rumen, They exhibit two basic forms, with nuclear migration throughout the hyphal mass for polycentric species and with concentration of nuclear material in a zoosporangium for monocentric species, Differentiation between isolates of these fungi is difficult using conventional techniques. In this study, DNA-based methodologies were used to examine the relationships within and between two genera of monocentric gut fungi gathered from various geographical locations and host animals. The ribosomal ITS1 sequence from 16 mono- and 4 polycentric isolates was PCR-amplified and sequenced; the sequences obtained were aligned with published sequences and phylogenetic analyses were performed, These analyses clearly differentiate between the two genera and reflect the previously published physiological conclusions that Neocallimastix spp. constitute a more closely related genus than the relatively divergent genus Piromyces. The analyses place two type species N, frontalis and N, hurleyensis together but, contrary to a recent suggestion in the literature, place them apart from the other agreed species N. patriciarum, In situ hybridization and slot-blotting were investigated as potential methods for detection of and differentiation between monocentric gut fungi. DNA slot-blot analysis using ribosomal sequences is able to differentiate between gut fungal genera and thus has considerable potential for use in ecological studies of these organisms.