Phylogeny of not-yet-cultured spirochetes from termite guts

被引:90
作者
Paster, BJ
Dewhirst, FE
Cooke, SM
Fussing, V
Poulsen, LK
Breznak, JA
机构
[1] DANISH VET LAB,DK-1790 COPENHAGEN V,DENMARK
[2] TECH UNIV DENMARK,DEPT MICROBIOL,DK-2800 LYNGBY,DENMARK
[3] MICHIGAN STATE UNIV,DEPT MICROBIOL,E LANSING,MI 48824
[4] MICHIGAN STATE UNIV,CTR MICROBIAL ECOL,E LANSING,MI 48824
关键词
D O I
10.1128/AEM.62.2.347-352.1996
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Comparisons of 16S rDNA sequences were used to determine the phylogeny of not-yet-cultured spirochetes from hindguts of the African higher termite, Nasutitermes lujae (Wasmann). The 16S rRNA genes were amplified directly from spirochete-rich hindguts by using universal primers, and the amplified products were cloned into Escherichia coli, Clones were screened with a spirochete-specific DNA probe, Analysis of 1,410 base positions of the 16S rDNA insert from one spirochete clone, designated NL1, supported its assignment to the genus Treponema, with average interspecies similarities of ca. 85%. The sequence of NL1 was most closely related (ca, 87 to 88% similarity) to sequences of Spirochaeta stenostrepta and Spirochaeta caldaria and to a previously published sequence (ca. 87% similarity) of spirochetal clone MDS1 from the Australian lower termite, Mastotermes darwiniensis (Froggatt), On the basis of 16S rRNA sequence comparisons and individual base signatures, clones NL1 and MDS1 clearly represent two novel species of Treponema, although specific epithets have not yet been proposed. The gross morphology of NL1 was determined from in situ hybridization experiments with an NL1-specific, fluorescently labeled oligonucleotide probe, Cells were approximately 0.3 to 0.4 by 30 mu m in size, with a wavelength and amplitude of about 10 mu m and 0.8 to 1.6 mu m, respectively, Moreover, electron microscopy of various undulate cells present in gut contents confirmed that they possessed ultrastructural features typical of spirochetes, i.e., a wavy protoplasmic cylinder, periplasmic flagella, and an outer sheath. The sequence data suggest that termite gut spirochetes may represent a separate line of descent from other treponemes and that they constitute a significant reservoir of previously unrecognized spirochetal biodiversity.
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页码:347 / 352
页数:6
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