Molecular phylogenetic identification of the intestinal anaerobic microbial community in the hindgut of the termite, Reticulitermes speratus, without cultivation

被引:35
作者
Kudo, T [1 ]
Ohkuma, M [1 ]
Moriya, S [1 ]
Noda, S [1 ]
Ohtoko, K [1 ]
机构
[1] RIKEN, Inst Phys & Chem Res, Wako, Saitama 35101, Japan
关键词
minimum size of anoxic habitat; termite hindgut; molecular phylogeny;
D O I
10.1007/s007920050055
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
A termite maintains an anaerobic microbial community in its hindgut, which seems to be the minimum size of an anaerobic habitat. This microbial community consists of bacteria and various anaerobic flagellates, and it is established that termites are totally dependent on the microbes for the utilization of their food. The molecular phylogenetic diversity of the intestinal microflora of a lower termite, Reticulitermes speratus, was examined by a strategy that does not rely on cultivation of the resident microorganisms. Small subunit ribosomal RNA (ssrRNA) genes were directly amplified from the mixed-population DNA of the termite gut by polymerase chain reaction (PCR) and clonally isolated. Most sequenced clones were phylogenetically affiliated with the four major groups of the domain Bacteria: the Proteobacteria group, the Spirochete group, the Bacteroides group, and the Low G + C gram-positive bacteria. The 16S rRNA sequence data show that the majority of the intestinal microflora of the termite consists of new species that are pet to be cultured. The phylogeny of a symbiotic methanogen inhabiting the gut of a lower termite (R. speratus) was analyzed without cultivation. The nucleotide sequence of the ssrDNA and the predicted amino acid sequence of the mcrA product were compared with those of the known methanogens, Both comparisons indicated that the termite symbiotic methanogen belonged to the order Methanobacteriales but was distinct from the known members of this order. The diversity of nitrogen-fixing organisms was also investigated without culturing the resident microorganisms. Fragments of the nifH gene, which encodes the dinitrogenase reductase, were directly amplified from the mixed-population DNA of the termite gut and were clonally isolated. The phylogenetic analysis of the nifH amino acid sequences showed that there was a remarkable diversity of nitrogenase genes in the termite gut: The molecular phylogeny of a symbiotic hypermastigote Trichonympha agilis (class Parabasalia; order Hypermastigida) in the hindgut of R. speratus was also examined by the same strategy. The whole-cell hybridization experiments indicated that the sequence originated from a large hypermastigote in the termite hindgut, Trichonympha agilis. According to the phylogenetic trees constructed, the hypermastigote represented one of the deepest branches of eukaryotes. The hypermastigote along with members of the order Trichomonadida formed a monophyletic lineage, indicating that the hypermastigote and trichomonads shared a recent common ancestry.
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页码:155 / 161
页数:7
相关论文
共 26 条
[1]   PHYLOGENETIC IDENTIFICATION AND IN-SITU DETECTION OF INDIVIDUAL MICROBIAL-CELLS WITHOUT CULTIVATION [J].
AMANN, RI ;
LUDWIG, W ;
SCHLEIFER, KH .
MICROBIOLOGICAL REVIEWS, 1995, 59 (01) :143-169
[2]   NITROGEN-FIXATION IN TERMITES [J].
BENEMANN, JR .
SCIENCE, 1973, 181 (4095) :164-165
[3]   CULTURE AND PHYLOGENETIC CHARACTERIZATION OF TRICHOMITUS-TRYPANOIDES DUBOSCQ AND GRASSE 1924, N-COMB - A TRICHOMONAD FLAGELLATE ISOLATED FROM THE HINDGUT OF THE TERMITE RETICULITERMES-SANTONENSIS FEYTAUD [J].
BERCHTOLD, M ;
BREUNIG, A ;
KONIG, H .
JOURNAL OF EUKARYOTIC MICROBIOLOGY, 1995, 42 (04) :388-391
[4]  
Berchtold Manfred, 1995, Systematic and Applied Microbiology, V18, P567
[5]   MORPHOLOGY AS A BASIS FOR TAXONOMY OF LARGE SPIROCHETES SYMBIOTIC IN WOOD-EATING COCKROACHES AND TERMITES - PILLOTINA GEN-NOV, NOM REV, PILLOTINA-CALOTERMITIDIS SP-NOV, NOM REV, DIPLOCALYX GEN-NOV, NOM REV, DIPLOCALYX-CALOTERMITIDIS SP-NOV, NOM REV, HOLLANDINA GEN-NOV, NOM REV, HOLLANDINA-PTEROTERMITIDIS SP-NOV, NOM REV, AND CLEVELANDINA-RETICULITERMITIDIS GEN-NOV, SP-NOVE [J].
BERMUDES, D ;
CHASE, D ;
MARGULIS, L .
INTERNATIONAL JOURNAL OF SYSTEMATIC BACTERIOLOGY, 1988, 38 (03) :291-302
[6]   GENESIS OF ACETATE AND METHANE BY GUT BACTERIA OF NUTRITIONALLY DIVERSE TERMITES [J].
BRAUMAN, A ;
KANE, MD ;
LABAT, M ;
BREZNAK, JA .
SCIENCE, 1992, 257 (5075) :1384-1387
[7]   SPOROMUSA-TERMITIDA SP-NOV, AN H2/CO2-UTILIZING ACETOGEN ISOLATED FROM TERMITES [J].
BREZNAK, JA ;
SWITZER, JM ;
SEITZ, HJ .
ARCHIVES OF MICROBIOLOGY, 1988, 150 (03) :282-288
[8]   ROLE OF MICROORGANISMS IN THE DIGESTION OF LIGNOCELLULOSE BY TERMITES [J].
BREZNAK, JA ;
BRUNE, A .
ANNUAL REVIEW OF ENTOMOLOGY, 1994, 39 :453-487
[9]   NITROGEN-FIXATION IN TERMITES [J].
BREZNAK, JA ;
BRILL, WJ ;
MERTINS, JW ;
COPPEL, HC .
NATURE, 1973, 244 (5418) :577-579
[10]   BACTERIA FROM GUT OF AUSTRALIAN TERMITES [J].
EUTICK, ML ;
OBRIEN, RW ;
SLAYTOR, M .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 1978, 35 (05) :823-828