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Volume 148, Issue 6

Molecular characterization and endosymbiotic localization of the gene encoding D-ribulose 1,5-bisphosphate carboxylase–oxygenase (RuBisCO) form II in the deep-sea vestimentiferan trophosome

The DDBJ accession numbers for the sequences reported in this paper are AB042416 [ST-Sym(16S)-1], AB032829 [ST-Sym(II)-1] and AB040509 [ST-Sym(II)-2].

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Abstract

To better understand the contribution of micro-organisms to the primary production in the deep-sea gutless tubeworm sp., the 16S-rDNA-based phylogenetic data would be complemented by knowledge of the genes that encode the enzymes relevant to chemoautotrophic carbon fixation, such as D-ribulose 1,5-bisphosphate carboxylase–oxygenase (RuBisCO; EC 4.1.1.39). To phylogenetically characterize the autotrophic endosymbiosis within the trophosome of the tubeworm sp., bulk trophosomal DNA was extracted and analysed based on the 16S-rRNA- and RuBisCO-encoding genes. The 16S-rRNA- and RuBisCO-encoding genes were amplified by PCR, cloned and sequenced. For the 16S rDNA, a total of 50 clones were randomly selected and analysed directly by sequencing. Only one operational taxonomic unit resulted from the 16S rDNA sequence analysis. This may indicate the occurrence of one endosymbiotic bacterial species within the trophosome of the sp. used in this study. Phylogenetic analysis of the 16S rDNA showed that the sp. endosymbiont was closely related to the genus , a member of the α-. For the RuBisCO genes, only the form II gene () was amplified by PCR. A total of 50 clones were sequenced, and these were grouped into two operational RuBisCO units (ORUs) based on their deduced amino acid sequences. The ORUs showed high amino acid identities with those recorded from the ambient sediment bacteria. To confirm the results of sequence analysis, the localization of the symbiont-specific 16S rRNA and sequences in the sp. trophosome was visualized by hybridization (ISH), using specific probes. Two types of cells, coccoid and filamentous, were observed at the peripheries of the trophosome lobules. Both the symbiont-specific 16S rDNA and probes hybridized at the same sites coincident with the location of the coccoid cells, whereas the filamentous cells showed no -specific signals. The RuBisCO form I gene () was neither amplified by PCR nor detected by ISH. This is the first demonstration of chemoautotrophic symbiosis in the deep-sea gutless tubeworm, based on sequence data and localization of both the 16S-rRNA- and RuBisCO-encoding genes.

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Keyword(s): 16S rDNA , cbbM , in situ hybridization , ISH, in situ hybridization , Lamellibrachia endosymbiont , NJ, neighbour-joining , ORU, operational RuBisCO unit , OTU, operational taxonomic unit , phylogenetic analysis and RuBisCO, D-ribulose 1,5-bisphosphate carboxylase–oxygenase
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2002-06-01
2024-04-16
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Molecular characterization and endosymbiotic localization of the gene encoding D-ribulose 1,5-bisphosphate carboxylase–oxygenase (RuBisCO) form II in the deep-sea vestimentiferan trophosomeThe DDBJ accession numbers for the sequences reported in this paper are AB042416 [ST-Sym(16S)-1], AB032829 [ST-Sym(II)-1] and AB040509 [ST-Sym(II)-2].
Microbiology 148, 1947 (2002); https://doi.org/10.1099/00221287-148-6-1947
/content/journal/micro/10.1099/00221287-148-6-1947
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