The present study is focused on the development of a cultivation-independent molecular approach for specific detection of bioluminescent bacteria within microbial communities by direct amplification of luxA gene from environmental DNA. An ewset of primers, specifically targeting free-living bioluminescent bacteria, was designed on the base of luxA sequences available from the public data base. Meso and bathypelagic sea water samples were collected from twostations in Tyrrhenian Sea at the depths of 500 and 2750m. The same sea water samples also were used to isolate bioluminescent bacteria that were further subjected to luxA and 16SrRNA gene sequencing. PCR products obtained by amplification with designed primers were cloned, and the phylogenetic affiliation of 40 clones was determined. All of them were clustered into three groups, only distantly related to the Photobacterium phosphoreum and Photobacterium kishitanii clades. The half of all clones formed at ight monophyletic clade, while the rest of clones were organized in ‘‘compartment’’-specific, meso-and bathypelagic ecotypes. No matches with luxA gene sequences off our bioluminescent strains, isolated from the same sea water samples, were observed. These findings indicate that the PCR-based approach developed in present manuscript, allowed us to detect the novel, ‘‘yet to be cultivated’’ lineages of bioluminescent bacteria, which are likely specific for distinct warm bathypelagic real ms of Mediterranean Sea.

PCR-Based Detection of Bioluminescent Microbial Populations in Tyrrhenian Sea

SMEDILE, FRANCESCO;DE DOMENICO, Emilio;DE FRANCESCO, Maria;
2009-01-01

Abstract

The present study is focused on the development of a cultivation-independent molecular approach for specific detection of bioluminescent bacteria within microbial communities by direct amplification of luxA gene from environmental DNA. An ewset of primers, specifically targeting free-living bioluminescent bacteria, was designed on the base of luxA sequences available from the public data base. Meso and bathypelagic sea water samples were collected from twostations in Tyrrhenian Sea at the depths of 500 and 2750m. The same sea water samples also were used to isolate bioluminescent bacteria that were further subjected to luxA and 16SrRNA gene sequencing. PCR products obtained by amplification with designed primers were cloned, and the phylogenetic affiliation of 40 clones was determined. All of them were clustered into three groups, only distantly related to the Photobacterium phosphoreum and Photobacterium kishitanii clades. The half of all clones formed at ight monophyletic clade, while the rest of clones were organized in ‘‘compartment’’-specific, meso-and bathypelagic ecotypes. No matches with luxA gene sequences off our bioluminescent strains, isolated from the same sea water samples, were observed. These findings indicate that the PCR-based approach developed in present manuscript, allowed us to detect the novel, ‘‘yet to be cultivated’’ lineages of bioluminescent bacteria, which are likely specific for distinct warm bathypelagic real ms of Mediterranean Sea.
2009
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11570/1889685
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