Combined Use of Cultivation-Dependent and Cultivation-Independent Methods Indicates that Members of Most Haloarchaeal Groups in an Australian Crystallizer Pond Are Cultivable
ABSTRACT Haloarchaea are the dominant microbial flora in hypersaline waters with near-saturating salt levels. The haloarchaeal diversity of an Australian saltern crystallizer pond was examined by use of a library of PCR-amplified 16S rRNA genes and by cultivation. High viable counts (10 6 CFU/ml) we...
Published in: | Applied and Environmental Microbiology |
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Online Access: | http://dx.doi.org/10.1128/aem.70.9.5258-5265.2004 https://journals.asm.org/doi/pdf/10.1128/AEM.70.9.5258-5265.2004 |
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crasmicro:10.1128/aem.70.9.5258-5265.2004 2024-06-23T07:47:34+00:00 Combined Use of Cultivation-Dependent and Cultivation-Independent Methods Indicates that Members of Most Haloarchaeal Groups in an Australian Crystallizer Pond Are Cultivable Burns, D. G. Camakaris, H. M. Janssen, P. H. Dyall-Smith, M. L. 2004 http://dx.doi.org/10.1128/aem.70.9.5258-5265.2004 https://journals.asm.org/doi/pdf/10.1128/AEM.70.9.5258-5265.2004 en eng American Society for Microbiology https://journals.asm.org/non-commercial-tdm-license Applied and Environmental Microbiology volume 70, issue 9, page 5258-5265 ISSN 0099-2240 1098-5336 journal-article 2004 crasmicro https://doi.org/10.1128/aem.70.9.5258-5265.2004 2024-06-10T04:07:09Z ABSTRACT Haloarchaea are the dominant microbial flora in hypersaline waters with near-saturating salt levels. The haloarchaeal diversity of an Australian saltern crystallizer pond was examined by use of a library of PCR-amplified 16S rRNA genes and by cultivation. High viable counts (10 6 CFU/ml) were obtained on solid media. Long incubation times (≥8 weeks) appeared to be more important than the medium composition for maximizing viable counts and diversity. Of 66 isolates examined, all belonged to the family Halobacteriaceae , including members related to species of the genera Haloferax , Halorubrum , and Natronomonas . In addition, isolates belonging to a novel group (the ADL group), previously detected only as 16S rRNA genes in an Antarctic hypersaline lake (Deep Lake), were cultivated for the first time. The 16S rRNA gene library identified the following five main groups: Halorubrum groups 1 and 2 (49%), the SHOW (square haloarchaea of Walsby) group (33%), the ADL group (16%), and the Natronomonas group (2%). There were two significant differences between the organisms detected in cultivation and 16S rRNA sequence results. Firstly, Haloferax spp. were frequently isolated on plates (15% of all isolates) but were not detected in the 16S rRNA sequences. Control experiments indicated that a bias against Haloferax sequences in the generation of the 16S rRNA gene library was unlikely, suggesting that Haloferax spp. readily form colonies, even though they were not a dominant group. Secondly, while the 16S rRNA gene library identified the SHOW group as a major component of the microbial community, no isolates of this group were obtained. This inability to culture members of the SHOW group remains an outstanding problem in studying the ecology of hypersaline environments. Article in Journal/Newspaper Antarc* Antarctic ASM Journals (American Society for Microbiology) Antarctic Saltern ENVELOPE(19.767,19.767,69.600,69.600) Applied and Environmental Microbiology 70 9 5258 5265 |
institution |
Open Polar |
collection |
ASM Journals (American Society for Microbiology) |
op_collection_id |
crasmicro |
language |
English |
description |
ABSTRACT Haloarchaea are the dominant microbial flora in hypersaline waters with near-saturating salt levels. The haloarchaeal diversity of an Australian saltern crystallizer pond was examined by use of a library of PCR-amplified 16S rRNA genes and by cultivation. High viable counts (10 6 CFU/ml) were obtained on solid media. Long incubation times (≥8 weeks) appeared to be more important than the medium composition for maximizing viable counts and diversity. Of 66 isolates examined, all belonged to the family Halobacteriaceae , including members related to species of the genera Haloferax , Halorubrum , and Natronomonas . In addition, isolates belonging to a novel group (the ADL group), previously detected only as 16S rRNA genes in an Antarctic hypersaline lake (Deep Lake), were cultivated for the first time. The 16S rRNA gene library identified the following five main groups: Halorubrum groups 1 and 2 (49%), the SHOW (square haloarchaea of Walsby) group (33%), the ADL group (16%), and the Natronomonas group (2%). There were two significant differences between the organisms detected in cultivation and 16S rRNA sequence results. Firstly, Haloferax spp. were frequently isolated on plates (15% of all isolates) but were not detected in the 16S rRNA sequences. Control experiments indicated that a bias against Haloferax sequences in the generation of the 16S rRNA gene library was unlikely, suggesting that Haloferax spp. readily form colonies, even though they were not a dominant group. Secondly, while the 16S rRNA gene library identified the SHOW group as a major component of the microbial community, no isolates of this group were obtained. This inability to culture members of the SHOW group remains an outstanding problem in studying the ecology of hypersaline environments. |
format |
Article in Journal/Newspaper |
author |
Burns, D. G. Camakaris, H. M. Janssen, P. H. Dyall-Smith, M. L. |
spellingShingle |
Burns, D. G. Camakaris, H. M. Janssen, P. H. Dyall-Smith, M. L. Combined Use of Cultivation-Dependent and Cultivation-Independent Methods Indicates that Members of Most Haloarchaeal Groups in an Australian Crystallizer Pond Are Cultivable |
author_facet |
Burns, D. G. Camakaris, H. M. Janssen, P. H. Dyall-Smith, M. L. |
author_sort |
Burns, D. G. |
title |
Combined Use of Cultivation-Dependent and Cultivation-Independent Methods Indicates that Members of Most Haloarchaeal Groups in an Australian Crystallizer Pond Are Cultivable |
title_short |
Combined Use of Cultivation-Dependent and Cultivation-Independent Methods Indicates that Members of Most Haloarchaeal Groups in an Australian Crystallizer Pond Are Cultivable |
title_full |
Combined Use of Cultivation-Dependent and Cultivation-Independent Methods Indicates that Members of Most Haloarchaeal Groups in an Australian Crystallizer Pond Are Cultivable |
title_fullStr |
Combined Use of Cultivation-Dependent and Cultivation-Independent Methods Indicates that Members of Most Haloarchaeal Groups in an Australian Crystallizer Pond Are Cultivable |
title_full_unstemmed |
Combined Use of Cultivation-Dependent and Cultivation-Independent Methods Indicates that Members of Most Haloarchaeal Groups in an Australian Crystallizer Pond Are Cultivable |
title_sort |
combined use of cultivation-dependent and cultivation-independent methods indicates that members of most haloarchaeal groups in an australian crystallizer pond are cultivable |
publisher |
American Society for Microbiology |
publishDate |
2004 |
url |
http://dx.doi.org/10.1128/aem.70.9.5258-5265.2004 https://journals.asm.org/doi/pdf/10.1128/AEM.70.9.5258-5265.2004 |
long_lat |
ENVELOPE(19.767,19.767,69.600,69.600) |
geographic |
Antarctic Saltern |
geographic_facet |
Antarctic Saltern |
genre |
Antarc* Antarctic |
genre_facet |
Antarc* Antarctic |
op_source |
Applied and Environmental Microbiology volume 70, issue 9, page 5258-5265 ISSN 0099-2240 1098-5336 |
op_rights |
https://journals.asm.org/non-commercial-tdm-license |
op_doi |
https://doi.org/10.1128/aem.70.9.5258-5265.2004 |
container_title |
Applied and Environmental Microbiology |
container_volume |
70 |
container_issue |
9 |
container_start_page |
5258 |
op_container_end_page |
5265 |
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1802651685848350720 |