Cargando…

Developing a genetic approach to target cyanobacterial producers of heterocyte glycolipids in the environment

Heterocytous cyanobacteria are important players in the carbon and nitrogen cycle. They can fix dinitrogen by using heterocytes, specialized cells containing the oxygen-sensitive nitrogenase enzyme surrounded by a thick polysaccharide and glycolipid layer which prevents oxygen diffusion and nitrogen...

Descripción completa

Detalles Bibliográficos
Autores principales: Pérez Gallego, Ruth, Bale, Nicole J., Sinninghe Damste, Jaap S., Villanueva, Laura
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10569477/
https://www.ncbi.nlm.nih.gov/pubmed/37840743
http://dx.doi.org/10.3389/fmicb.2023.1257040
_version_ 1785119553356824576
author Pérez Gallego, Ruth
Bale, Nicole J.
Sinninghe Damste, Jaap S.
Villanueva, Laura
author_facet Pérez Gallego, Ruth
Bale, Nicole J.
Sinninghe Damste, Jaap S.
Villanueva, Laura
author_sort Pérez Gallego, Ruth
collection PubMed
description Heterocytous cyanobacteria are important players in the carbon and nitrogen cycle. They can fix dinitrogen by using heterocytes, specialized cells containing the oxygen-sensitive nitrogenase enzyme surrounded by a thick polysaccharide and glycolipid layer which prevents oxygen diffusion and nitrogenase inactivation. Heterocyte glycolipids can be used to detect the presence of heterocytous cyanobacteria in present-day and past environments, providing insight into the functioning of the studied ecosystems. However, due to their good preservation throughout time, heterocyte glycolipids are not ideal to detect and study living communities, instead methods based on DNA are preferred. Currently cyanobacteria can be detected using untargeted genomic approaches such as metagenomics, or they can be specifically targeted by, for example, the use of primers that preferentially amplify their 16S rRNA gene or their nifH gene in the case of nitrogen fixing cyanobacteria. However, since not all cyanobacterial nitrogen fixers are heterocytous, there is currently no fast gene-based method to specifically detect and distinguish heterocytous cyanobacteria. Here, we developed a PCR-based method to specifically detect heterocytous cyanobacteria by designing primers targeting the gene (hglT) encoding the enzyme responsible for the last step in the biosynthesis of heterocyte glycolipid (i.e., a glycosyltransferase). We designed several primer sets using the publicly available sequences of 23 heterocytous cyanobacteria, after testing them on DNA extracts of 21 heterocyte-forming and 7 non-heterocyte forming freshwater cyanobacteria. The best primer set was chosen and successfully used to confirm the presence of heterocytous cyanobacteria in a marine environmental sample.
format Online
Article
Text
id pubmed-10569477
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher Frontiers Media S.A.
record_format MEDLINE/PubMed
spelling pubmed-105694772023-10-13 Developing a genetic approach to target cyanobacterial producers of heterocyte glycolipids in the environment Pérez Gallego, Ruth Bale, Nicole J. Sinninghe Damste, Jaap S. Villanueva, Laura Front Microbiol Microbiology Heterocytous cyanobacteria are important players in the carbon and nitrogen cycle. They can fix dinitrogen by using heterocytes, specialized cells containing the oxygen-sensitive nitrogenase enzyme surrounded by a thick polysaccharide and glycolipid layer which prevents oxygen diffusion and nitrogenase inactivation. Heterocyte glycolipids can be used to detect the presence of heterocytous cyanobacteria in present-day and past environments, providing insight into the functioning of the studied ecosystems. However, due to their good preservation throughout time, heterocyte glycolipids are not ideal to detect and study living communities, instead methods based on DNA are preferred. Currently cyanobacteria can be detected using untargeted genomic approaches such as metagenomics, or they can be specifically targeted by, for example, the use of primers that preferentially amplify their 16S rRNA gene or their nifH gene in the case of nitrogen fixing cyanobacteria. However, since not all cyanobacterial nitrogen fixers are heterocytous, there is currently no fast gene-based method to specifically detect and distinguish heterocytous cyanobacteria. Here, we developed a PCR-based method to specifically detect heterocytous cyanobacteria by designing primers targeting the gene (hglT) encoding the enzyme responsible for the last step in the biosynthesis of heterocyte glycolipid (i.e., a glycosyltransferase). We designed several primer sets using the publicly available sequences of 23 heterocytous cyanobacteria, after testing them on DNA extracts of 21 heterocyte-forming and 7 non-heterocyte forming freshwater cyanobacteria. The best primer set was chosen and successfully used to confirm the presence of heterocytous cyanobacteria in a marine environmental sample. Frontiers Media S.A. 2023-09-27 /pmc/articles/PMC10569477/ /pubmed/37840743 http://dx.doi.org/10.3389/fmicb.2023.1257040 Text en Copyright © 2023 Pérez Gallego, Bale, Sinninghe Damste and Villanueva. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Microbiology
Pérez Gallego, Ruth
Bale, Nicole J.
Sinninghe Damste, Jaap S.
Villanueva, Laura
Developing a genetic approach to target cyanobacterial producers of heterocyte glycolipids in the environment
title Developing a genetic approach to target cyanobacterial producers of heterocyte glycolipids in the environment
title_full Developing a genetic approach to target cyanobacterial producers of heterocyte glycolipids in the environment
title_fullStr Developing a genetic approach to target cyanobacterial producers of heterocyte glycolipids in the environment
title_full_unstemmed Developing a genetic approach to target cyanobacterial producers of heterocyte glycolipids in the environment
title_short Developing a genetic approach to target cyanobacterial producers of heterocyte glycolipids in the environment
title_sort developing a genetic approach to target cyanobacterial producers of heterocyte glycolipids in the environment
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10569477/
https://www.ncbi.nlm.nih.gov/pubmed/37840743
http://dx.doi.org/10.3389/fmicb.2023.1257040
work_keys_str_mv AT perezgallegoruth developingageneticapproachtotargetcyanobacterialproducersofheterocyteglycolipidsintheenvironment
AT balenicolej developingageneticapproachtotargetcyanobacterialproducersofheterocyteglycolipidsintheenvironment
AT sinninghedamstejaaps developingageneticapproachtotargetcyanobacterialproducersofheterocyteglycolipidsintheenvironment
AT villanuevalaura developingageneticapproachtotargetcyanobacterialproducersofheterocyteglycolipidsintheenvironment