Defining the cultured and uncultured bacterial fractions in Cannabis seeds

Abstract Background Seeds provide a unique environment shaped by co-evolutionary processes, hosting diverse microbial communities. While microbiome studies have uncovered an extensive diversity of microorganisms, culture-based approaches remain crucial for understanding microbial potential and funct...

Full description

Saved in:
Bibliographic Details
Main Authors: Carolina Lobato, Ahmed Abdelfattah, Gabriele Berg, Tomislav Cernava
Format: Article
Language:English
Published: BMC 2025-06-01
Series:Environmental Microbiome
Subjects:
Online Access:https://doi.org/10.1186/s40793-025-00731-4
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1849691335490011136
author Carolina Lobato
Ahmed Abdelfattah
Gabriele Berg
Tomislav Cernava
author_facet Carolina Lobato
Ahmed Abdelfattah
Gabriele Berg
Tomislav Cernava
author_sort Carolina Lobato
collection DOAJ
description Abstract Background Seeds provide a unique environment shaped by co-evolutionary processes, hosting diverse microbial communities. While microbiome studies have uncovered an extensive diversity of microorganisms, culture-based approaches remain crucial for understanding microbial potential and functional interactions. However, the factors influencing microbial culturability within seeds are not well understood. Results In this study, we investigated the culturing patterns of bacteria inside Cannabis seeds, assessing their phylogenetic diversity, abundance, and putative interactions. Bacteria were cultured from 54 different Cannabis accessions using germinated seeds and a range of nutrient media including those supplemented with Cannabis extracts. The cultured fraction consisted of taxa from five prominent classes—Gammaproteobacteria, Bacilli, Actinobacteria, Alphaproteobacteria, and Bacteroidia—encompassing 36 genera. Despite representing only 6.3% of the total microbiota, these cultured bacteria accounted for 89.2% of the microbial population. Almost 60% of the amplicon sequence variants (ASVs) were phylogenetically distant from cultured taxa. Rare bacterial groups such as Acidobacteriae and Verrucomicrobiae, known for their plant growth-promoting traits, were exclusively found in the uncultured fraction. Network analyses revealed that uncultured taxa are centralized and more connected to hubs, suggesting that interspecies interactions strongly influence culturability. Conclusion Our findings highlight the limitations of culture-based methods in capturing the full microbial diversity of Cannabis seeds and emphasize the importance of microbial interactions in determining culturability. The strong network connectivity of uncultured taxa suggests that interdependencies and competition within the seed microbiome may hinder the isolation of key bacterial groups. These insights provide a framework for refining cultivation strategies to recover ecologically significant microbes with potential agricultural applications.
format Article
id doaj-art-8910a5ca6d0c46af8b0c0d4f0060b087
institution DOAJ
issn 2524-6372
language English
publishDate 2025-06-01
publisher BMC
record_format Article
series Environmental Microbiome
spelling doaj-art-8910a5ca6d0c46af8b0c0d4f0060b0872025-08-20T03:21:03ZengBMCEnvironmental Microbiome2524-63722025-06-0120111310.1186/s40793-025-00731-4Defining the cultured and uncultured bacterial fractions in Cannabis seedsCarolina Lobato0Ahmed Abdelfattah1Gabriele Berg2Tomislav Cernava3Institute of Environmental Biotechnology, Graz University of TechnologyLeibniz Institute for Agricultural Engineering and BioeconomyInstitute of Environmental Biotechnology, Graz University of TechnologyInstitute of Environmental Biotechnology, Graz University of TechnologyAbstract Background Seeds provide a unique environment shaped by co-evolutionary processes, hosting diverse microbial communities. While microbiome studies have uncovered an extensive diversity of microorganisms, culture-based approaches remain crucial for understanding microbial potential and functional interactions. However, the factors influencing microbial culturability within seeds are not well understood. Results In this study, we investigated the culturing patterns of bacteria inside Cannabis seeds, assessing their phylogenetic diversity, abundance, and putative interactions. Bacteria were cultured from 54 different Cannabis accessions using germinated seeds and a range of nutrient media including those supplemented with Cannabis extracts. The cultured fraction consisted of taxa from five prominent classes—Gammaproteobacteria, Bacilli, Actinobacteria, Alphaproteobacteria, and Bacteroidia—encompassing 36 genera. Despite representing only 6.3% of the total microbiota, these cultured bacteria accounted for 89.2% of the microbial population. Almost 60% of the amplicon sequence variants (ASVs) were phylogenetically distant from cultured taxa. Rare bacterial groups such as Acidobacteriae and Verrucomicrobiae, known for their plant growth-promoting traits, were exclusively found in the uncultured fraction. Network analyses revealed that uncultured taxa are centralized and more connected to hubs, suggesting that interspecies interactions strongly influence culturability. Conclusion Our findings highlight the limitations of culture-based methods in capturing the full microbial diversity of Cannabis seeds and emphasize the importance of microbial interactions in determining culturability. The strong network connectivity of uncultured taxa suggests that interdependencies and competition within the seed microbiome may hinder the isolation of key bacterial groups. These insights provide a framework for refining cultivation strategies to recover ecologically significant microbes with potential agricultural applications.https://doi.org/10.1186/s40793-025-00731-4BacteriaCannabisCo-occurrence networkCulturingMicrobial dark matterMicrobial interactions
spellingShingle Carolina Lobato
Ahmed Abdelfattah
Gabriele Berg
Tomislav Cernava
Defining the cultured and uncultured bacterial fractions in Cannabis seeds
Environmental Microbiome
Bacteria
Cannabis
Co-occurrence network
Culturing
Microbial dark matter
Microbial interactions
title Defining the cultured and uncultured bacterial fractions in Cannabis seeds
title_full Defining the cultured and uncultured bacterial fractions in Cannabis seeds
title_fullStr Defining the cultured and uncultured bacterial fractions in Cannabis seeds
title_full_unstemmed Defining the cultured and uncultured bacterial fractions in Cannabis seeds
title_short Defining the cultured and uncultured bacterial fractions in Cannabis seeds
title_sort defining the cultured and uncultured bacterial fractions in cannabis seeds
topic Bacteria
Cannabis
Co-occurrence network
Culturing
Microbial dark matter
Microbial interactions
url https://doi.org/10.1186/s40793-025-00731-4
work_keys_str_mv AT carolinalobato definingtheculturedandunculturedbacterialfractionsincannabisseeds
AT ahmedabdelfattah definingtheculturedandunculturedbacterialfractionsincannabisseeds
AT gabrieleberg definingtheculturedandunculturedbacterialfractionsincannabisseeds
AT tomislavcernava definingtheculturedandunculturedbacterialfractionsincannabisseeds