Use of rpoB gene phylogenetic marker-based distinction of abiotic stress tolerant and plant-growth promoting Bacillus paralicheniformis isolates from their closely related Bacillus licheniformis

Bacillus paralicheniformis is a new identified species, which was distinguished from Bacillus licheniformis in 2015 through extensive phylogenomic and phylogenetic analyses. In this context, this study aimed to achieve a clear identification of the active plant-growth promoting rhizobacteria (PGP...

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Main Authors: Mohammed Ajdig, Bahia Rached, Ahlam Mbarki, Taha Chouati, Chouhra Talbi, Elmostafa El Fahime, Marouane Melloul
Format: Article
Language:English
Published: ResearchersLinks, Ltd 2024-05-01
Series:Novel Research in Microbiology Journal
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Online Access:https://nrmj.journals.ekb.eg/article_352759_55846fb8dc99a7dac641fe1a85912cd0.pdf
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author Mohammed Ajdig
Bahia Rached
Ahlam Mbarki
Taha Chouati
Chouhra Talbi
Elmostafa El Fahime
Marouane Melloul
author_facet Mohammed Ajdig
Bahia Rached
Ahlam Mbarki
Taha Chouati
Chouhra Talbi
Elmostafa El Fahime
Marouane Melloul
author_sort Mohammed Ajdig
collection DOAJ
description Bacillus paralicheniformis is a new identified species, which was distinguished from Bacillus licheniformis in 2015 through extensive phylogenomic and phylogenetic analyses. In this context, this study aimed to achieve a clear identification of the active plant-growth promoting rhizobacteria (PGPR) B. paralicheniformis isolates among the closely related B. licheniformis through molecular typing, helping for the development of clearly-identified PGPR isolates to be used as biofertilizers. A total of 15 rhizobacteria were isolated from the olive rhizosphere soils. These bacterial isolates exhibited various proprieties in terms of abiotic stress tolerance, biofilm formation under stress conditions, and enzyme activities (i.e., lipases, cellulases, and proteases). In addition, several PGP traits such as phosphate solubilization and the production of siderophores and indol-3 acetic acid were also observed. Molecular identification through 16S rRNA sequencing initially identified all the isolates as Bacillus spp. The multi-locus sequence typing (MLST) scheme, using six housekeeping genes (adk, ccpA, recF, rpoB, sucC, and spo0A) had unveiled distinct allelic profiles resulting in 13 unique sequence types (ST). Notably, a comprehensive analysis indicated no exact allele matches existed between the examined isolates and those documented in the PubMLST database. Among the six housekeeping genes, we noticed that rpoB gene (RNA polymerase, subunit beta) had multiple polymorphic sites that were bordered by conserved sequences. Furthermore, the phylogenetic analysis based on rpoB had identified clusters indicating distinct phylogenetic relationships between B. paralicheniformis and B. licheniformis, and successfully differentiated the two species from a pool of 90 strains. The rpoB partial sequence proved to be effective for accurate species discrimination.
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spelling doaj-art-864bf048c7be435f8677ec4ebf5c48ce2025-08-20T01:58:24ZengResearchersLinks, LtdNovel Research in Microbiology Journal2537-02862537-02942024-05-01832414243410.21608/NRMJ.2024.280135.1508Use of rpoB gene phylogenetic marker-based distinction of abiotic stress tolerant and plant-growth promoting Bacillus paralicheniformis isolates from their closely related Bacillus licheniformisMohammed Ajdig0Bahia Rached1Ahlam Mbarki2Taha Chouati3Chouhra Talbi4Elmostafa El Fahime5Marouane Melloul6Microbiology and Molecular Biology Team, Center of Plant and Microbial Biotechnology, Biodiversity and Environment, Faculty of Sciences, Mohammed V University, Rabat, MoroccoGenopath Research Center, ERNN, Faculty of Medicine and Pharmacy, University Mohammed V of Rabat, MoroccoFunctional Genomic Platform/National Centre for Scientific and Technical Research (CNRST), Rabat, MoroccoFunctional Genomic Platform/National Centre for Scientific and Technical Research (CNRST), Rabat, MoroccoMicrobiology and Molecular Biology Team, Center of Plant and Microbial Biotechnology, Biodiversity and Environment, Faculty of Sciences, Mohammed V University, Rabat, MoroccoLaboratory of Chemistry-Physics and Biotechnology of Biomolecules and Materials, Faculty of Science and Technology, Mohammedia, Hassan II University, Casablanca, MoroccoFunctional Genomic Platform/National Centre for Scientific and Technical Research (CNRST), Rabat, MoroccoBacillus paralicheniformis is a new identified species, which was distinguished from Bacillus licheniformis in 2015 through extensive phylogenomic and phylogenetic analyses. In this context, this study aimed to achieve a clear identification of the active plant-growth promoting rhizobacteria (PGPR) B. paralicheniformis isolates among the closely related B. licheniformis through molecular typing, helping for the development of clearly-identified PGPR isolates to be used as biofertilizers. A total of 15 rhizobacteria were isolated from the olive rhizosphere soils. These bacterial isolates exhibited various proprieties in terms of abiotic stress tolerance, biofilm formation under stress conditions, and enzyme activities (i.e., lipases, cellulases, and proteases). In addition, several PGP traits such as phosphate solubilization and the production of siderophores and indol-3 acetic acid were also observed. Molecular identification through 16S rRNA sequencing initially identified all the isolates as Bacillus spp. The multi-locus sequence typing (MLST) scheme, using six housekeeping genes (adk, ccpA, recF, rpoB, sucC, and spo0A) had unveiled distinct allelic profiles resulting in 13 unique sequence types (ST). Notably, a comprehensive analysis indicated no exact allele matches existed between the examined isolates and those documented in the PubMLST database. Among the six housekeeping genes, we noticed that rpoB gene (RNA polymerase, subunit beta) had multiple polymorphic sites that were bordered by conserved sequences. Furthermore, the phylogenetic analysis based on rpoB had identified clusters indicating distinct phylogenetic relationships between B. paralicheniformis and B. licheniformis, and successfully differentiated the two species from a pool of 90 strains. The rpoB partial sequence proved to be effective for accurate species discrimination.https://nrmj.journals.ekb.eg/article_352759_55846fb8dc99a7dac641fe1a85912cd0.pdfbacillus paralicheniformisbacillus licheniformisrpob genepgprmlst
spellingShingle Mohammed Ajdig
Bahia Rached
Ahlam Mbarki
Taha Chouati
Chouhra Talbi
Elmostafa El Fahime
Marouane Melloul
Use of rpoB gene phylogenetic marker-based distinction of abiotic stress tolerant and plant-growth promoting Bacillus paralicheniformis isolates from their closely related Bacillus licheniformis
Novel Research in Microbiology Journal
bacillus paralicheniformis
bacillus licheniformis
rpob gene
pgpr
mlst
title Use of rpoB gene phylogenetic marker-based distinction of abiotic stress tolerant and plant-growth promoting Bacillus paralicheniformis isolates from their closely related Bacillus licheniformis
title_full Use of rpoB gene phylogenetic marker-based distinction of abiotic stress tolerant and plant-growth promoting Bacillus paralicheniformis isolates from their closely related Bacillus licheniformis
title_fullStr Use of rpoB gene phylogenetic marker-based distinction of abiotic stress tolerant and plant-growth promoting Bacillus paralicheniformis isolates from their closely related Bacillus licheniformis
title_full_unstemmed Use of rpoB gene phylogenetic marker-based distinction of abiotic stress tolerant and plant-growth promoting Bacillus paralicheniformis isolates from their closely related Bacillus licheniformis
title_short Use of rpoB gene phylogenetic marker-based distinction of abiotic stress tolerant and plant-growth promoting Bacillus paralicheniformis isolates from their closely related Bacillus licheniformis
title_sort use of rpob gene phylogenetic marker based distinction of abiotic stress tolerant and plant growth promoting bacillus paralicheniformis isolates from their closely related bacillus licheniformis
topic bacillus paralicheniformis
bacillus licheniformis
rpob gene
pgpr
mlst
url https://nrmj.journals.ekb.eg/article_352759_55846fb8dc99a7dac641fe1a85912cd0.pdf
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