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Species Discrimination within the Metarhizium PARB Clade: Ribosomal Intergenic Spacer (rIGS)-Based Diagnostic PCR and Single Marker Taxonomy

(1) Background: The entomopathogenic fungus Metarhizium anisopliae sensu lato forms a species complex, comprising a tight cluster made up of four species, namely M. anisopliae sensu stricto, M. pinghaense, M. robertsii and M. brunneum. Unambiguous species delineation within this “PARB clade” that en...

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Autores principales: Schuster, Christina, Baró Robaina, Yamilé, Ben Gharsa, Haifa, Bobushova, Saikal, Manfrino, Romina Guadalupe, Gutierrez, Alejandra C., Lopez Lastra, Claudia C., Doolotkeldieva, Tinatin, Leclerque, Andreas
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10607842/
https://www.ncbi.nlm.nih.gov/pubmed/37888252
http://dx.doi.org/10.3390/jof9100996
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author Schuster, Christina
Baró Robaina, Yamilé
Ben Gharsa, Haifa
Bobushova, Saikal
Manfrino, Romina Guadalupe
Gutierrez, Alejandra C.
Lopez Lastra, Claudia C.
Doolotkeldieva, Tinatin
Leclerque, Andreas
author_facet Schuster, Christina
Baró Robaina, Yamilé
Ben Gharsa, Haifa
Bobushova, Saikal
Manfrino, Romina Guadalupe
Gutierrez, Alejandra C.
Lopez Lastra, Claudia C.
Doolotkeldieva, Tinatin
Leclerque, Andreas
author_sort Schuster, Christina
collection PubMed
description (1) Background: The entomopathogenic fungus Metarhizium anisopliae sensu lato forms a species complex, comprising a tight cluster made up of four species, namely M. anisopliae sensu stricto, M. pinghaense, M. robertsii and M. brunneum. Unambiguous species delineation within this “PARB clade” that enables both the taxonomic assignment of new isolates and the identification of potentially new species is highly solicited. (2) Methods: Species-discriminating primer pairs targeting the ribosomal intergenic spacer (rIGS) sequence were designed and a diagnostic PCR protocol established. A partial rIGS sequence, referred to as rIGS-ID800, was introduced as a molecular taxonomic marker for PARB species delineation. (3) Results: PARB species from a validation strain set not implied in primer design were clearly discriminated using the diagnostic PCR protocol developed. Using rIGS-ID800 as a single sequence taxonomic marker gave rise to a higher resolution and statistically better supported delineation of PARB clade species. (4) Conclusions: Reliable species discrimination within the Metarhizium PARB clade is possible through both sequencing-independent diagnostic PCR and sequencing-dependent single marker comparison, both based on the rIGS marker.
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spelling pubmed-106078422023-10-28 Species Discrimination within the Metarhizium PARB Clade: Ribosomal Intergenic Spacer (rIGS)-Based Diagnostic PCR and Single Marker Taxonomy Schuster, Christina Baró Robaina, Yamilé Ben Gharsa, Haifa Bobushova, Saikal Manfrino, Romina Guadalupe Gutierrez, Alejandra C. Lopez Lastra, Claudia C. Doolotkeldieva, Tinatin Leclerque, Andreas J Fungi (Basel) Article (1) Background: The entomopathogenic fungus Metarhizium anisopliae sensu lato forms a species complex, comprising a tight cluster made up of four species, namely M. anisopliae sensu stricto, M. pinghaense, M. robertsii and M. brunneum. Unambiguous species delineation within this “PARB clade” that enables both the taxonomic assignment of new isolates and the identification of potentially new species is highly solicited. (2) Methods: Species-discriminating primer pairs targeting the ribosomal intergenic spacer (rIGS) sequence were designed and a diagnostic PCR protocol established. A partial rIGS sequence, referred to as rIGS-ID800, was introduced as a molecular taxonomic marker for PARB species delineation. (3) Results: PARB species from a validation strain set not implied in primer design were clearly discriminated using the diagnostic PCR protocol developed. Using rIGS-ID800 as a single sequence taxonomic marker gave rise to a higher resolution and statistically better supported delineation of PARB clade species. (4) Conclusions: Reliable species discrimination within the Metarhizium PARB clade is possible through both sequencing-independent diagnostic PCR and sequencing-dependent single marker comparison, both based on the rIGS marker. MDPI 2023-10-08 /pmc/articles/PMC10607842/ /pubmed/37888252 http://dx.doi.org/10.3390/jof9100996 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Schuster, Christina
Baró Robaina, Yamilé
Ben Gharsa, Haifa
Bobushova, Saikal
Manfrino, Romina Guadalupe
Gutierrez, Alejandra C.
Lopez Lastra, Claudia C.
Doolotkeldieva, Tinatin
Leclerque, Andreas
Species Discrimination within the Metarhizium PARB Clade: Ribosomal Intergenic Spacer (rIGS)-Based Diagnostic PCR and Single Marker Taxonomy
title Species Discrimination within the Metarhizium PARB Clade: Ribosomal Intergenic Spacer (rIGS)-Based Diagnostic PCR and Single Marker Taxonomy
title_full Species Discrimination within the Metarhizium PARB Clade: Ribosomal Intergenic Spacer (rIGS)-Based Diagnostic PCR and Single Marker Taxonomy
title_fullStr Species Discrimination within the Metarhizium PARB Clade: Ribosomal Intergenic Spacer (rIGS)-Based Diagnostic PCR and Single Marker Taxonomy
title_full_unstemmed Species Discrimination within the Metarhizium PARB Clade: Ribosomal Intergenic Spacer (rIGS)-Based Diagnostic PCR and Single Marker Taxonomy
title_short Species Discrimination within the Metarhizium PARB Clade: Ribosomal Intergenic Spacer (rIGS)-Based Diagnostic PCR and Single Marker Taxonomy
title_sort species discrimination within the metarhizium parb clade: ribosomal intergenic spacer (rigs)-based diagnostic pcr and single marker taxonomy
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10607842/
https://www.ncbi.nlm.nih.gov/pubmed/37888252
http://dx.doi.org/10.3390/jof9100996
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