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<p>Two virulent <i>Metarhizium rileyi</i> isolates, SlMr-DOR and SfMr-DOR, were comparatively characterized using morphological, pathogenicity, molecular, phylogenetic, and whole-genome analyses to investigate genomic differences associated with their entomopathogenic characteristics. Both isolates exhibited typical morphological features of <i>M. rileyi</i> and caused high mortality against second-instar <i>Spodoptera litura</i> larvae, with cumulative mortalities of 95.2% (SlMr-DOR) and 91.0% (SfMr-DOR) at 10 days post-inoculation. Molecular identification based on ITS, β-tubulin, and translation elongation factor 1-alpha (TEF1-α) gene sequences, together with multilocus phylogenetic analysis, confirmed their identity as <i>M. rileyi</i> and their placement within the <i>Metarhizium</i> lineage. High-quality genome assemblies of 32,547,182&#xa0;bp (SlMr-DOR) and 36,078,628&#xa0;bp (SfMr-DOR) with high BUSCO completeness enabled comparative genome analysis against the reference genome <i>M. rileyi</i> RCEF_4871. Comparative annotation identified quantitative differences in predicted protein families, including fungal-specific transcription factors, Zn₂/Cys₆ transcription factors, cytochrome P450s, protein kinases, and transporter proteins. Both isolates also possessed expanded repertoires of predicted cuticle-degrading enzyme families, including serine proteases, subtilisin-like proteases, trypsin-like proteases, metalloproteases, chitinases, lipases, and glycoside hydrolases, relative to the reference genome. Genome-wide annotation against the PHI database identified conserved putative virulence-associated genes, including Egh16-like proteins and pectin lyase fold proteins, while biosynthetic gene cluster analysis predicted diverse secondary metabolite pathways comprising polyketide synthases, non-ribosomal peptide synthetases, RiPP-like clusters, and hybrid biosynthetic clusters. These findings provide comprehensive comparative genomic resources for <i>M. rileyi</i> and identify candidate genomic features that may contribute to fungal pathogenicity and environmental adaptation, providing a foundation for future functional genomic studies and supporting the continued evaluation of these isolates as potential biocontrol agents against lepidopteran pests.</p>

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Comparative genomic analysis of two virulent Metarhizium rileyi (Hypocreales: Clavicipitaceae) isolates reveals enhanced virulence potential

  • 期刊论文

  • 发表日期：

    2026年8月2日

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  • P. Duraimurugan,
  • D. Neethu Roy,
  • Divya Shree,
  • M. Rikvitha,
  • T. Meera Devi,
  • Pasumarthi Venkata Dinesh Kumar,
  • A. Chowdappa

摘要

Two virulent Metarhizium rileyi isolates, SlMr-DOR and SfMr-DOR, were comparatively characterized using morphological, pathogenicity, molecular, phylogenetic, and whole-genome analyses to investigate genomic differences associated with their entomopathogenic characteristics. Both isolates exhibited typical morphological features of M. rileyi and caused high mortality against second-instar Spodoptera litura larvae, with cumulative mortalities of 95.2% (SlMr-DOR) and 91.0% (SfMr-DOR) at 10 days post-inoculation. Molecular identification based on ITS, β-tubulin, and translation elongation factor 1-alpha (TEF1-α) gene sequences, together with multilocus phylogenetic analysis, confirmed their identity as M. rileyi and their placement within the Metarhizium lineage. High-quality genome assemblies of 32,547,182 bp (SlMr-DOR) and 36,078,628 bp (SfMr-DOR) with high BUSCO completeness enabled comparative genome analysis against the reference genome M. rileyi RCEF_4871. Comparative annotation identified quantitative differences in predicted protein families, including fungal-specific transcription factors, Zn₂/Cys₆ transcription factors, cytochrome P450s, protein kinases, and transporter proteins. Both isolates also possessed expanded repertoires of predicted cuticle-degrading enzyme families, including serine proteases, subtilisin-like proteases, trypsin-like proteases, metalloproteases, chitinases, lipases, and glycoside hydrolases, relative to the reference genome. Genome-wide annotation against the PHI database identified conserved putative virulence-associated genes, including Egh16-like proteins and pectin lyase fold proteins, while biosynthetic gene cluster analysis predicted diverse secondary metabolite pathways comprising polyketide synthases, non-ribosomal peptide synthetases, RiPP-like clusters, and hybrid biosynthetic clusters. These findings provide comprehensive comparative genomic resources for M. rileyi and identify candidate genomic features that may contribute to fungal pathogenicity and environmental adaptation, providing a foundation for future functional genomic studies and supporting the continued evaluation of these isolates as potential biocontrol agents against lepidopteran pests.