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Volume 11, Issue 11

Single-cell genome analysis of a feminizing in butterflies reveals a shared origin with male-killing Open Access

Abstract

is a ubiquitous endosymbiont in arthropods that is maternally transmitted and affects host reproduction in various ways. Among these, skewing the host sex ratio towards females, either by killing males (male killing) or producing exclusively functional females (feminization or parthenogenesis), is considered advantageous for . In the butterfly , individuals harbouring the strain Fem exclusively produce female offspring. This occurs through a two-step mechanism in which blocks the transmission of the Z chromosome from Z0 females and feminizes the resultant Z0 offspring. Given the unique characteristics of Fem, understanding its genomic features is crucial to uncover the evolution and mechanisms of -induced reproductive manipulation. However, technical challenges in isolating Fem from co-infecting, closely related, non-male-killing/non-feminizing CI strain have hindered genomic analyses of Fem. In this study, we established a closed circular genome of Fem by developing a series of purification, cell sorting and single-cell genome sequencing techniques. Fem genome, ~1.3 Mb in size, specifically encodes male-killing gene homologues ( and ) and other putative virulence factors that are absent in CI. In addition, Fem carried prophage elements that showed high similarity to previously characterized male-killing-associated prophages in strains. This study highlights the shared functional genomic features between feminizing and male-killing in Lepidoptera and suggests a mechanistic link between these two -induced reproductive phenotypes.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): feminization , oscar , phage , single-cell and Wolbachia
Funding
This study was supported by the:
  • Bio-oriented Technology Research Advancement Institution (Award JPJ009237)
    • Principal Award Recipient: DaisukeKageyama
  • Japan Society for the Promotion of Science (Award 23K27248)
    • Principal Award Recipient: TatsuroKonagaya
  • Japan Society for the Promotion of Science (Award 23H02557)
    • Principal Award Recipient: TatsuroKonagaya
  • Japan Society for the Promotion of Science (Award 24H02293)
    • Principal Award Recipient: DaisukeKageyama
  • Japan Society for the Promotion of Science (Award 23K26922)
    • Principal Award Recipient: DaisukeKageyama
  • Japan Society for the Promotion of Science (Award 23H02229)
    • Principal Award Recipient: DaisukeKageyama
  • Japan Society for the Promotion of Science (Award 22K14902)
    • Principal Award Recipient: HiroshiArai
  • Japan Society for the Promotion of Science (Award 21J00895)
    • Principal Award Recipient: HiroshiArai
  • Japan Society for the Promotion of Science (Award 19J13123)
    • Principal Award Recipient: HiroshiArai
© 2025 The Authors
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License. This article was made open access via a Publish and Read agreement between the Microbiology Society and the corresponding author’s institution.
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2025-11-28
2025-12-15

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Single-cell genome analysis of a feminizing Wolbachia in Eurema butterflies reveals a shared origin with male-killing Wolbachia
M Gen 11, 001578 (2025); https://doi.org/10.1099/mgen.0.001578
/content/journal/mgen/10.1099/mgen.0.001578
/content/journal/mgen/10.1099/mgen.0.001578
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