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PreprintCC BY-NDbioRxiv

Floral scent diversity and evolution in deceptive Cypripedium, and post-pollination effects on floral scent and its biosynthesis in C. henryi

  1. 1Staatliche Naturwissenschaftliche Sammlungen Bayerns (SNSB)

Abstract

Background and Aims: Floral scent is a primary driver of pollinator attraction and reproductive isolation in deceptive orchids. Yet, little is known about the variation and evolution of scent in a phylogenetic framework and how pollination affects the biosynthesis and emission of scents in these lineages. This study characterizes scent diversity across Cypripedium, a genus of deceptive orchids, examines phylogenetic constraints, and investigates post-pollination effects on scent composition in one of the species. Methods: Floral volatile organic compounds (VOCs) were sampled from 31 Cypripedium taxa using dynamic headspace techniques, and analysed by gas chromatography coupled to mass spectrometry (GC/MS). Uni- and multivariate methods were used to analyse intra- and interspecific scent variation and test for phylogenetic signal in floral scent. Post-pollination effects on scent composition were experimentally tested in C. henryi using four pollination treatments, combined with RNA-seq analyses of floral tissues to examine transcriptional responses in VOC biosynthetic pathways. Key results: We detected 258 floral VOCs, revealing a high chemical diversity dominated by terpenoids, aliphatics, and aromatic compounds. Floral scent composition was highly species-specific, and closely related taxa often shared only few compounds. Scent composition differed among pollination groups, but these groups explained only a limited proportion of overall scent variation. We detected a significant but weak phylogenetic signal, suggesting limited evolutionary constraint in floral scent. Pollination had limited effect on C. henryis floral scent and volatile biosynthesis gene expression, despite a significant decrease in total scent emission after pollinium deposition. Transcriptional changes predominantly occurred in the petals rather than the labellum and staminode. Conclusion: Floral scent evolution in Cypripedium is characterized by high chemical diversity, and a weak phylogenetic constraint, consistent with variable selection in food-deceptive systems. The pollination study further suggests the absence of a strong, directed post-pollination effect on the floral scent of C. henryi. Keywords: Cypripedium; deceptive pollination; floral scent; volatile organic compounds (VOCs); phylogenetic signal; linalool; post-pollination effects; transcriptome profiles; differential gene expression; floral organs

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BibTeX
@misc{chazalon2026floral,
  title = {{Floral scent diversity and evolution in deceptive Cypripedium, and post-pollination effects on floral scent and its biosynthesis in C. henryi}},
  author = {J.-B. Chazalon and S. Pfanzelt and C. Siadjeu and E. Y. Hu and S. Kotagal and S. Doetterl and G. Kadereit},
  howpublished = {bioRxiv},
  year = {2026},
  month = nov,
  doi = {10.64898/2026.11.04.760698},
  url = {https://www.biorxiv.org/content/10.64898/2026.11.04.760698v1},
  note = {CC BY-ND},
}
RIS
TY  - UNPB
TI  - Floral scent diversity and evolution in deceptive Cypripedium, and post-pollination effects on floral scent and its biosynthesis in C. henryi
AU  - J.-B. Chazalon
AU  - S. Pfanzelt
AU  - C. Siadjeu
AU  - E. Y. Hu
AU  - S. Kotagal
AU  - S. Doetterl
AU  - G. Kadereit
PY  - 2026
DA  - 2026/11/04
PB  - bioRxiv
DO  - 10.64898/2026.11.04.760698
UR  - https://www.biorxiv.org/content/10.64898/2026.11.04.760698v1
AB  - Background and Aims: Floral scent is a primary driver of pollinator attraction and reproductive isolation in deceptive orchids. Yet, little is known about the variation and evolution of scent in a phylogenetic framework and how pollination affects the biosynthesis and emission of scents in these lineages. This study characterizes scent diversity across Cypripedium, a genus of deceptive orchids, examines phylogenetic constraints, and investigates post-pollination effects on scent composition in one of the species. Methods: Floral volatile organic compounds (VOCs) were sampled from 31 Cypripedium taxa using dynamic headspace techniques, and analysed by gas chromatography coupled to mass spectrometry (GC/MS). Uni- and multivariate methods were used to analyse intra- and interspecific scent variation and test for phylogenetic signal in floral scent. Post-pollination effects on scent composition were experimentally tested in C. henryi using four pollination treatments, combined with RNA-seq analyses of floral tissues to examine transcriptional responses in VOC biosynthetic pathways. Key results: We detected 258 floral VOCs, revealing a high chemical diversity dominated by terpenoids, aliphatics, and aromatic compounds. Floral scent composition was highly species-specific, and closely related taxa often shared only few compounds. Scent composition differed among pollination groups, but these groups explained only a limited proportion of overall scent variation. We detected a significant but weak phylogenetic signal, suggesting limited evolutionary constraint in floral scent. Pollination had limited effect on C. henryis floral scent and volatile biosynthesis gene expression, despite a significant decrease in total scent emission after pollinium deposition. Transcriptional changes predominantly occurred in the petals rather than the labellum and staminode. Conclusion: Floral scent evolution in Cypripedium is characterized by high chemical diversity, and a weak phylogenetic constraint, consistent with variable selection in food-deceptive systems. The pollination study further suggests the absence of a strong, directed post-pollination effect on the floral scent of C. henryi. Keywords: Cypripedium; deceptive pollination; floral scent; volatile organic compounds (VOCs); phylogenetic signal; linalool; post-pollination effects; transcriptome profiles; differential gene expression; floral organs
ER  -