{"database":"biostudies-arrayexpress","file_versions":[],"scores":null,"additional":{"omics_type":["Unknown","Transcriptomics","Genomics","Proteomics"],"submitter":["Jean-Baptiste Chazalon"],"instrument_platform":["Illumina NovaSeq 6000"],"study_type":["RNA-seq of coding RNA"],"organism":["Cypripedium henryi"],"species":["Cypripedium henryi"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/E-MTAB-16751"],"description":["Floral scent serves as a primary mechanism for pollinator attraction, acting as a driver for reproductive isolation and speciation. Because shifts in the composition of floral Volatile Organic Compounds (VOCs) significantly influence a plant's attractiveness, and because the production of these compounds is metabolically costly, plants often reduce scent emissions following successful pollination. This resource-saving strategy is particularly expected in long-lasting fragrant flowers, such as those of the deceptive orchid genus Cypripedium. To investigate potential post-pollination effects on the floral scent of Cypripedium henryi, four pollination treatments were applied to flowers approximately two days post-anthesis (n = 3 replicates per treatment, 12 flowers total): • Treatment P: Removal of both pollinia. • Treatment G: Deposition of a donor flower’s pollinium onto the stigma. • Treatment GP: Simultaneous removal of pollinia and deposition of a donor pollinium. • Treatment K (Control): No artificial manipulation. Floral scent was sampled from each flower immediately before pollination treatment and again 48 hours later. Following the final scent collection, three specific tissue types per flower were harvested for RNA analysis: the labellum, staminode, and lateral petal. The resulting 36 RNA samples were extracted and sequenced to facilitate de novo transcriptome assembly. We quantified transcripts and performed differential expression analysis to compare the pollination treatments against the control group, as well as to identify variation across the different tissue types. Following gene annotation, we focused our investigation on candidate genes within the terpene and fatty acid biosynthesis pathways, which are responsible for producing the characteristic volatiles of C. henryi. Finally, the expression levels of these candidate genes were evaluated in direct relation to the volatile levels identified in the scent analysis to provide a comprehensive view of the molecular regulation of post-pollination scent emission."],"repository":["biostudies-arrayexpress"],"sample_protocol":["Sequencing - Eukaryotic mRNA sequencing was conducted by BMKGENE (Münster, Germany) on an Illumina NovaSeq 6000 system. Sequence data were generated as 150 bp paired-end reads.","Growth Protocol - Two multiflowered clonal individuals of Cypripedium henryi were maintained year-round according to species requirements under the care of expert Cypripedium horticulturalists.","Sample Collection - Sampling took place in the greenhouse 48 h (± 2 h) after pollination treatments. The labellum, staminode, and both lateral petals of each treated flower were collected using a thin razor blade and immediately flash-frozen in liquid nitrogen for transport to the laboratory.","Nucleic Acid Extraction - The whole staminode (around 15 mg), half of the labellum (around 60 mg) and a whole lateral petal (around 30 mg) were used for total RNA extraction using the protocol of innuPREP Plant RNA Kit (Innuscreen GmbH). Prior to the extraction, the tissue samples were crushed using a mechanical mortar. Total RNA was eluted in 20 μl RNAse free H2O. Total RNA concentration and purity were determined by RNA screen tape analysis the Tapestation (Agilent Genomic Screentape, Agilent Technologies) performed on the Genomic Screen Tape device (Agilent Technologies) according to the manufacturer’s instructions. RIN number of samples ranged between 5.8 and 8.4.","Library Construction - Strand-specific RNA-seq libraries were prepared by BMKGENE (Münster, Germany). Following poly(A) enrichment of the mRNA fraction, libraries were constructed using the dUTP method.","Sample Treatment - Three biological replicates were used for each of the four treatments (n=12 flowers total). To minimize potential bias, replicates were distributed evenly across the two clonal individuals. The experimental treatments were executed as follows: • Treatment P (Pollen Removal): Manual removal of both pollinia.   • Treatment G (Cross-Pollination): Deposition of pollinia from a donor flower onto the stigma.   • Treatment GP (Combined): Manual removal of both pollinia followed by the deposition of donor pollinia onto the stigma.   • Treatment K (Control): Untouched flowers served as the negative control."],"figure_sub":["Organization","MINSEQE Score","Assays and Data","MAGE-TAB Files"],"pubmed_authors":["Jean-Baptiste Chazalon"],"additional_accession":[]},"is_claimable":false,"name":"RNA-seq analysis of floral volatile gene expression in Cypripedium henryi in response to pollination treatments","description":"Floral scent serves as a primary mechanism for pollinator attraction, acting as a driver for reproductive isolation and speciation. Because shifts in the composition of floral Volatile Organic Compounds (VOCs) significantly influence a plant's attractiveness, and because the production of these compounds is metabolically costly, plants often reduce scent emissions following successful pollination. This resource-saving strategy is particularly expected in long-lasting fragrant flowers, such as those of the deceptive orchid genus Cypripedium. To investigate potential post-pollination effects on the floral scent of Cypripedium henryi, four pollination treatments were applied to flowers approximately two days post-anthesis (n = 3 replicates per treatment, 12 flowers total): • Treatment P: Removal of both pollinia. • Treatment G: Deposition of a donor flower’s pollinium onto the stigma. • Treatment GP: Simultaneous removal of pollinia and deposition of a donor pollinium. • Treatment K (Control): No artificial manipulation. Floral scent was sampled from each flower immediately before pollination treatment and again 48 hours later. Following the final scent collection, three specific tissue types per flower were harvested for RNA analysis: the labellum, staminode, and lateral petal. The resulting 36 RNA samples were extracted and sequenced to facilitate de novo transcriptome assembly. We quantified transcripts and performed differential expression analysis to compare the pollination treatments against the control group, as well as to identify variation across the different tissue types. Following gene annotation, we focused our investigation on candidate genes within the terpene and fatty acid biosynthesis pathways, which are responsible for producing the characteristic volatiles of C. henryi. Finally, the expression levels of these candidate genes were evaluated in direct relation to the volatile levels identified in the scent analysis to provide a comprehensive view of the molecular regulation of post-pollination scent emission.","dates":{"release":"2026-03-19T00:00:00Z","modification":"2026-03-19T02:03:21.324Z","creation":"2026-03-12T13:28:28.23Z"},"accession":"E-MTAB-16751","cross_references":{"ENA":["ERP190725"],"EFO":["EFO_0002944","EFO_0004170","EFO_0003789","EFO_0005518","EFO_0003738","EFO_0004184","EFO_0003969"]}}