Defensive fungal symbiosis on insect hindlegs
Data files
Aug 23, 2025 version files 14.87 MB
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Data_For_Defensive_fungal_symbiosis_on_insect_hindlegs.zip
14.85 MB
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README.md
10.93 KB
Abstract
Tympanal organs, as “insect ears,” have evolved repeatedly. Dinidorid stinkbugs were reported to possess a conspicuous tympanal organ on females’ hind legs. Here we report an unexpected discovery that the stinkbug’s “tympanal organ” is actually an external symbiotic organ. The organ’s surface is not membranous but a porous cuticle, where each pore connects to glandular secretory cells. In reproductive females, the hind leg organ is covered with fungal hyphae growing out of the pores. Upon oviposition, the females skillfully transfer the fungi from the organ to the eggs. The eggs are covered with fungal hyphae and physically protected against wasp parasitism. The fungi are mostly benign Cordycipitaceae fungi and show considerable diversity among insect individuals and populations, indicating environmental acquisition of the specific fungal associates.
Dataset DOI: 10.5061/dryad.66t1g1kcp
Description of the data and file structure
The results of SEM observations of the insect materials, amplicon sequence analyses of fungal microbiota of the stink bugs, RNAseq analyses of the insect hindlegs, the insect morphometries, phylogenetic analyses of the insect mitochondria.
Files and variables
File: Data_For_Defensive_fungal_symbiosis_on_insect_hindlegs.zip
Description:
The ZIP-compressed file contains folders corresponding to the figures in the manuscript. Each folder includes the digital images and/or source data files containing the raw data used to prepare the figures.
Folder: Fig1
SEM images and their metadata for Fig. 1.
- Mg_Ad_Mal_Hind_R20200311.000004.tif – SEM image of the hind tibia of a mature adult male M. gracilicorne.
- Mg_Ad_Mal_Hind_R20200311.000004.txt – Metadata of equipment settings when capturing the above image.
- Mg_Ad_Mal_Hind_R20200311.000007.tif – Magnified SEM image of the hind tibia of a mature adult male M. gracilicorne.
- Mg_Ad_Mal_Hind_R20200311.000007.txt – Metadata of equipment settings when capturing the above image.
- Mg_Te_Fem_Hind_R20151026.000185.tif – SEM image of the hind tibia of an immature adult female M. gracilicorne.
- Mg_Te_Fem_Hind_R20151026.000185.txt – Metadata of equipment settings when capturing the above image.
- Mg_Te_Fem_Hind_R20151026.000187.tif – Magnified SEM image of the hind tibia of an immature adult female M. gracilicorne.
- Mg_Te_Fem_Hind_R20151026.000187.txt – Metadata of equipment settings when capturing the above image.
- Mg_Ad_Fem_Hind_R20200311.000001.tif – SEM image of the hind tibia of a mature adult female M. gracilicorne.
- Mg_Ad_Fem_Hind_R20200311.000001.txt – Metadata of equipment settings when capturing the above image.
- Mg_Ad_Fem_Hind_R20200311.000004.tif – Magnified SEM image of the hind tibia of a mature adult female M. gracilicorne.
- Mg_Ad_Fem_Hind_R20200311.000004.txt – Metadata of equipment settings when capturing the above image.
Folder: Fig2
ITS amplicon seq data for Fig. 2 and Fig. S3.
- Fig2abcd_S3abcdef.tsv – Number of reads for each OTU in the M. gracilicorne ITS amplicon experiment.
- First column (id): OTU IDs.
- Second to 76th columns: Sample names (headers); rows contain the number of reads for each OTU ID.
- Last column (taxon): Taxon assignments for each OTU ID
Folder: Fig4
Data of suppression of wasp parasitism by egg-covering fungal hyphae for Fig. 4.
- Fig4e_parasitized rate in wild.csv - Parasitism rates of eggs in wild.
- egg no: Egg ID.
- egg mass group: Egg mass ID.
- parasitized: Egg parasitized (1) or not (0).
- fertilized: Egg fertilized (1) or not (0).
- disappeared: Egg disappeared (1) or not (0).
- unfertilized: Egg unfertilized (1) or not (0).
- Fig4f_parasitism success rate.csv - Parasitism success rates of fungus-covered or fungus-removed eggs.
- egg no: Egg ID.
- egg mass group: Egg mass ID.
- parasitized: Egg parasitized (1) or not (0).
- fertilized: Egg fertilized (1) or not (0).
- unfertilized: Egg unfertilized (1) or not (0).
- treatment: "Fungus-covered eggs" or
"Fungus-removed eggs".
- Fig4g_oviposition trials.csv - Oviposition trials by parasitic wasps for fungus-covered or fungus-supressed eggs.
- oviposition: Yes (1) or no (0).
- abandon: Yes (1) or no (0).
- treatment: "Fungus-covered eggs" or "Fungus-supressed eggs".
- Fig4h_parasitism success rate.csv - Parasitism success rate of fungus-covered or fungus-supressed eggs.
- parasitized eggs: Egg parasitized (1) or not (0).
- hatched eggs: Egg hatched (1) or not (0).
- unhatched eggs: Egg unhatched (1) or not (0).
- treatment: "Fungus-covered eggs" or "Fungus-supressed eggs".
- Fig4i_wasp survival.csv - Survival of T. brevinotaulus experimentally forced to contact with conidia of the hindleg-derived fungi.
- survival: Wasp survival days
- treatment: "Untreated wasps" or "Fungus-treated wasps".
Folder: FigS03
Number of isolates for each OTU in the M. gracinicorne fungal isolate experiment for Fig. S3.
- FigS3abcdef.tsv - Number of isolates for each OTU in the M. gracinicorne fungal isolate experiment.
- First column (OTU): OTU IDs.
- Second column (id): Corresponding IDs of Amplicon-seq OTUs.
- Third to last columns: Sample names (headers). rows contain the number of isolates.
Folder: FigS04
Read distribution assigned to each taxonomic class in the fungal isolate and Amplicon-seq experiment for Fig. S4
- FigS4.tsv - Read distribution assigned to each taxonomic class in the fungal isolate and Amplicon-seq experiment.
- Phylum and Class columns: Taxonomic names of each rank.
- TotalIsolate and %: Number and percentage of isolates in each taxonomic group.
- TotalRead and %: Number and percentage of reads in each taxonomic group.
Folder: FigS05
Multiple alignment of ITS sequences for Fig. S5.
- FigS5.fasta - aligned sequences used for the phylogenetic analysis in Fig S5.
Folder: FigS06
TPM expression levels from the RNAseq experments for Fig. S6.
- FigS6ab.tsv - TPM expression levels from the RNAseq experments.
Folder: FigS07
SEM images and their metadata for Fig. S7.
- Mg_egg_0day20160309.000231.tif – SEM image of egg surface at Day 0 (immediately post-oviposition).
- Mg_egg_0day20160309.000231.txt – Metadata of equipment settings when capturing the above image.
- Mg_egg_0day20160309.000236.tif – Magnified SEM image of egg surface at Day 0.
- Mg_egg_0day20160309.000236.txt – Metadata of equipment settings when capturing the above image.
- Mg_egg_7day20180607.000007.tif – SEM image of egg surface at Day 7 (7 days post-oviposition).
- Mg_egg_7day20180607.000007.txt – Metadata of equipment settings when capturing the above image.
- Mg_egg_7day20180607.000011.tif – Magnified SEM image of egg surface at Day 7.
- Mg_egg_7day20180607.000011.txt – Metadata of equipment settings when capturing the above image.
Folder: FigS08
SEM images and their metadata for Fig. S8.
- Mg_egg_ddw20180122.000001.tif – SEM image of egg surface after extraction with distilled water.
- Mg_egg_ddw20180122.000001.txt – Metadata of equipment settings when capturing the above image.
- Mg_egg_MerUre20180406.000008.tif – SEM image of egg surface after extraction with 8 M urea + 1% 2-mercaptoethanol.
- Mg_egg_MerUre20180406.000008.txt – Metadata of equipment settings when capturing the above image.
- Mg_egg_SamBuf20180420.000002.tif – SEM image of egg surface after extraction with 10% SDS + 1% 2-mercaptoethanol.
- Mg_egg_SamBuf20180420.000002.txt – Metadata of equipment settings when capturing the above image.
Folder: FigS09
Survaival data of M. gracilicorne and P. stali populations after inoculation for Fig S9.
- survival_rate.csv: - Counts of surviving M. gracilicorne and P. stali populations at each days post-inoculation, used for constructing survival curves.
Folder: FigS10
Fungal growth level on eggs for Fig. S10.
- FigS10h_fungal growth on eggs.csv - Fungal growth level on eggs under each treatment.
- level: Fungal growth level; 3 (+++), 2 (++), 1 (+).
- treatment: Female treatments; "No treatment", "Midlegs cut", or "Hindlegs cut".
Folder: FigS11-12
Field experiment data on the effects of fungal cover on parasitization susceptibility for Figs S11 and S12.
- FigS11j_S12abcdef.tsv - Original data from field experiments that examined the effects of fungal cover on parasitization susceptibility.
- Oviposition: Date of oviposition.
- Set: Date of field settlement.
- Retrieve: Date of retrieval of the sample from the field
- Site: Field site IDs
- Sheet: Sheet IDs
- Position: Position IDs on the sheet
- Treat: N indicates intact, while R indicates fungal removal
- Retrieved: Retrieved (1) or lost (0)
- Egg_N: Total number of eggs in a mass
- Predated: Number of predated eggs
- Hatch: Number of hatched eggs
- Embryonic: Number of dead eggs with a developing embryo
- Non-embryonic: Number of dead eggs without a visible embryo
- Ooencyrtus: Number of eggs parasitized by Ooencyrtus wasp
- Trissolcus: Number of eggs parasitized by Trissolcus wasp
- Unknown: Number of eggs parasiteized by wasps that could not be identified due to developmental failure
Folder: FigS13
Morphometric data of diverse dinidorid stinkbugs.
- FigS13ef.tsv - prothorax widths, hind tibia widths and associated metadata.
- Sampler: Name of samplers
- ID1 and ID2: insect identifiers
- Species: Cc = Coridius chinensis, Cp = Cyclopelta parva, Mb = Megymenumbrevicorne, Mg = M. gracilicorne, MgTW = M. gracilicorne (Taiwan), Ep = Eumenotes pacao
- Sex: Sex of individuals
- TW: Prothorax widths (mm)
- HLW: Hindleg tibia widths (mm)
Folder: FigS14
Multiple alignment and partition information for Fig. S14.
- figS14_alignment.phy - PHYLIP-formatted amino acid sequence alignment used for phylogenetic inference. The file was generated by concatenating alignments of all 13 mitochondrial protein-coding genes. Sequence names are 8 or 9 characters, each of which corresponds to the accession number shown in the brackets next to the OTU name of Fig. S14.
- figS14_part.nex - NEXUS-formatted partition file for IQ-TREE analysis. The file shows gene alignment regions, partition assignments, and evolutionary models.
Folder: FigS15
ITS-amplicon data of various Dinidorinid stinkbugs for Fig. S15.
- FigS15abcd.tsv - Number of reads for each OTU in the ITS-Amplicon experiment.
- First column (id): OTU IDs
- Second to 58th columns: Sample names (headers); rows contains number of reads for each OTU ID
- The last column (taxon): taxon assignments
Code/software
- For text files, conventional text editors, spreadsheet softwares.
- For SEM images, conventional imaging softwares.
Access information
Other publicly accessible locations of the data:
- Nucleotide sequence data obtained in this study were deposited in the DNA Data Bank of Japan (DDBJ) with the following accession numbers: fungal ITS sequence data, LC792825 –LC793396; Fungal ITS amplicon sequencing data, DRX509842 – DRX509916 and DRX637795-DRX637852; RNA sequencing data, DRX509810 – DRX509841; insect mitochondrial genome data, LC670767 and PV423521-PV423524.