Original article accessible at: https://doi.org/10.1111/syen.70016

Description of the data and file structure

Sample collection

Colonies of Termes aff. fatalis were collected in French Guiana and Trinidad. Specimens were preserved in 100% ethanol for genetic analyses, in 80% ethanol for morphological studies, and extracted in n-hexane for chemical analyses.

Additional reference material from the University of Florida collection was used for genetic analyses of T. hispaniolae and T. panamaensis.

Type material from the American Museum of Natural History and the Smithsonian National Museum of Natural History was examined for morphological comparisons with T. fatalis, T. medioculatus, and T. panamaensis.

DNA sequencing

DNA was extracted from the head of one soldier per colony collected in French Guiana and Trinidad (n = 236). The mitochondrial cytochrome c oxidase subunit II (COII) gene was sequenced using Sanger technology.

Illumina sequencing was also performed to obtain the complete mitochondrial genome of at least one individual from each T. aff. fatalis lineage from French Guiana, as well as from the reference colonies of the University of Florida.

Phylogenetic analyses

Phylogenetic relationships were first inferred from all COII sequences, including those obtained from Illumina-sequenced mitogenomes and additional Termitinae sequences (for rooting and phylogenetic context). Sequences were aligned using MUSCLE in CodonCodeAligner, and the phylogenetic tree was reconstructed with MrBayes.

A second phylogeny was generated using complete mitogenomes, including newly sequenced data and previously published mitogenomes of Termes, other Termitinae, and representative termite taxa. The concatenated alignment was partitioned into five categories: rRNA, tRNA, and the first, second, and third codon positions of protein-coding genes. Analyses were conducted with and without the third codon positions using BEAST.

Chemical analyses

For 22 colonies, 4–50 workers and soldiers were extracted in hexane and analyzed by gas chromatography. This allowed recovery of cuticular hydrocarbons from workers and defensive secretions from soldiers.

Morphological analyses

A total of 172 soldiers from 39 colonies, 55 workers from 27 colonies, and 68 reproductives from 9 colonies were measured using a micrometer mounted on a stereomicroscope. Workers’ mandibles were dissected and measured to calculate the mandibular index following Emerson (1960). A linear discriminant analysis (LDA) was performed on the measurements of soldiers and workers.

Files and variables

File: Aligned_COII_sequences.nex

Description: This Nexus file contains the alignment of (1) newly generated COII sequences, (2) COII sequences extracted from newly assembled mitogenomes, and (3) COII sequences retrieved from GenBank. Sequences were aligned in CodonCodeAligner using the MUSCLE algorithm.

File: Aligned_mitogenomes_with_3rd_codon.nexus

Description: This Nexus file contains the whole mitogenome alignment, partitioned into rRNA, tRNA, and the first, second, and third codon positions.

File: BEAST_configuration_without_3rd_codon.xml

Description: This Nexus file contains the whole mitogenome alignment, partitioned into rRNA, tRNA, and the first and second codon positions (excluding third positions)

File: BEAST_configuration_with_3rd_codon.xml

Description: This XML file was generated with BEAUti and contains the parameter configuration for the BEAST analysis including third codon positions.

File: Aligned_mitogenomes_without_3rd_codon.nexus

Description: This XML file was generated with BEAUti and contains the parameter configuration for the BEAST analysis excluding third codon positions.

File: BEAST_consensus_tree_with_3rd_codon.trees

Description: This file contains the consensus tree from the BEAST analysis including third codon positions. The tree was summarized in TreeAnnotator as a maximum clade credibility tree.

File: BEAST_consensus_tree_without_3rd_codon.trees

Description: This file contains the consensus tree from the BEAST analysis excluding third codon positions. The tree was summarized in TreeAnnotator as a maximum clade credibility tree.

File: MrBayes_consensus_tree.tre

Description: This file contains the consensus tree generated with MrBayes using the sumt command.

File: COII_sequences_(fasta).zip

Description: This compressed archive contains the new cytochrome c oxidase subunit II (COII) sequences generated in this study, provided in FASTA format. Each sequence is labeled with its colony code, with corresponding details available in Samples_information.xlsx. Sequences were obtained from DNA extractions of soldier heads and sequenced using Sanger technology. COII sequences extracted from Illumina-sequenced mitogenomes are not included in this folder.

File: BEAST_raw_trees_with_3rd_codon.trees

Description: This file contains the raw BEAST output (10,000 trees) including third codon positions.

File: BEAST_raw_trees_without_3rd_codon.trees.trees

Description: This file contains the raw BEAST output (10,000 trees) excluding third codon positions.

File: Raw_measurements.xlsx

Description: This Excel file contains raw morphometric measurements of termites collected in French Guiana and of type specimens. Data are organized in three sheets, one for each caste (soldiers, workers, reproductives). Measurements are given in millimetres, except for the mandibular index (IM), which is unitless. Missing values are indicated with “NA”.

Variables:

• Colony: colony code of the measured specimen
• Species_name: scientific name of identified type specimens (T. fatalis, T. medioculatus, T. panamaensis), and temporary name given to new specimens (T. aff.* fatalis* 1–3).
• Sex: sex of the individual identified through the disposition of sternites. An additionnal (Q) or (K) indicate that the specimen is a queen or a king.
• Head lenght to the tip of the frontal process (HLT): measured according to Miller 1991
• Head lenght to the anterior part of the genae (HLG): measured according to Miller 1991
• Head depth (HD): measured according to Miller 1991
• Head width (HW): measured according to Roonwal (1969), measure number 17
• Pronotum width (PW): measured according to Roonwal (1969), measure number 68
• Mandible lenght (ML): measured according to Miller 1991
• Minimal width of the gula (Gwmin): measured according to Miller 1991
• Maximal width of the gula (Gwmax): measured according to Miller 1991
• Tibia lenght (TL): measured according to Roonwal (1969), measure number 85
• Mandibular index (IM): measured according to Emerson (1960)
• Head lenght (HL): measured according to Roonwal (1969), measure number 8
• Pronotum lenght (PL): measured according to Roonwal (1969), measure number 66
• Inter-eyes distance (EyesDist): measured according to Roonwal (1969), measure number 52
• Diameter of the eye (EyeDiam): measured according to Roonwal (1969), measure number 48
• Diameter of the ocellus (OcDiam): measured according to Roonwal (1969), measure number 55
• Eye-ocellus distance (EyeOcDist): measured according to Roonwal (1969), measure number 57

Cited references:

Emerson, A.E. (1960) New genera of termites related to Subulitermes from the Oriental, Malagasy, and Australian regions (Isoptera, Termitidae, Nasutitermitinae). American Museum Novitates, 1986, 1–28.

Miller, L.R. (1991) A revision of the Termes-Capritermes branch of the Termitinae in Australia (Isoptera: Termitidae). Invertebrate Systematics, 4 (6), 1147–282. https://doi.org/10.1071/IT9901147

Roonwal, M.L. (1969) Measurements of termites for taxonomic purposes. Journal of the Zoological Society of India, 21, 9–66

File: Chemical_identities.xlsx

Description: This Excel file contains raw results from chemical extractions of termite cuticular hydrocarbons (sheet “workers”) and defensive products (sheet “soldiers”). Results are expressed as relative abundances (%), and are colored using conditional formatting along a green (low values) to red (high values) gradient. Colonies G18-10c, G18-21, G18-22, G18-27, G18-28, G18-33, G18-34, G18-36A, G18-44, G18-45 are T. fatalis, G18-02, G18-03, G18-06, G18-16, G18-24, G18-37, G18-42 are T. medioculatus and G18-01, G18-11, G18-13, G18-19, G18-29 are T. incognitus.

Variables:

• Peak label: letter identifying the chromatographic peak of each compound
• Retention index: retention index of the compound
• Identity: chemical name of the compound identified from mass spectral analysis and retention index, confirmed by comparison with the NIST 20 library.
• Chemical class: category of the identified compound
• Retention time (min): elution time in minutes of the compound

File: LDA_soldiers.Rmd

Description: R Markdown file containing the code used for the LDA analysis of the soldiers dataset. Results are presented in the file "LDA_soldiers_markdown.pdf".

File: LDA_soldiers_markdown.pdf

Description: This PDF was produced using the R Markdown file "LDA_soldiers.Rmd". It integrates the code used for the LDA analysis of the soldiers dataset with contextual text for explanation and organization, and displays the corresponding outputs alongside the code

File: LDA_workers.Rmd

Description: R Markdown file containing the code used for the LDA analysis of the workers dataset. Results are presented in the file "LDA_workers_markdown.pdf".

File: LDA_workers_markdown.pdf

Description: This PDF was produced using the R Markdown file "LDA_workers.Rmd". It integrates the code used for the LDA analysis of the worers dataset with contextual text for explanation and organization, and displays the corresponding outputs alongside the code.

File: Samples_information.xlsx

Description: This Excel file contains detailed data on all samples used in the study. Missing values are indicated by “NA”.

Variables:

• Colony code: unique identifier assigned to each colony by the collector or the institution of deposit
• Country: country where the specimens were collected
• Locality: region and eventual exact locality in which the specimens were collected
• GPS (North) / GPS (East): geographic coordinates (in decimal degrees) of the collection site
• Date of collect: date when the specimens were collected, formatted as “yyyy-mm-dd”
• Collector(s): names of the person(s) who collected the specimens
• Collection: institution where the specimens are deposited
• Species name: final scientific name assigned to the specimens
• Temporary name: provisional name used in the article or supplementary material (e.g., T. aff. fatalis 1)
• Type status: type status of the specimens, if applicable
• Presence of alates: an “x” indicates the presence of alates in the nest at the time of collection
• Soldiers measured: an “x” indicates that soldiers from this colony were used for morphological analyses
• Workers measured and/or dissected: an “x” indicates that workers from this colony were used for morphological analyses
• Alates measured: an “x” indicates that reproductives (alates and eventually the queen/king) from this colony were used for morphological analyses
• Mitogenome accession: GenBank accession numbers of mitogenome sequences
• COII accession: GenBank accession numbers of COII gene sequences
• COII haplotype: “Identical to …” indicates correspondence between the colony (if barcoded) and another haplotype whose sequence is either deposited in GenBank or listed in the previous column. All sequences are available in the file COII_sequences_(fasta).zip