Data for: "Convergence in carnivorous pitcher plants reveals mechanism for composite trait evolution" by Guillaume Chomicki, Gustavo Burin, Lucas Busta, Jedrzej Gozdzik, Reinhard Jetter, Beth Mortimer, Ulrike Bauer

Abstract: The evolution of complex, composite traits remains a puzzle. Composite traits involve multiple components that, only when combined, gain a new synergistic function. Using field experiments, microscopy, chemical analyses, and laser Doppler vibrometry, we show that two carnivorous Nepenthes pitcher plants independently evolved identical adaptations in three distinct traits to acquire a new, complex trapping mechanism. Comparative phylogenetic analysis revealed that this composite trait arose convergently via spontaneous coincidence of independently evolving components with high pre-existing phenotypic variation, rather than gradual directional selection in one or more component traits. We show that high pre-existing phenotypic variability can generate evolutionary novelty by increasing the chance of a spontaneous beneficial combination of previously non-functional traits.

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Summary description: Data were collected from 55 Nepenthes pitcher plant species from Botanical collections in the UK, Germany and Switzerland, as well as from two species (N. gracilis and N. pervillei) from natural populations in Borneo and the Seychelles. Fotos and high-speed videos were used to assess two macromorphological traits: (1) lid orientation (deviation from horizontal in °), and (2) lid loading/impact response (qualitatively from Fotos, and quantitatively from high-speed video recordings). Laser Doppler vibrometry was used for detailed analysis of lid oscillations after a drop impact on the lid. Scanning electron microscopy was used to assess surface microtopography (images are shown in the article supplementary material, Figures S1 and S2, and are not included in this dataset). Gas-chromatography mass-spectrometry was used to investigate the chemical composition of surface waxes. For detailed descriptions of the experimental methods and sampling techniques see online supplementary material, Materials and Methods.

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Files (alphabetical):

Comparative_dataset_55species_summary.csv
Context: Testing the independent origin of springboard trapping; Testing for correlated evolution of the component traits of springboard trapping; Figs.3, S3-S7.
Summary of the semi-quantitative trait data used to probe the evolutionary independence of springboard trapping in its three component traits in N. gracilis and N. pervillei.
Data table.
Rows: (1) header; (2-56) data.
Columns: (A) species names; (B-G) qualitative load responses determined from manual loading of the lid tip (see online supplementary material, Materials and Methods, section Assessment of lid position and loading response); (B) number of individual pitchers showing a lid pivoting response; (C) number of individual pitchers showing a pivot-like response; (D) number of individual pitchers showing a lid bending response; (E) number of individual pitchers showing a lid buckling response; (F) number of individual pitchers showing a mixed bending-buckling response; (G) number of individual pitchers showing a complex mixture of response types, e.g. bending-buckling-twisting or pivoting-buckling; (H) sample sizes for each species; (I-K) three-state coding of lid loading responses for the ancestral state reconstructions and tests of independent evolution (see online supplementary material, Materials and Methods, section Testing the independent origins of springboard trapping); (I) equals (B); (J) equals (C); (K) equals SUM(D:G); (L-Q) proportion of each loading response category in percent; (L) proportion of pivoting lids in percent; (M) proportion of pivot-like lids in percent; (N) proportion of bending lids in percent; (O) proportion of buckling lids in percent; (P) proportion of bending-buckling lids in percent; (Q) proportion of complex response lids in percent; (R) predominant response (shown by >50% of all investigated pitchers per species); if no single category reached 50% prevalence, the predominant response is ‘variable’; (S) predominant response using three-state coding: pivot = >50% pivoting, pivot-like = >50% pivoting OR pivot-like, non-pivot = pivoting + pivot-like < 50%; (T) mean lid angle (°) = deviation from horizontal (see online supplementary material, Materials and Methods, section Assessment of lid position and loading response) for each species; (U) three-state coding of lid orientation for the ancestral state reconstructions and tests of independent evolution (see online supplementary material, Materials and Methods, section Testing the independent origins of springboard trapping): horizontal = ± 10° deviation from horizontal, near-horizontal = ± 20° deviation from horizontal, non-horizontal = > ± 20° deviation from horizontal; (V) Three-state coding of wax crystal presence: wax = pillar-shaped wax crystals on the lower lid surface, weak wax = sparse or patchy wax crystals of variable shape, no wax = smooth lower lid surface, devoid of crystals; (W) Plant source (botanical collection), multiple sources are separated by commas.

Custom_matlab_script_for_laser_Doppler_vibrometry.txt
Context: Laser Doppler vibrometry of impact-induced lid oscillations; Fig.1, F and L.
Matlab code used to analyze the laser Doppler vibrometry data (included as *.csv files)

Custom_matlab_script_for_tracking_lid_movement.txt
Context: High-speed videography of the impact response of the pitcher lid; Fig.1, E and K.
Matlab code used to track the lid movement in highspeed video files (included as *.avi files)

Highspeed_video_analysis_mass_area_frequency.csv
Context: High-speed videography of the impact response of the pitcher lid; Fig.1, E and K.
Masses, areas and resonant frequencies of N. gracilis and N. pervilei lids tracked in highspeed videos (included as *.avi files). Area determined from included *tif files.
Data table.
Rows: (1) header; (2-17) data.
Columns: (A) species; (B) pitcher identifier; (C) oscillation frequency of a tracked point near the lid attachment point (lid base), in s-1; (D) oscillation frequency of a tracked point halfway between lid attachment and distal lid tip (midpoint), in s-1; (E) oscillation frequency of a tracked point near the distal lid tip (tip), in s-1; (F) fresh lid mass in mg; (G) lid area in mm2; (H) mean oscillation frequency in (averaged over the frequencies in rows B-D), in s-1; see also online supplementary material, Materials and Methods, section High-speed videography of the impact response of the pitcher lid.

Laser_Doppler_vibrometry_data_N_gracilis.csv
Context: Laser Doppler vibrometry of impact-induced lid oscillations; Fig.1, F and L.
Raw data for 3 N. gracilis lids.
Data table.
Rows: (1) header; (2-4800) data.
Columns: (A,C,E) time steps normalized for 5 oscillation cycles (dimensionless); (B,D,F) velocity normalized for lid size (dimensionless); (A,B) data for pitcher 3; (C,D) data for pitcher 2; (E,F) data for pitcher 1; see also online supplementary material, Materials and Methods, section Laser Doppler vibrometry of impact-induced lid oscillations.

Laser_Doppler_vibrometry_data_N_pervillei.csv
Context: Laser Doppler vibrometry of impact-induced lid oscillations; Fig.1, F and L.
Raw data for 4 N. pervillei lids.
Data table.
Rows: (1) header; (2-4800) data.
Columns: (A,C,E,G) time steps normalized for 5 oscillation cycles (dimensionless); (B,D,F,H) velocity normalized for lid size (dimensionless); (A,B) data for pitcher 7; (C,D) data for pitcher 5; (E,F) data for pitcher 6; (G,H) data for pitcher 4; see also online supplementary material, Materials and Methods, section Laser Doppler vibrometry of impact-induced lid oscillations.

Laser_Doppler_vibrometry_mass_area_frequency.csv
Context: Laser Doppler vibrometry of impact-induced lid oscillations; Fig.1, F and L.
Masses, areas and resonant frequencies of N. gracilis and N. pervilei lids investigated using laser Doppler vibrometry.
Data table.
Rows: (1) header; (2-8) data.
Columns: (A) species; (B) pitcher identifier; (C) fresh lid mass (mg); (D) lid area (mm²); (E) lid oscillation frequency measured with a laser Doppler vibrometer, in s-1; see also online supplementary material, Materials and Methods, section Laser Doppler vibrometry of impact-induced lid oscillations.

Lid_angles_raw_data.csv
Context: Testing the independent origin of springboard trapping; Testing for correlated evolution of the component traits of springboard trapping; Quantification of variation in lid angles and lid loading behavior in 42 Nepenthes species; Figs.3, 5, S3-S8.
Data table.
Rows: (1) header; (2-74) data: angle measurements in (°).
Columns (A-CK): Nepenthes species.

Lid_loading_response_raw_data.csv
Context: Testing the independent origin of springboard trapping; Testing for correlated evolution of the component traits of springboard trapping; Quantification of variation in lid angles and lid loading behavior in 42 Nepenthes species; Figs.3-5, S3-S9.
Data table.
Rows: (1) header; (2-77) data: qualitative load responses determined from manual loading of the lid tip (see online supplementary material, Materials and Methods, section Assessment of lid position and loading response).
Columns (A-CK): Nepenthes species

Lid_wax_raw_data.csv
Context: Testing the independent origin of springboard trapping; Testing for correlated evolution of the component traits of springboard trapping; Figs.3, 5, S1-S7.
Data table.
Rows: (1-86): data.
Columns: (A) Nepenthes species names; (B) description of wax crystal presence and morphology as determined from scanning electron micrographs (one image per species; see also online supplementary material, Materials and Methods, section Scanning electron microscopy of lower lid surfaces)

N_gracilis_lid_area_P02.tif
N_gracilis_lid_area_P05.tif
N_gracilis_lid_area_P06.tif
N_gracilis_lid_area_P07.tif
N_gracilis_lid_area_P08.tif
N_gracilis_lid_area_P09.tif
N_gracilis_lid_area_P10.tif
N_gracilis_lid_area_P11.tif
N_gracilis_lid_area_P12.tif
N_gracilis_lid_area_P13.tif
N_gracilis_lid_area_P14.tif
Context: High-speed videography of the impact response of the pitcher lid; Fig.1E.
Photographs of abscised N. gracilis lids on graph paper. Numbers in the file name identify individual pitchers.
Image files.

N_gracilis_P02-1.avi
N_gracilis_P02-2.avi
N_gracilis_P02-3.avi
N_gracilis_P05-1.avi
N_gracilis_P05-2.avi
N_gracilis_P05-3.avi
N_gracilis_P06-1.avi
N_gracilis_P06-2.avi
N_gracilis_P06-3.avi
N_gracilis_P07-1.avi
N_gracilis_P07-2.avi
N_gracilis_P07-3.avi
N_gracilis_P08-1.avi
N_gracilis_P08-2.avi
N_gracilis_P08-3.avi
N_gracilis_P09-1.avi
N_gracilis_P09-2.avi
N_gracilis_P09-3.avi
N_gracilis_P10-1.avi
N_gracilis_P10-2.avi
N_gracilis_P10-3.avi
N_gracilis_P11-1.avi
N_gracilis_P11-2.avi
N_gracilis_P11-3.avi
N_gracilis_P12-1.avi
N_gracilis_P12-2.avi
N_gracilis_P12-3.avi
N_gracilis_P13-1.avi
N_gracilis_P13-2.avi
N_gracilis_P13-3.avi
N_gracilis_P14-1.avi
N_gracilis_P14-2.avi
N_gracilis_P14-3.avi
Context: High-speed videography of the impact response of the pitcher lid; Fig.1E.
Highspeed video recordings of simulated rain drop impacts on N. gracilis lids. Numbers P02 to P14 identify individual pitchers. Numbers -1 to -3 identify repeated drop impacts on the same lid.
Video files.

N_pervillei_lid_area_P06.TIF
N_pervillei_lid_area_P07.TIF
N_pervillei_lid_area_P08.TIF
N_pervillei_lid_area_P09.TIF
N_pervillei_lid_area_P10.TIF
Context: High-speed videography of the impact response of the pitcher lid; Fig.1K.
Photographs of abscised N. pervillei lids on graph paper. Numbers in the file name identify individual pitchers.
Image files.

N_pervillei_P06-1.avi
N_pervillei_P06-2.avi
N_pervillei_P06-3.avi
N_pervillei_P06-4.avi
N_pervillei_P06-5.avi
N_pervillei_P07-1.avi
N_pervillei_P07-2.avi
N_pervillei_P07-3.avi
N_pervillei_P07-4.avi
N_pervillei_P07-5.avi
N_pervillei_P08-1.avi
N_pervillei_P08-2.avi
N_pervillei_P08-3.avi
N_pervillei_P08-4.avi
N_pervillei_P08-5.avi
N_pervillei_P09-1.avi
N_pervillei_P09-2.avi
N_pervillei_P09-3.avi
N_pervillei_P09-4.avi
N_pervillei_P09-5.avi
N_pervillei_P10-1.avi
N_pervillei_P10-2.avi
N_pervillei_P10-3.avi
N_pervillei_P10-4.avi
N_pervillei_P10-5.avi
Context: High-speed videography of the impact response of the pitcher lid; Fig.1K.
Highspeed video recordings of simulated rain drop impacts on N. pervillei lids. Numbers P06 to P10 identify individual pitchers. Numbers -1 to -5 identify repeated drop impacts on the same lid.
Video files.

Nepenthes_wax_GC-MS_data.csv
Context: Chemical analysis of surface waxes; Fig.2.
Data table.
Rows: (1) header; (2-197) data.
Columns: (A) species name; (B) pitcher surface; (C) carbon-chain length of the identified wax compound; (D) compound class; (E) compound identifier; (F) compound abundance in µg cm^2, replicate sample 1; (G) compound abundance in µg cm^2, replicate sample 2; (H) compound abundance in µg cm^2, replicate sample 3; (I) compound abundance in µg cm^2, replicate sample 4.

Trait_variability_dataset_42species_summary.csv
Context: Quantification of variation in lid angles and lid loading behavior in 42 Nepenthes species; Figs. 3-5, S8-9.
Data table.
Rows: (1) header; (3-44) data.
Columns: (A) Nepenthes species; (B-H) lid angle measurements (see online supplementary material, Materials and Methods, section Assessment of lid position and loading response); (B) sample size; (C) mean lid angle (°); (D) standard error of the mean (±°); (E) median lid angle (°); (F) minimum lid angle (°); (G) maximum lid angle (°); (H) normal distribution of data (determined from Shapiro-Wilks tests if n ≤ 10 and Kolmogoroff-Smirnov tests if n > 10), Y = yes, N = no; (I-AC) qualitative load responses determined from manual loading of the lid tip (see online supplementary material, Materials and Methods, section Assessment of lid position and loading response); (I) number of individual pitchers showing a lid pivoting response; (J) number of individual pitchers showing a pivot-like response; (K) number of individual pitchers showing a lid bending response; (L) number of individual pitchers showing a lid buckling response; (M) number of individual pitchers showing a mixed bending-buckling response; (N) number of individual pitchers showing a complex mixture of response types, e.g. bending-buckling-twisting or pivoting-buckling; (O) sample size; (P) average number of pitchers per lid loading category (= expected number per category under equal distribution); (Q) variance of data distribution across categories; (R) index of dispersion (variance-to-mean ratio); this is 1 for a perfectly random distribution, <1 for an equal distribution, and >1 for an aggregated (clumped) distribution; (S) chi-square statistics for the index of dispersion; (T) degrees of freedom for the chi-square statistics; (U) data distribution determined from chi-square statistics with an error probability of <5%; (V) data distribution determined from chi-square statistics with an error probability of <1%; (W) proportion of pivoting lids in percent; (X) proportion of pivot-like lids in percent; (Y) proportion of bending lids in percent; (Z) proportion of buckling lids in percent; (AA) proportion of bending-buckling lids in percent; (AB) proportion of complex response lids in percent; (AC) predominant response (shown by >50% of all investigated pitchers per species); if no single category reached 50% prevalence, the predominant response is ‘variable’

The code used for the corHMM analysis can be found at https://github.com/gburin/nepenthes-springboard.