Data from: Separate and synergistic anti-herbivore effects of non-glandular trichomes and leaf chemistry in a desert plant
Data files
Dec 16, 2025 version files 130.15 KB
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canopus_compound_summary.tsv
28.20 KB
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choice_assay_caterpillar_data.csv
1.86 KB
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choice_assay_herbivory_data.csv
4.16 KB
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compound_feature_list.csv
1.48 KB
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factorial_diet_assay_data.csv
40.30 KB
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mandible_damage_measurements.csv
3.59 KB
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mzMine_steps_parameters.txt
1.54 KB
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no_choice_leaf_assay_data.csv
3.88 KB
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README.md
9.88 KB
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SIRIUS_steps_parameters.txt
1.17 KB
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Synergistic_Effects_of_Trichomes_and_Chemistry_stats.R
34.08 KB
Abstract
Plant defense phenotypes commonly integrate physical and chemical traits that may act synergistically against herbivores, but empirical evidence for synergy as a defense strategy remains limited. We experimentally tested the separate and combined effects of two common plant defenses, non-glandular trichomes and leaf chemistry, on the feeding, behavior, and performance of a generalist herbivore. We conducted choice and no-choice assays that exposed beet armyworm caterpillars (Spodoptera exigua, Noctuidae) to desert stingbush (Eucnide urens, Loasaceae) leaves with manipulated trichome cover and raised caterpillars on a diet containing factorial combinations of E. urens trichomes and leaf extracts. We assessed differences in caterpillar behavior, performance, and survival across treatments. We found a significant interactive effect of trichomes and leaf chemistry that markedly decreased caterpillar growth, providing evidence for a defense synergism. Both defenses also reduced caterpillar performance individually. Leaf extracts contained a high proportion of putatively defensive compounds, including iridoid glycosides, which were associated with reduced caterpillar growth and high mortality before pupation. Barbed trichomes impeded caterpillar movement and, when consumed, caused mandible damage and reduced eclosion rates compared to controls. Our results indicate that two commonly co-expressed plant defenses can synergistically reduce herbivore performance. These findings provide a rare example of trait synergy as a defense strategy and represent a step forward in the effort to study plant defense holistically, as suites or syndromes of defenses, rather than isolated traits.
Description of the data and file structure
These are the data and code to accompany the submitted manuscript entitled “Separate and Synergistic Anti-Herbivore Effects of Non-Glandular Trichomes and Leaf Chemistry in a Desert Plant”. Using a series of experiments with the desert stingbush (Eucnide urens: Loasaceae), we experimentally tested for synergisms between trichomes and chemistry on the feeding, behavior, and performance of a generalist herbivore. We conducted choice and no-choice assays that exposed beet armyworm caterpillars (Spodoptera exigua Hübner, Noctuidae) to E.urens leaves with manipulated trichome cover and raised caterpillars on a diet containing factorial combinations of trichomes and leaf metabolite extracts.
Files and variables
File: choice_assay_caterpillar_data.csv
Description: Data from an assay where S. exigua caterpillars were given a choice between E. urens leaf discs with different levels of trichome cover (shaved vs. unshaved).
Variables
- dish_ID: Replicate ID
- starved_mass_g: Mass, in grams, of caterpillars at the start of the assay. They were starved for 24 hours prior.
- loc_after_2hrs: Location of each caterpillar two hours into the
assay (S = shaved leaf disc, U = unshaved leaf disc, NA = not on
leaf). - loc_after_18hrs: Location of each caterpillar 18 hours into the
assay (S = shaved leaf disc, U = unshaved leaf disc, NA = not on
leaf). - trapped_after_18_hrs: Whether or not the caterpillar was trapped by trichomes on the leaf disc after 18 hours (Y = trapped, N = not trapped)
- notes: notes about the location of trapped caterpillars
File: choice_assay_herbivory_data.csv
Description: Leaf herbivory data from an assay where S. exigua
caterpillars were given a choice between E. urens leaf discs with different levels of trichome cover (shaved vs. unshaved).
Variables
- dish_ID: Replicate ID
- leaf_disc_treatment: Whether the herbivory data refer to the shaved or unshaved leaf disc in each replicate.
- proportion_consumed: Proportion of the leaf disc eaten by the caterpillars, estimated using the R package herbvar.
- leaf_area_consumed_mm2: Area of the leaf disc, in square millimeters, eaten by the caterpillar, estimated using the R package herbvar.
- total_leaf_area_mm2: Total area of the leaf disc, in square millimeters, estimated using the R package herbvar.
File: factorial_diet_assay_data.csv
Description: Data from an assay in which neonate S. exigua
caterpillars were divided into four groups (n=35 per group) and raised to eclosion on one of four artificial diets: diet supplemented with trichomes isolated from Eucnide urens leaves, diet supplemented with
E. urens leaf extract, a diet containing both trichomes and leaf extract, or a control diet with no trichomes or leaf extract added.
Variables
- individual: individual caterpillar ID
- treatment: whether the caterpillar was fed a control diet, diet with added trichomes, diet with added leaf extract, or diet with both trichomes and leaf extract.
- has_trichomes: binary variable indicating whether the caterpillar received a diet that contained trichomes (1 = yes, 0 = no)
- has_metabolites: binary variable indicating whether the caterpillar received a diet that contained leaf metabolite extract (1 = yes, 0 =
no) - tray_number: Tray in which the cup containing the caterpillar was placed during the assay. Cups were alternated by treatment within each tray, and trays were kept in a single layer within a growth chamber.
- cat_mass_MONTH_DAY: Mass, in grams, of the caterpillar at a given time point. Caterpillars were weighed every 48 hours. NA = individual was not weighed (typically because it had died).
- weighed_by_MONTH_DAY: letter (A or B) identifying which researcher performed the weighing. NA = individual was not weighed (typically because it had died).
- alive_MONTH_DAY: Whether or not the caterpillar was alive that day. Once a caterpillar died, we stopped weighing it.
- life_stage_MONTH_DAY: Life stage of the insect, recorded once they started pupating. NA = individual died or pupated.
- Sex: Pupal sex, NA for individuals that did not survive to
pupation. - alive_after_30_days: whether an individual was alive after 30
days. - Pupated: Whether an individual successfully pupated.
- pupal_mass: The mass, in grams, of each pupa.
- emerged: Whether a living adult moth emerged from the pupa (1 = yes,
0 = no) - notes: Various notes on caterpillar behavior, mortality, and appearance.
File: mandible_damage_measurements.csv
Description: Damage to exuvial mandibles of S. exigua caterpillars fed control and trichome-augmented diet in the trichomes and metabolites assay. Damage was quantified by measuring (using the objectJ plugin in ImageJ) the length of the scars left by broken mandible teeth.
Variables
- individual: caterpillar ID
- side: Whether measurements were taken from the left or right mandible.
- chip_length_um: The length, in um, of the scar left by a chipped mandible tooth.
File: no_choice_leaf_assay_data.csv
Description: Data from an assay in which S. exigua larvae were raised on either shaved or unshaved E. urens leaves for 20 hours.
Variables
- dish_ID: Replicate ID
- treatment: Whether the caterpillar was given a shaved or unshaved leaf.
- cat_starting_mass_g: The starting mass, in grams, of each caterpillar.
- cat_mass_g_20hrs: The mass, in grams, of each caterpillar at the conclusion of the assay.
- cat_status_20hrs: Description of whether the caterpillar was alive and if/where it was trapped at the conclusion of the assay.
- Trapped: Binary variable indicating whether the caterpillar was trapped (Y) or not trapped (N) on the leaf at the conclusion of the assay.
- proportion_consumed: Proportion of the leaf consumed by the end of the assay, estimated using the R package herbvar.
- total_leaf_area_mm2: Total leaf area, in mm^2, estimated using
the R package herbvar. - leaf_area_consumed_mm2: Area, in mm^2, of the leaf consumed by the end of the assay, estimated using the R package herbvar.
File: mzMine_steps_parameters.txt
Description: Parameters used to group spectral features of E. urens extracts into compounds using mzMine
File: SIRIUS_steps_parameters.txt
Description: Parameters used for compound structure prediction using SIRIUS
File: compound_feature_list.csv
Description: Features of compound peaks identified from E. urens leaf extracts
Variables
- rowID: compound number (unique identifier)
- row m/z: mass to charge ratio of compound peak (positive ionization mode)
- row retention time: elution time of compound peak in minutes
- E120.mzXML Peak height: total ion current for compound in the pooled sample
File: canopus_compound_summary.tsv
Description: Summary of structure predictions and annotations from SIRIUS and CANOPUS
Variables
- id: numerical order of compounds in the input file, followed by the input file name and the compound number
- molecularFormula: molecular formula for MS peak predicted by SIRIUS
- adduct: most likely adduct for predicted molecular formula (positive ionization)
- precursorFormula: molecular formula of parent or precursor mass for MS2 data predicted by SIRIUS
- NPC#pathway: natural products pathway predicted by CANOPUS
- NPC#pathway Probability: probability/confidence of CANOPUS natural products pathway prediction
- NPC#superclass: natural products superclass predicted by CANOPUS
- NPC#superclassProbability: probability/confidence of CANOPUS natural products superclass prediction
- NPC#class: natural products class predicted by CANOPUS
- NPC#class Probability: = probability/confidence of CANOPUS natural products class prediction
- ClassyFire#most specific class: Classyfire most specific chemical class predicted by CANOPUS
- ClassyFire#most specific class Probability: probability/confidence of CANOPUS Classyfire most specific chemical class prediction
- ClassyFire#subclass: Classyfire chemical subclass predicted by CANOPUS
- ClassyFire#subclass Probability: probability/confidence of CANOPUS Classyfire chemical subclass prediction
- ClassyFire#class: Classyfire chemical class predicted by CANOPUS
- ClassyFire#class Probability: probability/confidence of CANOPUS Classyfire chemical class prediction
- ClassyFire#superclass: Classyfire chemical superclass predicted by CANOPUS
- ClassyFire#superclass Probability: probability/confidence of CANOPUS Classyfire chemical superclass prediction
- ClassyFire#all classifications: all Classyfire chemical classifications separated by ;
- featureId: compound number (unique identifier)
Code/software
File: Synergistic_Effects_of_Trichomes_and_Chemistry_stats.R
Description: R script used for statistical analysis and to generate manuscript figures.
Statistical analyses
(“Synergistic_Effects_of_Trichomes_and_Chemistry_stats.R”) were
performed in RStudio using R version 4.4.2 and the following packages:
tidyr - data wrangling
plyr - data wrangling
dplyr - data wrangling
ggplot2 - general plotting
hrbrthemes - ggplot themes
PairedData - to plot paired data in the leaf choice assay
broom - summaries of statistical tests
plotrix - calculating standard error
car - QQplot while testing ANOVA assumptions
survival - Kaplan-Meier survival analysis
survminer - pairwise log-rank tests comparing Kaplan-Meier curves
Effects - checking LMM model assumptions
sjPlot - checking LMM model assumptions
stringr - manipulating character strings during compound annotation
