DOI: 10.5061/dryad.8w9ghx3vp
title: Anthocyanin impacts multiple plant-insect interactions in a carnivorous plant

description: Although there are many hypothesized ecological functions of plant
 coloration, they have only partly been resolved by examining ecological
 hypotheses in isolation. Multiple ecological interactions may act in
 concert or in opposition to fix or maintain variation in plant coloration,
 i.e. via ecological pleiotropy. To investigate the adaptive value of red
 plant pigment (anthocyanin) in a carnivorous plant, we compared insect
 prey capture, herbivore damage, and recruitment of specialist insect
 larvae in naturally-occurring, sympatric red and green color morphs of the
 pitcher plant Sarracenia purpurea. We integrated field and laboratory
 bioassays, visual modeling, chemical analysis of anthocyanins, and a
 long-term demographic study to investigate multiple ways anthocyanins
 mediate plant-insect interactions. In support of ecological pleiotropy,
 each morph performed better in one or more ecological context, providing
 evidence for ecological interactions exerting opposing selection on plant
 color and thus maintaining variation. The mixture of both ecological
 benefits and costs to anthocyanin production is further supported by
 stable color polymorphism and seed set data consistent with balancing
 selection. More broadly, this work reveals the impacts of a single
 anthocyanin compound on multiple key plant-insect interactions,
 demonstrating evidence for ecological pleiotropy maintaining intraspecific
 diversity in plant color.
description: # Anthocyanin impacts multiple plant-insect interactions in a carnivorous
 plant
 [https://doi.org/10.5061/dryad.8w9ghx3vp](https://doi.org/10.5061/dryad.8w9ghx3vp) Data for "Anthocyanin impacts multiple plant-insect interactions in a carnivorous plant." Describes data on arthropod prey capture, herbivory, specialist insect larvae (inquiline) recruitment, seed set, and germination of anthocyanin-producing (red) and anthocyanin-free (green) morphs of the pitcher plant *Sarracenia purpurea*. Also includes data on LCMS characterization of these anthocyanin compounds, UV-vis spectrometer measurements from each color morph, data on change in color morph ratios over time, and a longitudinal study of prey capture of red morph plants. Change in color morph ratio data includes historical data from Case, Frederick W. "Some Michigan records for Sarracenia purpurea forma heterophylla." Rhodora 58.692 (1956): 203-207. ## Description of the data and file structure Files Raw data files: ``` FAHSprey.csv -- Abundance of each prey morphospecies LongitudinalStudy.csv -- weekly insect counts of different prey (order level) at a monomorphic (red morph) bog MorphRatios.csv -- Historical and recent data on morph ratios at three polymorphic bogs seedGerm.csv -- the germination rate from each individual that produced seed SpurLC-MS_MS.csv -- mass spec response values for cyanidin-3-glucoside and cyanidin for young and old leaves of red and green morphs MorphSpectra.csv -- spectral reflectance data from green and red morphs (both red and green regions of the leaf) ``` Cleaned data files: ``` CombinedDataset.csv -- Combined, pre-cleaned field and experimental data on herbivory, prey capture, and inquiline abundance ``` Glossary: CombinedDataset.csv pairID -- unique pair ID for one red and one green morph growing in close proximity morph -- morph, either red (anthocyanin producing) or green (anthocyanin free) perforation -- 0 = leaf intact below water line; 1 = leaf punctured below water line (does not include tears) height -- height of experimental leaf, in cm seedCount -- the number of intact seeds produced by the plant, NA indicates no attempt to flower; 0 indicates unsuccessful flowering attempt or no intact / viable seed seedMass -- the total combined mass of all intact seeds, in g; NA indicates no attempt to flower; 0 indicates unsuccessful flowering attempt or no intact / viable seed flowering -- 1 if flowering, 0 if not flowering; attempts to flower (e.g. aborted but, bud eaten) were scored as 1 insituProp -- the proportion of leaf area removed by (wild) herbivores from 1 leaf per plant in situ in the bog; NA indicates leaf too senesced to measure herbivory insitumm2 -- the area of leaf removed (mm2) by (wild) herbivores from 1 leaf per plant in situ in the bog; NA indicates scan was not made when leaf was fresh (proportion herbivory is a post hoc estimate) cupIDp -- the cup number (1-5) where a caterpillar was reared for the performance (P) experiment, NA = leaf too senesced for herbivory bioassays areaEatenP -- the area consumed by a caterpillar from a 2 cm2 square of tissue during the performance experiment, NA = leaf too senesced for herbivory bioassays propEatenP -- the proportion of the leaf consumed by a caterpillar from a 2 cm2 square of tissue during the performance experiment, NA = leaf too senesced for herbivory bioassays preweightP -- the initial weight (in g) of each caterpillar used in the performance experiment, NA = leaf too senesced for herbivory bioassays postweightP -- the final weight (in g) of each caterpillar used in the performance experiment, NA = leaf too senesced for herbivory bioassays rgr -- relative growth rate of each caterpillar; (log(postweight) - log(preweight))/1.5 #experiment was run for 1.5 days, NA = leaf too senesced for herbivory bioassays preweightC -- the initial weight (in g) of each caterpillar used in the choice experiment, NA = leaf too senesced for herbivory bioassays cupIDc -- the cup number (1-5) where a caterpillar was reared for the choice (C) experiment, NA = leaf too senesced for herbivory bioassays propEatenC -- the proportion of the leaf consumed by a caterpillar from a 2 cm2 square of tissue during the choice experiment, NA = leaf too senesced for herbivory bioassays areaEatenC -- the area consumed by a caterpillar from a 2 cm2 square of tissue during the choice experiment, NA = leaf too senesced for herbivory bioassays preyCount -- the number of individual prey items in each pitcher, NA indicates pitcher contained no fluid and could not captured prey or host inquilines preyMass -- the mass (g) of prey insects (whole and in pieces) collected from each pitcher, NA indicates pitcher contained no fluid and could not captured prey or host inquilines wymTotal -- the number of Wyeomyia smithii larvae in each pitcher, NA indicates pitcher contained no fluid and could not captured prey or host inquilines met -- the number of Metriocnemius knabi larvae in each pitcher, NA indicates pitcher contained no fluid and could not captured prey or host inquilines sarc -- the number of Fletcherimyia fletcheri larvae in each pitcher, NA indicates pitcher contained no fluid and could not captured prey or host inquilines FAHSprey.csv pairID -- unique pair ID for one red and one green morph growing in close proximity morph -- morph, either red (anthocyanin producing) or green (anthocyanin free) tube -- 1 == tube of prey contents was created at time of collection; 0 = no tube (no prey present when leaf collected) mass -- the mass (g) of prey contents; "missing" refers to lines where a tube was recorded as collected, but could not be located for weighing, NP = tube collected but contained no prey, and NA the pitcher contained no fluid and could not capture prey beemass -- for those pitchers that captured a Bombus impatiens bumblebee, the total dry mass of all bees captured, in g unknown:mite -- morpho species or undifferentiated members of an order (e.g. "hym" is any Hymenopteran not in a morphospecies category); empty and NA cells refer to 0 counts of that taxon for that pitcher, and are corrected to zeros on lines 65 to 75 of Anthocyanin_loss.RMD (subsection "Rarefied diversity calculation" in knitted RMD) met, wym, wym_pupa, sarc -- counts of inquiline larvae (see CombinedDataset for species names), empty and NA cells refer to 0 counts of that taxon for that pitcher, and are corrected to zeros on lines 65 to 75 of Anthocyanin_loss.RMD (subsection "Rarefied diversity calculation" in knitted RMD) eggs_binary -- 1 == small round eggs present in sample; 0 == eggs absent LongitudinalStudy.csv plantID -- unique plant ID for each individual plant (collected prey from 1 leaf per plant). All plants were red morphs (anthocyanin-producing) date -- sample collection date month/day/year springtail:snail -- counts of morphospecies, families, or undifferentiated members of an order ("Wmite" = water mite) frass -- counts of frass (separate chunks) in leaf leaf:seed -- counts of plant material; some plant material is subdivided into more specific categories Metriocnemius:Sarcophagid -- counts of each of the three insect inquiline species; "Sarcophagid" refers to Fletcherimyia fletcheri in paper spideParasite -- 1 indicates spider parasitizing pitcher (building web across opening) algage -- 1 indicates the presence of algage unknown -- counts of unknown animal mass no_fluid -- 1 indicates pitcher was empty of fluid, making prey capture impossible damage -- 1 indicates pitcher was damaged, preventing fluid acumulation mL -- number of mL of water accumulated in control tubes of water (50mL falcon tubes) left in the bog pitcherTotal -- the total number of pitchers per plant; entered only for September 1st senescing -- 1 indicates pitcher is beginning to senece on that day putrified -- 1 indicates pitcher fluid is anoxic, smells of decay, and/or is opaque or viscous with biofilm; 0 indicates clear, scentless fluid, no putrefaction notes -- notes, especially on prey ID MorphRatios.csv year -- year of data collection; 1954 data from Case 1955 and 2022 data are presented in this paper year_col -- "year color", placeholder column for graphing bog -- a numeric assesion for each bog assigned by Case 1955 plot -- ID number for each 9x9 foot plot (per Case 1955); two plots per bog red -- number of red plants (includes seedlings and mature plants) in each plot green -- number of green morph (anthocyanin-free) plants (includes seedlings and mature plants) in each plot density -- the total number of plants in each standardized area plot; sum of red and green columns ratio -- the ratio between red and green counts in each plot; red/green columns seedGerm.csv plantID -- combined pairID and morph (g = green, r = red) for each unique plant that bore seed potID -- integer identification for each pot with seed from a particular mother (e.g. 16r pot 1, 16r pot 2, etc); 1:n_pots seeds -- number of seeds planted in a given pot seedlings -- number of seedlings germinated in given pot sum -- sum of the gr_seedling, rd_seedling, and dead columns (see below); redundant with "seedlings" column; created as a double check gr_seedlings -- number of green morph seedlings rd_seedlings -- number of red morph seedlings dead -- number of seedlings that germinated and then died before scoring of seedling morph tray -- integer identification for each tray containing 10 pots date -- planting date month/day/year morph -- long-form morph of mother rate -- germination rate (seedlings column divided by seeds column) parent -- long-form pairID of mother SpurLC-MS_MS.csv morph -- morph, either red (anthocyanin producing) or green (anthocyanin free) age -- leaf age, either young (tender, pale green) or mature ("old", fully expanded and lignified) sample -- sample number; replicates within the same leaf C3G_MS -- the mass spec intensity for cyanidin-3-glucoside Cy_MS -- the mass spec intensity for cyanidin MorphSpectra.csv wl -- wavelength of reflectance (nm) All other column names take the form \[tissue name] [plant ID] [leaf a,b, or c] [replicate] ex: grSpot 001 a .1 Tissues: bodyRed -- the main 'vase' part of a red morph leaf rdSpot -- a red area on the hood of a red morph grSpot -- a green area on the hood of a green morph hood -- any part of the hood of a green morph plantIDs: 001, 002, and 003 -- red morphs 004, 005, and 006 -- green morphs leaf: a,b,c -- the first, second, and third leaf measured on a plant, respectively replicate: " ", ".1", ".2" -- the first, second, and third measurement taken for single area of leaf ## Sharing/Access information Historical color morph ratio data from Case, Frederick W. "Some Michigan records for Sarracenia purpurea forma heterophylla." Rhodora 58.692 (1956): 203-207. ## Code/Software Required R packages: tidyverse, glmmTMB, performance, DHARMa, vegan, pavo ('see' and 'patchwork' are dependencies) Code: ``` Anthocyanin_loss.Rmd -- Code for data analysis and making plots from pre-cleaned, combined morph field and experimental data Anthocyanin_loss.html -- Knitted version of Anthocyanin_loss.Rmd file VisualModels.Rmd -- Code for cleaning spectra data, maping onto pre-built pavo visual models, and plotting in vision model colorspace VisualModels.html -- Knitted version of Visual Models RMD ```