DOI: 10.5061/dryad.3xsj3txqt
title: Data from: A fungal endophyte alters poplar leaf chemistry, deters insect feeding, and shapes insect community assembly

description: The endophytic fungi of certain grasses and other herbaceous plants have
 long been known to provide plants with anti-herbivore defense compounds,
 but there is little information about whether the endophytes of trees also
 engage in such mutualisms. We investigated the influence of the endophytic
 fungus Cladosporium sp. on the chemical defenses of black poplar (Populus
 nigra) trees and the consequences for feeding preference and fitness of
 herbivorous insects and insect community assembly. Endophyte colonization
 increased both constitutive- and induced poplar defenses. Generalist
 Lymantria dispar larvae preferred and performed better on uninfected over
 endophyte-infected poplar leaves, most likely due to higher concentrations
 of salicinoids in endophytic leaves and the endophyte-produced alkaloid
 stachydrine. Under field conditions, the endophytic fungus also shapes
 insect community assembly in young black poplar trees. Our results show
 that endophytic fungi can play a major role both in defending trees
 against herbivorous insects and in structuring insect communities of
 poplar endophytes.
description: # Data from: A fungal endophyte alters poplar leaf chemistry, deters
 insect feeding, and shapes insect community assembly Corresponding author
 Sybille B. Unsicker
 [sunsicker@bot.uni-kiel.de](mailto:sunsicker@bot.uni-kiel.de) ## General
 description This README file describes the data package accompanying the
 above-mentioned publication. Files used in the analysis are either 1.
 .xlsx and .csv files for R.scripts 2. R. scripts 3. Excel file with a
 summary of raw data and data used for statistics in spss 4. MSMS data as
 .mzML files 5. Metaboanalyst.zip containing .csv files, R-history and
 Report.pdf obtained from metaboanalyst 6. IBM SPSS-syntax file ## Code/
 software 1. .xlsx and .csv files for R.scripts R 4.3.2 2. R. scripts:
 Files can be opened using R 4.3.2 3. Excel file with a summary of raw data
 and data used for statistics in IBM SPSS statistics 25 4. MSMS data as
 .mzML files: Files can be opened using open software e.g., ProteoWizard or
 OpenMS. 5. Metaboanalyst.zip containing .csv files, R-history, and
 Report.pdf obtained from metaboanalyst and can be opened with
 metaboanalyst 6. Spss-syntax file can be opened using IBM SPSS statistics
 25 ## File list and description .xlsx and .csv files for R.scripts ####
 "Walther et al. 2024\_volcano.xlsx" A file containing results of
 feature intensities done by metaboscape from endophyte-free plants
 (control) and endophyte-inoculated black poplar trees from the lab
 experiment Columns contain the following information: * Feature: feature
 name given by metaboscape * p-Value: p-Value obtained by metaboscape *
 p-Value(Rank): rank of p-Value obtained by metaboscape * Average Ratio
 "c"/"e": average ratio between control and endophyte *
 Fold Change "c"/"e": fold change between control and
 endophyte * p-Value(FWER): p-value for family-wise error rate obtained by
 metaboscape * p-Value(FDR): p-value for false discovery rate obtained by
 metaboscape * Max. Intensity: Maximum intensity obtained by metaboscape *
 CV "c": Coefficient of variation for control treatment obtained
 by metaboscape * CV "e": Coefficient of variation for endophyte
 treatment obtained by metaboscape * Value Count "c": number of
 replicates for control * Value Count "e": number of replicates
 for endophyte * n/a: not available, under detection limit ####
 "Walther et al. 2024\_bipartite network.csv" A file containing a
 summary of counted arthropods observed on endophyte-free control plants
 and endophyte-inoculated plants from the field experiment Columns contain
 the following information: * Treatment: black poplar trees were either
 inoculated with the endophyte *Cladosporium* sp. (e) or were treated with
 a control solution * Acari: sum of counted individuals observed during the
 time period of the experiment belonging to this order * Araneae: sum of
 counted individuals observed during the time period of the experiment
 belonging to this order * Coleoptera: sum of counted individuals observed
 during the time period of the experiment belonging to this order *
 Dermaptera: sum of counted individuals observed during the time period of
 the experiment belonging to this order * Diptera: sum of counted
 individuals observed during the time period of the experiment belonging to
 this order * Ephemeroptera: sum of counted individuals observed during the
 time period of the experiment belonging to this order * Hemiptera: sum of
 counted individuals observed during the time period of the experiment
 belonging to this order * Hymenoptera: sum of counted individuals observed
 during the time period of the experiment belonging to this order *
 Ixodida: sum of counted individuals observed during the time period of the
 experiment belonging to this order * Leaf miner: sum of counted
 individuals observed during the time period of the experiment belonging to
 this order * Lepidoptera: sum of counted individuals observed during the
 time period of the experiment belonging to this order * Neuroptera: sum of
 counted individuals observed during the time period of the experiment
 belonging to this order * Orthoptera: sum of counted individuals observed
 during the time period of the experiment belonging to this order *
 Thysanoptera: sum of counted individuals observed during the time period
 of the experiment belonging to this order * unknown: sum of counted
 individuals observed during the time period of the experiment, which could
 not be classified #### "Walther et al.
 2024\_Field\_stachydrine.csv" File containing stachydrine
 concentration from black poplar trees standing in the field Columns
 contain the following information: * sample.ID: replicate * compound: name
 of the measured compound * concentration: amount of the measured compound
 stachydrine in nmol/g * endophyte: describes the treatment, c (control,
 tree with no endophyte inoculation), e (endophyte, tree with endophyte
 inoculation) * bereich: area of the tree which was observed * tree: unique
 number of tree * herbivory: leaf with damage by herbivory * damage.perc:
 leaf area loss in percentages * damage: leaf area loss in cm² ####
 "Walther et al. 2024\_heatmap\_neg\_annotade.csv" A file
 containing significant features (measured in negative mode) annotated by
 Michael Reichelt. Significant features were obtained by a two-way-ANOVA
 done in MetaboAnalyst (but see Metaboanalyst.zip) Columns contain the
 following information: * c1..c4: control, uninfected black poplar trees (4
 replicates), containing information about the feature intensity *
 ch1...ch4: control + herbivory, uninfected black poplar trees which
 experienced herbivory by *L. dispar* larvae (4 replicates), containing
 information about the feature intensity * e1...e4: endophyte, with
 *Cladosporium* sp. inoculated black poplar trees (4 replicates),
 containing information about the feature intensity * eh1...eh4: endophyte
 + herbivory, with *Cladosporium* sp. inoculated black poplar trees which
 experienced herbivory by *L. dispar* larvae (4 replicates), containing
 information about the feature intensity * Name: feature name and
 annotation given by Michael Reichelt * Formular: sum formula of chemical
 structure * bucket: feature name given by metaboscape * RT..min: Retention
 time in min * m.z..Da.: mass to ratio in Dalton * putative.identification:
 annotation by Michael Reichelt * NA in column Formular: chemical formula
 could not be generated * NA in column putative.identification: feature
 could not be annotated #### "Walther et al.
 2024\_heatmap\_neg\_anova.csv" The file contains results of a two-way
 ANOVA done by metaboanalyst features obtained by an LCMS measured in
 negative polarity from endophyte-free plants (control) and
 endophyte-inoculated black poplar trees from the lab experiment. The
 corresponding R.history, Report.pdf file was obtained from MetaboAnalyst
 and CSV. data used for the analysis can be found in the folder
 Metaboanalyst.zip. Columns contain the following information: * X.1:
 continuous number of individual feature * X: Features *
 endophyte.F.val:F-value for endophyte (two-way ANOVA) * endophyte.raw.p.:
 raw p-value for endophyte * endophyte.adj.p.: adjusted p-value for
 endophyte * herbivory.F.val.: F-value for herbivory (two-way ANOVA) *
 herbivory.raw.p.: raw p-value for herbivory * herbivory.adj.p.: adjusted
 p-value for herbivory * interaction.F.val.: F-value for
 interaction(two-way ANOVA) * interaction.raw.p.: raw p-value for
 interaction * interaction.adj.p.: adjusted p-value for interaction Related
 files from the folder Metaboanalyst.zip: *
 Walther_et_al._2024_heatmap_metaboanalyst_metadata.csv *
 Walther_et_al._2024_heatmap_metaboanalyst_neg.mode_feat1-200.csv *
 Walther_et_al._2024_heatmap_metaboanalyst_neg.mode_feat201-400.csv *
 Walther_et_al._2024_heatmap_metaboanalyst_neg.mode_feat401-492.csv *
 Walther_et_al._2024_heatmap_metaboanalyst_neg.mode_feat1-200_Rhistory.R *
 Walther_et_al._2024_heatmap_metaboanalyst_neg.mode_feat201-400_Rhistory.R
 *
 Walther_et_al._2024_heatmap_metaboanalyst_neg.mode_feat401-492_Rhistory.R
 * Walther_et_al._2024_heatmap_metaboanalyst_neg.mode_feat1-200_Report.pdf
 *
 Walther_et_al._2024_heatmap_metaboanalyst_neg.mode_feat201-400_Report.pdf
 *
 Walther_et_al._2024_heatmap_metaboanalyst_neg.mode_feat401-492_Report.pdf
 #### "Walther et al. 2024\_heatmap\_neg\_intensities.csv" The
 file containing feature obtained by an LCMS measured in negative polarity
 from endophyte-free plants (control) and endophyte-inoculated black poplar
 trees from the lab experiment Columns contain the following information: *
 sample: feature name * c..c4: control, uninfected black poplar trees (4
 replicates), containing information about the feature intensity *
 ch...ch4: control + herbivory, uninfected black poplar trees which
 experienced herbivory by *L. dispar* larvae (4 replicates), containing
 information about the feature intensity * e...e4: endophyte, with
 *Cladosporium* sp. inoculated black poplar trees (4 replicates),
 containing information about the feature intensity * eh...eh4: endophyte +
 herbivory, with *Cladosporium* sp. inoculated black poplar trees that
 experienced herbivory by *L. dispar* larvae (4 replicates), containing
 information about the feature intensity #### "Walther et al.
 2024\_heatmap\_pos\_annotade.csv" A file containing significant
 features (measured in positive mode) annotated by Michael Reichelt.
 Significant features were obtained by a two-way-ANOVA done in
 MetaboAnalyst (but see Metaboanalyst.zip) Columns contain the following
 information: * c1..c4: control, uninfected black poplar trees (4
 replicates), containing information about the feature intensity *
 ch1...ch4: control + herbivory, uninfected black poplar trees which
 experienced herbivory by *L. dispar* larvae (4 replicates), containing
 information about the feature intensity * e1...e4: endophyte, with
 *Cladosporium* sp. inoculated black poplar trees (4 replicates),
 containing information about the feature intensity * eh1...eh4: endophyte
 + herbivory, with *Cladosporium* sp. inoculated black poplar trees that
 experienced herbivory by *L. dispar* larvae (4 replicates), containing
 information about the feature intensity * Name: feature name and
 annotation given by Michael Reichelt * Formular: sum formula of chemical
 structure * bucket: feature name given by metabscape * RT..min: Retention
 time in min * m.z..Da.: mass to ratio in Dalton * putative.identification:
 annotation by Michael Reichelt * NA in column Formular: chemical formula
 could not be generated * NA in column putative.identification: feature
 could not be annotated #### "Walther et al.
 2024\_heatmap\_pos\_anova.csv" The file contains results of a two-way
 ANOVA done by metaboanalyst features obtained by an LCMS measured in
 positive polarity from endophyte-free plants (control) and
 endophyte-inoculated black poplar trees from the lab experiment. The
 corresponding R.history, Report.pdf file was obtained from MetaboAnalyst
 and CSV. data used for the analysis in MetaboAnalyst can be found in the
 folder Metaboanalyst.zip. Columns contain the following information: *
 X.1: continuous number of individual feature * X: Features *
 endophyte.F.val:F-value for endophyte (two-way ANOVA) * endophyte.raw.p.:
 raw p-value for endophyte * endophyte.adj.p.: adjusted p-value for
 endophyte * herbivory.F.val.: F-value for herbivory (two-way ANOVA) *
 herbivory.raw.p.: raw p-value for herbivory * herbivory.adj.p.: adjusted
 p-value for herbivory * interaction.F.val.: F-value for
 interaction(two-way ANOVA) * interaction.raw.p.: raw p-value for
 interaction * interaction.adj.p.: adjusted p-value for interaction Related
 files from the folder Metaboanalyst.zip: *
 Walther_et_al._2024_heatmap_metaboanalyst_metadata.csv *
 Walther_et_al._2024_heatmap_metaboanalyst_pos.mode_feat1-200.csv *
 Walther_et_al._2024_heatmap_metaboanalyst_pos.mode_feat201-400.csv *
 Walther_et_al._2024_heatmap_metaboanalyst_pos.mode_feat401-600.csv *
 Walther_et_al._2024_heatmap_metaboanalyst_pos.mode_feat601-698.csv *
 Walther_et_al._2024_heatmap_metaboanalyst_pos.mode_feat1-200_Rhistory.R *
 Walther_et_al._2024_heatmap_metaboanalyst_pos.mode_feat201-400_Rhistory.R
 *
 Walther_et_al._2024_heatmap_metaboanalyst_pos.mode_feat401-600_Rhistory.R
 *
 Walther_et_al._2024_heatmap_metaboanalyst_pos.mode_feat601-698_Rhistory.R
 * Walther_et_al._2024_heatmap_metaboanalyst_pos.mode_feat1-200_Report.pdf
 *
 Walther_et_al._2024_heatmap_metaboanalyst_pos.mode_feat201-400_Report.pdf
 *
 Walther_et_al._2024_heatmap_metaboanalyst_pos.mode_feat401-600_Report.pdf
 *
 Walther_et_al._2024_heatmap_metaboanalyst_pos.mode_feat601-698_Report.pdf
 #### "Walther et al. 2024\_heatmap\_pos\_intensities.csv" File
 containing features obtained by an LCMS measured in negative polarity from
 endophyte-free plants (control) and endophyte-inoculated black poplar
 trees from the lab experiment Columns contain the following information: *
 sample: feature name * c..c4: control, uninfected black poplar trees (4
 replicates), containing information about the feature intensity *
 ch...ch4: control + herbivory, uninfected black poplar trees which
 experienced herbivory by *L. dispar* larvae (4 replicates), containing
 information about the feature intensity * e...e4: endophyte, with
 *Cladosporium* sp. inoculated black poplar trees (4 replicates),
 containing information about the feature intensity * eh...eh4: endophyte +
 herbivory, with *Cladosporium* sp. inoculated black poplar trees that
 experienced herbivory by *L. dispar* larvae (4 replicates), containing
 information about the feature intensity #### "Walther et al.
 2024\_performance.csv" A file containing results of a performance
 experiment. Larvae of *L. dispar* (above) and *C. tremulae* (below) fed
 either on endophyte-inoculated leaves or control leaves. Columns contain
 the following information: * unique.no.Of.insect: biological replicate
 (insect species feeding on black poplar leaves) * insect.species: *L.
 dispar* larvae (moth) or *C. tremulae* (beetle) feeding on black poplar
 tree * treatment: black poplar trees were either inoculated with the
 endophyte *Cladosporium* sp. (e) or were treated with a control solution
 (c) * T0...T10: weight (mg) of insect species on that time point *
 W1...W10: weight gain (mg) calculated by T(x)-T(x-1) * delta1....delta10:
 weight gain (mg) calculated by T(x)-T(0) * NA: data not available, as
 replicate 3 (moth) died or experiment finished (insect.species: *C.
 tremulae* (beetle)) ## R code: #### "Walther et al.
 2024\_heatmap.R" Metabolomic investigations of black poplar (*P.
 nigra*) leaves after fungal endophyte inoculation and herbivory by *L.
 dispar* larvae. R code used to create a heatmap of the significant
 features obtained by a two-way ANOVA. #### "Walther et al.
 2024\_bipartite network.R" R code used to visualize plant-arthropod
 interactions comparing black poplar trees infected with a fungal endophyte
 to control uninfected trees placed in the field. #### "Walther et al.
 2024\_performance.R" R code used to estimate the effect of time,
 endophyte, and the interaction on insect performance. #### "Walther
 et al. 2024\_volcano.R" Detection and quantification of the alkaloid
 stachydrine R code used to create a for volcano plot which illustrates
 metabolite features from leaf tissue inoculated with the endophyte and
 those of control plants. Features were obtained from an untargeted LC-MS
 analysis with a QTOF mass spectrometer in positive ionization mode. ####
 "Walther et al. 2024\_stachydrine field.R" R code to test
 differences in stachydrine concentration of black poplar trees in the
 field experiment. ## Excel file with a summary of raw data ####
 "Walther et al. 2024 raw data 240909.xlsx" Summary of all data
 used in the study and used for statistical analysis run in IBM SPSS
 statistics 25 Sheet: Figure 1 raw data File containing an overview of raw
 data about feature intensities obtained from LCMS. Data used for PCA and
 heatmap Columns contain the following information: * feature
 (metaboscape): feature name given by Metaboscape * mode: positive or
 negative ionization mode run on an LCMS * c1-c4: control, uninfected black
 poplar trees * ch1-ch4: control + herbivory, uninfected black poplar trees
 which experienced herbivory by *L. dispar* larvae * e1-e4: endophyte, with
 *Cladosporium* sp. inoculated black poplar trees * eh1-eh4: endophyte +
 herbivory, with *Cladosporium* sp. inoculated black poplar trees that
 experienced herbivory by *L. dispar* larvae * chemical formular: chemical
 formula obtained from metaboscape * mass to charge ratio: mass to charge
 ratio of the specific feature * retention time: retention time in minutes
 * n/a: no data available, no chemical formula could be generated Sheet:
 Figure 1 PCA A file containing raw data about PCA loadings Columns contain
 the following information: * mode: positive or negative ionization mode
 run on an LCMS * feature (metaboscape): feature name given by Metaboscape
 * PC1-16: PCA axis Sheet: Figure 1 heatmap.1 A file containing results of
 the 2way ANOVA of significant features obtained from metaboanalyst used
 for heatmap Columns contain the following information: * feature
 (metaboscape): feature name given by Metaboscape * mode: positive or
 negative ionization mode run on an LCMS * endophyte: with *Cladosporium*
 sp. inoculated black poplar trees * herbivory: black poplar trees which
 experienced herbivory by *L. dispar* larvae * interaction: interaction of
 endophyte infection and herbivory on black poplar trees *
 endophyte(F.val): F-value for endophyte (two-way ANOVA) *
 endophyte(raw.p): raw p-value for endophyte * endophyte(adj.p): adjusted
 p-value for endophyte * herbivory(F.val): F-value for herbivory (two-way
 ANOVA) * herbivory(raw.p): raw p-value for herbivory * herbivory(adj.p):
 adjusted p-value for herbivory * Interaction(F.val): F-value for
 interaction(two-way ANOVA) * Interaction(raw.p): raw p-value for
 interaction * Interaction(adj.p): adjusted p-value for interaction Sheet:
 Figure 1 heatmap.2 A file containing significant features used for the
 heatmap. Columns contain the following information: * c: control,
 uninfected black poplar trees * ch: control + herbivory, uninfected black
 poplar trees that experienced herbivory by *L. dispar* larvae * e:
 endophyte, with *Cladosporium* sp. inoculated black poplar trees * eh:
 endophyte + herbivory, with *Cladosporium* sp. inoculated black poplar
 trees that experienced herbivory by *L. dispar* larvae * RT (min):
 retention time in minutes * m/z: mass to ratio in Dalton * mode: positive
 or negative ionization mode run on an LCMS * formular: chemical subformula
 * feature (metaboscape): feature name given by Metaboscape * feature
 (final): feature name after adjustment * n/a: no data available, no
 chemical formula could be generated Sheet: Figure 2 File containing
 results of the effect of endophyte inoculation and herbivory on salicinoid
 concentrations in leaves of black Columns contain the following
 information: * c: control, uninfected black poplar trees * ch: control +
 herbivory, uninfected black poplar trees that experienced herbivory by *L.
 dispar* larvae * e: endophyte, with *Cladosporium* sp. inoculated black
 poplar trees * eh: endophyte + herbivory, with *Cladosporium* sp.
 inoculated black poplar trees that experienced herbivory by *L. dispar*
 larvae * treatment: treatment of black poplar trees in the assays *
 endophyte: with *Cladosporium* sp. inoculated black poplar trees *
 herbivory: black poplar trees that experienced herbivory by *L. dispar*
 larvae * concentration: content of the metabolite * unit: unit of
 metabolite * compound: measured metabolite Sheet: Figure 3 File containing
 results of the effect of fungal endophyte inoculation on the levels of
 non-salicinoid phenolic defense Columns contain the following information:
 * c: control, uninfected black poplar trees * ch: control + herbivory,
 uninfected black poplar trees which experienced herbivory by *L. dispar*
 larvae * e: endophyte, with *Cladosporium* sp. inoculated black poplar
 trees * eh: endophyte + herbivory, with *Cladosporium* sp. inoculated
 black poplar trees which experienced herbivory by *L. dispar* larvae *
 treatment: treatment of black poplar trees in the assays * endophyte: with
 *Cladosporium* sp. inoculated black poplar trees * herbivory: black poplar
 trees that experienced herbivory by *L. dispar* larvae * concentration:
 content of the metabolite * unit: unit of metabolite * compound: measured
 metabolite Sheet: Figure 4b A file containing results of the effect of
 fungal endophyte inoculation on the preference of insect herbivores.
 Columns contain the following information: * c: control, uninfected black
 poplar trees * e: endophyte, with *Cladosporium* sp. inoculated black
 poplar trees * tree: individual tree number * caterpillar: individual
 caterpillar (*L. dispar*) feeding on black poplar leafs * damage (cm²):
 consumed leaf area by *L. dispar* larvae * leaf area loss %: consumed leaf
 area in percentages by *L. dispar* larvae Sheet: Figure 4c A file
 containing results of the effect of fungal endophyte inoculation on the
 performance of insect herbivores. Columns contain the following
 information: * c: control, uninfected black poplar trees * e: endophyte,
 with *Cladosporium* sp. inoculated black poplar trees * moth: *L. dispar*
 larvae feeding on black poplar tree * beetle: *C. tremulae* larvae feeding
 on black poplar tree * sample ID: biological replicate (insect species
 feeding on black poplar leaves) * insect species: *L. dispar* larvae
 (moth) or *C. tremulae* (beetle) feeding on black poplar tree * treatment:
 treatment of black poplar used in the assays * time (d): time in days
 after insects were placed on the black poplar trees * weight gain (mg):
 weight gain in mg of observed insect Sheet: Figure 5a The volcano plot
 illustrates metabolite features from leaf tissue inoculated with the
 endophyte and those of control plants. Columns contain the following
 information: * c: control, uninfected black poplar trees * e: endophyte,
 with *Cladosporium* sp. inoculated black poplar trees * Feature: feature
 name given by metaboscape * p-Value: p-Value obtained by metaboscape *
 p-Value(Rank): rank of p-Value obtained by metaboscape * Average Ratio
 "c"/"e": average ratio between control and endophyte *
 Fold Change "c"/"e": fold change between control and
 endophyte * p-Value(FWER): p-value family-wise error rate obtained by
 metaboscape * p-Value(FDR): p-value false discovered rate obtained by
 metaboscape * Max. Intensity: Maximum intensity obtained by metaboscape *
 CV "c": Coefficient of variation for control treatment obtained
 by metaboscape * CV "e": Coefficient of variation for endophyte
 treatment obtained by metaboscape * Value Count "c": number of
 replicates for control * Value Count "e": number of replicates
 for endophyte * n/a: no data available, under the detection limit Sheet:
 Figure 5b File containing data on stachydrine concentration from
 *Cladosporium* sp. and black poplar leaves from the lab experiment Columns
 contain the following information: * sample ID: replicate * tissue: tissue
 used for chemical analysis * endophyte: with *Cladosporium* sp. inoculated
 black poplar trees * herbivory: black poplar trees that experienced
 herbivory by *L. dispar* larvae * treatment: treatment of black poplar
 used in the assays * concentration: content of the metabolite * unit: unit
 of metabolite * compound: measured metabolite Abbreviation: * c: control,
 uninfected black poplar trees * ch: control + herbivory, uninfected black
 poplar trees which experienced herbivory by *L. dispar* larvae * e:
 endophyte, with *Cladosporium* sp. inoculated black poplar trees * eh:
 endophyte + herbivory, with *Cladosporium* sp. inoculated black poplar
 trees which experienced herbivory by *L. dispar* larvae * n/a: not
 available, collected mycelium, no treatment was applied Sheet: Figure 5d A
 file containing data on leaf area loss from the stachydrine choice
 experiment Columns contain the following information: * replicate:
 biological replicate * insect: insect species used for the assay * damage
 stachydrine (cm²): leaf area consumed by an insect, leaf discs treated
 with a solution containing stachydrine * damage control cm2: leaf area
 consumed by an insect, leaf discs treated with a control solution Sheet:
 Figure 6 A file containing raw data of plant-arthropod interactions
 comparing black poplar trees infected with a fungal endophyte to control
 uninfected trees placed in the field. Columns contain the following
 information: * date: observation date * time: time of observation * datum:
 real date * treatment: treatment of black poplar trees ( c: control,
 uninfected black poplar trees, e: endophyte, with *Cladosporium* sp.
 inoculated black poplar trees) * tree: individual tree * Acari ...
 Thysanoptera: observed Arthropod on a specific observation point *
 unknown: not identified arthropod Sheet: Table S1A A file containing data
 on endophyte abundance from different experiments Columns contain the
 following information: * c: control, uninfected black poplar trees * ch:
 control + herbivory, uninfected black poplar trees which experienced
 herbivory by *L. dispar* larvae * e: endophyte, with *Cladosporium* sp.
 inoculated black poplar trees * eh: endophyte + herbivory, with
 *Cladosporium* sp. inoculated black poplar trees which experienced
 herbivory by *L. dispar* larvae * d: damage * experiment: type of
 experiment * treatment: treatment of black poplar trees * treat: number
 abbreviation of treatment of black poplar trees * target: target sequence
 to quantify the abundance of the endophyte * Sample: biological replicate
 * expression: Relative endophyte abundance quantified with qPCR (relative
 expression ∆∆ Ct) Sheet: Table S1B A file containing data on stachydrine
 concentrations from different experiments Columns contain the following
 information: * c: control, uninfected black poplar trees * ch: control +
 herbivory, uninfected black poplar trees which experienced herbivory by
 *L. dispar* larvae * e: endophyte, with *Cladosporium* sp. inoculated
 black poplar trees * eh: endophyte + herbivory, with Cladosporium sp.
 inoculated black poplar trees which experienced herbivory by *L. dispar*
 larvae * experiment: type of experiment * treatment: treatment of black
 poplar trees in the assays * endophyte: with *Cladosporium* sp. inoculated
 black poplar trees * herbivory: black poplar trees that experienced
 herbivory by *L. dispar* larvae * concentration: content of the metabolite
 * unit: unit of metabolite * compound: measured metabolite * tree: unique
 number of individual tree Sheet: Table S1D A file containing data on
 stachydrine concentrations from stachydrine preference assay Columns
 contain the following information: * c: control, uninfected black poplar
 trees * stach: leaves coated with stachydrine solution * experiment: type
 of experiment * treatment: treatment of black poplar trees in the assays *
 concentration: content of the metabolite * unit: unit of metabolite *
 compound: measured metabolite Sheet: Table S2 A file containing data on
 leaf chemical analysis from the laboratory experiment Columns contain the
 following information: * c: control, uninfected black poplar trees * ch:
 control + herbivory, uninfected black poplar trees which experienced
 herbivory by *L. dispar* larvae * e: endophyte, with *Cladosporium* sp.
 inoculated black poplar trees * eh: endophyte + herbivory, with
 *Cladosporium* sp. inoculated black poplar trees which experienced
 herbivory by *L. dispar* larvae * experiment: type of experiment *
 treatment: treatment of black poplar trees in the assays * treat: number
 abbreviation of treatment of black poplar trees * endophyte: with
 *Cladosporium* sp. inoculated black poplar trees * herbivory: black poplar
 trees that experienced herbivory by *L. dispar* larvae * concentration:
 content of the metabolite * unit: unit of metabolite * compound: measured
 metabolite Sheet: Table S4 A file containing data on arthropods attracted
 during the field experiment Columns contain the following information: *
 c: control, uninfected black poplar trees * e: endophyte, with
 *Cladosporium* sp. inoculated black poplar trees * date: observation date
 * time: time of observation * treatment: treatment of black poplar trees *
 tree: individual tree * count: number of observed insects counted on that
 day during that time on the tree * class: insect was identified to class
 level * order: insect was identified to order level * suborder: insect was
 identified to suborder level * family: insect was identified at the family
 level Sheet: Table S5 A file containing data on leaf chemical analysis
 from the field experiment Columns contain the following information: * c:
 control, uninfected black poplar trees * e: endophyte, with *Cladosporium*
 sp. inoculated black poplar trees * treatment: treatment of black poplar
 trees in the assays * concentration: content of the metabolite * unit:
 unit of metabolite * compound: measured metabolite Sheet: Table S9 A file
 containing data on the germination tests of *Cladosporium* sp. spores for
 the lab experiment, the preference assay, and the field experiment.
 Columns contain the following information: * experiment: type of
 experiment * replicate: 100µl of diluted spore solution were pipetted on a
 single petri dish (one petri dish = one replicate) * germinated spores:
 number of germinated spores Sheet: Figure S5 File containing data of
 herbivore damage to endophytic fungus-inoculated (fungus) and uninoculated
 control (control) trees from the experiment conducted in the lab Columns
 contain the following information: * treatment: treatment of black poplar
 trees in the assays * treat: treatment of black poplar trees in the assays
 * tree: unique number of individual tree * sum of damage: sum of damage of
 the whole tree * leaf area loss %: leaf area loss in percentage
 Abbreviations: * c: control, uninfected black poplar trees * e: endophyte,
 with Cladosporium sp. inoculated black poplar trees * 1: c * 2: e Sheet:
 Figure S6 File containing data on the effect of fungal endophyte
 inoculation on concentrations of phytohormones of black poplar Columns
 contain the following information: * treatment: treatment of black poplar
 trees in the assays * treat: number abbreviation of treatment of black
 poplar trees * endophyte: with *Cladosporium* sp. inoculated black poplar
 trees * herbivory: black poplar trees that experienced herbivory by *L.
 dispar* larvae * concentration: content of the metabolite * unit: unit of
 metabolite * compound: measured metabolite Abbreviations: * c: control,
 uninfected black poplar trees * ch: control + herbivory, uninfected black
 poplar trees which experienced herbivory by *L. dispar* larvae * e:
 endophyte, with *Cladosporium* sp. inoculated black poplar trees * eh:
 endophyte + herbivory, with *Cladosporium* sp. inoculated black poplar
 trees which experienced herbivory by *L. dispar* larvae Sheet: Figure S7b
 File containing data on herbivore damage from control and inoculated
 plants from the field experiment Columns contain the following
 information: * tree: unique number of individual tree * treatment:
 treatment of black poplar trees in the assays * treat: number abbreviation
 of treatment of black poplar trees * sum leaf area (cm²): reconstructed
 area of a whole plant * damage (cm²): leaf area loss in cm² * damage %:
 leaf area loss in % Abbreviation: * 1: control * 2: endophyte Sheet:
 Figure S8 A file containing data on stachydrine concentration in aphid
 honeydew. Columns contain the following information: * treatment:
 treatment of black poplar trees in the assays * treat: number abbreviation
 of treatment of black poplar trees * tree: unique number of individual
 tree * concentration: content of the metabolite * unit: unit of metabolite
 * compound: measured metabolite Abbreviation: * 1: control * 2: endophyte
 ## MSMS data as .mzML files #### "Walther et al. 2024\_MSMS.zip"
 Zip-folder containing data of mass spectra from nontarget analysis. Files
 can be opened using open software e.g., ProteoWizard or OpenMS. Files
 .mzML are named after sample name: continues number - treatment -
 replicate - ionization mode * c: control, uninfected black poplar trees *
 ch: control + herbivory, uninfected black poplar trees which experienced
 herbivory by *L. dispar* larvae * e: endophyte, with *Cladosporium* sp.
 inoculated black poplar trees * eh: endophyte + herbivory, with
 *Cladosporium* sp. inoculated black poplar trees which experienced
 herbivory by *L. dispar* larvae ## Metaboanalyst.zip containing .csv
 files, R-history and Report.pdf ####
 "metaboanalyst.zip" Zip-folder containing peak intensities of
 features obtained from untarget LCMS analysis and proceed in metaboanlayst
 to create a PCA and to obtain significant features as a result of a
 2wayAnova run in MetaboAnalyst to create a heatmap The folder contains
 .csv files, which were uploaded to metaboanalyst (including the metadata
 file), and R-history and a Report.pdf file obtained from metaboanalyst
 after running the statistics. File name describes: author_the purpose for
 the analysis_software_ionization mode_feature range Rhistory.R and
 Report.pdf files are named accordingly. .csv files used to create PCA and
 files obtained from Metaboanalyst: *
 Walther_et_al._2024_PCA_metaboanalyst_neg.mode.csv *
 Walther_et_al._2024_PCA_metaboanalyst_neg.mode_Rhistory.R *
 Walther_et_al._2024_PCA_metaboanalyst_neg.mode_Report.pdf *
 Walther_et_al._2024_PCA_metaboanalyst_pos.mode.csv *
 Walther_et_al._2024_PCA_metaboanalyst_pos.mode_Rhistory.R *
 Walther_et_al._2024_PCA_metaboanalyst_pos.mode_Report.pdf Due to
 computational limitations in metaboanalyst, only the top 200 features
 (based on variance) are tested. Therefore .csv files with features were
 split into several .csv files for 2wayanova in order to analyse all the
 data. .csv files used for a two-way-ANOVA and files obtained from
 Metaboanalyst: * Walther_et_al._2024_heatmap_metaboanalyst_metadata.csv *
 Walther_et_al._2024_heatmap_metaboanalyst_neg.mode_feat1-200.csv *
 Walther_et_al._2024_heatmap_metaboanalyst_neg.mode_feat201-400.csv *
 Walther_et_al._2024_heatmap_metaboanalyst_neg.mode_feat401-492.csv *
 Walther_et_al._2024_heatmap_metaboanalyst_neg.mode_feat1-200_Rhistory.R *
 Walther_et_al._2024_heatmap_metaboanalyst_neg.mode_feat201-400_Rhistory.R
 *
 Walther_et_al._2024_heatmap_metaboanalyst_neg.mode_feat401-492_Rhistory.R
 * Walther_et_al._2024_heatmap_metaboanalyst_neg.mode_feat1-200_Report.pdf
 *
 Walther_et_al._2024_heatmap_metaboanalyst_neg.mode_feat201-400_Report.pdf
 *
 Walther_et_al._2024_heatmap_metaboanalyst_neg.mode_feat401-492_Report.pdf
 - Walther_et_al._2024_heatmap_metaboanalyst_metadata.csv -
 Walther_et_al._2024_heatmap_metaboanalyst_pos.mode_feat1-200.csv -
 Walther_et_al._2024_heatmap_metaboanalyst_pos.mode_feat201-400.csv -
 Walther_et_al._2024_heatmap_metaboanalyst_pos.mode_feat401-600.csv -
 Walther_et_al._2024_heatmap_metaboanalyst_pos.mode_feat601-698.csv -
 Walther_et_al._2024_heatmap_metaboanalyst_pos.mode_feat1-200_Rhistory.R -
 Walther_et_al._2024_heatmap_metaboanalyst_pos.mode_feat201-400_Rhistory.R
 -
 Walther_et_al._2024_heatmap_metaboanalyst_pos.mode_feat401-600_Rhistory.R
 -
 Walther_et_al._2024_heatmap_metaboanalyst_pos.mode_feat601-698_Rhistory.R
 - Walther_et_al._2024_heatmap_metaboanalyst_pos.mode_feat1-200_Report.pdf
 -
 Walther_et_al._2024_heatmap_metaboanalyst_pos.mode_feat201-400_Report.pdf
 -
 Walther_et_al._2024_heatmap_metaboanalyst_pos.mode_feat401-600_Report.pdf
 -
 Walther_et_al._2024_heatmap_metaboanalyst_pos.mode_feat601-698_Report.pdf
 Abbreviation used: * feat: features * c: control, uninfected black poplar
 trees * ch: control + herbivory, uninfected black poplar trees which
 experienced herbivory by *L. dispar* larvae * e: endophyte, with
 *Cladosporium* sp. inoculated black poplar trees * eh: endophyte +
 herbivory, with *Cladosporium* sp. inoculated black poplar trees which
 experienced herbivory by *L. dispar* larvae ## IBM SPSS-syntax file
 **"Walther_et_al._2024_spss_syntax.sps"** The file
 contains the Syntax of Statistical analysis run by IBM SPSS 25.
 Corresponding Excel file: * Walther_et_al._2024_raw_data_240909.xlsx
 (description see above). ## Version changes ####  26-sept-2024 1\.
 R-scripts: * the used package version is now included in each R-script *
 corrected the name of all .csv-files in the script for proper data loading
 2\. Walther_et_al._2024_raw_data_240909: * Sheets have been updated (table
 structure changed) for proper data loading in SPSS using the SPSS syntax
 file Walther_et_al._2024_spss_syntax.sps: * “Figure 5d”: split the column
 “damage (cm²)” by column “treatment” to obtain two columns “damage
 stachydrine cm2” and “damage control cm2”, which fit the table design for
 paired t-test in SPSS * “Figure 6”: changed table structure to now have
 arthropod orders as columns, which matches the table design for
 statistical analysis in SPSS * “Figure S5”: summed up the leaf damage and
 calculated the leaf area loss (%) for all leaves per tree to analyze
 herbivore damage on whole poplar trees, which was used to compare damage
 between control and endophyte-infected trees from the lab experiment,
 non-herbivory treatments were dropped * “Figure S7b”: summed up the leaf
 damage and calculated the leaf area loss (%) for all leaves per tree to
 analyze herbivore damage on whole poplar trees, which was used to compare
 damage between control and endophyte-infected trees from the field
 experiment 3\. Walther_et_al._2024_spss_syntax.sps: * uploaded the Syntax
 of the statistical analysis done in SPSS 4\. metaboanalyst.zip: uploaded
 statistical analysis done in MetaboAnalyst * File contains R-history and
 Report.pdf obtained from MetaboAnalyst and corresponding .csv files used
 to analyze peak intensities of features obtained from untargeted LCMS
 analysis to create a PCA and to obtain significant features as a result of
 a 2wayAnova run in MetaboAnalyst to create a heatmap.