Replication data for: Implications of the existence of separated sexual forms of Opuntia robusta for plant defense hypotheses (doi:10.7910/DVN/LERCFK)

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Replication data for: Implications of the existence of separated sexual forms of Opuntia robusta for plant defense hypotheses

doi:10.7910/DVN/LERCFK

Harvard Dataverse

2020-08-14

2

Janczur, Mariusz Krzysztof; González-Camarena, Emilio; Jenner, Bartosz, 2020, "Replication data for: Implications of the existence of separated sexual forms of Opuntia robusta for plant defense hypotheses", https://doi.org/10.7910/DVN/LERCFK, Harvard Dataverse, V2, UNF:6:GJKng/DxExj81K4HQurspg== [fileUNF]

Replication data for: Implications of the existence of separated sexual forms of Opuntia robusta for plant defense hypotheses

Secondary metabolites in separated sexual forms of Opuntia robusta

doi:10.7910/DVN/LERCFK

Janczur, Mariusz Krzysztof (Universidad Autónoma del Estado de México)

González-Camarena, Emilio (Universidad Autónoma del Estado de México)

Jenner, Bartosz (Actelion Pharmaceuticals Ltd, Gewerbestrasse 16, CH-4123 Allschwil, Switzerland)

Janczur, Mariusz Krzysztof

Janczur, Mariusz Krzysztof

Janczur, Mariusz Krzysztof

González-Camarena, Emilio

Jenner, Bartosz

Janczur, Mariusz Krzysztof

2020-08-09

180694

Harvard Dataverse

Janczur, Mariusz Krzysztof

Janczur, Mariusz Krzysztof

Janczur, Mariusz Krzysztof

2020-08-09

2020-08-09

https://doi.org/10.7910/DVN/LERCFK

Medicine, Health and Life Sciences, Opuntia robusta, plant sexual forms, trioecious population, herbivory, secondary metabolites, 4-hydroxybenzoic acid, chlorogenic acid, quercetin, plant defense, plant defence, plant chemical defense, plant chemical defence

Biology, Ecology, Evolutionary ecology, Plant sciences, Botany, Chemical ecology, Chemistry

Plant optimal defense hypothesis (ODH) predicts higher levels of defenses, lower damage, and lower investment in vegetative biomass in more costly sexual form and/or younger plant parts. The resource competition hypothesis (RCH) predicts slower growth, lower concentrations of defenses, and greater herbivory levels in more costly sexual form. In our study, hermaphrodite flowers Opuntia robusta grew significantly faster and were less damaged than female flowers. Female cladodes contained 4-hydroxybenzoic acid (4-HBA) and quercetin (QUE) with higher probability than hermaphrodite cladodes did, but not the chlorogenic acid (CGA). Cladodes from all age classes were smaller in female than in hermaphrodite plants. There was a negative relationship between several cladode age estimators and the proportion of cladodes bearing 4-HBA. The probability of detection of either CGA or QUE depended only of the number of daughter cladodes and cladode length, respectively. The concentration of 4-HBA was higher in hermaphrodite than in female cladodes. Some but not all relationships between the probability of detection or the concentration of a given secondary metabolite and some life-history traits confirmed the predictions of ODH. The effect of the sexual form of the concentration of 4-HBA was parallel with the predictions of RCH. The results obtained here show that each of the three secondary metabolites fallowed their own fate. There is no universal rule concerning the production of secondary metabolites. The term “higher levels of defense” used by the hypotheses mentioned is not accurate, since it does not precise if it refers to the extent of plant defended, or to the concentration in some plant tissues.

2014-03-01-2017-06-30

2014-03-01-2017-06-30

Content of secondary metabolites

Harvard Dataverse only

Not embargoed

Supplementary Table S1_2014.xlsx - raw data Supplementary Table S2_2014.xlsx - meteorological data and SEM analysis exploring the effect of meteorological factor on the concentration and incidence of secondary compounds Supplementary Table S3_2014.doc - Descriptive statistics (exploratory analysis) concerning the incidence of secondary compounds Supplementary Table S4_2014.docx - GLIMMIX results concerning the incidence of secondary compounds Supplementary Table S5.docx - GLM results concerning the concentrations of secondary compounds Supplementary Table S6_2014.xlsx - Least square linear regression functions for the relationships between the proportions of cladodes bearing 4-HBA, CGA, and QUE for cladodes arranged by age or by month of sapling. Supplementary Table S7_2014.xlsx - Least square linear regression functions for the relationship between cladode length and cladode order from soil or cladode age. Supplementary Table S8_2014.pdf - Relationship between cladode length and cladode order from soil, and between cladode length and cladode age class, together with slope comparisons for female and hermaphrodite individuals. Supplementary Table S9_2014.xlsx - Least square linear regression functions for the relationship between cladode order from soil or nr of cladodes above, and cladode age estimator based on the color of the spines.

This is the complete data gathered during the part of the study concerning the effect production of secondary compounds by two sexual forms of Opuntia robusta (hermaphrodite and female individuals).

As far as I know, the use of this data does not affect interests of any third party person or organization. It may affect only the authors of the data if not used in accordance of this Term of use.

N/A

If used for commercial purposes, the authors should be contacted and asked the permission. For scientific use: You can use the data for statistical analysis, to generate figure derived from statistical analysis, to generate figure, always when you cite the data together with authors.

Mariusz Krzysztof Janczur majmx@interia.pl majmx@yahoo.com.mx majmx@biolevolmex.com

You should cite the data as provided by Harvard Dataverse.

Do not use this data for publication without citing them. If the data are used extensively in your study, please, consider including the owner of the data as coauthor. Do not provide the data to other persons: each interested person should download them personally from Harvard Dataverse.

Standard Harvard Dataverse conditions

I whereby declare that I financed the collection of the data uploaded here and participated in the fieldwork. Also I declare that I was the project owner.

You can use the data for statistical analysis, to generate figure derived from statistical analysis, to generate figure derived from community ecology methods, always when you cite the data together with authors.

On request

• Number of cases: 4

• No. of variables per record: 1

• Type of File: text/tab-separated-values

UNF:6:1olteIpxz9qqtQ6OSX72Xg==

Morphometric traits of Opuntia robusta from San Nicolas Tecoaco, Municipality of Singuilucan, Hidalgo State, Central Mexico, cladode order counted from soil, number of cladodes above a given clacode, date of sampling, spine color (cladode age estimator) absorbanse (surface belowe the absorbance curve), calibrations equations, and concentration of 4-hydroxybenzoic acid (4-HBA), quercetine (QUE), salicilic acid (SAL), and chlorogenic acid (CGA) obtained from the calibration equations for females and hermafrodite cladodes

• Number of cases: 9

• No. of variables per record: 1

• Type of File: text/tab-separated-values

UNF:6:x7SZC7Cc8qGNqVNPZsXumw==

Results of Structural equation models (SEM) for the relationship between meteorological variables and concentrations of three secondary metabolites or proportions of cladodes bering them in female and hermaphrodite individuals of Opuntia robusta from San Nicolas Tecoaco, Municipality of Singuilucan, Hidalgo State, Mexico 4-HBA - 4-hydroxybenzoic acid CGA - chlorogenic acid QUE - quercetin * - P < 0.05, ** - P < 0.01, and *** - P< 0.001. ** - P < 0.01 *** - P< 0.001. ________________________________ Meteorological data for Singuilucan Municipality, 2014 Meteorological Data 2014 - Averege or additive per-month data Grapphics - Histograms for average and additive variables Graphics Per-Day - Graphics for sampling day Coef Determ - Coefficients of determination and t-test probabolity for correlation coefficiens; per-month average or additive variables Coef Determ_Per-Day - Coefficients of determination and t-test probabolity for correlation coefficiens; averages or sums for additive variables for the samplig day Correl. Per-Month vs. Per-Day - correlations coefficients, coefficients of determination, and t-test probability, fore the relationship among per-month and per-sampling-day data March-October - meteorological data for March through October 2014 Lackind data that were adjusted, are marked with red fonts Data obtained from: clima.inifap.gob.mx

• Number of cases: 6

• No. of variables per record: 7

• Type of File: text/tab-separated-values

UNF:6:kJhCD8HdFzyby9H8hfiRuQ==

Least square linear regression functions for the relationships between the age-dependent proportions of cladodes bearing 4-HBA, CGA, and QUE for cladodes arranged by age or by month of sapling. Cladode age classes corresponded to the color of the spines in the fallowing manner: 1 – yellowish, 2 – yellow, white base, 3 – white yellowish, 4 – white, 5 – grayish, 6 – black, being “1” the youngest, and “6”, the oldest age class. 4-HBA - 4-Hydroxybenzoic acid CGA - Chlorogenic acid QUE - Quercetin

• Number of cases: 5

• No. of variables per record: 1

• Type of File: text/tab-separated-values

UNF:6:2qs54oCSszU2DmkBAAQu9g==

Least square linear regression functions for the relationship between Opuntia robusta's cladode length and cladode order from soil or cladode age. Age estimator is based on the color of the spines. Cladode age classes corresponded to the color of the spines in the fallowing manner: 1 – yellowish, 2 – yellow, white base, 3 – white yellowish, 4 – white, 5 – grayish, 6 – black, being “1” the youngest, and “6”, the oldest age class. We compared sloepes between sexual forms, for these relationships.

• Number of cases: 6

• No. of variables per record: 1

• Type of File: text/tab-separated-values

UNF:6:11UV9Y0u94v9Dhc46xeehg==

Least square linear regression functions for the relationship between cladode order from soil or nr of cladodes above, and cladode age estimator based on the color of the spines.

• Morphometrictraits,cladodeordercountedfromsoil,numberofcladodesaboveagivenclacode,dateofsampling, - Morphometrictraits,cladodeordercountedfromsoil,numberofcladodesaboveagivenclacode,dateofsampling,
• MeteorologicaldataforSinguilucanMunicipality,2014 - MeteorologicaldataforSinguilucanMunicipality,2014
• Leastsquarelinearregressionfunctionsfortherelationshipsbetweentheproportionsofcladodesbearing4-HBA,CGA,andQUE - Leastsquarelinearregressionfunctionsfortherelationshipsbetweentheproportionsofcladodesbearing4-HBA,CGA,andQUE
• B - B
• C - C
• D - D
• E - E
• F - F
• G - G
• Leastsquarelinearregressionfunctionsfortherelationshipbetweencladodelength - Leastsquarelinearregressionfunctionsfortherelationshipbetweencladodelength
• Leastsquarelinearregressionfunctionsfortherelationshipbetweencladodeorderfromsoilornrofcladodesabove, - Leastsquarelinearregressionfunctionsfortherelationshipbetweencladodeorderfromsoilornrofcladodesabove,

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Fig.3.tif

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Monthly dynamics of the of the proportion of cladodes of female and hermaphrodite individuals of Opuntia robusta containing 4-Hydroxybenzoic acid, Chlorogenic acid, and Quercetine, as well as of the concentrations of these metabolites.

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Supplementary Table S3 Exploratory analysis.doc

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Descriptive statistics (exploratory analysis). We obtained the descriptive statistics for the proportion of cladodes bearing 4-Hydroxybenzoic acid (4-HBA), chlorogenic acid (CGA), and Quercetin (QUE), and we looked for the relationship between each dependent and independent variable: we used chi-squared test (sex, month), Cochran-Armitage (C-A) test (cladode age class, number of cladodes above a given cladodes, cladode order above ground), and t-test (size estimators). With the C-A test we looked at how the probability of production of secondary metabolites depended on cladode age, order and the number of cladodes the plant bore. We treated the variables of sex and month as categorical variables, and estimated least square proportions for each cladode order. We considered the other variables to be continuous.

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Supplementary Table S4.docx

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Results of GLMM model for the proportion of cladodes bearing 4-Hydroxybenzoic acid (4-HBA), chlorogenic acid (CGA), and Quercetin (QUE), and we looked for the relationship between each dependent and independent variable. We applied the generalized linear mixed model (GLMM) with logit link function (ln(P/(1-P)), where P – probability of detection of a given metabolite), binomial response distribution, maximum likelihood estimation technique, Newton-Raphson optimization algorithm, and Person Chi-Square/df fit criterion. We used GLIMMIX procedure of the SAS statistical software (SAS Institute Inc., 2017).

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Supplementary Table S5.docx

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Supplementary Table S5. Model statistics for the best models and post-hoc contrasts. The effect of month, cladode size, cladode age cladode order from soil, number of daughter cladodes above a given cladode on the concentrations of 4-Hydroxybenzoic acid (4-HBA), Chlorogenic acid (CGA) and Quercetin (QUE) in female and hermaphrodite cladodes of individuals of Opuntia robusta from Central Mexico. To analyze how concentrations of the different substances were related to cladode length, width, thickness, months, age, cladodes order from soil, and cladodes above a given cladode, we used generalized linear models (GLMs) created in R (R Core Team, 2020). Since many concentrations were null, we analyzed only the positive concentrations. The full model contained the interaction between each individual variable and sex. For each response variable, we used different error distribution and link functions that gave the best fit of the models. To analyze the effect of months and age on 4-HBA concentrations, we used the Gamma error distribution (GED) and the inverse link function. For the effect of months on CGA concentrations, we used GED and log link functions. For the effect of cladode age on CGA concentrations, we used GED and inverse link functions. For the effect of months on QUE concentration, we used GED and log link functions. For the effect of cladode age on QUE concentration, we used GED and inverse link function. For the models that related substances with cladode length, width and thickness, we used different models: for 4-HBA concentrations we used a GED with square root link function. For CGA concentrations, we used GED and identity link function; for QUE concentrations, we used GED and inverse link function. To test the effect of cladodes order from soil on the concentrations of the three substances, we used GED and square root link function. To test the effect of the number of cladodes above a given cladode on the concentrations of the three substances, we used GED and inverse link function. We checked the goodness of fit of the linear models (GLM or GLMM) by plotting the standardized residuals against fitted values and by normal QQ‐plots, and revised the assumptions of the homoscedasticity, proper distribution used, and independence. We chose the best GLM model used in this study according to the lowest AIC (Akaike, 1987) following the recommendations of Zuur et al. (2010). Table 1. The effect of months on the concentrations of 4-Hydroxybenzoic acid (4-HBA) in female and hermaphrodite cladodes of the individuals of Opuntia robusta from San Nicolás Tecoaco (trioecious population), Central Mexico. Model statistics and post-hoc contrasts. Table 2. The effect of months on the concentration of Chlorogenic acid (CGA) in female and hermaphrodite cladodes of the individuals of Opuntia robusta from San Nicolás Tecoaco (trioecious population), Central Mexico. Model statistics and post-hoc contrasts. Table 3. The effect of months on the concentration of Quercetin (QUE) in female and hermaphrodite cladodes of the individuals of Opuntia robusta from San Nicolás Tecoaco (trioecious population), Central Mexico. Model statistics and post-hoc contrasts. Table 4. The effect of cladode age class determined by the color of spines on the concentration of Hydroxybenzoic acid (4-HBA) in female and hermaphrodite cladodes of the individuals of Opuntia robusta from San Nicolás Tecoaco (trioecious population), Central Mexico. Model statistics and post-hoc contrasts. Table 5. The effect of cladode age class determined by the color of spines on the concentration of Chlorogenic acid (CGA) in female and hermaphrodite cladodes of the individuals of Opuntia robusta from San Nicolás Tecoaco (trioecious population), Central Mexico. Model statistics and post-hoc contrasts. Table 6. The effect of cladode age class determined by the color of spines on the concentration of Quercetin (QUE) in female and hermaphrodite cladodes of the individuals of Opuntia robusta from San Nicolás Tecoaco (trioecious population), Central Mexico. Model statistics and post-hoc contrasts. Table 7. The effect of cladode length, width and thickness on the concentrations of 4-Hydroxybenzoic acid (4-HBA), Chlorogenic acid (CGA) and Quercetin (QUE) in female and hermaphrodite cladodes of the individuals of Opuntia robusta from San Nicolás Tecoaco (trioecious population), Central Mexico. Model statistics and post-hoc contrasts. Table 8. The effect of cladode order from soil on the concentrations of 4-Hydroxybenzoic acid (4-HBA), Chlorogenic acid (CGA) and Quercetin (QUE) in female and hermaphrodite cladodes of the individuals of Opuntia robusta from San Nicolás Tecoaco (trioecious population), Central Mexico. Model statistics and post-hoc contrasts. Table 9. The effect of the number of levels of daughter cladodes above a given cladode on the concentrations of 4-Hydroxybenzoic acid (4-HBA), Chlorogenic acid (CGA) and Quercetin (QUE) in female and hermaphrodite cladodes of the individuals of Opuntia robusta from San Nicolás Tecoaco (trioecious population), Central Mexico. Model statistics and post-hoc contrasts.

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Supplementary Table S8_2014.pdf

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Relationship between cladode length and cladode order from soil, and between cladode length and cladode age class, together with slope comparisons for female and hermaphrodite individuals.

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