From 6a15cc06c5187ed54b232adcd37d6b4b03faa7cd Mon Sep 17 00:00:00 2001 From: Stephen Wood Date: Tue, 6 Aug 2019 17:57:47 -0400 Subject: [PATCH] Modify README --- README.md | 69 ++-- .../unnamed-chunk-10-1.png | Bin 38236 -> 0 bytes .../unnamed-chunk-10-2.png | Bin 42276 -> 0 bytes .../unnamed-chunk-10-3.png | Bin 36987 -> 0 bytes .../unnamed-chunk-10-4.png | Bin 36209 -> 0 bytes data-analysis.Rmd | 387 +++++++++++++++++- data/icp.xlsx | Bin 23564 -> 23912 bytes 7 files changed, 408 insertions(+), 48 deletions(-) delete mode 100644 README_files/figure-markdown_strict/unnamed-chunk-10-1.png delete mode 100644 README_files/figure-markdown_strict/unnamed-chunk-10-2.png delete mode 100644 README_files/figure-markdown_strict/unnamed-chunk-10-3.png delete mode 100644 README_files/figure-markdown_strict/unnamed-chunk-10-4.png diff --git a/README.md b/README.md index 25e27c6..55ea979 100644 --- a/README.md +++ b/README.md @@ -1,14 +1,6 @@ Salinas Organic Cropping Systems Experiment ------------------------------------------- -This document walks through the statistical models used by Wood and -Brennan in their analysis of soil carbon fractions at the Salinas -Organic Cropping Systems Experiment. For more information on the methods -and results, please see the paper. For visualizations, see -data-analysis.R. We did not generate a markdown for the visualizations -because not all of the final figures in the manuscript were generated in -R. - ### Read in the data files The data are stored in separate files. usda\_soil\_data has the soil @@ -295,16 +287,16 @@ models. We'll also select a smaller set of predictor variables that might be of special interest. leg_rye <- all_data %>% - filter(cover_crop=="legume-rye") %>% - select( - Replicate, cover_crop, Compost, Cover_crop_freq, organic_matter, water_holding_capacity, - substrate_induced_respiration:POM_CN, `POM C`, `MAOM C`, pom.stock, pom.n.stock, maom.stock, maom.n.stock, - total_C_no_roots_no_exudates, CEC, pH, `X-Ca` - ) + filter(cover_crop=="legume-rye") %>% + select(Replicate, cover_crop, Compost, Cover_crop_freq, organic_matter, + water_holding_capacity,substrate_induced_respiration:POM_CN,pom.stock, + pom.n.stock, maom.stock, maom.n.stock, + total_C_no_roots_no_exudates, CEC, pH, `X-Ca` + ) # Organic matter lmer(organic_matter ~ Compost + Cover_crop_freq + (1|Replicate), - data = leg_rye + data = leg_rye ) -> om.model summary(om.model) @@ -351,7 +343,7 @@ might be of special interest. # Particulate C lmer(pom.stock ~ Compost + Cover_crop_freq + (1|Replicate), - data = leg_rye + data = leg_rye ) -> pom.model ## boundary (singular) fit: see ?isSingular @@ -403,7 +395,7 @@ might be of special interest. # Particulate N lmer(pom.n.stock ~ Compost + Cover_crop_freq + (1|Replicate), - data = leg_rye + data = leg_rye ) -> pom.n.model summary(pom.n.model) @@ -450,7 +442,7 @@ might be of special interest. # Mineral-associated C lmer(maom.stock ~ Compost + Cover_crop_freq + (1|Replicate), - data = leg_rye + data = leg_rye ) -> maom.model summary(maom.model) @@ -495,7 +487,7 @@ might be of special interest. # Mineral-associated N lmer(maom.n.stock ~ Compost + Cover_crop_freq + (1|Replicate), - data = leg_rye + data = leg_rye ) -> maom.n.model summary(maom.n.model) @@ -724,6 +716,9 @@ our outcomes of interest ) ) + ## Warning in VSURF_pred.default(x = x, y = y, ntree = ntree, err.interp = interp$err.interp, : Unable to perform prediction step, because the interpretation step + ## did not eliminate variables + Now we can look at each of these model results and see which variables are the most relevant. This Random Forest procedure generates lists of variables most suitable for both interpretatoin and prediction. Briefly, @@ -736,14 +731,14 @@ identified for interpretation. # Print the response variable print(rf.res[[i]]$call$y) # Print the predictor variables selected - all_data %>% - select(total_compost:total_C_no_roots_no_exudates) %>% + all_data %>% + select(total_compost:total_C_no_roots_no_exudates) %>% select(vars) %>% names() %>% print() } ## all_data$organic_matter - ## [1] "total_C_no_roots_no_exudates" "fresh_om" - ## [3] "total_om" "total_cc_shoot" + ## [1] "total_C_no_roots_no_exudates" "total_om" + ## [3] "fresh_om" "total_cc_shoot" ## [5] "fresh_om_perc" "total_compost" ## [7] "compost_n" ## all_data$pom.stock @@ -753,14 +748,12 @@ identified for interpretation. ## all_data$substrate_induced_respiration ## [1] "annual_cc_shoot" - rm(vars) - Some of these variables are highly correlated with each other so we'll select out the ones that we know to be somewhat independent and fit models with these. # Organic matter - lm.om <- lmer(organic_matter ~ total_C_no_roots_no_exudates + (1|Replicate), data=all_data) + lm.om <- lmer(organic_matter ~ total_C_no_roots_no_exudates + (1 | Replicate), data = all_data) ## Warning: Some predictor variables are on very different scales: consider ## rescaling @@ -815,7 +808,7 @@ models with these. rm(lm.om) # Particulate - lm.pc <- lmer(pom.stock ~ fresh_om + (1|Replicate), data=all_data) + lm.pc <- lmer(pom.stock ~ fresh_om + (1 | Replicate), data = all_data) ## Warning: Some predictor variables are on very different scales: consider ## rescaling @@ -867,7 +860,7 @@ models with these. rm(lm.pc) # Mineral - lm.mc <- lmer(maom.stock ~ total_veg_residue_shoot + annual_legume_cc_shoot + (1|Replicate), data=all_data) + lm.mc <- lmer(maom.stock ~ total_veg_residue_shoot + annual_legume_cc_shoot + (1 | Replicate), data = all_data) ## Warning: Some predictor variables are on very different scales: consider ## rescaling @@ -916,7 +909,7 @@ models with these. rm(lm.mc) # SIR - lm.sir <- lmer(substrate_induced_respiration ~ annual_cc_shoot + (1|Replicate), data=all_data) + lm.sir <- lmer(substrate_induced_respiration ~ annual_cc_shoot + (1 | Replicate), data = all_data) summary(lm.sir) ## Linear mixed model fit by REML. t-tests use Satterthwaite's method [ @@ -990,7 +983,7 @@ were capturing with new fixed effects. ) # Fit model with best management predictors - mgmt.model <- lm(maom.stock ~ Compost+Cover_crop_freq+total_veg_residue_shoot, data=all_data) + mgmt.model <- lm(maom.stock ~ Compost + Cover_crop_freq + total_veg_residue_shoot, data = all_data) mgmt.model %>% summary() ## @@ -1028,7 +1021,7 @@ model does. soil.prop <- VSURF( y = mgmt.model$residuals, x = all_data %>% - select(perc_sand:perc_clay,EC:`Fe (DTPA)`), + select(perc_sand:perc_clay, EC:`Fe (DTPA)`), parallel = TRUE ) @@ -1036,10 +1029,10 @@ model does. ## did not eliminate variables # Create data frame of resulting variables, also including clay - resid.data <- data.frame(mgmt.model$residuals,all_data$perc_clay,all_data$`X-K`,all_data$`Ca (SP)`,all_data$`Mn (DTPA)`) - names(resid.data) <- c('model_residuals','perc_clay','K','Ca','Mn') + resid.data <- data.frame(mgmt.model$residuals, all_data$perc_clay, all_data$`X-K`, all_data$`Ca (SP)`, all_data$`Mn (DTPA)`) + names(resid.data) <- c("model_residuals", "perc_clay", "K", "Ca", "Mn") - resid.model <- lm(model_residuals ~ perc_clay + K + Ca + Mn, data=resid.data) + resid.model <- lm(model_residuals ~ perc_clay + K + Ca + Mn, data = resid.data) resid.model %>% summary() ## @@ -1064,19 +1057,19 @@ model does. ## Multiple R-squared: 0.5442, Adjusted R-squared: 0.4226 ## F-statistic: 4.477 on 4 and 15 DF, p-value: 0.01403 - effect_plot(model=resid.model, pred = Ca, interval = TRUE, rug=TRUE, plot.points = TRUE,partial.residuals=TRUE) + effect_plot(model = resid.model, pred = Ca, interval = TRUE, rug = TRUE, plot.points = TRUE, partial.residuals = TRUE) ![](data-analysis_files/figure-markdown_strict/unnamed-chunk-10-1.png) - effect_plot(model=resid.model, pred = K, interval = TRUE, rug=TRUE, plot.points = TRUE,partial.residuals=TRUE) + effect_plot(model = resid.model, pred = K, interval = TRUE, rug = TRUE, plot.points = TRUE, partial.residuals = TRUE) ![](data-analysis_files/figure-markdown_strict/unnamed-chunk-10-2.png) - effect_plot(model=resid.model, pred = perc_clay, interval = TRUE, rug=TRUE, plot.points = TRUE,partial.residuals=TRUE) + effect_plot(model = resid.model, pred = perc_clay, interval = TRUE, rug = TRUE, plot.points = TRUE, partial.residuals = TRUE) ![](data-analysis_files/figure-markdown_strict/unnamed-chunk-10-3.png) - effect_plot(model=resid.model, pred = Mn, interval = TRUE, rug=TRUE, plot.points = TRUE,partial.residuals=TRUE) + effect_plot(model = resid.model, pred = Mn, interval = TRUE, rug = TRUE, plot.points = TRUE, partial.residuals = TRUE) ![](data-analysis_files/figure-markdown_strict/unnamed-chunk-10-4.png) 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zMSqHBnz7mhBOaBu77pgWZkX3*FMsr@zuNZl>qmV%gzr37#_Kt)?juBu&>PQ6okX~) zVhl*u+Bn+Esj+v7Po>oJb=LfI^?KXV5a{;ny~lrjD6pyGIujOj3q!)A{~5C&L$KXV z{wyZ^&m`6ZllcFeL(tjX-RfJw&<2DdC8}-fX`CDcMlSYNny&;H1F>?|{2<$jmEhv9^AN*K9Fw+eJ$S#f)M72DII{mcFV|gGY5*keEJU(4| zr94l}1fT`#Pr*4TT$oW{bg)4#9{7D2wj}A(dHJGtSTBT7%I>^)Rta-eE*$4e0$1!` z@-g}Yj4qylGXa1(l!=q?m|4662OKeR5#Hjcf{^dzuUFuR0`Il)-47?ey^fnCO%Cc9uRQw}_?iZG z;g3P!)PobT4bMyw*mMwaH*#NRPaI$hg>!ApB^qA^wm-i1y!yUw$X^R{itf50ME(!P8}9+8DeF{bm0$!&d!`. +### Read in the data files -When you click the **Knit** button a document will be generated that includes both content as well as the output of any embedded R code chunks within the document. You can embed an R code chunk like this: +The data are stored in separate files. usda\_soil\_data has the soil +carbon fractionation data analyzed by Wood, icp has the data analyzed by +UC Davis, and the other site-level descriptors are in the other files. -```{r cars, echo = FALSE} +```{r} # READ IN ALL DATA FILES usda_soil_data <- read_excel("data/usda-soil-data.xlsx") %>% mutate( @@ -48,12 +61,366 @@ inputs <- read_excel("data/site-data.xlsx", icp <- read_excel("data/icp.xlsx") ``` -## Including Plots +### Data manipulation + +Now we need to do some slight manipulations to rename variables, merge +the data sets together, and select the variables that we'll want to used +for analysis. + +```{r} +inputsLabels <- colnames(inputs) +colnames(inputs)[4:18] <- c( + "total_compost", + "annual_cc_shoot", + "total_cc_shoot", + "annual_legume_cc_shoot", + "total_legume_cc_shoot", + "n_fixation", + "compost_n", + "fertilizer_n", + "total_n", + "perc_n_from_fixation", + "total_veg_residue_shoot", + "total_om", + "fresh_om", + "fresh_om_perc", + "total_C_no_roots_no_exudates" +) + +inputs$RepTreat <- paste(inputs$trt, inputs$rep) +usda_soil_data$RepTreat <- paste(usda_soil_data$Trt_id, usda_soil_data$Replicate) +bd$RepTreat <- paste(bd$trt, bd$rep) +icp$RepTreat <- paste(icp$Trt_id, icp$Replicate) + +all_data <- merge(usda_soil_data, bd, by = "RepTreat") %>% + select(RepTreat, System, Replicate, Trt_id, cover_crop, Compost:POM_CN, blkden) %>% + mutate( + pom.stock = (`POM C`*blkden*30)/10, + pom.n.stock = (`POM N`*blkden*30)/10, + maom.stock = (`MAOM C`*blkden*30)/10, + maom.n.stock = (`MAOM N`*blkden*30)/10, + totC.stock = pom.stock + maom.stock + ) + +all_data <- all_data %>% + merge(inputs, by = "RepTreat") %>% + select(RepTreat,System,Replicate:totC.stock, total_compost:total_C_no_roots_no_exudates) + +all_data$CC <- recode(all_data$Trt_id, + leg3x="Legume-rye every year", + mus1x="Mustard every year", + nocc="Legume-rye every four years", + noccnocp="Legume-rye every four years", + rye1x="Rye every year") + +all_data <- all_data %>% + merge(icp %>% select(IC,EC:RepTreat), by="RepTreat") + +rm(bd); rm(inputs); rm(usda_soil_data); rm(icp) + ``` + +### Models of management effect on soil fractions + +Now that we have the data, let's look through some models. We are going +to used hierarchical models with a random intercept for replicate block. +This is to capture random differences among plots within a block. + +Let's start with a series of models, using all of the data, and looking +at the impact on particulate and mineral-associated C and microbial +biomass. + +```{r} +# Particulate carbon +lmer(pom.stock ~ Compost + Cover_crop_freq + cover_crop + (1|Replicate), + data = all_data +) -> pom.model +summary(pom.model) +r.squaredGLMM(pom.model) +rm(pom.model) -You can also embed plots, for example: +# Mineral-associated carbon +lmer(maom.stock ~ Compost + Cover_crop_freq + cover_crop + (1|Replicate), + data = all_data +) -> maom.model +summary(maom.model) +r.squaredGLMM(maom.model) +rm(maom.model) -```{r pressure, echo=FALSE} -plot(pressure) +# Microbial biomass +lmer(substrate_induced_respiration ~ Compost + Cover_crop_freq + cover_crop + (1|Replicate), + data = all_data +) -> sir.model +summary(sir.model) +r.squaredGLMM(sir.model) +rm(sir.model) ``` -Note that the `echo = FALSE` parameter was added to the code chunk to prevent printing of the R code that generated the plot. +Looking at the model estimates, there does not seem to be a relationship +between cover crop type and our variables of interest. We could continue +by dropping that variable from the model and re-running, but that would +take the average effect of the other variables across systems with +different cover crops. Even though cover crop type does not have an +effect in our statistical model, for the remaining comparisons we felt +that it makes more sense to only look at the variables that we know have +the same cover crop type. + +Let's filter out observations that are not legume rye, and re-run the +models. We'll also select a smaller set of predictor variables that +might be of special interest. + +```{r} +leg_rye <- all_data %>% +filter(cover_crop=="legume-rye") %>% +select(Replicate, cover_crop, Compost, Cover_crop_freq, organic_matter, + water_holding_capacity,substrate_induced_respiration:POM_CN,pom.stock, + pom.n.stock, maom.stock, maom.n.stock, + total_C_no_roots_no_exudates, CEC, pH, `X-Ca` +) + +# Organic matter +lmer(organic_matter ~ Compost + Cover_crop_freq + (1|Replicate), +data = leg_rye +) -> om.model +summary(om.model) +r.squaredGLMM(om.model) +rm(om.model) + +# Particulate C +lmer(pom.stock ~ Compost + Cover_crop_freq + (1|Replicate), + data = leg_rye +) -> pom.model +summary(pom.model) +r.squaredGLMM(pom.model) +rm(pom.model) + +# Particulate N +lmer(pom.n.stock ~ Compost + Cover_crop_freq + (1|Replicate), + data = leg_rye +) -> pom.n.model +summary(pom.n.model) +r.squaredGLMM(pom.n.model) +rm(pom.n.model) + +# Mineral-associated C +lmer(maom.stock ~ Compost + Cover_crop_freq + (1|Replicate), + data = leg_rye +) -> maom.model +summary(maom.model) +r.squaredGLMM(maom.model) +rm(maom.model) + +# Mineral-associated N +lmer(maom.n.stock ~ Compost + Cover_crop_freq + (1|Replicate), + data = leg_rye +) -> maom.n.model +summary(maom.n.model) +r.squaredGLMM(maom.n.model) +rm(maom.n.model) +``` + +For our model of microbial biomass--measured by substrate induced +respiration--we're going to take a slightly different approach. Whereas +for the previous models we were mainly interested in how soil fractions +respond to management, now we think that microbial biomass could depend +on the stock sizes themselves, which we might want to control for. So we +can look at including stocks as predictor variables. For this we're +going to actually keep all of the data, rather than drop cover crop +type, because visual inspection suggested that microbial biomass might +be slightly higher with leguminous cover crops. + +```{r} +# SIR +lmer(substrate_induced_respiration ~ cover_crop + Compost + Cover_crop_freq + (1|Replicate), + data = all_data +) -> sir.model +summary(sir.model) +r.squaredGLMM(sir.model) +rm(sir.model) + +# SIR, depending on N input to microbes +lmer(substrate_induced_respiration ~ cover_crop + Compost + Cover_crop_freq + pom.n.stock + (1|Replicate), + data = all_data +) -> sir.model.cn +car::vif(sir.model.cn) +summary(sir.model.cn) +r.squaredGLMM(sir.model.cn) +rm(sir.model.cn) +``` + +### Predicting soil fractions by continuous inputs + +There is a certain feedback between some of the treatments and the +drivers of soil fraction. For instance, cover crops increase soil carbon +pools partially because of root exudation. More cover crop biomass can +lead to more root inputs. On plots with compost, cover crop biomass can +be greater, which can mean different plots have different amounts of +carbon going into the soil through cover crops. In other words, cover +crop frequency is not a perfect categorical variable to capture the +amount of carbon going belowground. + +We estimated the quantitative amount of nitrogen and carbon inputs going +into the system. Let's fit some models to explore if any of these +variables predict soil fractions. + +First, we're going to use a machine learning algorithm called Random +Forest to see which of the quantitative input variables are the most +closely related to the fractions that we're interested in. We're going +to create a list of all of the different Random Forest models for all of +our outcomes of interest + +```{r} +rf.res <- list( + VSURF( + y = all_data$organic_matter, + x = all_data %>% + select(total_compost:total_C_no_roots_no_exudates), + parallel = TRUE + ), + VSURF( + y = all_data$pom.stock, + x = all_data %>% + select(total_compost:total_C_no_roots_no_exudates), + parallel = TRUE + ), + VSURF( + y = all_data$maom.stock, + x = all_data %>% + select(total_compost:total_C_no_roots_no_exudates), + parallel = TRUE + ), + VSURF( + y = all_data$substrate_induced_respiration, + x = all_data %>% + select(total_compost:total_C_no_roots_no_exudates), + parallel = TRUE + ) +) +``` + +Now we can look at each of these model results and see which variables +are the most relevant. This Random Forest procedure generates lists of +variables most suitable for both interpretatoin and prediction. Briefly, +prediction is more strict in the number of variables. We're not using +our data for predictive modeling, so we'll look at the variables +identified for interpretation. + +```{r} +for(i in 1:length(rf.res)){ + vars = rf.res[[i]]$varselect.interp + # Print the response variable + print(rf.res[[i]]$call$y) + # Print the predictor variables selected + all_data %>% + select(total_compost:total_C_no_roots_no_exudates) %>% + select(vars) %>% names() %>% print() +} +``` + +Some of these variables are highly correlated with each other so we'll +select out the ones that we know to be somewhat independent and fit +models with these. + +```{r} +# Organic matter +lm.om <- lmer(organic_matter ~ total_C_no_roots_no_exudates + (1 | Replicate), data = all_data) +summary(lm.om) +r.squaredGLMM(lm.om) +rm(lm.om) + +# Particulate +lm.pc <- lmer(pom.stock ~ fresh_om + (1 | Replicate), data = all_data) +summary(lm.pc) +r.squaredGLMM(lm.pc) +rm(lm.pc) + +# Mineral +lm.mc <- lmer(maom.stock ~ total_veg_residue_shoot + annual_legume_cc_shoot + (1 | Replicate), data = all_data) +summary(lm.mc) +r.squaredGLMM(lm.mc) +rm(lm.mc) + +# SIR +lm.sir <- lmer(substrate_induced_respiration ~ annual_cc_shoot + (1 | Replicate), data = all_data) +summary(lm.sir) +r.squaredGLMM(lm.sir) +rm(lm.sir) +``` + +A surprising part of these models is how poorly the fixed effects +predict mineral-associated C, while how well the random effects do. This +says that there's an important component of among-replicate variation +that we are not capturing with fixed effects. Since mineral-associated C +is known to be maintained by physical and chemical properties of the +soil, there might be some soil properties that explain this variation. +We would not expect soil properties to be as important to particulate C, +because that C is mainly in the form of partially broken down plant +material and it is not as reactive with soil surfaces. + +To explore the potential for soil properties to impact +mineral-associated carbon we used a tool called two-stage least squares +regression. This approach, which is more commonly used by economists, +first models the effect of the management practices on +mineral-associated carbon (we've already done this). Then we want to +know how much of the left-over variation we can explain with other soil +properties. To do that, we extract the residuals from the first +regression and regress those against soil properties we think might be +important. + +Let's start with the management model, even though we've already done +this above. First we're going to use Random Forest to identify the most +important management properties for mineral-associated C. Then we'll +used those predictors in a linear model. We're not using random effects +because we want to try to explain the variance that the random effects +were capturing with new fixed effects. + +```{r} +# Determine best management predictors of MAOM +mgmt <- VSURF( + y = all_data$maom.stock, + x = all_data %>% + select(total_compost:total_C_no_roots_no_exudates), + parallel = TRUE +) + +# Fit model with best management predictors +mgmt.model <- lm(maom.stock ~ Compost + Cover_crop_freq + total_veg_residue_shoot, data = all_data) +mgmt.model %>% summary() +``` + +Now that we have our management model, we can use a Random Forest +approach to identify the variables related to the residuals of the +previous model. We can then use those in a model. We'll then plot the +effect of each of the final selected variables on the residuals. This +will show which variables are the most important to explaining the +variation in mineral-associated C that is not explained by management. +We will also look at model fit statistics (like R2) to see how well this +model does. + +```{r} +# Find best non-management predictors of model residuals +soil.prop <- VSURF( + y = mgmt.model$residuals, + x = all_data %>% + select(perc_sand:perc_clay, EC:`Fe (DTPA)`), + parallel = TRUE +) + +# Create data frame of resulting variables, also including clay +resid.data <- data.frame(mgmt.model$residuals, all_data$perc_clay, all_data$`X-K`, all_data$`Ca (SP)`, all_data$`Mn (DTPA)`) +names(resid.data) <- c("model_residuals", "perc_clay", "K", "Ca", "Mn") + +resid.model <- lm(model_residuals ~ perc_clay + K + Ca + Mn, data = resid.data) +resid.model %>% summary() + +effect_plot(model = resid.model, pred = Ca, interval = TRUE, rug = TRUE, plot.points = TRUE, partial.residuals = TRUE) + +effect_plot(model = resid.model, pred = K, interval = TRUE, rug = TRUE, plot.points = TRUE, partial.residuals = TRUE) + +effect_plot(model = resid.model, pred = perc_clay, interval = TRUE, rug = TRUE, plot.points = TRUE, partial.residuals = TRUE) + +effect_plot(model = resid.model, pred = Mn, interval = TRUE, rug = TRUE, plot.points = TRUE, partial.residuals = TRUE) +``` + +The adjusted R2 of this model is over 0.4! 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