From e40d5f406fd093eeb95c5c6dea4691c82e8e35b3 Mon Sep 17 00:00:00 2001 From: Cursor Agent Date: Fri, 25 Sep 2026 11:00:03 +0000 Subject: [PATCH 1/3] Add a composition fan tree, ANCOM-BC2 volcano, and MicrobiotaProcess difftree. The default differential tree is now an abundance heatmap. df2volcano fits ANCOMBC::ancombc2, and engine = "microbiota" draws the biomarker tree when MicrobiotaProcess is installed. Co-authored-by: dvsmutin --- .../aRchiteutis-plot-composition/SKILL.md | 5 +- .../skills/aRchiteutis-plot-difftree/SKILL.md | 8 +- .../aRchiteutis-plot-ordination/SKILL.md | 2 +- DESCRIPTION | 3 + NAMESPACE | 1 + R/plots_composition.R | 125 +++++ R/plots_harness.R | 492 ++++++++++++++++-- R/plots_ordination.R | 234 ++++++--- R/report.R | 7 +- README.md | 12 +- img/composition_tree.png | Bin 0 -> 86094 bytes img/difftree.png | Bin 82477 -> 110309 bytes img/difftree_microbiota.png | Bin 0 -> 180274 bytes img/volcano.png | Bin 82337 -> 27544 bytes man/archi_ancombc_volcano.Rd | 12 + man/archi_composition_tree.Rd | 12 + man/archi_difftree_ggtree.Rd | 14 +- man/archi_difftree_heat.Rd | 12 + man/archi_difftree_microbiota.Rd | 14 + man/archi_geom_fruit.Rd | 19 +- man/archi_legend_groups.Rd | 12 + man/archi_mp_diff.Rd | 12 + man/archi_mpse_abundance.Rd | 12 + man/archi_mpse_from_df.Rd | 12 + man/archi_mpse_taxonomy.Rd | 12 + man/archi_plot_mp_diff.Rd | 12 + man/archi_position_boxx.Rd | 12 + man/archi_report.Rd | 3 +- man/df2composition_tree.Rd | 46 ++ man/df2difftree.Rd | 28 +- man/df2volcano.Rd | 29 +- skills/aRchiteutis-plot-composition/SKILL.md | 5 +- skills/aRchiteutis-plot-difftree/SKILL.md | 8 +- skills/aRchiteutis-plot-ordination/SKILL.md | 2 +- tests/testthat/test-composition.R | 7 + tests/testthat/test-harness-viz.R | 30 ++ tests/testthat/test-ordination.R | 25 +- vignettes.md | 6 +- 38 files changed, 1070 insertions(+), 175 deletions(-) create mode 100644 img/composition_tree.png create mode 100644 img/difftree_microbiota.png create mode 100644 man/archi_ancombc_volcano.Rd create mode 100644 man/archi_composition_tree.Rd create mode 100644 man/archi_difftree_heat.Rd create mode 100644 man/archi_difftree_microbiota.Rd create mode 100644 man/archi_legend_groups.Rd create mode 100644 man/archi_mp_diff.Rd create mode 100644 man/archi_mpse_abundance.Rd create mode 100644 man/archi_mpse_from_df.Rd create mode 100644 man/archi_mpse_taxonomy.Rd create mode 100644 man/archi_plot_mp_diff.Rd create mode 100644 man/archi_position_boxx.Rd create mode 100644 man/df2composition_tree.Rd diff --git a/.cursor/skills/aRchiteutis-plot-composition/SKILL.md b/.cursor/skills/aRchiteutis-plot-composition/SKILL.md index a379a44..0ffaaf0 100644 --- a/.cursor/skills/aRchiteutis-plot-composition/SKILL.md +++ b/.cursor/skills/aRchiteutis-plot-composition/SKILL.md @@ -1,6 +1,6 @@ --- name: aRchiteutis-plot-composition -description: Draw aRchiteutis composition plots (donut, stacked bar, box plot) with df2donut, df2composition, and df2barplot from a tidy count table. Use when the user wants taxonomic composition figures. +description: Draw aRchiteutis composition plots (donut, stacked bar, box plot, fan tree) with df2donut, df2composition, df2barplot, and df2composition_tree from a tidy count table. Use when the user wants taxonomic composition figures. --- # Composition plots @@ -16,8 +16,11 @@ df2composition(dfT) # samples ordered by FPC (PC1) df2composition(dfT, order_samples = "hclust") # or abundance, alpha, none df2barplot(dfC) # box plot per taxon df2barplot(dfC, style = "raincloud") # needs ggviolinbox +df2composition_tree(dfC, top = 20) # ggtree fan + ggtreeExtra abundance boxplots ``` +`df2composition_tree()` follows the ggtree / ggtreeExtra fruit-boxplot layout (`layout = "fan"`, `open.angle = 10`, `ggtree::rotate_tree()`). Pass `tax` with a `phylum` column to colour tips by phylum; otherwise tips are coloured by genus. + Images: `vignettes.md` (donut, composition, barplot). All three return ggplot objects. If the user started from Python / phyloseq / a matrix, load first (`python2r`, `from_phyloseq`, `from_abundance`) then plot the same way. diff --git a/.cursor/skills/aRchiteutis-plot-difftree/SKILL.md b/.cursor/skills/aRchiteutis-plot-difftree/SKILL.md index 812e7a1..1ac30df 100644 --- a/.cursor/skills/aRchiteutis-plot-difftree/SKILL.md +++ b/.cursor/skills/aRchiteutis-plot-difftree/SKILL.md @@ -1,11 +1,11 @@ --- name: aRchiteutis-plot-difftree -description: Draw a differential-abundance tree with aRchiteutis df2difftree. Use for ggtree fruit bars, a metacoder heat tree of log2 fold change, or two-group taxon trees. +description: Draw a differential-abundance tree with aRchiteutis df2difftree. Use for a ggtree abundance heatmap, a fold-change bar, a metacoder heat tree, or a MicrobiotaProcess biomarker tree. --- # Differential tree -`df2difftree()` computes log2 fold change between two levels of a grouping column and draws the largest changes on a genus/species formula tree (`ape::as.phylo`, not `hclust`). The default engine is ggtree: a circular cladogram (`layout = "rectangular"` is the other layout) and a ggtreeExtra fruit bar at each tip. `engine = "metacoder"` draws `metacoder::heat_tree` coloured by that fold change and stops when metacoder is not installed. Pass `contrast` when the grouping column has more than two levels. +`df2difftree()` computes log2 fold change between two levels of a grouping column and draws the largest changes on a genus/species formula tree (`ape::as.phylo`, not `hclust`). The default engine is ggtree: a circular cladogram (`layout = "rectangular"` is the other layout) and a ggtreeExtra heatmap of relative abundance. Tip colour is the log2 fold change. `fruit = "bar"` draws that fold change as a column instead. `engine = "metacoder"` draws `metacoder::heat_tree` and stops when metacoder is not installed. `engine = "microbiota"` runs `MicrobiotaProcess::mp_diff_analysis` and draws the ggtree / ggtreeExtra biomarker tree (phylum highlight, abundance stars, LDA, FDR). It stops when MicrobiotaProcess is not installed. Pass `contrast` when the grouping column has more than two levels. ```r library(aRchiteutis) @@ -14,8 +14,10 @@ legend <- system.file("extdata", "legend.csv", package = "aRchiteutis") df <- get_counts(path, pattern = "decont_b", legend = legend, trim_char = "_") df2difftree(df[df$clade == "S", ], group = "stage", max_tips = 20) -df2difftree(df[df$clade == "S", ], group = "stage", +df2difftree(df[df$clade == "S", ], group = "stage", fruit = "bar", contrast = c("larvae", "pupa"), layout = "rectangular") +df2difftree(df[df$clade == "G", ], group = "stage", engine = "microbiota", + contrast = c("larvae", "pupa"), tax = tax) ``` Pass `tree =` an `ape::phylo` whose tip labels match `taxa` when a taxonomy tree is already built with `ranks_to_tree()` or `taxids_to_tree()`. diff --git a/.cursor/skills/aRchiteutis-plot-ordination/SKILL.md b/.cursor/skills/aRchiteutis-plot-ordination/SKILL.md index c712f09..62a060b 100644 --- a/.cursor/skills/aRchiteutis-plot-ordination/SKILL.md +++ b/.cursor/skills/aRchiteutis-plot-ordination/SKILL.md @@ -22,7 +22,7 @@ Long-table plots (not untidy): ```r df2clust2d(df[df$clade == "G", ], legend_detect = "pupa", top = 40, k_means = 10) -df2volcano(df[df$clade == "G", ], legend_detect = c("pupa", "larvae")) +df2volcano(df[df$clade == "G", ], legend_detect = c("pupa", "larvae")) # ANCOMBC::ancombc2 df2pca_sample( df_untidy(df, clade = "S", scale = "scale", keep_sample_name = FALSE), scale = FALSE, detect = "pupa") diff --git a/DESCRIPTION b/DESCRIPTION index 531521f..09a9493 100644 --- a/DESCRIPTION +++ b/DESCRIPTION @@ -46,10 +46,13 @@ Imports: utils, viridis Suggests: + ANCOMBC, biomformat, ComplexUpset, + ggstar, ggviolinbox, metacoder, + MicrobiotaProcess, phyloseq, qiime2R, reticulate, diff --git a/NAMESPACE b/NAMESPACE index f5fc341..7dcd392 100644 --- a/NAMESPACE +++ b/NAMESPACE @@ -16,6 +16,7 @@ export(df2chord) export(df2clust2d) export(df2cluster) export(df2composition) +export(df2composition_tree) export(df2corrplot) export(df2difftree) export(df2donut) diff --git a/R/plots_composition.R b/R/plots_composition.R index 67bef8e..610629d 100644 --- a/R/plots_composition.R +++ b/R/plots_composition.R @@ -182,3 +182,128 @@ df2barplot <- function(df, ..., style = c("box", "raincloud")) { axis.text.y = ggplot2::element_text(size = 8, face = "italic"), plot.margin = ggplot2::margin(8, 12, 8, 8)) } + +#' Taxonomic composition on a fan tree +#' +#' A fan cladogram drawn with \pkg{ggtree}, tip colour marking a higher rank, +#' and a \pkg{ggtreeExtra} boxplot ring of per-sample relative abundance. The +#' layout is the ggtreeExtra fruit-boxplot example (itself built on ggtree): +#' `layout = "fan"`, `open.angle = 10`, then [ggtree::rotate_tree()]. +#' +#' @param df A tidy table from [get_counts()]. +#' @param tax Optional rank table with a `taxa` column (or row names) matching +#' `df$taxa`, plus Linnaean columns. When `NULL`, genus and species are +#' parsed from the labels and tips are coloured by genus. +#' @param color_rank Rank used to colour tips and boxes. `"phylum"` when that +#' column exists, otherwise genus. +#' @param top Integer. Most abundant taxa to keep. +#' +#' @return A [ggplot2::ggplot] object. +#' +#' @references +#' Yu G, Smith DK, Zhu H, Guan Y, Lam TTY (2017). "ggtree: an R package for +#' visualization and annotation of phylogenetic trees with their covariates +#' and other associated data." *Methods in Ecology and Evolution*, 8, 28–36. +#' \doi{10.1111/2041-210X.12628} +#' +#' Xu S, Dai Z, Guo P, Fu X, Liu S, Zhou L, Tang W, Feng T, Chen M, Zhan L, +#' Wu T, Hu E, Jiang Y, Bo X, Yu G (2021). "ggtreeExtra: Compact Visualization +#' of Richly Annotated Phylogenetic Data." *Molecular Biology and Evolution*, +#' 38, 4039–4042. \doi{10.1093/molbev/msab166} +#' +#' @examples +#' path <- system.file("extdata", package = "aRchiteutis") +#' df <- get_counts(path = path, pattern = "m1[124]_", trim_char = "_") +#' df2composition_tree(df[df$clade == "G", ], top = 12) +#' +#' @export +#' @importFrom rlang .data +df2composition_tree <- function(df, tax = NULL, color_rank = "phylum", top = 25L) { + if (is.null(df)) stop("df2composition_tree needs a tidy table", call. = FALSE) + df <- df_tidy_drop_unclassified(df) + means <- dplyr::summarise(df, m = mean(.data$amount), .by = "taxa") + means <- means[order(-means$m), , drop = FALSE] + means <- utils::head(means, min(as.integer(top), nrow(means))) + if (nrow(means) < 2L) stop("df2composition_tree needs at least two taxa", call. = FALSE) + df <- df[df$taxa %in% means$taxa, , drop = FALSE] + built <- archi_composition_tree(df, tax, color_rank) + archi_prepare_ggtree() + p <- ggtree::ggtree(built$tree, layout = "fan", open.angle = 10, branch.length = "none") + p <- ggtree::`%<+%`(p, built$meta) + p <- p + ggtree::geom_tippoint( + ggplot2::aes(color = .data$rank_color), size = 1.5, show.legend = FALSE + ) + p <- suppressMessages(ggtree::rotate_tree(p, -90)) + boxes <- dplyr::summarise( + df, val = mean(.data$amount) * 100, .by = c("taxa", "sample") + ) + boxes <- boxes[boxes$taxa %in% built$tree$tip.label, , drop = FALSE] + levels <- unique(built$meta$rank_color) + pal <- stats::setNames(viridis::viridis(length(levels)), levels) + p + + archi_geom_fruit( + data = boxes, + mapping = ggplot2::aes( + y = taxa, x = val, group = taxa, fill = rank_color + ), + offset = 0.12, + pwidth = 0.45, + geom = ggplot2::geom_boxplot, + geomname = "geom_boxplot", + position = archi_position_boxx(), + linewidth = 0.2, + outlier.size = 0.5, + outlier.stroke = 0.08, + outlier.shape = 21 + ) + + ggplot2::scale_fill_manual( + values = pal, name = built$rank_title, + guide = ggplot2::guide_legend(keywidth = 0.8, keyheight = 0.8, ncol = 1) + ) + + ggplot2::scale_color_manual(values = pal, guide = "none") + + ggplot2::xlab("Relative abundance (%)") + + ggplot2::theme( + legend.title = ggplot2::element_text(size = 9), + legend.text = ggplot2::element_text(size = 7) + ) +} + +#' Fan-tree tips and the rank used to colour them +#' @keywords internal +archi_composition_tree <- function(df, tax, color_rank) { + labels <- unique(as.character(df$taxa)) + labels <- labels[!is.na(labels) & nzchar(labels)] + if (is.null(tax)) { + genus <- ifelse(grepl(" ", labels), sub(" .*", "", labels), labels) + meta <- data.frame( + label = labels, rank_color = genus, stringsAsFactors = FALSE + ) + return(list(tree = archi_label_tree(labels), meta = meta, rank_title = "Genus")) + } + tax <- as.data.frame(tax, stringsAsFactors = FALSE) + if (!"taxa" %in% names(tax)) tax$taxa <- rownames(tax) + tax$taxa <- as.character(tax$taxa) + names(tax) <- tolower(names(tax)) + tax <- tax[tax$taxa %in% labels, , drop = FALSE] + ranks <- intersect(archi_rank_cols(), names(tax)) + color_rank <- tolower(color_rank) + if (!color_rank %in% names(tax) || all(is.na(tax[[color_rank]]) | !nzchar(tax[[color_rank]]))) { + color_rank <- if ("genus" %in% names(tax)) "genus" else ranks[[length(ranks)]] + } + if (length(ranks) >= 2L && nrow(tax) >= 2L) { + tax$tip_name <- tax$taxa + tree <- ranks_to_tree(tax) + } else { + tree <- archi_label_tree(intersect(labels, tax$taxa)) + } + color <- as.character(tax[[color_rank]][match(tree$tip.label, tax$taxa)]) + missing <- is.na(color) | !nzchar(color) + color[missing] <- ifelse( + grepl(" ", tree$tip.label[missing]), + sub(" .*", "", tree$tip.label[missing]), + tree$tip.label[missing] + ) + meta <- data.frame(label = tree$tip.label, rank_color = color, stringsAsFactors = FALSE) + title <- paste0(toupper(substr(color_rank, 1, 1)), substr(color_rank, 2, nchar(color_rank))) + list(tree = tree, meta = meta, rank_title = title) +} diff --git a/R/plots_harness.R b/R/plots_harness.R index b0a8ccd..f691a42 100644 --- a/R/plots_harness.R +++ b/R/plots_harness.R @@ -272,11 +272,15 @@ archi_optional <- function(pkg, what) { #' Differential abundance tree #' #' Selects the taxa with the largest absolute log2 fold change between two -#' groups. The default engine follows the harness ggtree script: a cladogram -#' (`branch.length = "none"`, circular unless `layout = "rectangular"`) and a -#' [ggtreeExtra::geom_fruit] bar of log2 fold change at each tip. `engine = -#' "metacoder"` colours a metacoder heat tree by the same fold change and stops -#' when \pkg{metacoder} is not installed. +#' groups. The default engine is a \pkg{ggtree} cladogram +#' (`branch.length = "none"`, circular unless `layout = "rectangular"`) with a +#' [ggtreeExtra::geom_fruit] heatmap of relative abundance at each tip. Tip +#' colour is the log2 fold change. `fruit = "bar"` restores the single +#' fold-change column. `engine = "metacoder"` colours a metacoder heat tree by +#' the same fold change and stops when \pkg{metacoder} is not installed. +#' `engine = "microbiota"` runs \pkg{MicrobiotaProcess} `mp_diff_analysis` and +#' draws the ggtree / ggtreeExtra biomarker tree from that vignette; it stops +#' when MicrobiotaProcess is not installed. #' #' @param df A tidy table from [get_counts()]. #' @param group Column index or name with at least two levels (for example the @@ -284,15 +288,16 @@ archi_optional <- function(pkg, what) { #' @param contrast Character vector of length 2 naming the groups. The fold #' change is `log2(contrast[2] / contrast[1])`. #' @param max_tips Integer. How many taxa to keep. -#' @param layout `"circular"` (default, as in the harness) or `"rectangular"`. -#' Ignored when `engine = "metacoder"`. +#' @param layout `"circular"` (default) or `"rectangular"`. Ignored when +#' `engine` is `"metacoder"` or `"microbiota"`. #' @param tree Optional [ape::phylo]. Tips must match `taxa` values. When #' `NULL`, a genus/species formula tree is built from the labels. Used only #' by the ggtree engine. -#' @param engine `"ggtree"` or `"metacoder"`. +#' @param engine `"ggtree"`, `"metacoder"`, or `"microbiota"`. +#' @param fruit `"heatmap"` (default) or `"bar"`. Used only by the ggtree engine. #' @param tax Optional rank table (`kingdom` … `species`, plus `taxa`). Used -#' only by the metacoder engine. When `NULL`, genus and species are parsed -#' from the taxon labels. +#' by the metacoder and microbiota engines. When `NULL`, genus and species +#' are parsed from the taxon labels. #' #' @return A [ggplot2::ggplot] object. #' @@ -307,10 +312,15 @@ archi_optional <- function(pkg, what) { df2difftree <- function(df, group, contrast = NULL, max_tips = 20L, layout = c("circular", "rectangular"), tree = NULL, - engine = c("ggtree", "metacoder"), + engine = c("ggtree", "metacoder", "microbiota"), + fruit = c("heatmap", "bar"), tax = NULL) { layout <- match.arg(layout) engine <- match.arg(engine) + fruit <- match.arg(fruit) + if (identical(engine, "microbiota")) { + return(archi_difftree_microbiota(df, group, contrast, tax, max_tips)) + } lfc <- archi_group_lfc(df, group, contrast = contrast) lfc$abs_lfc <- abs(lfc$log2_lfc) lfc <- lfc[order(-lfc$abs_lfc), , drop = FALSE] @@ -332,35 +342,38 @@ df2difftree <- function(df, group, contrast = NULL, max_tips = 20L, lfc <- lfc[match(keep, lfc$id), , drop = FALSE] if (nrow(lfc) < 2L) stop("Tree and taxa share fewer than two tips", call. = FALSE) tree <- ape::keep.tip(tree, lfc$id) - archi_difftree_ggtree(tree, lfc, layout) + groups <- archi_sample_group(df, group, unique(as.character(df$sample))) + archi_difftree_ggtree(tree, lfc, layout, df, groups, fruit) } -#' ggtree differential tree with a fold-change fruit bar at each tip +#' ggtree differential tree #' -#' Same order as the harness `plot_diff_ggtree`: cladogram, italic tip labels, -#' then [ggtreeExtra::geom_fruit] columns. ggtreeExtra looks up `geom_col` with -#' `do.call` inside its own namespace, where that geom is not imported, so the -#' layer is built from the imported `geom_text` name and then swapped for -#' `ggplot2::geom_col`. +#' Default fruit is a heatmap of relative abundance (samples in the two +#' groups, or the two group means when there are many samples). Tip colour is +#' log2 fold change. `fruit = "bar"` keeps a single fold-change column. +#' ggtreeExtra looks geoms up with `do.call` inside its own namespace, so the +#' layer is built from the imported `geom_text` name and then swapped. #' #' @keywords internal -archi_difftree_ggtree <- function(tree, lfc, layout) { +archi_difftree_ggtree <- function(tree, lfc, layout, df, groups, fruit) { lim <- max(abs(lfc$log2_lfc), na.rm = TRUE) if (!is.finite(lim) || lim <= 0) lim <- 1 is_circ <- identical(layout, "circular") - tip_offset <- if (is_circ) 0.5 else 0.2 - fruit_offset <- if (is_circ) 0 else 0.05 - fruit_pwidth <- if (is_circ) 0.35 else 0.40 + tip_offset <- if (is_circ) 0.35 else 0.15 + fruit_offset <- if (is_circ) 0.08 else 0.05 + fruit_pwidth <- if (identical(fruit, "heatmap")) { + if (is_circ) 0.55 else 0.45 + } else if (is_circ) { + 0.35 + } else { + 0.40 + } display <- as.character(lfc$id) tip_meta <- data.frame( label = display, display = display, is_italic = grepl(" ", display, fixed = TRUE), - stringsAsFactors = FALSE - ) - fruit <- data.frame( - id = display, log2_lfc = as.numeric(lfc$log2_lfc), stringsAsFactors = FALSE ) @@ -373,6 +386,9 @@ archi_difftree_ggtree <- function(tree, lfc, layout) { p <- ggtree::ggtree(tree, layout = "rectangular", branch.length = "none") } p <- ggtree::`%<+%`(p, tip_meta) + p <- p + ggtree::geom_tippoint( + ggplot2::aes(color = log2_lfc), size = 1.6, show.legend = TRUE + ) if (any(tip_meta$is_italic)) { p <- p + ggtree::geom_tiplab( ggplot2::aes(label = display, subset = is_italic), @@ -387,20 +403,45 @@ archi_difftree_ggtree <- function(tree, lfc, layout) { fontface = "plain" ) } + + if (identical(fruit, "bar")) { + bars <- data.frame( + id = display, + lfc = as.numeric(lfc$log2_lfc), + stringsAsFactors = FALSE + ) + p <- p + + archi_geom_fruit( + data = bars, + mapping = ggplot2::aes(y = id, x = lfc, fill = lfc), + offset = fruit_offset, + pwidth = fruit_pwidth + ) + + ggplot2::scale_fill_gradient2( + low = "#1B9E77", mid = "gray80", high = "#D81B60", + midpoint = 0, limits = c(-lim, lim), name = "log2 LFC" + ) + } else { + heat <- archi_difftree_heat(df, groups, lfc) + p <- p + + archi_geom_fruit( + data = heat, + mapping = ggplot2::aes(y = id, x = sample, fill = amount), + offset = fruit_offset, + pwidth = fruit_pwidth, + geom = ggplot2::geom_tile, + geomname = "geom_tile" + ) + + ggplot2::scale_fill_viridis_c(name = "Relative abundance", option = "cividis") + } p <- p + - archi_geom_fruit( - data = fruit, - mapping = ggplot2::aes(y = id, x = log2_lfc, fill = log2_lfc), - offset = fruit_offset, - pwidth = fruit_pwidth - ) + - ggplot2::scale_fill_gradient2( + ggplot2::scale_color_gradient2( low = "#1B9E77", mid = "gray80", high = "#D81B60", midpoint = 0, limits = c(-lim, lim), name = "log2 LFC" ) + ggplot2::theme(legend.position = "right") if (is_circ) { - p <- p + ggplot2::expand_limits(x = c(0, 3)) + p <- p + ggplot2::expand_limits(x = c(0, 4)) } else if (requireNamespace("ggtree", quietly = TRUE) && exists("hexpand", envir = asNamespace("ggtree"), inherits = FALSE)) { p <- p + ggtree::hexpand(0.85) @@ -408,6 +449,30 @@ archi_difftree_ggtree <- function(tree, lfc, layout) { p } +#' Sample or group abundance matrix for the differential-tree heatmap +#' @keywords internal +archi_difftree_heat <- function(df, groups, lfc) { + lev <- c(lfc$group_low[[1]], lfc$group_high[[1]]) + use <- names(groups)[groups %in% lev] + use <- use[order(match(unname(groups[use]), lev), use)] + sub <- df[as.character(df$taxa) %in% lfc$id & as.character(df$sample) %in% use, , drop = FALSE] + if (!nrow(sub)) stop("No samples in the two contrast groups", call. = FALSE) + if (length(use) > 12L) { + sub$sample <- unname(groups[as.character(sub$sample)]) + use <- lev + } + heat <- dplyr::summarise( + sub, amount = mean(.data$amount), .by = c("taxa", "sample") + ) + heat$sample <- factor(as.character(heat$sample), levels = unique(as.character(use))) + data.frame( + id = as.character(heat$taxa), + sample = heat$sample, + amount = as.numeric(heat$amount), + stringsAsFactors = FALSE + ) +} + #' ggplot2 4 renamed is.waive; released ggtree still calls it while drawing #' @keywords internal archi_prepare_ggtree <- function() { @@ -420,31 +485,348 @@ archi_prepare_ggtree <- function() { invisible(NULL) } -#' Fruit column that ggtreeExtra can construct from inside another package +#' Fruit layer that ggtreeExtra can construct from inside another package +#' +#' `geom_text` is imported by ggtreeExtra, so its name check succeeds. +#' `geom_col`, `geom_tile` and `geom_boxplot` are not, and `do.call` of those +#' names inside that namespace fails. The layer is built under the imported +#' name and then swapped. `axis.params` and `grid.params` must be literal +#' `list()` calls: `geom_fruit` captures them with `enquo`. +#' #' @keywords internal -archi_geom_fruit <- function(data, mapping, offset, pwidth) { - # geom_text is imported by ggtreeExtra, so its name check succeeds. - # geom_col is not, and do.call("geom_col") inside that namespace fails. - # axis.params and grid.params must be literal list() calls: geom_fruit - # captures them with enquo and does not evaluate a variable. - fruit <- ggtreeExtra::geom_fruit( - data = data, - geom = geom_text, - mapping = mapping, - offset = offset, - pwidth = pwidth, - orientation = "y", - axis.params = list( - axis = "x", text.size = 1.8, hjust = 1, vjust = 0.5, nbreak = 3 - ), - grid.params = list() - ) - fruit$geom <- ggplot2::geom_col - fruit$geomname <- "geom_col" - fruit$params$position <- ggtreeExtra::position_stackx() +archi_geom_fruit <- function(data = NULL, mapping, offset, pwidth, + geom = ggplot2::geom_col, + geomname = "geom_col", + position = NULL, + ...) { + # Two calls, not do.call: geom_fruit captures axis.params with enquo. + if (is.null(data)) { + fruit <- ggtreeExtra::geom_fruit( + geom = geom_text, + mapping = mapping, + offset = offset, + pwidth = pwidth, + orientation = "y", + axis.params = list( + axis = "x", text.size = 1.8, hjust = 1, vjust = 0.5, nbreak = 3 + ), + grid.params = list() + ) + } else { + fruit <- ggtreeExtra::geom_fruit( + data = data, + geom = geom_text, + mapping = mapping, + offset = offset, + pwidth = pwidth, + orientation = "y", + axis.params = list( + axis = "x", text.size = 1.8, hjust = 1, vjust = 0.5, nbreak = 3 + ), + grid.params = list() + ) + } + fruit$geom <- geom + fruit$geomname <- geomname + # ggtreeExtra forwards orientation = "y". geom_tile and geom_star warn on it. + if (geomname %in% c("geom_tile", "geom_star")) { + fruit$geom <- function(...) { + args <- list(...) + args$orientation <- NULL + do.call(geom, args) + } + } + if (!is.null(position)) { + fruit$params$position <- position + } else if (identical(geomname, "geom_col")) { + fruit$params$position <- ggtreeExtra::position_stackx() + } + extra <- list(...) + if (length(extra)) fruit$params <- utils::modifyList(fruit$params, extra) fruit } +#' Horizontal boxplot position that shifts xmin/xmax, not a missing x column +#' @keywords internal +archi_position_boxx <- function() { + ggplot2::ggproto( + NULL, + getFromNamespace("PositionIdentityx", "ggtreeExtra"), + compute_layer = function(self, data, params, layout) { + if (!is.na(params$hexpand)) { + for (col in c( + "x", "xmin", "xmax", "xlower", "xmiddle", "xupper", + "xmin_final", "xmax_final" + )) { + if (col %in% names(data)) data[[col]] <- data[[col]] + params$hexpand + } + } + data + } + ) +} + +#' MicrobiotaProcess biomarker tree (mp_diff_analysis + ggtree) +#' +#' The layout follows the MicrobiotaProcess biomarker vignette, which is drawn +#' with ggtree and ggtreeExtra: radial taxonomy tree, phylum highlight, +#' per-sample abundance stars, LDA bars and FDR points. +#' +#' @keywords internal +archi_difftree_microbiota <- function(df, group, contrast, tax, max_tips) { + archi_optional("MicrobiotaProcess", "df2difftree(engine = \"microbiota\")") + archi_optional("ggstar", "df2difftree(engine = \"microbiota\")") + archi_optional("phyloseq", "df2difftree(engine = \"microbiota\")") + mpse <- archi_mpse_from_df(df, group, contrast, tax, max_tips) + group_name <- "archi_group" + prepared <- archi_mpse_abundance(mpse) + mpse <- archi_mp_diff(prepared$mpse, prepared$relative, group_name) + tree <- MicrobiotaProcess::mp_extract_tree(mpse, type = "taxatree") + archi_prepare_ggtree() + archi_plot_mp_diff(tree, group_name) +} + +#' Counts, taxonomy and a two-or-more group column as an MPSE +#' @keywords internal +archi_mpse_from_df <- function(df, group, contrast, tax, max_tips) { + df <- df_tidy_drop_unclassified(df) + groups <- archi_sample_group(df, group, unique(as.character(df$sample))) + if (!is.null(contrast)) { + if (length(contrast) != 2L || !all(contrast %in% groups)) { + stop("contrast must be two group levels present in the table", call. = FALSE) + } + groups <- groups[groups %in% contrast] + } + lev <- unique(as.character(groups)) + if (length(lev) < 2L) { + stop("df2difftree(engine = \"microbiota\") needs at least two groups", call. = FALSE) + } + df <- df[as.character(df$sample) %in% names(groups), , drop = FALSE] + totals <- tapply(as.numeric(df$N), as.character(df$taxa), sum) + totals <- sort(totals, decreasing = TRUE) + cap <- max(40L, as.integer(max_tips)) + keep_taxa <- names(totals)[seq_len(min(cap, length(totals)))] + df <- df[as.character(df$taxa) %in% keep_taxa, , drop = FALSE] + mat <- df_untidy(df, amount_from = "N", drop_unclassified = TRUE) + mat <- round(mat) + mat[mat < 0 | !is.finite(mat)] <- 0 + storage.mode(mat) <- "integer" + mat <- mat[, intersect(names(groups), colnames(mat)), drop = FALSE] + tax_mat <- archi_mpse_taxonomy(rownames(mat), tax) + sam <- data.frame( + archi_group = factor(unname(groups[colnames(mat)]), levels = lev), + row.names = colnames(mat), + stringsAsFactors = FALSE + ) + ps <- phyloseq::phyloseq( + phyloseq::otu_table(mat, taxa_are_rows = TRUE), + phyloseq::tax_table(as.matrix(tax_mat)), + phyloseq::sample_data(sam) + ) + MicrobiotaProcess::as.MPSE(ps) +} + +#' Capitalised rank table indexed by taxon label +#' @keywords internal +archi_mpse_taxonomy <- function(taxa, tax) { + ranks <- c("Kingdom", "Phylum", "Class", "Order", "Family", "Genus", "Species") + out <- as.data.frame( + matrix(NA_character_, nrow = length(taxa), ncol = length(ranks), + dimnames = list(taxa, ranks)), + stringsAsFactors = FALSE + ) + if (is.null(tax)) { + genus <- ifelse(grepl(" ", taxa), sub(" .*", "", taxa), taxa) + out$Kingdom <- "Bacteria" + out$Genus <- genus + } else { + tax <- as.data.frame(tax, stringsAsFactors = FALSE) + if (!"taxa" %in% names(tax)) tax$taxa <- rownames(tax) + names(tax) <- tolower(names(tax)) + tax <- tax[match(taxa, as.character(tax$taxa)), , drop = FALSE] + for (rk in tolower(ranks)) { + if (!rk %in% names(tax)) next + out[[paste0(toupper(substr(rk, 1, 1)), substr(rk, 2, nchar(rk)))]] <- as.character(tax[[rk]]) + } + if (all(is.na(out$Genus))) { + out$Genus <- ifelse(grepl(" ", taxa), sub(" .*", "", taxa), taxa) + } + if (all(is.na(out$Kingdom))) out$Kingdom <- "Bacteria" + } + # convert_to_treedata uses the rank strings as node ids, then adds the row + # name as the tip. A tip that repeats a rank name makes an invalid phylo. + # Rank prefixes keep those ids apart. Empty cells stay NA so fillNAtax can + # fill them without copying the parent name. + for (rk in ranks) { + x <- as.character(out[[rk]]) + ok <- !is.na(x) & nzchar(x) + x[ok] <- paste0(substr(tolower(rk), 1, 1), "__", x[ok]) + out[[rk]] <- x + } + as.matrix(out) +} + +#' mp_diff_analysis, relaxed once if the first pass finds no biomarkers +#' @keywords internal +archi_mp_diff <- function(mpse, relative, group_name) { + # A failed edge check inside tidytree prints this even when the taxatree + # that mp_extract_tree returns is a usable phylo. + old <- options(check.tbl_tree.verbose = FALSE) + on.exit(options(old), add = TRUE) + run <- function(alpha, lda) { + rlang::inject(MicrobiotaProcess::mp_diff_analysis( + mpse, + .abundance = !!rlang::sym(relative), + .group = !!rlang::sym(group_name), + first.test.alpha = alpha, + filter.p = "pvalue", + strict = FALSE, + cl.test = FALSE, + subcl.test = FALSE, + cl.min = 2, + subcl.min = 2, + ldascore = lda, + bootnums = 20 + )) + } + fit <- run(0.05, 2) + tree <- tryCatch( + MicrobiotaProcess::mp_extract_tree(fit, type = "taxatree"), + error = function(e) NULL + ) + has_lda <- !is.null(tree) && "LDAmean" %in% names(tibble::as_tibble(tree)) && + any(!is.na(tibble::as_tibble(tree)$LDAmean)) + if (!has_lda) { + fit2 <- tryCatch(run(0.2, 1), error = function(e) NULL) + if (!is.null(fit2)) fit <- fit2 + } + fit +} + +#' Rarefy when possible, then store a relative-abundance assay +#' @keywords internal +archi_mpse_abundance <- function(mpse) { + rare <- tryCatch( + MicrobiotaProcess::mp_rrarefy(mpse, action = "add"), + error = function(e) NULL + ) + if (!is.null(rare)) mpse <- rare + assays <- names(SummarizedExperiment::assays(mpse)) + raw <- if ("RareAbundance" %in% assays) "RareAbundance" else "Abundance" + mpse <- rlang::inject(MicrobiotaProcess::mp_cal_abundance( + mpse, .abundance = !!rlang::sym(raw), action = "add" + )) + mpse <- rlang::inject(MicrobiotaProcess::mp_cal_abundance( + mpse, .abundance = !!rlang::sym(raw), .group = archi_group, action = "add" + )) + assays <- names(SummarizedExperiment::assays(mpse)) + relative <- assays[grepl("^Rel", assays) & grepl("Sample", assays)] + if (!length(relative)) relative <- assays[grepl("^Rel", assays)] + if (!length(relative)) { + stop("MicrobiotaProcess did not store a relative-abundance assay", call. = FALSE) + } + list(mpse = mpse, relative = relative[[1]]) +} + +#' ggtree drawing of an mp_diff_analysis taxatree +#' @keywords internal +archi_plot_mp_diff <- function(tree, group_name) { + td <- tibble::as_tibble(tree) + sign_col <- paste0("Sign_", group_name) + if (!sign_col %in% names(td)) { + sign_col <- grep("^Sign_", names(td), value = TRUE) + sign_col <- if (length(sign_col)) sign_col[[1]] else NA_character_ + } + nested <- grep("^RareAbundanceBySample$", names(td), value = TRUE) + if (!length(nested)) nested <- grep("^RareAbundance", names(td), value = TRUE)[[1]] + stars <- tidyr::unnest(tibble::as_tibble(td), tidyr::all_of(nested)) + rel_col <- grep("^RelRare", names(stars), value = TRUE) + if (!length(rel_col)) rel_col <- grep("Rel", names(stars), value = TRUE) + rel_col <- rel_col[[1]] + grp_col <- if (group_name %in% names(stars)) { + group_name + } else { + setdiff(names(stars), c(names(td), rel_col, "Sample", nested))[[1]] + } + stars$rel <- as.numeric(stars[[rel_col]]) + stars$grp <- as.character(stars[[grp_col]]) + if (!"Sample" %in% names(stars)) { + sample_col <- setdiff(names(stars), c(names(td), rel_col, "rel", "grp", nested)) + sample_col <- sample_col[vapply(stars[sample_col], is.atomic, logical(1))] + names(stars)[names(stars) == sample_col[[1]]] <- "Sample" + } + stars <- stars[, c("label", "Sample", "rel", "grp"), drop = FALSE] + levels <- unique(stars$grp) + levels <- levels[!is.na(levels) & nzchar(levels)] + pal <- stats::setNames( + rep_len(c("#1B9E77", "#D95F02", "#7570B3", "#E7298A", "#66A61E"), length(levels)), + levels + ) + p <- ggtree::ggtree(tree, layout = "radial", size = 0.3, branch.length = "none") + internal <- p$data[!p$data$isTip, , drop = FALSE] + p <- p + ggtree::geom_point(data = internal, fill = "white", size = 1, shape = 21) + phyla <- p$data[!is.na(p$data$nodeClass) & p$data$nodeClass == "Phylum", , drop = FALSE] + if (nrow(phyla)) { + phyla$phylum <- phyla$label + phyla$label <- NULL + p <- p + ggtree::geom_hilight( + data = phyla, + ggplot2::aes(node = node, fill = phylum) + ) + } + p <- p + + ggnewscale::new_scale_fill() + + archi_geom_fruit( + data = stars, + mapping = ggplot2::aes( + y = label, x = Sample, size = rel, fill = grp, subset = rel > 0 + ), + offset = 0.04, + pwidth = 0.8, + geom = ggstar::geom_star, + geomname = "geom_star", + starshape = 13, + starstroke = 0.25 + ) + + ggplot2::scale_size_continuous(name = "Relative abundance (%)", range = c(0.5, 3)) + + ggplot2::scale_fill_manual(values = pal, name = group_name) + + ggtree::geom_tiplab(size = 2, offset = 7.2) + if ("LDAmean" %in% names(td) && !is.na(sign_col) && sign_col %in% names(td)) { + p <- p + + ggnewscale::new_scale_fill() + + archi_geom_fruit( + data = NULL, + mapping = rlang::inject(ggplot2::aes( + x = LDAmean, + fill = !!rlang::sym(sign_col), + subset = !is.na(LDAmean) + )), + offset = 0.3, + pwidth = 0.5, + geom = ggplot2::geom_col, + geomname = "geom_col" + ) + + ggnewscale::new_scale("size") + + ggtree::geom_point( + data = p$data[!is.na(p$data[[sign_col]]), , drop = FALSE], + mapping = rlang::inject(ggplot2::aes( + size = -log10(pmax(fdr, .Machine$double.xmin)), + fill = !!rlang::sym(sign_col) + )), + shape = 21 + ) + + ggplot2::scale_size_continuous(range = c(1, 3), name = expression(-log[10]~FDR)) + + ggplot2::scale_fill_manual(values = pal, name = "Biomarker") + } + p + ggplot2::theme( + legend.key.height = ggplot2::unit(0.3, "cm"), + legend.key.width = ggplot2::unit(0.3, "cm"), + legend.spacing.y = ggplot2::unit(0.02, "cm"), + legend.text = ggplot2::element_text(size = 7), + legend.title = ggplot2::element_text(size = 9) + ) +} + #' metacoder heat tree coloured by log2 fold change #' @keywords internal archi_difftree_metacoder <- function(df, lfc, tax) { diff --git a/R/plots_ordination.R b/R/plots_ordination.R index 85eba4b..281299f 100644 --- a/R/plots_ordination.R +++ b/R/plots_ordination.R @@ -515,92 +515,186 @@ df2tsne <- function(df, color = "clust", k_means = 10, text_top = FALSE, ggplot2::theme(plot.margin = ggplot2::margin(14, 14, 14, 14)) } -#' Volcano plot of abundance change between two sample groups +#' Volcano plot of ANCOM-BC2 log fold change #' -#' Runs a per-taxon t-test between the two groups defined by `legend_detect` and -#' plots the log abundance change against the t-test p-value. +#' Fits [ANCOMBC::ancombc2] on raw counts (`N`) for the two groups matched by +#' `legend_detect` (sample names or legend fields). Counts are passed as a +#' \pkg{phyloseq} object, which ANCOM-BC2 converts to a tree experiment. The +#' x axis is the bias-corrected log2 fold change of the second group against +#' the first, and the y axis is `-log10` of the adjusted q-value. Taxa with +#' no counts in one of the two groups are omitted: ANCOM-BC2 cannot estimate +#' a sampling variance for them. The function stops when \pkg{ANCOMBC} or +#' \pkg{phyloseq} is not installed. #' #' @param df A tidy `tibble` from [get_counts()] with legend columns. #' @param legend_detect Length-2 character vector of patterns identifying the -#' two groups. -#' @param treshhold_logAC Numeric. Absolute log abundance-change threshold for +#' two groups. The first pattern is the ANCOM-BC reference level. +#' @param treshhold_logAC Numeric. Absolute log2 fold-change threshold for #' labelling. -#' @param treshhold_p Numeric. P-value threshold for labelling. +#' @param treshhold_p Numeric. Adjusted q-value threshold for labelling. #' #' @return A [ggplot2::ggplot] object. #' #' @examples #' path <- system.file("extdata", package = "aRchiteutis") #' legend <- system.file("extdata", "legend.csv", package = "aRchiteutis") -#' df <- get_counts(path = path, pattern = "m[1345][0-9]?_", legend = legend, -#' trim_char = "_") -#' df2volcano(df[df$clade == "G", ], legend_detect = c("pupa", "larvae")) +#' df <- get_counts(path = path, pattern = "m(11|12|13|18|4|39)_", +#' legend = legend, trim_char = "_") +#' if (requireNamespace("ANCOMBC", quietly = TRUE) && +#' requireNamespace("phyloseq", quietly = TRUE)) { +#' g <- df[df$clade == "G", ] +#' keep <- names(sort(tapply(g$N, g$taxa, sum), decreasing = TRUE))[seq_len(30)] +#' df2volcano(g[g$taxa %in% keep, ], legend_detect = c("larvae", "pupa")) +#' } #' #' @export #' @importFrom rlang .data df2volcano <- function(df, legend_detect, treshhold_logAC = 0.5, treshhold_p = 0.05) { + archi_optional("ANCOMBC", "df2volcano") + archi_optional("phyloseq", "df2volcano") + if (length(legend_detect) != 2L) { + stop("legend_detect must name two groups", call. = FALSE) + } + res <- archi_ancombc_volcano(df, legend_detect) + res$neglog10q <- -log10(pmax(res$q, .Machine$double.xmin)) + res$diffexpressed <- "NS" + ok <- !is.na(res$log2_lfc) & !is.na(res$q) + res$diffexpressed[ok & res$q < treshhold_p & res$log2_lfc >= treshhold_logAC] <- "Up" + res$diffexpressed[ok & res$q < treshhold_p & res$log2_lfc <= -treshhold_logAC] <- "Down" + res$label <- res$taxa + res$label[res$diffexpressed == "NS"] <- NA_character_ + + ggplot2::ggplot(res, ggplot2::aes(.data$log2_lfc, .data$neglog10q, + color = .data$diffexpressed)) + + ggplot2::geom_point(alpha = 0.75, size = 1.6) + + ggplot2::geom_hline(yintercept = -log10(treshhold_p), linetype = 2, + linewidth = 0.3) + + ggplot2::geom_vline(xintercept = c(-treshhold_logAC, treshhold_logAC), + linetype = 2, linewidth = 0.3) + + ggrepel::geom_text_repel( + ggplot2::aes(label = .data$label), size = 2.4, max.overlaps = 15, + show.legend = FALSE, na.rm = TRUE, seed = 1 + ) + + ggplot2::scale_color_manual( + values = c(Up = "#D81B60", Down = "#1B9E77", NS = "grey70"), + name = NULL + ) + + ggplot2::labs( + x = expression(log[2]~fold~change), + y = expression(-log[10]~adjusted~italic(q)) + ) + + ggplot2::theme_bw(base_size = 11) + + ggplot2::theme(legend.position = "top") +} - df_un <- df %>% df_untidy(keep_sample_name = FALSE) - df1 <- df_un[, stringr::str_detect(colnames(df_un), legend_detect[1]), - drop = FALSE] - df2 <- df_un[, stringr::str_detect(colnames(df_un), legend_detect[2]), - drop = FALSE] - - t_df <- vapply(seq_len(nrow(df_un)), function(i) { - if ((sum(df1[i, ]) != 0) && (sum(df2[i, ]) != 0)) { - tryCatch(stats::t.test(df1[i, ], df2[i, ], paired = FALSE)$p.value, - error = function(e) NA_real_) - } else { - NA_real_ +#' ANCOM-BC2 log2 fold change for two legend groups +#' @keywords internal +archi_ancombc_volcano <- function(df, legend_detect) { + grouped <- archi_legend_groups(df, legend_detect) + if (length(grouped$samples1) < 2L || length(grouped$samples2) < 2L) { + stop( + "ANCOM-BC2 needs at least two samples in each group (found ", + length(grouped$samples1), " and ", length(grouped$samples2), ")", + call. = FALSE + ) + } + keep <- c(grouped$samples1, grouped$samples2) + sub <- df[as.character(df$sample) %in% keep, , drop = FALSE] + mat <- df_untidy(sub, amount_from = "N", drop_unclassified = TRUE) + mat <- mat[, intersect(keep, colnames(mat)), drop = FALSE] + mat <- round(mat) + mat[mat < 0 | !is.finite(mat)] <- 0 + storage.mode(mat) <- "integer" + in1 <- colnames(mat) %in% grouped$samples1 + # A taxon missing from an entire group has a zero sampling variance, and + # ANCOM-BC2 stops in the bias step. Keep taxa seen in both groups. + seen <- rowSums(mat[, in1, drop = FALSE] > 0) > 0 & + rowSums(mat[, !in1, drop = FALSE] > 0) > 0 + mat <- mat[seen & apply(mat, 1, stats::var) > 0, , drop = FALSE] + if (nrow(mat) < 2L) { + stop("ANCOM-BC2 needs taxa present in both groups", call. = FALSE) + } + group <- factor( + ifelse(colnames(mat) %in% grouped$samples1, legend_detect[[1]], legend_detect[[2]]), + levels = legend_detect + ) + sam <- phyloseq::sample_data(data.frame( + group = group, row.names = colnames(mat), stringsAsFactors = FALSE + )) + fit <- NULL + for (attempt in seq_len(8L)) { + ps <- phyloseq::phyloseq( + phyloseq::otu_table(mat, taxa_are_rows = TRUE), sam + ) + fit <- tryCatch( + ANCOMBC::ancombc2( + data = ps, + fix_formula = "group", + p_adj_method = "fdr", + prv_cut = 0.1, + lib_cut = 0, + group = "group", + struc_zero = FALSE, + neg_lb = FALSE, + pseudo = 0, + pseudo_sens = FALSE, + global = FALSE, + pairwise = FALSE, + verbose = FALSE, + n_cl = 1L + ), + error = function(e) e + ) + if (!inherits(fit, "error")) break + msg <- conditionMessage(fit) + if (!grepl("Zero variances have been detected", msg, fixed = TRUE)) { + stop(msg, call. = FALSE) } - }, numeric(1)) - - t_df <- data.frame(taxa = row.names(df_un), - p = -log10(t_df), - sd = apply(df_un, 1, stats::sd)) - - df_str <- apply(df[, -c(1:6)], 1, - function(z) stringr::str_c(z, collapse = "_")) - df1mean <- dplyr::summarise( - df[stringr::str_detect(df_str, legend_detect[1]), ], - x = mean(amount), .by = "taxa") - df2mean <- dplyr::summarise( - df[stringr::str_detect(df_str, legend_detect[2]), ], - y = mean(amount), .by = "taxa") - - res <- as.data.frame( - dplyr::left_join(dplyr::full_join(df1mean, df2mean, by = "taxa"), - t_df, by = "taxa")) - res[is.na(res)] <- 0 - res$logAC <- log10(res$y / res$x) - res$logAC[!is.finite(res$logAC)] <- NA_real_ - res$p[!is.finite(res$p)] <- NA_real_ - - res$taxa[(abs(res$logAC) < treshhold_logAC) | - (res$p < -log10(treshhold_p))] <- NA - - ggplot2::ggplot(res, ggplot2::aes(.data$logAC, .data$p)) + - ggplot2::geom_point(ggplot2::aes(color = abs(.data$logAC * .data$p), - size = .data$x + .data$y), - show.legend = FALSE) + - ggplot2::geom_hline(yintercept = -log10(treshhold_p), linetype = 3, - alpha = 0.5, color = "red") + - ggplot2::geom_vline(xintercept = treshhold_logAC, linetype = 3, - alpha = 0.5, color = "red") + - ggplot2::geom_vline(xintercept = -treshhold_logAC, linetype = 3, - alpha = 0.5, color = "red") + - ggrepel::geom_label_repel(ggplot2::aes(label = .data$taxa), - size = 2.8, max.overlaps = 25, - box.padding = 0.4, min.segment.length = 0, - seed = 1, na.rm = TRUE) + - ggplot2::scale_color_gradient("logAC", high = "blue", low = "gray", - na.value = "blue") + - ggplot2::scale_x_continuous(expand = ggplot2::expansion(mult = 0.08)) + - ggplot2::scale_y_continuous(expand = ggplot2::expansion(mult = 0.08)) + - ggplot2::coord_cartesian(clip = "off") + - ggplot2::xlab("log10 amount change") + - ggplot2::ylab("p-value by t.test") + - ggplot2::theme_minimal(base_size = 11) + - ggplot2::theme(plot.margin = ggplot2::margin(14, 14, 14, 14)) + inner <- sub(".*following taxa:\\s*", "", msg) + inner <- sub("\\s*Please remove these taxa.*", "", inner) + bad <- trimws(strsplit(inner, ",\\s*")[[1]]) + bad <- bad[nzchar(bad)] + mat <- mat[!rownames(mat) %in% bad, , drop = FALSE] + if (nrow(mat) < 2L || !length(bad)) stop(msg, call. = FALSE) + } + if (inherits(fit, "error")) stop(conditionMessage(fit), call. = FALSE) + res <- fit$res + if (is.null(res) || !nrow(res)) stop("ANCOM-BC2 returned no taxa", call. = FALSE) + lfc_col <- grep("^lfc_", names(res), value = TRUE) + lfc_col <- setdiff(lfc_col, "lfc_(Intercept)") + if (!length(lfc_col)) stop("ANCOM-BC2 returned no log-fold-change column", call. = FALSE) + term <- sub("^lfc_", "", lfc_col[[1]]) + q_col <- paste0("q_", term) + if (!q_col %in% names(res)) q_col <- paste0("p_", term) + taxon <- if ("taxon" %in% names(res)) res$taxon else res[[1]] + data.frame( + taxa = as.character(taxon), + log2_lfc = as.numeric(res[[lfc_col[[1]]]]), + q = as.numeric(res[[q_col]]), + stringsAsFactors = FALSE + ) +} + +#' Samples matching two legend patterns +#' @keywords internal +archi_legend_groups <- function(df, legend_detect) { + meta_cols <- setdiff(names(df), c( + "taxa", "clade", "sample", "N", "amount", "amount_cl" + )) + samples <- unique(as.character(df$sample)) + if (length(meta_cols)) { + meta <- unique(df[, c("sample", meta_cols), drop = FALSE]) + key <- apply(meta[, meta_cols, drop = FALSE], 1, function(z) paste(z, collapse = "_")) + names(key) <- as.character(meta$sample) + } else { + key <- stats::setNames(samples, samples) + } + key[is.na(key)] <- "" + samples1 <- names(key)[stringr::str_detect(key, legend_detect[[1]]) | + stringr::str_detect(names(key), legend_detect[[1]])] + samples2 <- names(key)[stringr::str_detect(key, legend_detect[[2]]) | + stringr::str_detect(names(key), legend_detect[[2]])] + samples2 <- setdiff(samples2, samples1) + list(samples1 = unique(samples1), samples2 = unique(samples2)) } diff --git a/R/report.R b/R/report.R index a6f3edb..3f9c442 100644 --- a/R/report.R +++ b/R/report.R @@ -222,7 +222,8 @@ archi_report_catalog <- function(beta_method, order_samples, style) { rarefaction = "Observed, Shannon and Simpson rarefaction. Thin line per sample, loess by target.", heattree = "metacoder heat tree. Node size is the number of taxa; colour is mean relative abundance. Empty ranks are dropped so the taxonomy is one tree. Requires the optional metacoder package.", upset = "Taxon presence intersections across target levels, drawn with ComplexUpset. Requires the optional ComplexUpset package.", - difftree = "Log2 fold change between two target levels on a ggtree layout. A ggtreeExtra fruit bar at each tip shows the log2 fold change. engine = \"metacoder\" uses metacoder::heat_tree and requires the optional metacoder package." + composition_tree = "Fan cladogram (ggtree) with a ggtreeExtra boxplot of relative abundance, coloured by phylum.", + difftree = "Log2 fold change between two target levels on a ggtree layout. The default fruit is a heatmap of relative abundance; tip colour is the fold change. engine = \"metacoder\" uses metacoder::heat_tree and requires the optional metacoder package. engine = \"microbiota\" uses MicrobiotaProcess::mp_diff_analysis and requires the optional MicrobiotaProcess package." ) } @@ -256,6 +257,7 @@ archi_report_draw <- function(id, df, tax, target_col, beta_method, order_sample df, split_by = "target", depths = rarefaction_depths, n_reps = rarefaction_reps, top = top ), + composition_tree = df2composition_tree(trimmed, tax = tax, top = top), heattree = df2heattree(trimmed, tax = tax, top = top), upset = df2upset(df, group = "target"), difftree = df2difftree( @@ -400,7 +402,8 @@ archi_write_report_html <- function(path, dropped, sections, disclaimer) { #' @param curve_fraction Terminal rarefaction fraction used by the curve check. #' @param curve_reps Replicates used by the curve check. #' @param plots Character vector of plot ids. Any of `composition`, `donut`, -#' `barplot`, `alpha`, `beta`, `rarefaction`, `heattree`, `upset`, `difftree`. +#' `barplot`, `alpha`, `beta`, `rarefaction`, `composition_tree`, `heattree`, +#' `upset`, `difftree`. #' @param beta_method Name passed to [df2beta()], including phyloseq distances #' and `"aitchison"` when the matching package is installed. #' @param order_samples Sample order for [df2composition()]. diff --git a/README.md b/README.md index a6c4a02..80598e2 100644 --- a/README.md +++ b/README.md @@ -82,6 +82,10 @@ Figures below are rendered from the bundled honey-bee brood reports. `df2barplot()` — amount per taxon. `style = "raincloud"` draws a ggviolinbox raincloud. +`df2composition_tree()` — fan cladogram from [ggtree](https://doi.org/10.1111/2041-210X.12628) (Yu et al. 2017) with a [ggtreeExtra](https://doi.org/10.1093/molbev/msab166) boxplot of relative abundance (Xu et al. 2021). Tips and boxes are coloured by phylum when a rank table is available. + +Fan tree of composition + Raincloud of taxon amounts ### Diversity @@ -112,10 +116,14 @@ Figures below are rendered from the bundled honey-bee brood reports. Taxonomic heat tree -`df2difftree()` — log2 fold change between two groups on a ggtree layout, with a \pkg{ggtreeExtra} fruit bar at each tip. With more than two groups, pass `contrast`. `engine = "metacoder"` draws the same fold changes with `metacoder::heat_tree` and stops when that package is not installed. +`df2difftree()` — log2 fold change between two groups on a ggtree layout. The default fruit is a heatmap of relative abundance; tip colour is the fold change. `fruit = "bar"` draws the fold change as a column. With more than two groups, pass `contrast`. `engine = "metacoder"` draws `metacoder::heat_tree` and stops when that package is not installed. `engine = "microbiota"` runs MicrobiotaProcess `mp_diff_analysis` and stops when that package is not installed. Differential abundance tree +`engine = "microbiota"` draws the MicrobiotaProcess biomarker tree (radial ggtree, phylum highlight, abundance stars, LDA and FDR) when that package is installed. + +MicrobiotaProcess differential tree + `df2upset()` — taxon presence as a ComplexUpset plot. The function stops when \pkg{ComplexUpset} is not installed. UpSet of taxon presence @@ -148,7 +156,7 @@ Figures below are rendered from the bundled honey-bee brood reports. t-SNE of taxa -`df2volcano()` — log abundance change between two legend groups. +`df2volcano()` — ANCOM-BC2 log2 fold change against adjusted q. The first `legend_detect` pattern is the reference group. The function stops when \pkg{ANCOMBC} is not installed. 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z-#Db;w|@yH3Z6@K+n8h~-?`t##pN&y2%2G4C2)?_dqrqQg^5T(LBZ>ITf}}=+{5G4 z`4}6*Im6AD$qH$riB=Bs2a$tkeii>fV<+ffk0`r%pxi3A zyoT;`x4F{e>7?qnj|wnMOJ7bknFxrz!8now7V z9I)VNThfHPcz6bWmxq99g@o!F8%0O<+Ko#t&yNi~mf}Io<|XHp_dy>}|9uU;n+d1-L5{0!v*^aF_DdGDE3Q_bmN zGzH!=G=)G8(870oeB9!>!Cm{J8?&&tJKT^r;~is`k%mCboNm|c>|>bD!QMLEgOCb4 zqs5I6pn4FKNtwA=2FwOhT^>Gq1Q;np6)q$wn4{i!k3=-p8d5#`mnVEcwnvoT#R9-V zhPt{N&VqMvacygs5`rjw$)0rH!SCmV= -### `df2volcano` — log-amount change between groups +### `df2volcano` — ANCOM-BC2 log fold change + +The first `legend_detect` pattern is the reference group. Requires the optional packages ANCOMBC and phyloseq. ```r -df2volcano(df[df$clade == "G", ], legend_detect = c("pupa", "larvae")) +df2volcano(df[df$clade == "G", ], legend_detect = c("larvae", "pupa")) ``` df2volcanodf2volcano (second dataset) From 80001cb278b5319731627689eb886abe3ae5b14d Mon Sep 17 00:00:00 2001 From: Cursor Agent Date: Fri, 25 Sep 2026 15:15:00 +0000 Subject: [PATCH 2/3] Pass ANCOM-BC2 a count matrix so checks do not need microbiome. Current ancombc2 pulls microbiome only for phyloseq input. Older releases still receive a phyloseq object. Co-authored-by: dvsmutin --- DESCRIPTION | 1 + R/plots_harness.R | 6 ++- R/plots_ordination.R | 85 +++++++++++++++++++++++----------------- man/archi_ancombc_fit.Rd | 12 ++++++ man/df2volcano.Rd | 18 ++++----- vignettes.md | 2 +- 6 files changed, 77 insertions(+), 47 deletions(-) create mode 100644 man/archi_ancombc_fit.Rd diff --git a/DESCRIPTION b/DESCRIPTION index 09a9493..28f452c 100644 --- a/DESCRIPTION +++ b/DESCRIPTION @@ -57,6 +57,7 @@ Suggests: qiime2R, reticulate, robCompositions, + SummarizedExperiment, testthat (>= 3.0.0), vegan, withr diff --git a/R/plots_harness.R b/R/plots_harness.R index f691a42..2657c82 100644 --- a/R/plots_harness.R +++ b/R/plots_harness.R @@ -551,7 +551,7 @@ archi_geom_fruit <- function(data = NULL, mapping, offset, pwidth, archi_position_boxx <- function() { ggplot2::ggproto( NULL, - getFromNamespace("PositionIdentityx", "ggtreeExtra"), + utils::getFromNamespace("PositionIdentityx", "ggtreeExtra"), compute_layer = function(self, data, params, layout) { if (!is.na(params$hexpand)) { for (col in c( @@ -1226,7 +1226,9 @@ archi_parse_phyloseq <- function(ps, to_relative = TRUE) { # parse_phyloseq looks it up from the metacoder namespace and misses it # until the package is attached. if (!"package:metacoder" %in% search()) { - suppressPackageStartupMessages(library("metacoder")) + # Attach so ranks_ref is on the search path. library() is a check warning + # for a suggested package; attachNamespace runs the same hook. + attachNamespace(loadNamespace("metacoder")) } tryCatch(metacoder::parse_phyloseq(ps), error = function(e) NULL) } diff --git a/R/plots_ordination.R b/R/plots_ordination.R index 281299f..a388c0e 100644 --- a/R/plots_ordination.R +++ b/R/plots_ordination.R @@ -518,13 +518,14 @@ df2tsne <- function(df, color = "clust", k_means = 10, text_top = FALSE, #' Volcano plot of ANCOM-BC2 log fold change #' #' Fits [ANCOMBC::ancombc2] on raw counts (`N`) for the two groups matched by -#' `legend_detect` (sample names or legend fields). Counts are passed as a -#' \pkg{phyloseq} object, which ANCOM-BC2 converts to a tree experiment. The -#' x axis is the bias-corrected log2 fold change of the second group against -#' the first, and the y axis is `-log10` of the adjusted q-value. Taxa with -#' no counts in one of the two groups are omitted: ANCOM-BC2 cannot estimate -#' a sampling variance for them. The function stops when \pkg{ANCOMBC} or -#' \pkg{phyloseq} is not installed. +#' `legend_detect` (sample names or legend fields). Current ANCOMBC receives +#' the count matrix and a sample-metadata data frame. Older ANCOMBC, which +#' has no `meta_data` argument, receives a \pkg{phyloseq} object instead. +#' The x axis is the bias-corrected log2 fold change of the second group +#' against the first, and the y axis is `-log10` of the adjusted q-value. +#' Taxa with no counts in one of the two groups are omitted: ANCOM-BC2 cannot +#' estimate a sampling variance for them. The function stops when +#' \pkg{ANCOMBC} is not installed. #' #' @param df A tidy `tibble` from [get_counts()] with legend columns. #' @param legend_detect Length-2 character vector of patterns identifying the @@ -540,8 +541,7 @@ df2tsne <- function(df, color = "clust", k_means = 10, text_top = FALSE, #' legend <- system.file("extdata", "legend.csv", package = "aRchiteutis") #' df <- get_counts(path = path, pattern = "m(11|12|13|18|4|39)_", #' legend = legend, trim_char = "_") -#' if (requireNamespace("ANCOMBC", quietly = TRUE) && -#' requireNamespace("phyloseq", quietly = TRUE)) { +#' if (requireNamespace("ANCOMBC", quietly = TRUE)) { #' g <- df[df$clade == "G", ] #' keep <- names(sort(tapply(g$N, g$taxa, sum), decreasing = TRUE))[seq_len(30)] #' df2volcano(g[g$taxa %in% keep, ], legend_detect = c("larvae", "pupa")) @@ -552,7 +552,6 @@ df2tsne <- function(df, color = "clust", k_means = 10, text_top = FALSE, df2volcano <- function(df, legend_detect, treshhold_logAC = 0.5, treshhold_p = 0.05) { archi_optional("ANCOMBC", "df2volcano") - archi_optional("phyloseq", "df2volcano") if (length(legend_detect) != 2L) { stop("legend_detect must name two groups", call. = FALSE) } @@ -619,33 +618,9 @@ archi_ancombc_volcano <- function(df, legend_detect) { ifelse(colnames(mat) %in% grouped$samples1, legend_detect[[1]], legend_detect[[2]]), levels = legend_detect ) - sam <- phyloseq::sample_data(data.frame( - group = group, row.names = colnames(mat), stringsAsFactors = FALSE - )) fit <- NULL for (attempt in seq_len(8L)) { - ps <- phyloseq::phyloseq( - phyloseq::otu_table(mat, taxa_are_rows = TRUE), sam - ) - fit <- tryCatch( - ANCOMBC::ancombc2( - data = ps, - fix_formula = "group", - p_adj_method = "fdr", - prv_cut = 0.1, - lib_cut = 0, - group = "group", - struc_zero = FALSE, - neg_lb = FALSE, - pseudo = 0, - pseudo_sens = FALSE, - global = FALSE, - pairwise = FALSE, - verbose = FALSE, - n_cl = 1L - ), - error = function(e) e - ) + fit <- tryCatch(archi_ancombc_fit(mat, group), error = function(e) e) if (!inherits(fit, "error")) break msg <- conditionMessage(fit) if (!grepl("Zero variances have been detected", msg, fixed = TRUE)) { @@ -676,6 +651,46 @@ archi_ancombc_volcano <- function(df, legend_detect) { ) } +#' Call ancombc2 with a count matrix, or phyloseq on older ANCOMBC +#' @keywords internal +archi_ancombc_fit <- function(mat, group) { + meta <- data.frame( + group = group, row.names = colnames(mat), stringsAsFactors = FALSE + ) + formals_nms <- names(formals(ANCOMBC::ancombc2)) + args <- list( + fix_formula = "group", + p_adj_method = "fdr", + prv_cut = 0.1, + lib_cut = 0, + group = "group", + struc_zero = FALSE, + neg_lb = FALSE, + pseudo = 0, + pseudo_sens = FALSE, + global = FALSE, + pairwise = FALSE, + verbose = FALSE, + n_cl = 1L + ) + args <- args[names(args) %in% formals_nms] + # Current ANCOMBC turns a phyloseq object into abundances via microbiome, + # which is only a Suggests of ANCOMBC and is absent in a clean check. + # A count matrix plus meta_data skips that path. + if ("meta_data" %in% formals_nms) { + args$data <- mat + args$meta_data <- meta + if ("taxa_are_rows" %in% formals_nms) args$taxa_are_rows <- TRUE + } else { + archi_optional("phyloseq", "df2volcano") + args$data <- phyloseq::phyloseq( + phyloseq::otu_table(mat, taxa_are_rows = TRUE), + phyloseq::sample_data(meta) + ) + } + do.call(ANCOMBC::ancombc2, args) +} + #' Samples matching two legend patterns #' @keywords internal archi_legend_groups <- function(df, legend_detect) { diff --git a/man/archi_ancombc_fit.Rd b/man/archi_ancombc_fit.Rd new file mode 100644 index 0000000..8125c64 --- /dev/null +++ b/man/archi_ancombc_fit.Rd @@ -0,0 +1,12 @@ +% Generated by roxygen2: do not edit by hand +% Please edit documentation in R/plots_ordination.R +\name{archi_ancombc_fit} +\alias{archi_ancombc_fit} +\title{Call ancombc2 with a count matrix, or phyloseq on older ANCOMBC} +\usage{ +archi_ancombc_fit(mat, group) +} +\description{ +Call ancombc2 with a count matrix, or phyloseq on older ANCOMBC +} +\keyword{internal} diff --git a/man/df2volcano.Rd b/man/df2volcano.Rd index f0bb77a..8fb02f1 100644 --- a/man/df2volcano.Rd +++ b/man/df2volcano.Rd @@ -22,21 +22,21 @@ A \link[ggplot2:ggplot]{ggplot2::ggplot} object. } \description{ Fits \link[ANCOMBC:ancombc2]{ANCOMBC::ancombc2} on raw counts (\code{N}) for the two groups matched by -\code{legend_detect} (sample names or legend fields). Counts are passed as a -\pkg{phyloseq} object, which ANCOM-BC2 converts to a tree experiment. The -x axis is the bias-corrected log2 fold change of the second group against -the first, and the y axis is \code{-log10} of the adjusted q-value. Taxa with -no counts in one of the two groups are omitted: ANCOM-BC2 cannot estimate -a sampling variance for them. The function stops when \pkg{ANCOMBC} or -\pkg{phyloseq} is not installed. +\code{legend_detect} (sample names or legend fields). Current ANCOMBC receives +the count matrix and a sample-metadata data frame. Older ANCOMBC, which +has no \code{meta_data} argument, receives a \pkg{phyloseq} object instead. +The x axis is the bias-corrected log2 fold change of the second group +against the first, and the y axis is \code{-log10} of the adjusted q-value. +Taxa with no counts in one of the two groups are omitted: ANCOM-BC2 cannot +estimate a sampling variance for them. The function stops when +\pkg{ANCOMBC} is not installed. } \examples{ path <- system.file("extdata", package = "aRchiteutis") legend <- system.file("extdata", "legend.csv", package = "aRchiteutis") df <- get_counts(path = path, pattern = "m(11|12|13|18|4|39)_", legend = legend, trim_char = "_") -if (requireNamespace("ANCOMBC", quietly = TRUE) && - requireNamespace("phyloseq", quietly = TRUE)) { +if (requireNamespace("ANCOMBC", quietly = TRUE)) { g <- df[df$clade == "G", ] keep <- names(sort(tapply(g$N, g$taxa, sum), decreasing = TRUE))[seq_len(30)] df2volcano(g[g$taxa \%in\% keep, ], legend_detect = c("larvae", "pupa")) diff --git a/vignettes.md b/vignettes.md index a1d016f..4cdb300 100644 --- a/vignettes.md +++ b/vignettes.md @@ -133,7 +133,7 @@ df2tsne(matG, k_means = 10, text_top = 20) ### `df2volcano` — ANCOM-BC2 log fold change -The first `legend_detect` pattern is the reference group. Requires the optional packages ANCOMBC and phyloseq. +The first `legend_detect` pattern is the reference group. Requires the optional package ANCOMBC. Older ANCOMBC builds also need phyloseq. ```r df2volcano(df[df$clade == "G", ], legend_detect = c("larvae", "pupa")) From 7fbbc7a5725f36798b273d1ec26b70a663bec798 Mon Sep 17 00:00:00 2001 From: Cursor Agent Date: Fri, 25 Sep 2026 15:25:49 +0000 Subject: [PATCH 3/3] Keep ANCOM-BC2 sample metadata as a data.frame. A one-column metadata table drops to a vector when ANCOMBC subsets samples, so the group column was missing. Co-authored-by: dvsmutin --- R/plots_ordination.R | 7 ++++++- 1 file changed, 6 insertions(+), 1 deletion(-) diff --git a/R/plots_ordination.R b/R/plots_ordination.R index a388c0e..1325314 100644 --- a/R/plots_ordination.R +++ b/R/plots_ordination.R @@ -654,8 +654,13 @@ archi_ancombc_volcano <- function(df, legend_detect) { #' Call ancombc2 with a count matrix, or phyloseq on older ANCOMBC #' @keywords internal archi_ancombc_fit <- function(mat, group) { + # ANCOMBC subsets with meta_data[samples, ]. A one-column data.frame + # drops to a vector, so the group column disappears. Keep a second column. meta <- data.frame( - group = group, row.names = colnames(mat), stringsAsFactors = FALSE + sample = colnames(mat), + group = group, + row.names = colnames(mat), + stringsAsFactors = FALSE ) formals_nms <- names(formals(ANCOMBC::ancombc2)) args <- list(

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