#!/usr/bin/env Rscript # ===================================================== # YuanSeq Snakemake - 火山图 # ===================================================== suppressPackageStartupMessages({ library(ggplot2) library(dplyr) }) args <- commandArgs(trailingOnly = TRUE) parse_arg <- function(name, default = NULL) { idx <- grep(paste0("^--", name, "="), args) if (length(idx) == 0) { idx <- match(paste0("--", name), args) if (!is.na(idx) && idx < length(args)) return(args[idx + 1]) return(default) } sub(paste0("^--", name, "="), "", args[idx][1]) } deg_file <- parse_arg("deg") output <- parse_arg("output") pval_cutoff <- as.numeric(parse_arg("pval_cutoff", "0.05")) log2fc_cutoff <- as.numeric(parse_arg("log2fc_cutoff", "1")) if (is.null(deg_file) || is.null(output)) { stop("Usage: Rscript 02_volcano_plot.R --deg=DEG.csv --output=volcano.pdf [--pval_cutoff=0.05] [--log2fc_cutoff=1]") } deg <- read.csv(deg_file, stringsAsFactors = FALSE) pval_col <- if ("padj" %in% colnames(deg)) "padj" else "pvalue" deg <- deg %>% mutate( neglog10p = -log10(.data[[pval_col]]), sig = case_when( .data[[pval_col]] < pval_cutoff & log2FoldChange > log2fc_cutoff ~ "Up", .data[[pval_col]] < pval_cutoff & log2FoldChange < -log2fc_cutoff ~ "Down", TRUE ~ "NS" ) ) label_col <- if ("SYMBOL" %in% colnames(deg) && !all(is.na(deg$SYMBOL))) "SYMBOL" else "GeneID" top_n <- 20 deg_label <- deg %>% filter(sig != "NS") %>% arrange(desc(abs(log2FoldChange))) %>% head(top_n) p <- ggplot(deg, aes(x = log2FoldChange, y = neglog10p, color = sig)) + geom_point(alpha = 0.6, size = 2) + scale_color_manual(values = c(Up = "#e74c3c", Down = "#3498db", NS = "gray70")) + geom_hline(yintercept = -log10(pval_cutoff), linetype = "dashed", color = "gray40") + geom_vline(xintercept = c(-log2fc_cutoff, log2fc_cutoff), linetype = "dashed", color = "gray40") + labs(x = "log2 Fold Change", y = paste0("-log10(", pval_col, ")"), color = "Status") + theme_bw(base_size = 12) + theme(legend.position = "top") if (nrow(deg_label) > 0 && requireNamespace("ggrepel", quietly = TRUE)) { p <- p + ggrepel::geom_text_repel( data = deg_label, aes(label = .data[[label_col]]), size = 3, max.overlaps = 20, box.padding = 0.5 ) } dir.create(dirname(output), recursive = TRUE, showWarnings = FALSE) ggsave(output, p, width = 8, height = 6) message("火山图已保存: ", output)