2502 lines
85 KiB
Python
Executable File
2502 lines
85 KiB
Python
Executable File
#!/usr/bin/env python3
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"""
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Synteny阶段：对基因家族进行种内共线性分析
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输入：
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- Identify阶段的蛋白序列文件
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- GFF3注释文件
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- 基因组文件
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输出：
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- {prefix}.blast (BLAST比对结果)
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- {prefix}.gff (过滤后的GFF)
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- {prefix}.dup_class.log (复制基因分类)
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- {prefix}.collinearity (MCScanX共线性结果)
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- {prefix}.synteny_report.md (分析报告)
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- {prefix}.synteny.manifest.json (机器清单)
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"""
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import argparse
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import gzip
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import json
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import os
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import subprocess
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import sys
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import time
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import shutil
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from datetime import datetime
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from pathlib import Path
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from typing import Any, Dict, List, Optional
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import hashlib
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import re
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try:
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from gfa_core.gene_family_scope import read_scope, sanitize_token
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except Exception:
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def read_scope(scope_path):
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return json.loads(Path(scope_path).read_text(encoding="utf-8"))
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def sanitize_token(value: str) -> str:
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token = re.sub(r"[^A-Za-z0-9_.-]+", "-", str(value or "").strip())
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token = token.strip(".-_")
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return token or "NA"
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# 添加conda环境bin目录到PATH
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CONDA_PREFIX = Path(sys.executable).resolve().parent.parent
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CONDA_BIN = str(CONDA_PREFIX / "bin")
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if CONDA_BIN not in os.environ.get("PATH", "").split(os.pathsep):
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os.environ["PATH"] = CONDA_BIN + os.pathsep + os.environ.get("PATH", "")
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def compute_file_hash(filepath: Path) -> str:
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"""计算文件的SHA256哈希值"""
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sha256_hash = hashlib.sha256()
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try:
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with open(filepath, "rb") as f:
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for byte_block in iter(lambda: f.read(4096), b""):
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sha256_hash.update(byte_block)
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return sha256_hash.hexdigest()
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except Exception:
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return ""
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def _relative_to_workspace(path: Path, workspace: Path) -> str:
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try:
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return str(path.resolve().relative_to(workspace.resolve()))
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except Exception:
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return str(path)
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def _scoped_copy_destination(
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source: Path,
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workspace: Path,
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prefix: str,
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scope_id: str,
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artifact_name: str,
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) -> Path:
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scope_token = sanitize_token(scope_id)
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suffix = source.suffix
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lower_parts = {part.lower() for part in source.parts}
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if "final" in lower_parts:
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dest_dir = workspace / "final"
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else:
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dest_dir = workspace / "synteny" / "analysis"
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return dest_dir / f"{prefix}.{scope_token}.{sanitize_token(artifact_name)}{suffix}"
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def materialize_scoped_artifacts(
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artifacts: Dict[str, Dict],
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workspace: Path,
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prefix: str,
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scope_id: str,
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) -> Dict[str, Dict]:
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"""Copy scope-specific outputs to stable names so later species runs cannot overwrite them."""
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if not scope_id:
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return artifacts
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scoped: Dict[str, Dict] = {}
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for name, info in artifacts.items():
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if not isinstance(info, dict):
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scoped[name] = info
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continue
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if name == "manifest":
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scoped[name] = dict(info)
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continue
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source_text = info.get("full_path") or info.get("path") or ""
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source = Path(source_text)
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if not source.is_absolute():
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source = workspace / source
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if not source.exists():
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scoped[name] = dict(info)
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continue
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dest = _scoped_copy_destination(source, workspace, prefix, scope_id, name)
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dest.parent.mkdir(parents=True, exist_ok=True)
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if source.absolute() != dest.absolute():
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shutil.copy2(source, dest)
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copied = dict(info)
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copied["path"] = _relative_to_workspace(dest, workspace)
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copied["full_path"] = str(dest.absolute())
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copied["size"] = dest.stat().st_size
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copied["sha256"] = compute_file_hash(dest)
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scoped[name] = copied
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return scoped
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def current_input_dir(workspace: Path) -> Path:
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return workspace.parent.parent / "input" / workspace.name
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def rebase_current_job_path(workspace: Path, raw_path: Any) -> Optional[Path]:
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"""Map copied-job paths from old scope/manifest records back into this job."""
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path_text = str(raw_path or "").strip()
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if not path_text:
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return None
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path = Path(path_text)
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job_id = workspace.name
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input_dir = current_input_dir(workspace)
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parts = list(path.parts)
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for marker, root in (("input", input_dir), ("workspace", workspace)):
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if marker in parts:
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idx = parts.index(marker)
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if idx + 1 < len(parts) and parts[idx + 1] == job_id:
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suffix = parts[idx + 2 :]
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candidate = root.joinpath(*suffix)
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if candidate.exists():
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return candidate
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if not path.is_absolute():
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for candidate in (Path.cwd() / path, workspace / path):
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if candidate.exists():
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return candidate
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by_name_candidates = [
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input_dir / path.name,
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workspace / "final" / path.name,
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workspace / "synteny" / "analysis" / path.name,
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]
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for candidate in by_name_candidates:
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if candidate.exists():
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return candidate
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if path.exists():
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return path
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return None
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def rebase_job_path_text(workspace: Path, value: str) -> str:
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job_id = re.escape(workspace.name)
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pattern = re.compile(rf"/[^\s\"']*?(?:input|workspace)/{job_id}/[^\s\"']+")
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def replace_match(match: re.Match[str]) -> str:
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original = match.group(0).rstrip(",)")
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suffix = match.group(0)[len(original):]
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rebased = rebase_current_job_path(workspace, original)
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if rebased:
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return str(rebased.absolute()) + suffix
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return match.group(0)
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return pattern.sub(replace_match, value)
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def rebase_manifest_payload_paths(workspace: Path, value: Any) -> Any:
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if isinstance(value, dict):
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return {key: rebase_manifest_payload_paths(workspace, item) for key, item in value.items()}
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if isinstance(value, list):
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return [rebase_manifest_payload_paths(workspace, item) for item in value]
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if isinstance(value, str):
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return rebase_job_path_text(workspace, value)
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return value
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def get_tool_versions() -> Dict[str, str]:
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"""获取所需工具版本信息"""
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versions = {}
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# diamond
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try:
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result = subprocess.run(
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["diamond", "--version"],
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capture_output=True,
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text=True,
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check=False,
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timeout=5
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)
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output = result.stdout.strip() or result.stderr.strip()
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versions["diamond"] = output.split('\n')[0] if output else "unknown"
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except Exception:
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versions["diamond"] = "not_found"
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# blastp
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try:
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result = subprocess.run(
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["blastp", "-version"],
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capture_output=True,
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text=True,
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check=False,
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timeout=5
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)
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output = result.stdout.strip() or result.stderr.strip()
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versions["blastp"] = output.split('\n')[0] if output else "unknown"
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except Exception:
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versions["blastp"] = "not_found"
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# MCScanX
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try:
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result = subprocess.run(
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["MCScanX", "-h"],
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capture_output=True,
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text=True,
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check=False,
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timeout=5
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)
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versions["MCScanX"] = "available" if result.returncode in [0, 1] else "not_found"
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except Exception:
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versions["MCScanX"] = "not_found"
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# duplicate_gene_classifier
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try:
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result = subprocess.run(
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["duplicate_gene_classifier", "-h"],
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capture_output=True,
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text=True,
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check=False,
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timeout=5
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)
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versions["duplicate_gene_classifier"] = "available" if result.returncode in [0, 1] else "not_found"
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except Exception:
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versions["duplicate_gene_classifier"] = "not_found"
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# circos
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try:
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result = subprocess.run(
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["circos", "-version"],
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capture_output=True,
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text=True,
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check=False,
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timeout=5
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)
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output = result.stdout.strip() or result.stderr.strip()
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versions["circos"] = output.split('\n')[0] if output else "available"
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except Exception:
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versions["circos"] = "not_found"
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return versions
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def validate_inputs(
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workspace: Path,
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prefix: str,
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input_dir: Path,
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gff3: Optional[Path] = None,
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genome: Optional[Path] = None,
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protein_fasta: Optional[Path] = None,
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) -> Dict[str, Any]:
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"""验证输入文件"""
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print(f"正在验证输入文件...")
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results = {
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"protein_fasta": None,
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"gff3": None,
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"genome": None,
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"gene_id_file": None
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}
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# 1. 查找蛋白序列文件。Synteny/MCScanX 需要目标物种全蛋白组，
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# scope 模式或 input 中的 *.pep.* 应优先于 identify 的家族蛋白。
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if protein_fasta:
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protein_path = Path(protein_fasta)
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if not protein_path.exists():
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raise FileNotFoundError(f"蛋白组文件不存在: {protein_fasta}")
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results["protein_fasta"] = protein_path
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species_prefix = prefix.split(".")[0] if "." in prefix else prefix
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if not results["protein_fasta"]:
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for pattern in [
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f"{species_prefix}.pep.fasta",
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f"{species_prefix}.pep.fa",
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"*.pep.fasta",
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"*.pep.fa",
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f"{prefix}*.fa",
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f"{prefix}*.fasta",
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"*.fasta"
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]:
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files = list(input_dir.glob(pattern))
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if files:
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results["protein_fasta"] = files[0]
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break
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if not results["protein_fasta"]:
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identify_manifest = workspace / "final" / f"{prefix}.identify.manifest.json"
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if identify_manifest.exists():
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with open(identify_manifest, 'r') as f:
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m = json.load(f)
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arts = m.get("artifacts", {})
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if isinstance(arts, dict):
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protein_path = arts.get("final_proteins", {}).get("full_path")
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if protein_path and Path(protein_path).exists():
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results["protein_fasta"] = Path(protein_path)
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if not results["protein_fasta"]:
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raise FileNotFoundError("找不到目标物种全蛋白组 FASTA")
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# 2. GFF3 文件
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if gff3:
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gff3_path = Path(gff3)
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if not gff3_path.exists():
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raise FileNotFoundError(f"GFF3文件不存在: {gff3}")
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results["gff3"] = gff3_path
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else:
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# 自动查找
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for pattern in ["*.gff3", "*.gff"]:
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files = list(input_dir.glob(pattern))
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if files:
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results["gff3"] = files[0]
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break
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if not results["gff3"]:
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raise FileNotFoundError("找不到GFF3注释文件")
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print(f" GFF3: {results['gff3']}")
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# 3. 基因组文件
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if genome:
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genome_path = Path(genome)
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if not genome_path.exists():
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raise FileNotFoundError(f"基因组文件不存在: {genome}")
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results["genome"] = genome_path
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else:
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# 自动查找
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for pattern in ["*.genome.fasta", "*.genome.fa", "*.fa", "*.fasta"]:
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files = list(input_dir.glob(pattern))
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if files:
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results["genome"] = files[0]
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break
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if not results["genome"]:
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||
raise FileNotFoundError("找不到基因组文件")
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print(f" 基因组: {results['genome']}")
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# 4. 基因ID列表文件（可选）
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final_id = workspace / "final" / f"{prefix}.final_members.id"
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||
if final_id.exists():
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results["gene_id_file"] = final_id
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for pattern in ["*.geneID*", "*.gene_id*"]:
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if results["gene_id_file"]:
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break
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files = list(input_dir.glob(pattern))
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||
if files:
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results["gene_id_file"] = files[0]
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||
break
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if results["gene_id_file"]:
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||
print(f" 基因ID文件: {results['gene_id_file']}")
|
||
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||
return results
|
||
|
||
|
||
def first_existing_scope_file(
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workspace: Path,
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scope_data: Dict[str, Any],
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species: str,
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file_types: List[str],
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||
) -> Optional[Path]:
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species_files = (scope_data.get("files") or {}).get(species) or {}
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||
for file_type in file_types:
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||
for record in species_files.get(file_type, []) or []:
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||
path_text = record.get("path") or record.get("full_path") or ""
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||
if path_text:
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||
path = rebase_current_job_path(workspace, path_text)
|
||
if path and path.exists():
|
||
return path
|
||
return None
|
||
|
||
|
||
def first_existing_path(candidates: List[Path]) -> Optional[Path]:
|
||
for candidate in candidates:
|
||
if candidate.exists() and candidate.is_file() and candidate.stat().st_size > 0:
|
||
return candidate
|
||
return None
|
||
|
||
|
||
def prefer_representative_target_file(
|
||
input_dir: Path,
|
||
species: str,
|
||
file_type: str,
|
||
fallback: Optional[Path],
|
||
) -> tuple[Optional[Path], str]:
|
||
"""Prefer genome_prepare/standardized representative-transcript files for MCScanX.
|
||
|
||
Scope records preserve uploaded raw files, which may include multiple
|
||
isoforms per locus. MCScanX should receive one representative protein/GFF
|
||
per gene whenever the standardized prepare/genome_prepare outputs exist.
|
||
"""
|
||
species = str(species or "").strip()
|
||
if not species:
|
||
return fallback, "scope" if fallback else "missing"
|
||
|
||
if file_type == "pep":
|
||
candidates = [
|
||
input_dir / f"{species}.pep.fasta",
|
||
input_dir / f"{species}.pep.fa",
|
||
input_dir / species / f"{species}.pep.fasta",
|
||
input_dir / species / f"{species}.pep.fa",
|
||
input_dir / "genome_prepare" / f"{species}_extracted.pep.fa",
|
||
input_dir / "genome_prepare" / f"{species}.pep.fasta",
|
||
input_dir / "genome_prepare" / f"{species}.pep.fa",
|
||
]
|
||
elif file_type == "gff":
|
||
candidates = [
|
||
input_dir / f"{species}_final.gff3",
|
||
input_dir / f"{species}_final.gff",
|
||
input_dir / f"{species}.gff3",
|
||
input_dir / f"{species}.gff",
|
||
input_dir / species / f"{species}.gff3",
|
||
input_dir / species / f"{species}.gff",
|
||
input_dir / "genome_prepare" / f"{species}.gff3.tmp",
|
||
input_dir / "genome_prepare" / f"{species}_final.gff3",
|
||
]
|
||
elif file_type == "genome":
|
||
candidates = [
|
||
input_dir / f"{species}.genome.fasta",
|
||
input_dir / f"{species}.genome.fa",
|
||
input_dir / f"{species}.genome.fasta.gz",
|
||
input_dir / f"{species}.genome.fa.gz",
|
||
input_dir / f"{species}.fasta",
|
||
input_dir / f"{species}.fa",
|
||
input_dir / species / f"{species}.genome.fasta",
|
||
input_dir / species / f"{species}.genome.fa",
|
||
input_dir / species / f"{species}.genome.fasta.gz",
|
||
input_dir / species / f"{species}.genome.fa.gz",
|
||
input_dir / species / f"{species}.fasta",
|
||
input_dir / species / f"{species}.fa",
|
||
input_dir / "genome_prepare" / f"{species}.genome.fasta",
|
||
input_dir / "genome_prepare" / f"{species}.genome.fa",
|
||
input_dir / "genome_prepare" / f"{species}.fasta",
|
||
input_dir / "genome_prepare" / f"{species}.fa",
|
||
]
|
||
else:
|
||
return fallback, "scope" if fallback else "missing"
|
||
|
||
representative = first_existing_path(candidates)
|
||
if representative:
|
||
return representative, "representative"
|
||
return fallback, "scope_raw_fallback" if fallback else "missing"
|
||
|
||
|
||
def resolve_synteny_scope_inputs(
|
||
workspace: Path,
|
||
scope_file: Optional[Path],
|
||
) -> tuple[Dict[str, Any], Optional[Path], Optional[Path], Optional[Path]]:
|
||
if not scope_file:
|
||
return {}, None, None, None
|
||
scope_data = read_scope(scope_file)
|
||
targets = [str(x).strip() for x in (scope_data.get("target_species") or []) if str(x).strip()]
|
||
if len(targets) != 1:
|
||
raise RuntimeError(
|
||
"Synteny 阶段是种内共线性，一次只能分析一个目标物种；"
|
||
f"当前 scope target_species={targets or '空'}。请在弹窗中只选择一个 target，或分别运行每个目标物种。"
|
||
)
|
||
species = targets[0]
|
||
protein_fasta = first_existing_scope_file(workspace, scope_data, species, ["pep"])
|
||
gff3_file = first_existing_scope_file(workspace, scope_data, species, ["gff"])
|
||
genome_file = first_existing_scope_file(workspace, scope_data, species, ["genome"])
|
||
if not protein_fasta:
|
||
raise RuntimeError(f"Synteny scope 未找到目标物种 {species} 的全蛋白组 pep FASTA")
|
||
if not gff3_file:
|
||
raise RuntimeError(f"Synteny scope 未找到目标物种 {species} 的 GFF/GFF3 文件")
|
||
if not genome_file:
|
||
raise RuntimeError(f"Synteny scope 未找到目标物种 {species} 的 genome FASTA 文件")
|
||
return scope_data, protein_fasta, gff3_file, genome_file
|
||
|
||
|
||
def open_text_maybe_gzip(path: Path):
|
||
if str(path).endswith(".gz"):
|
||
return gzip.open(path, "rt", encoding="utf-8", errors="replace")
|
||
return path.open("r", encoding="utf-8", errors="replace")
|
||
|
||
|
||
def write_blast_safe_protein_fasta(input_fasta: Path, output_fasta: Path) -> tuple[Path, Dict[str, int]]:
|
||
"""Write a DIAMOND/BLAST-safe protein FASTA.
|
||
|
||
Whole proteomes produced by different preparation tools may contain terminal
|
||
stop markers (`*` or `.`) or other non-amino-acid characters. DIAMOND rejects
|
||
those records during makedb, causing the pipeline to fall back to very slow
|
||
BLASTP. For synteny, remove non-letter characters while keeping the first
|
||
FASTA header token as the stable protein ID.
|
||
"""
|
||
output_fasta.parent.mkdir(parents=True, exist_ok=True)
|
||
records = 0
|
||
removed_chars = 0
|
||
total_aa = 0
|
||
header = ""
|
||
seq_parts: list[str] = []
|
||
|
||
def flush_record(out_handle) -> None:
|
||
nonlocal records, removed_chars, total_aa, header, seq_parts
|
||
if not header:
|
||
return
|
||
raw_seq = "".join(seq_parts)
|
||
clean_seq = "".join(ch for ch in raw_seq.upper() if ch.isalpha())
|
||
removed_chars += len(raw_seq) - len(clean_seq)
|
||
if clean_seq:
|
||
out_handle.write(f">{header}\n")
|
||
for i in range(0, len(clean_seq), 80):
|
||
out_handle.write(clean_seq[i:i + 80] + "\n")
|
||
records += 1
|
||
total_aa += len(clean_seq)
|
||
header = ""
|
||
seq_parts = []
|
||
|
||
with open_text_maybe_gzip(input_fasta) as in_handle, output_fasta.open("w", encoding="utf-8") as out_handle:
|
||
for raw in in_handle:
|
||
line = raw.strip()
|
||
if not line:
|
||
continue
|
||
if line.startswith(">"):
|
||
flush_record(out_handle)
|
||
header = line[1:].split()[0]
|
||
seq_parts = []
|
||
else:
|
||
seq_parts.append(line)
|
||
flush_record(out_handle)
|
||
|
||
if records == 0:
|
||
raise RuntimeError(f"No valid protein records after cleaning: {input_fasta}")
|
||
return output_fasta, {
|
||
"records": records,
|
||
"removed_chars": removed_chars,
|
||
"total_aa": total_aa,
|
||
}
|
||
|
||
|
||
def prepare_directories(workspace: Path) -> Dict[str, Path]:
|
||
"""创建Synteny阶段的标准目录结构"""
|
||
paths = {
|
||
"analysis_dir": workspace / "synteny" / "analysis",
|
||
"final_dir": workspace / "final",
|
||
"logs_dir": workspace / "logs",
|
||
}
|
||
|
||
print("创建目录结构:")
|
||
for name, path in paths.items():
|
||
path.mkdir(parents=True, exist_ok=True)
|
||
print(f" {name}: {path}")
|
||
|
||
return paths
|
||
|
||
|
||
def run_diamond_blastp(
|
||
input_fasta: Path,
|
||
output_blast: Path,
|
||
cpu: int = 8,
|
||
evalue: float = 1e-10,
|
||
max_target_seqs: int = 5
|
||
) -> Dict[str, Any]:
|
||
"""运行diamond blastp比对"""
|
||
print(f"运行Diamond BLASTP: {input_fasta} -> {output_blast}")
|
||
|
||
# 构建数据库
|
||
db_path = input_fasta.with_suffix('')
|
||
db_cmd = [
|
||
"diamond", "makedb",
|
||
"--in", str(input_fasta),
|
||
"--db", str(db_path)
|
||
]
|
||
|
||
start_time = time.time()
|
||
|
||
# 创建数据库
|
||
try:
|
||
result = subprocess.run(
|
||
db_cmd,
|
||
capture_output=True,
|
||
text=True,
|
||
encoding="utf-8",
|
||
errors="replace",
|
||
check=False
|
||
)
|
||
except FileNotFoundError:
|
||
print(f" Diamond未安装，无法使用")
|
||
return {
|
||
"success": False,
|
||
"command": " ".join(db_cmd),
|
||
"stderr": "diamond not found"
|
||
}
|
||
|
||
if result.returncode != 0:
|
||
print(f" Diamond makedb 失败: {result.stderr}")
|
||
return {
|
||
"success": False,
|
||
"command": " ".join(db_cmd),
|
||
"stderr": result.stderr
|
||
}
|
||
|
||
# 运行比对
|
||
blast_cmd = [
|
||
"diamond", "blastp",
|
||
"--query", str(input_fasta),
|
||
"--db", str(db_path),
|
||
"--out", str(output_blast),
|
||
"--outfmt", "6",
|
||
"--max-target-seqs", str(max_target_seqs),
|
||
"--evalue", str(evalue),
|
||
"--threads", str(cpu)
|
||
]
|
||
|
||
try:
|
||
result = subprocess.run(
|
||
blast_cmd,
|
||
capture_output=True,
|
||
text=True,
|
||
encoding="utf-8",
|
||
errors="replace",
|
||
check=False
|
||
)
|
||
except FileNotFoundError:
|
||
print(f" Diamond未安装，无法使用")
|
||
return {
|
||
"success": False,
|
||
"command": " ".join(blast_cmd),
|
||
"stderr": "diamond not found"
|
||
}
|
||
|
||
elapsed = time.time() - start_time
|
||
|
||
if result.returncode == 0:
|
||
print(f" Diamond成功完成，耗时: {elapsed:.1f}秒")
|
||
return {
|
||
"success": True,
|
||
"command": " ".join(blast_cmd),
|
||
"returncode": result.returncode,
|
||
"stderr": result.stderr,
|
||
"output_file": output_blast,
|
||
"elapsed_sec": elapsed
|
||
}
|
||
else:
|
||
print(f" Diamond失败，返回码: {result.returncode}")
|
||
return {
|
||
"success": False,
|
||
"command": " ".join(blast_cmd),
|
||
"returncode": result.returncode,
|
||
"stderr": result.stderr,
|
||
"output_file": None,
|
||
"elapsed_sec": elapsed
|
||
}
|
||
|
||
|
||
def run_blastp(
|
||
input_fasta: Path,
|
||
output_blast: Path,
|
||
cpu: int = 8,
|
||
evalue: float = 1e-10,
|
||
max_alignments: int = 5
|
||
) -> Dict[str, Any]:
|
||
"""运行 NCBI blastp 比对（备选方案）"""
|
||
print(f"运行NCBI BLASTP: {input_fasta} -> {output_blast}")
|
||
|
||
# 构建数据库
|
||
db_path = input_fasta.with_suffix('')
|
||
db_cmd = [
|
||
"makeblastdb",
|
||
"-in", str(input_fasta),
|
||
"-dbtype", "prot",
|
||
"-out", str(db_path)
|
||
]
|
||
|
||
start_time = time.time()
|
||
|
||
# 创建数据库
|
||
try:
|
||
result = subprocess.run(
|
||
db_cmd,
|
||
capture_output=True,
|
||
text=True,
|
||
encoding="utf-8",
|
||
errors="replace",
|
||
check=False
|
||
)
|
||
except FileNotFoundError:
|
||
print(f" makeblastdb未安装，无法使用NCBI blastp")
|
||
return {
|
||
"success": False,
|
||
"command": " ".join(db_cmd),
|
||
"stderr": "makeblastdb not found - NCBI BLAST+ not installed"
|
||
}
|
||
|
||
if result.returncode != 0:
|
||
print(f" makeblastdb 失败: {result.stderr}")
|
||
return {
|
||
"success": False,
|
||
"command": " ".join(db_cmd),
|
||
"stderr": result.stderr
|
||
}
|
||
|
||
# 运行比对
|
||
blast_cmd = [
|
||
"blastp",
|
||
"-query", str(input_fasta),
|
||
"-db", str(db_path),
|
||
"-outfmt", "6",
|
||
"-out", str(output_blast),
|
||
"-num_threads", str(cpu),
|
||
"-num_alignments", str(max_alignments),
|
||
"-evalue", str(evalue)
|
||
]
|
||
|
||
result = subprocess.run(
|
||
blast_cmd,
|
||
capture_output=True,
|
||
text=True,
|
||
encoding="utf-8",
|
||
errors="replace",
|
||
check=False
|
||
)
|
||
|
||
elapsed = time.time() - start_time
|
||
|
||
if result.returncode == 0:
|
||
print(f" BLASTP成功完成，耗时: {elapsed:.1f}秒")
|
||
return {
|
||
"success": True,
|
||
"command": " ".join(blast_cmd),
|
||
"returncode": result.returncode,
|
||
"stderr": result.stderr,
|
||
"output_file": output_blast,
|
||
"elapsed_sec": elapsed
|
||
}
|
||
else:
|
||
print(f" BLASTP失败，返回码: {result.returncode}")
|
||
return {
|
||
"success": False,
|
||
"command": " ".join(blast_cmd),
|
||
"returncode": result.returncode,
|
||
"stderr": result.stderr,
|
||
"output_file": None,
|
||
"elapsed_sec": elapsed
|
||
}
|
||
|
||
|
||
def read_fasta_ids_for_matching(fasta_path: Path) -> set[str]:
|
||
"""Read FASTA primary IDs used by BLAST/MCScanX."""
|
||
ids: set[str] = set()
|
||
try:
|
||
with open_text_maybe_gzip(fasta_path) as handle:
|
||
for line in handle:
|
||
if line.startswith(">"):
|
||
token = line[1:].strip().split()[0]
|
||
if token:
|
||
ids.add(token)
|
||
except Exception:
|
||
pass
|
||
return ids
|
||
|
||
|
||
def parse_gff3_attributes(attr_text: str) -> Dict[str, List[str]]:
|
||
attrs: Dict[str, List[str]] = {}
|
||
for item in attr_text.strip().split(";"):
|
||
item = item.strip()
|
||
if not item:
|
||
continue
|
||
if "=" in item:
|
||
key, value = item.split("=", 1)
|
||
elif " " in item:
|
||
key, value = item.split(" ", 1)
|
||
else:
|
||
continue
|
||
key = key.strip()
|
||
values = [v.strip() for v in value.strip().split(",") if v.strip()]
|
||
if key and values:
|
||
attrs.setdefault(key, []).extend(values)
|
||
return attrs
|
||
|
||
|
||
def gff_id_candidates(attrs: Dict[str, List[str]]) -> List[str]:
|
||
ordered_keys = [
|
||
"Name",
|
||
"transcript_id",
|
||
"protein_id",
|
||
"locus_tag",
|
||
"gene_id",
|
||
"gene",
|
||
"ID",
|
||
"Parent",
|
||
]
|
||
candidates: List[str] = []
|
||
for key in ordered_keys:
|
||
for value in attrs.get(key, []):
|
||
candidates.append(value)
|
||
for prefix in ("gene:", "transcript:", "rna:", "mRNA:", "CDS:", "protein:"):
|
||
if value.startswith(prefix):
|
||
candidates.append(value[len(prefix):])
|
||
for prefix in ("gene-", "rna-", "cds-", "mrna-", "protein-"):
|
||
if value.lower().startswith(prefix):
|
||
candidates.append(value[len(prefix):])
|
||
|
||
seen = set()
|
||
unique: List[str] = []
|
||
for candidate in candidates:
|
||
if candidate and candidate not in seen:
|
||
unique.append(candidate)
|
||
seen.add(candidate)
|
||
return unique
|
||
|
||
|
||
def choose_gff_feature_id(attrs: Dict[str, List[str]], preferred_ids: Optional[set[str]]) -> str:
|
||
candidates = gff_id_candidates(attrs)
|
||
if preferred_ids:
|
||
for candidate in candidates:
|
||
if candidate in preferred_ids:
|
||
return candidate
|
||
preferred_base = {item.split(".")[0]: item for item in preferred_ids}
|
||
for candidate in candidates:
|
||
match = preferred_base.get(candidate.split(".")[0])
|
||
if match:
|
||
return match
|
||
if candidates:
|
||
return candidates[0]
|
||
return ""
|
||
|
||
|
||
def filter_gff3(
|
||
gff3_file: Path,
|
||
output_gff: Path,
|
||
feature_type: str = "mRNA",
|
||
preferred_ids: Optional[set[str]] = None,
|
||
) -> Dict[str, Any]:
|
||
"""Filter GFF3 and use IDs compatible with the stage protein FASTA."""
|
||
print(f"过滤GFF3文件: {gff3_file} -> {output_gff}")
|
||
|
||
start_time = time.time()
|
||
|
||
try:
|
||
gene_count = 0
|
||
matched_count = 0
|
||
with open_text_maybe_gzip(gff3_file) as src, open(output_gff, "w", encoding="utf-8") as dst:
|
||
for raw in src:
|
||
if not raw.strip() or raw.startswith("#"):
|
||
continue
|
||
parts = raw.rstrip("\n").split("\t")
|
||
if len(parts) < 9 or parts[2] != feature_type:
|
||
continue
|
||
attrs = parse_gff3_attributes(parts[8])
|
||
feature_id = choose_gff_feature_id(attrs, preferred_ids)
|
||
if not feature_id:
|
||
continue
|
||
if preferred_ids and feature_id in preferred_ids:
|
||
matched_count += 1
|
||
dst.write(f"{parts[0]}\t{feature_id}\t{parts[3]}\t{parts[4]}\n")
|
||
gene_count += 1
|
||
|
||
elapsed = time.time() - start_time
|
||
|
||
print(f" 成功过滤 {gene_count} 个基因，匹配FASTA ID {matched_count} 个，耗时: {elapsed:.1f}秒")
|
||
|
||
return {
|
||
"success": True,
|
||
"output_file": output_gff,
|
||
"gene_count": gene_count,
|
||
"matched_fasta_id_count": matched_count,
|
||
"elapsed_sec": elapsed,
|
||
}
|
||
|
||
except Exception as e:
|
||
return {
|
||
"success": False,
|
||
"stderr": str(e),
|
||
}
|
||
|
||
def run_duplicate_classifier(
|
||
work_dir: Path,
|
||
prefix: str,
|
||
blast_file: Path,
|
||
gff_file: Path
|
||
) -> Dict[str, Any]:
|
||
"""运行duplicate_gene_classifier进行复制基因分类"""
|
||
print(f"运行Duplicate Gene Classifier...")
|
||
|
||
start_time = time.time()
|
||
|
||
try:
|
||
result = subprocess.run(
|
||
f"duplicate_gene_classifier {prefix}",
|
||
capture_output=True,
|
||
text=True,
|
||
cwd=str(work_dir),
|
||
check=False,
|
||
shell=True
|
||
)
|
||
|
||
elapsed = time.time() - start_time
|
||
|
||
# 输出文件
|
||
log_file = work_dir / f"{prefix}.dup_class.log"
|
||
gene_type_file = work_dir / "gene_type"
|
||
|
||
# 写入日志
|
||
with open(log_file, 'w') as f:
|
||
f.write(result.stdout)
|
||
if result.stderr:
|
||
f.write("\n=== STDERR ===\n")
|
||
print(f" 完成，耗时: {elapsed:.1f}秒")
|
||
print(f" 日志文件: {log_file}")
|
||
|
||
return {
|
||
"success": result.returncode == 0,
|
||
"returncode": result.returncode,
|
||
"stdout": result.stdout,
|
||
"stderr": result.stderr,
|
||
"log_file": log_file,
|
||
"gene_type_file": gene_type_file,
|
||
"elapsed_sec": elapsed
|
||
}
|
||
|
||
except Exception as e:
|
||
return {
|
||
"success": False,
|
||
"stderr": str(e)
|
||
}
|
||
|
||
|
||
|
||
def run_mcscanx(
|
||
work_dir: Path,
|
||
prefix: str
|
||
) -> Dict[str, Any]:
|
||
"""运行MCScanX进行共线性分析"""
|
||
print(f"运行MCScanX...")
|
||
|
||
# 命令格式: MCScanX ./
|
||
# 需要在包含 {prefix}.gff 和 {prefix}.blast 的目录中运行
|
||
|
||
cmd = [
|
||
"MCScanX",
|
||
prefix
|
||
]
|
||
|
||
start_time = time.time()
|
||
|
||
try:
|
||
result = subprocess.run(
|
||
cmd,
|
||
capture_output=True,
|
||
text=True,
|
||
cwd=str(work_dir),
|
||
check=False
|
||
)
|
||
|
||
elapsed = time.time() - start_time
|
||
|
||
# MCScanX 可能输出到 .collinearity 或 {prefix}.collinearity
|
||
# 检查两种可能性
|
||
collinearity_file = work_dir / f"{prefix}.collinearity"
|
||
if not collinearity_file.exists():
|
||
collinearity_file = work_dir / ".collinearity"
|
||
|
||
print(f" 完成，耗时: {elapsed:.1f}秒")
|
||
print(f" 共线性文件: {collinearity_file}")
|
||
|
||
return {
|
||
"success": result.returncode == 0,
|
||
"returncode": result.returncode,
|
||
"stdout": result.stdout,
|
||
"stderr": result.stderr,
|
||
"collinearity_file": collinearity_file,
|
||
"elapsed_sec": elapsed
|
||
}
|
||
|
||
except Exception as e:
|
||
return {
|
||
"success": False,
|
||
"stderr": str(e)
|
||
}
|
||
|
||
|
||
|
||
|
||
def find_scope_gene_id_file(workspace: Path, prefix: str, scope_data: Dict[str, Any]) -> Optional[Path]:
|
||
"""Find family member IDs for scoped target-species synteny highlighting."""
|
||
if not scope_data:
|
||
return None
|
||
final_dir = workspace / "final"
|
||
family = str(scope_data.get("family") or "").strip()
|
||
targets = [str(x).strip() for x in (scope_data.get("target_species") or []) if str(x).strip()]
|
||
candidates: List[Path] = []
|
||
if targets and family:
|
||
for species in targets:
|
||
candidates.extend([
|
||
final_dir / f"Target.{species}.{family}.final_members.id",
|
||
final_dir / f"{species}.{family}.final_members.id",
|
||
final_dir / f"{species}.{prefix.split('.')[-1]}.final_members.id",
|
||
])
|
||
candidates.append(final_dir / f"{prefix}.final_members.id")
|
||
for candidate in candidates:
|
||
if candidate.exists() and candidate.stat().st_size > 0:
|
||
return candidate
|
||
return None
|
||
|
||
|
||
def _read_id_set(path: Optional[Path]) -> set[str]:
|
||
ids: set[str] = set()
|
||
if not path or not path.exists():
|
||
return ids
|
||
with path.open("r", encoding="utf-8", errors="replace") as handle:
|
||
for raw in handle:
|
||
token = raw.strip().split()[0] if raw.strip() else ""
|
||
if token:
|
||
ids.add(token)
|
||
return ids
|
||
|
||
|
||
def _parse_mcscanx_gff(gff_file: Path) -> tuple[Dict[str, tuple[str, int, int]], Dict[str, int]]:
|
||
genes: Dict[str, tuple[str, int, int]] = {}
|
||
chrom_lengths: Dict[str, int] = {}
|
||
with gff_file.open("r", encoding="utf-8", errors="replace") as handle:
|
||
for raw in handle:
|
||
if not raw.strip() or raw.startswith("#"):
|
||
continue
|
||
parts = raw.rstrip("\n").split("\t")
|
||
if len(parts) < 4:
|
||
continue
|
||
chrom, gene_id = parts[0], parts[1]
|
||
try:
|
||
start, end = int(float(parts[2])), int(float(parts[3]))
|
||
except ValueError:
|
||
continue
|
||
if start > end:
|
||
start, end = end, start
|
||
genes[gene_id] = (chrom, start, end)
|
||
chrom_lengths[chrom] = max(chrom_lengths.get(chrom, 0), end)
|
||
return genes, chrom_lengths
|
||
|
||
|
||
def _parse_collinearity_pairs(collinearity_file: Path) -> list[tuple[str, str]]:
|
||
pairs: list[tuple[str, str]] = []
|
||
with collinearity_file.open("r", encoding="utf-8", errors="replace") as handle:
|
||
for raw in handle:
|
||
line = raw.strip()
|
||
if not line or line.startswith("#"):
|
||
continue
|
||
parts = line.split()
|
||
if len(parts) >= 3:
|
||
pairs.append((parts[1], parts[2]))
|
||
return pairs
|
||
|
||
|
||
def generate_synteny_fallback_plot(
|
||
gff_file: Path,
|
||
collinearity_file: Path,
|
||
output_prefix: Path,
|
||
gene_id_file: Optional[Path] = None,
|
||
max_background_links: int = 3000,
|
||
) -> Dict[str, Any]:
|
||
"""Generate a chromosome-link synteny plot when Circos is unavailable.
|
||
|
||
This is a fallback visualization from the same MCScanX GFF/collinearity
|
||
outputs: chromosome models from gene coordinates, grey collinear gene-pair
|
||
links, and red links/ticks for pairs touching target family members.
|
||
"""
|
||
genes, chrom_lengths = _parse_mcscanx_gff(gff_file)
|
||
pairs = [(a, b) for a, b in _parse_collinearity_pairs(collinearity_file) if a in genes and b in genes]
|
||
family_ids = _read_id_set(gene_id_file)
|
||
if not genes or not chrom_lengths or not pairs:
|
||
return {
|
||
"success": False,
|
||
"reason": "fallback_plot_missing_gff_or_collinearity_pairs",
|
||
"genes": len(genes),
|
||
"chromosomes": len(chrom_lengths),
|
||
"pairs": len(pairs),
|
||
}
|
||
|
||
import matplotlib
|
||
matplotlib.use("Agg")
|
||
import matplotlib.pyplot as plt
|
||
from matplotlib.path import Path as MplPath
|
||
from matplotlib.patches import PathPatch
|
||
|
||
def chrom_key(name: str):
|
||
m = re.search(r"(\d+)$", name)
|
||
return (0, int(m.group(1))) if m else (1, name)
|
||
|
||
chroms = sorted(chrom_lengths, key=chrom_key)
|
||
n_chrom = len(chroms)
|
||
max_len = max(chrom_lengths.values()) or 1
|
||
fig_h = max(6.0, min(18.0, n_chrom * 0.45 + 2.2))
|
||
fig, ax = plt.subplots(figsize=(11.5, fig_h), dpi=220)
|
||
y_for = {chrom: n_chrom - i for i, chrom in enumerate(chroms)}
|
||
|
||
for chrom in chroms:
|
||
y = y_for[chrom]
|
||
length = chrom_lengths[chrom]
|
||
ax.plot([0, length], [y, y], color="#263238", linewidth=4, solid_capstyle="round", zorder=3)
|
||
ax.text(-max_len * 0.015, y, chrom, ha="right", va="center", fontsize=8, color="#263238")
|
||
|
||
highlighted = []
|
||
background = []
|
||
for gene_a, gene_b in pairs:
|
||
is_family = gene_a in family_ids or gene_b in family_ids
|
||
(highlighted if is_family else background).append((gene_a, gene_b))
|
||
|
||
if len(background) > max_background_links:
|
||
step = max(1, len(background) // max_background_links)
|
||
background = background[::step][:max_background_links]
|
||
|
||
def draw_pair(gene_a: str, gene_b: str, color: str, alpha: float, lw: float, z: int):
|
||
chr_a, start_a, end_a = genes[gene_a]
|
||
chr_b, start_b, end_b = genes[gene_b]
|
||
x1 = (start_a + end_a) / 2
|
||
x2 = (start_b + end_b) / 2
|
||
y1 = y_for[chr_a]
|
||
y2 = y_for[chr_b]
|
||
if chr_a == chr_b:
|
||
lift = 0.18 + min(0.75, abs(x2 - x1) / max_len * 2.0)
|
||
verts = [(x1, y1), ((x1 + x2) / 2, y1 + lift), (x2, y2)]
|
||
else:
|
||
lift = 0.22
|
||
verts = [(x1, y1), (x1, (y1 + y2) / 2 + lift), (x2, y2)]
|
||
patch = PathPatch(
|
||
MplPath(verts, [MplPath.MOVETO, MplPath.CURVE3, MplPath.CURVE3]),
|
||
facecolor="none",
|
||
edgecolor=color,
|
||
linewidth=lw,
|
||
alpha=alpha,
|
||
zorder=z,
|
||
)
|
||
ax.add_patch(patch)
|
||
|
||
for gene_a, gene_b in background:
|
||
draw_pair(gene_a, gene_b, "#9E9E9E", 0.18, 0.35, 1)
|
||
for gene_a, gene_b in highlighted:
|
||
draw_pair(gene_a, gene_b, "#D32F2F", 0.75, 0.85, 2)
|
||
|
||
plotted_family = 0
|
||
for gene_id in sorted(family_ids):
|
||
if gene_id not in genes:
|
||
continue
|
||
chrom, start, end = genes[gene_id]
|
||
y = y_for[chrom]
|
||
x = (start + end) / 2
|
||
ax.scatter([x], [y], s=10, color="#B71C1C", zorder=4)
|
||
plotted_family += 1
|
||
if plotted_family <= 80:
|
||
ax.text(x, y + 0.10, gene_id, rotation=60, ha="left", va="bottom", fontsize=4.5, color="#B71C1C")
|
||
|
||
ax.set_xlim(-max_len * 0.08, max_len * 1.03)
|
||
ax.set_ylim(0.4, n_chrom + 0.9)
|
||
ax.set_xlabel("Genomic position (bp)", fontsize=9)
|
||
ax.set_yticks([])
|
||
title = "Intraspecies synteny (MCScanX chromosome-link fallback)"
|
||
subtitle = f"{len(pairs)} collinear gene pairs"
|
||
if family_ids:
|
||
subtitle += f"; {len(highlighted)} pairs involving target-family genes"
|
||
ax.set_title(f"{title}\n{subtitle}", fontsize=11, pad=12)
|
||
ax.spines[["left", "right", "top"]].set_visible(False)
|
||
ax.grid(axis="x", color="#EEEEEE", linewidth=0.6)
|
||
fig.tight_layout()
|
||
|
||
png_file = output_prefix.with_suffix(".synteny.png")
|
||
pdf_file = output_prefix.with_suffix(".synteny.pdf")
|
||
fig.savefig(png_file, bbox_inches="tight")
|
||
fig.savefig(pdf_file, bbox_inches="tight")
|
||
plt.close(fig)
|
||
return {
|
||
"success": png_file.exists() and pdf_file.exists(),
|
||
"png_file": png_file,
|
||
"pdf_file": pdf_file,
|
||
"total_pairs_plotted_source": len(pairs),
|
||
"background_pairs_plotted": len(background),
|
||
"highlighted_pairs_plotted": len(highlighted),
|
||
"family_genes_plotted": plotted_family,
|
||
"fallback_reason": "circos_output_unavailable",
|
||
}
|
||
|
||
|
||
def _is_primary_chromosome_name(chrom: str) -> bool:
|
||
name = str(chrom or "").strip()
|
||
if re.fullmatch(r"\d+", name):
|
||
return True
|
||
return bool(re.fullmatch(r"chr(?:omosome)?[\s_.-]*\d+[A-Za-z]?", name, flags=re.IGNORECASE))
|
||
|
||
|
||
def _is_obvious_non_nuclear_sequence(chrom: str) -> bool:
|
||
name = str(chrom or "").strip().lower()
|
||
if re.search(r"(chloroplast|plastid|mitochond|mitoch|scaffold|contig|unplaced|unlocalized|random)", name):
|
||
return True
|
||
return bool(re.fullmatch(r"(chrm|chrmt|mt|mito|chrc|chrpt|pt|cp)", name))
|
||
|
||
|
||
def _select_circos_chromosomes(parsed: list[tuple[str, str]]) -> tuple[list[tuple[str, str]], Dict[str, Any]]:
|
||
"""Keep the nuclear chromosome background while dropping organelles/tiny accessions.
|
||
|
||
The fallback to all parsed records is intentional: species with unusual naming
|
||
should not fail synteny visualization just because a conservative filter cannot
|
||
identify the main chromosome set.
|
||
"""
|
||
records: list[tuple[str, int]] = []
|
||
for chrom, length_text in parsed:
|
||
try:
|
||
length = int(float(length_text))
|
||
except (TypeError, ValueError):
|
||
continue
|
||
if chrom and length > 0:
|
||
records.append((chrom, length))
|
||
|
||
total = len(parsed)
|
||
if not records:
|
||
return parsed, {
|
||
"circos_chromosomes_total": total,
|
||
"circos_chromosomes_plotted": len(parsed),
|
||
"circos_chromosomes_filtered": 0,
|
||
"circos_chromosomes_filter_mode": "fallback_unparsed_lengths",
|
||
"circos_chromosomes_filtered_names": [],
|
||
}
|
||
|
||
selected: list[tuple[str, int]] = []
|
||
mode = ""
|
||
named_primary = [(chrom, length) for chrom, length in records if _is_primary_chromosome_name(chrom)]
|
||
if named_primary:
|
||
selected = named_primary
|
||
mode = "primary_name"
|
||
else:
|
||
nuclear_candidates = [
|
||
(chrom, length)
|
||
for chrom, length in records
|
||
if not _is_obvious_non_nuclear_sequence(chrom)
|
||
]
|
||
length_source = nuclear_candidates or records
|
||
sorted_lengths = sorted((length for _, length in length_source), reverse=True)
|
||
median_length = sorted_lengths[len(sorted_lengths) // 2]
|
||
max_length = sorted_lengths[0]
|
||
threshold = max(1_000_000, int(median_length * 0.10), int(max_length * 0.02))
|
||
selected = [(chrom, length) for chrom, length in length_source if length >= threshold]
|
||
mode = "length_distribution"
|
||
|
||
if not selected:
|
||
selected = records
|
||
mode = "fallback_all_records"
|
||
|
||
selected_names = {chrom for chrom, _ in selected}
|
||
filtered_names = [chrom for chrom, _ in records if chrom not in selected_names]
|
||
selected_text = [(chrom, str(length)) for chrom, length in selected]
|
||
return selected_text, {
|
||
"circos_chromosomes_total": total,
|
||
"circos_chromosomes_plotted": len(selected_text),
|
||
"circos_chromosomes_filtered": max(0, total - len(selected_text)),
|
||
"circos_chromosomes_filter_mode": mode,
|
||
"circos_chromosomes_filtered_names": filtered_names,
|
||
}
|
||
|
||
def _format_circos_points(value: Any, default: float) -> str:
|
||
if value is None or value == "":
|
||
value = default
|
||
text = str(value).strip()
|
||
if text.endswith("p"):
|
||
return text
|
||
try:
|
||
return f"{float(text):g}p"
|
||
except ValueError:
|
||
return f"{default:g}p"
|
||
|
||
|
||
def _format_circos_radius(value: Any, default: float) -> str:
|
||
if value is None or value == "":
|
||
value = default
|
||
text = str(value).strip()
|
||
if "r" in text or "p" in text:
|
||
return text
|
||
try:
|
||
return f"{float(text):.2f}r"
|
||
except ValueError:
|
||
return f"{default:.2f}r"
|
||
|
||
|
||
def run_circos_prepare(
|
||
gff_file: Path,
|
||
collinearity_file: Path,
|
||
gene_id_file: Path,
|
||
output_prefix: Path,
|
||
genome_file: Path,
|
||
gene_label_size: Any = 38,
|
||
gene_track_r0: Any = 1.03,
|
||
gene_track_r1: Any = 1.30,
|
||
chrom_label_radius: Any = "1r - 60p",
|
||
chrom_label_size: Any = 40,
|
||
ideogram_thickness: Any = 90,
|
||
) -> Dict[str, Any]:
|
||
"""准备Circos绘图数据"""
|
||
print(f"准备Circos绘图数据...")
|
||
gene_label_size_text = _format_circos_points(gene_label_size, 38)
|
||
gene_track_r0_text = _format_circos_radius(gene_track_r0, 1.03)
|
||
gene_track_r1_text = _format_circos_radius(gene_track_r1, 1.30)
|
||
chrom_label_radius_text = _format_circos_radius(chrom_label_radius, 1.0)
|
||
chrom_label_size_text = _format_circos_points(chrom_label_size, 40)
|
||
ideogram_thickness_text = _format_circos_points(ideogram_thickness, 90)
|
||
|
||
# 1. 计算染色体长度
|
||
genome_len_file = output_prefix.parent / "genome.len"
|
||
cmd = [
|
||
"seqkit", "fx2tab", "-l", "-n", "-i", str(genome_file)
|
||
]
|
||
|
||
result = subprocess.run(
|
||
cmd,
|
||
capture_output=True,
|
||
text=True,
|
||
check=False
|
||
)
|
||
|
||
if result.returncode != 0:
|
||
return {"success": False, "stderr": result.stderr}
|
||
|
||
lines = result.stdout.strip().split('\n')
|
||
parsed = []
|
||
for line in lines:
|
||
parts = line.split('\t')
|
||
if len(parts) >= 2:
|
||
parsed.append((parts[0], parts[1]))
|
||
chromosomes, chromosome_metrics = _select_circos_chromosomes(parsed)
|
||
with open(genome_len_file, 'w') as f:
|
||
for chrom, length in chromosomes:
|
||
f.write(f"{chrom}\t{length}\n")
|
||
|
||
# 2. 生成核型文件
|
||
karyotype_file = output_prefix.with_suffix('.karyotype.circos.txt')
|
||
cmd2 = [
|
||
"awk",
|
||
'{print "chr\t-\t"$1"\t"$1"\t0\t"$2"\tchr"NR}',
|
||
str(genome_len_file)
|
||
]
|
||
|
||
result2 = subprocess.run(
|
||
cmd2,
|
||
capture_output=True,
|
||
text=True,
|
||
check=False
|
||
)
|
||
|
||
if result2.returncode == 0:
|
||
with open(karyotype_file, 'w') as f:
|
||
f.write(result2.stdout)
|
||
|
||
# 3. 使用get_mcscanx_circos.py生成绘图数据
|
||
circos_script = Path("scripts/get_mcscanx_circos.py")
|
||
|
||
if circos_script.exists() and gene_id_file.exists():
|
||
cmd3 = [
|
||
sys.executable,
|
||
str(circos_script),
|
||
str(gff_file),
|
||
str(collinearity_file),
|
||
str(gene_id_file),
|
||
str(output_prefix)
|
||
]
|
||
|
||
result3 = subprocess.run(
|
||
cmd3,
|
||
capture_output=True,
|
||
text=True,
|
||
check=False
|
||
)
|
||
|
||
print(f" Circos数据准备完成")
|
||
|
||
# 4. 创建Circos配置文件并绘图
|
||
circos_dir = output_prefix.parent / "circos_config"
|
||
circos_dir.mkdir(exist_ok=True)
|
||
|
||
# 创建简化版circos.conf配置文件
|
||
circos_conf = circos_dir / "circos.conf"
|
||
karyotype_rel = karyotype_file.name
|
||
links_rel = f"{output_prefix.name}.links.txt"
|
||
gene_text_rel = f"{output_prefix.name}.geneText.txt"
|
||
gene_block_links_rel = f"{output_prefix.name}.geneBlocklinks.txt"
|
||
|
||
# 检查SPL基因相关文件是否存在
|
||
gene_text_file = Path(f"{output_prefix}.geneText.txt")
|
||
gene_block_links_file = Path(f"{output_prefix}.geneBlocklinks.txt")
|
||
has_spl_data = gene_text_file.exists() and gene_block_links_file.exists()
|
||
|
||
with open(circos_conf, 'w') as f:
|
||
f.write(f"""# Circos configuration file
|
||
# Generated by BotanicalBaseAgent Synteny pipeline
|
||
|
||
karyotype = {karyotype_rel}
|
||
chromosomes_units = 1000000
|
||
chromosomes_display_default = yes
|
||
|
||
<<include etc/colors_fonts_patterns.conf>>
|
||
|
||
<ideogram>
|
||
<spacing>
|
||
default = 0.01r
|
||
</spacing>
|
||
radius = 0.5r
|
||
thickness = {ideogram_thickness_text}
|
||
fill = yes
|
||
fill_color = black
|
||
stroke_color = dgrey
|
||
stroke_thickness = 1p
|
||
show_label = yes
|
||
label_font = default
|
||
label_radius = {chrom_label_radius_text}
|
||
label_size = {chrom_label_size_text}
|
||
label_parallel = yes
|
||
</ideogram>
|
||
|
||
<ticks>
|
||
radius = 1r
|
||
color = black
|
||
thickness = 2p
|
||
multiplier = 1e-6
|
||
orientation = out
|
||
format = %d
|
||
<tick>
|
||
spacing = 10u
|
||
size = 15p
|
||
show_label = no
|
||
format = %d
|
||
</tick>
|
||
</ticks>
|
||
""")
|
||
# 添加plots区块显示基因ID
|
||
if has_spl_data:
|
||
f.write(f"""
|
||
<plots>
|
||
<plot>
|
||
type = text
|
||
file = {gene_text_rel}
|
||
color = red
|
||
r1 = {gene_track_r1_text}
|
||
r0 = {gene_track_r0_text}
|
||
label_font = default
|
||
label_size = {gene_label_size_text}
|
||
label_snuggle = yes
|
||
show_links = yes
|
||
link_dims = 5p,4p,60p,2p,2p
|
||
link_thickness = 2p
|
||
link_color = green
|
||
</plot>
|
||
</plots>
|
||
""")
|
||
# 添加links区块
|
||
f.write(f"""
|
||
<links>
|
||
<link>
|
||
file = {links_rel}
|
||
radius = 0.9r
|
||
bezier_radius = 0r
|
||
color = lgrey
|
||
thickness = 2
|
||
ribbon = yes
|
||
</link>
|
||
""")
|
||
# 添加SPL基因高亮连线
|
||
if has_spl_data:
|
||
f.write(f"""<link>
|
||
file = {gene_block_links_rel}
|
||
radius = 0.9r
|
||
bezier_radius = 0r
|
||
color = red
|
||
thickness = 2
|
||
ribbon = yes
|
||
</link>
|
||
""")
|
||
f.write(f"""</links>
|
||
|
||
<image>
|
||
dir = .
|
||
file = {output_prefix.name}.circos
|
||
png = yes
|
||
svg = yes
|
||
radius = 3000p
|
||
background = white
|
||
angle_offset = -90
|
||
</image>
|
||
|
||
<<include etc/housekeeping.conf>>
|
||
""")
|
||
|
||
# 复制Circos输入文件到circos_config目录。Circos以cwd运行，
|
||
# 配置中使用相对文件名，必须覆盖旧的空文件并同步links/text输入。
|
||
import shutil
|
||
for src in (
|
||
karyotype_file,
|
||
Path(f"{output_prefix}.links.txt"),
|
||
Path(f"{output_prefix}.geneText.txt"),
|
||
Path(f"{output_prefix}.geneBlocklinks.txt"),
|
||
):
|
||
if src.exists():
|
||
shutil.copy2(src, circos_dir / src.name)
|
||
|
||
# 运行Circos绘图
|
||
print(f" 运行Circos绘图...")
|
||
cmd4 = [
|
||
"circos",
|
||
"-conf", str(circos_conf.resolve())
|
||
]
|
||
|
||
circos_env = os.environ.copy()
|
||
conda_bin = str(CONDA_PREFIX / "bin")
|
||
circos_env["PATH"] = conda_bin + os.pathsep + circos_env.get("PATH", "")
|
||
circos_env.pop("PERL5LIB", None)
|
||
result4 = subprocess.run(
|
||
cmd4,
|
||
capture_output=True,
|
||
text=True,
|
||
check=False,
|
||
cwd=str(circos_dir), # 在circos_config目录中运行
|
||
env=circos_env
|
||
)
|
||
|
||
# 检查输出文件（在circos_config目录中）
|
||
svg_file_in_config = circos_dir / f"{output_prefix.name}.circos.svg"
|
||
png_file_in_config = circos_dir / f"{output_prefix.name}.circos.png"
|
||
|
||
# 输出文件目标位置（analysis目录）
|
||
svg_file = output_prefix.parent / f"{output_prefix.name}.circos.svg"
|
||
png_file = output_prefix.parent / f"{output_prefix.name}.circos.png"
|
||
|
||
# 移动输出文件到analysis目录
|
||
if svg_file_in_config.exists():
|
||
shutil.move(svg_file_in_config, svg_file)
|
||
if png_file_in_config.exists():
|
||
shutil.move(png_file_in_config, png_file)
|
||
|
||
# 如果只有SVG没有PNG，尝试转换
|
||
if svg_file.exists() and not png_file.exists():
|
||
print(f" 转换SVG为PNG...")
|
||
convert_cmd = [
|
||
"convert",
|
||
"-density", "300",
|
||
str(svg_file),
|
||
str(png_file)
|
||
]
|
||
subprocess.run(convert_cmd, capture_output=True, check=False)
|
||
|
||
circos_success = result4.returncode == 0
|
||
print(f" Circos绘图完成" if circos_success else f" Circos绘图失败: {result4.stderr[:200]}")
|
||
|
||
return {
|
||
"success": circos_success,
|
||
"circos_returncode": result4.returncode,
|
||
"circos_stdout": result4.stdout,
|
||
"circos_stderr": result4.stderr,
|
||
"genome_len_file": genome_len_file,
|
||
"karyotype_file": karyotype_file,
|
||
"links_file": f"{output_prefix}.links.txt",
|
||
"genepair_file": f"{output_prefix}.genepair.txt",
|
||
"circos_conf": circos_conf,
|
||
"svg_file": svg_file if svg_file.exists() else None,
|
||
"png_file": png_file if png_file.exists() else None,
|
||
**chromosome_metrics,
|
||
}
|
||
|
||
return {
|
||
"success": True,
|
||
"genome_len_file": genome_len_file,
|
||
"karyotype_file": karyotype_file,
|
||
**chromosome_metrics,
|
||
}
|
||
|
||
|
||
def _create_circos_config_files(config_dir: Path):
|
||
"""创建Circos所需的配置文件"""
|
||
# ideogram.conf - 使用内联配置，避免嵌套include
|
||
with open(config_dir / "ideogram.conf", 'w') as f:
|
||
f.write("""<ideogram>
|
||
<spacing>
|
||
default = 0.01r
|
||
break = 0.5r
|
||
</spacing>
|
||
|
||
radius = 0.90r
|
||
thickness = 100p
|
||
fill = yes
|
||
fill_color = black
|
||
|
||
show_label = yes
|
||
label_font = default
|
||
label_radius = 1.075r
|
||
label_size = 10p
|
||
label_parallel = yes
|
||
|
||
show_bands = yes
|
||
fill_bands = yes
|
||
thickness = 2p
|
||
</ideogram>
|
||
""")
|
||
|
||
# ticks.conf
|
||
with open(config_dir / "ticks.conf", 'w') as f:
|
||
f.write("""show_ticks = yes
|
||
show_tick_labels = yes
|
||
|
||
<ticks>
|
||
radius = 1r
|
||
color = black
|
||
thickness = 2p
|
||
|
||
<tick>
|
||
spacing = 5u
|
||
size = 10p
|
||
show_label = no
|
||
</tick>
|
||
|
||
<tick>
|
||
spacing = 25u
|
||
size = 15p
|
||
show_label = yes
|
||
label_size = 10p
|
||
label_offset = 5p
|
||
format = %d
|
||
</tick>
|
||
</ticks>
|
||
""")
|
||
|
||
# image.conf
|
||
with open(config_dir / "image.conf", 'w') as f:
|
||
f.write("""dir = .
|
||
file = circos
|
||
png = yes
|
||
svg = yes
|
||
radius = 1500p
|
||
background = white
|
||
angle_offset = -90
|
||
""")
|
||
|
||
# colors_fonts_patterns.conf - 使用系统默认配置
|
||
with open(config_dir / "colors_fonts_patterns.conf", 'w') as f:
|
||
f.write("""# 使用Circos系统默认颜色配置
|
||
# 这些文件在Circos安装目录的etc/下
|
||
<<include etc/colors.ucsc.conf>>
|
||
<<include etc/colors.brewer.conf>>
|
||
<<include etc/colors.hsv.conf>>
|
||
|
||
<<include etc/fonts.conf>>
|
||
|
||
<<include etc/patterns.conf>>
|
||
""")
|
||
|
||
# housekeeping.conf
|
||
with open(config_dir / "housekeeping.conf", 'w') as f:
|
||
f.write("""anglestep = 0.5
|
||
minslicestep = 10
|
||
beziersamples = 40
|
||
|
||
warnings = 1
|
||
debug_group = summary
|
||
debug_verbosity = 5
|
||
|
||
text_snuggle_method = max
|
||
snuggle_tolerance = 1
|
||
snuggle_sampling = 2
|
||
snuggle_refine = yes
|
||
snuggle_link_overlap_tolerance = 1
|
||
|
||
offsets = 0
|
||
|
||
file_delim = \\s
|
||
file_delim_collapse = yes
|
||
|
||
list_record_delim = \\s*[;,]\\s*
|
||
list_field_delim = \\s*[:=]\\s*
|
||
options_record_delim = \\s*[;,]\\s*
|
||
options_field_delim = \\s*[:=]\\s*
|
||
""")
|
||
|
||
|
||
def compute_blast_stats(blast_file: Path) -> Dict[str, Any]:
|
||
"""计算BLAST比对统计"""
|
||
try:
|
||
with open(blast_file, 'r') as f:
|
||
lines = f.readlines()
|
||
|
||
# 统计比对数量
|
||
return {
|
||
"total_alignments": len(lines),
|
||
"unique_queries": len(set(l.split('\t')[0] for l in lines if l.strip())),
|
||
"unique_subjects": len(set(l.split('\t')[1] for l in lines if l.strip()))
|
||
}
|
||
except Exception:
|
||
return {}
|
||
|
||
|
||
def compute_collinearity_stats(collinearity_file: Path) -> Dict[str, Any]:
|
||
"""计算共线性统计"""
|
||
try:
|
||
with open(collinearity_file, 'r') as f:
|
||
content = f.read()
|
||
|
||
# 统计共线性块数量
|
||
blocks = content.count('##')
|
||
|
||
# 统计基因对数量
|
||
gene_pairs = 0
|
||
for line in content.split('\n'):
|
||
if line.startswith('#') or not line.strip():
|
||
continue
|
||
parts = line.split('\t')
|
||
if len(parts) >= 3:
|
||
gene_pairs += 1
|
||
|
||
return {
|
||
"synteny_blocks": blocks,
|
||
"gene_pairs": gene_pairs
|
||
}
|
||
except Exception:
|
||
return {}
|
||
|
||
|
||
def write_report(
|
||
workspace: Path,
|
||
prefix: str,
|
||
metrics: Dict[str, Any],
|
||
parameters: Dict[str, Any],
|
||
tool_versions: Dict[str, str],
|
||
diamond_result: Dict[str, Any],
|
||
gff_filter_result: Dict[str, Any],
|
||
classifier_result: Dict[str, Any],
|
||
mcscanx_result: Dict[str, Any],
|
||
artifacts: Dict[str, Dict]
|
||
) -> Path:
|
||
"""写入人可读的分析报告"""
|
||
report_path = workspace / "final" / f"{prefix}.synteny_report.md"
|
||
|
||
print(f"生成分析报告: {report_path}")
|
||
|
||
with open(report_path, "w") as f:
|
||
f.write(f"# 共线性分析报告 - {prefix}\n\n")
|
||
f.write(f"**生成时间**: {datetime.now().isoformat()}\n\n")
|
||
|
||
f.write("## 1. 分析概述\n")
|
||
f.write(f"- **分析阶段**: Synteny\n")
|
||
f.write(f"- **分析状态**: {'成功' if mcscanx_result.get('success', False) else '部分成功'}\n")
|
||
f.write(f"- **基因数量**: {metrics.get('gene_count', 'N/A')}\n")
|
||
f.write(f"- **BLAST比对数**: {metrics.get('blast_alignments', 'N/A')}\n")
|
||
f.write(f"- **共线性块数**: {metrics.get('synteny_blocks', 'N/A')}\n\n")
|
||
|
||
f.write("## 2. 工具版本\n")
|
||
for tool, version in tool_versions.items():
|
||
f.write(f"- **{tool}**: {version}\n")
|
||
f.write("\n")
|
||
|
||
f.write("## 3. 执行参数\n")
|
||
for param, value in parameters.items():
|
||
f.write(f"- **{param}**: {value}\n")
|
||
f.write("\n")
|
||
|
||
f.write("## 4. 执行命令\n")
|
||
f.write("### BLAST比对\n")
|
||
if diamond_result.get('command'):
|
||
f.write(f"```bash\n{diamond_result.get('command')}\n```\n\n")
|
||
|
||
f.write("### GFF过滤\n")
|
||
f.write(f"```bash\nawk -F '\\t' ...\n```\n\n")
|
||
|
||
f.write("### 复制基因分类\n")
|
||
if classifier_result.get('returncode') == 0:
|
||
f.write("✅ **复制基因分类成功**\n\n")
|
||
|
||
f.write("### MCScanX共线性分析\n")
|
||
if mcscanx_result.get('success'):
|
||
f.write("✅ **共线性分析成功**\n\n")
|
||
|
||
f.write("### Circos共线性绘图\n")
|
||
if artifacts.get('circos_svg') or artifacts.get('circos_png'):
|
||
f.write("✅ **Circos共线性绘图成功**\n\n")
|
||
f.write("Circos图展示了基因在染色体间的共线性关系，包括：\n")
|
||
f.write("- **染色体圈图**: 外圈显示染色体及其长度\n")
|
||
f.write("- **共线性连线**: 弧线连接具有共线性关系的基因对\n")
|
||
f.write("- **基因标签**: 显示关键基因的名称和位置\n")
|
||
if metrics.get('circos_chromosomes_total') is not None:
|
||
f.write(
|
||
"- **染色体筛选**: 圈图保留主核染色体背景，"
|
||
"过滤掉疑似细胞器/非主序列 "
|
||
f"({metrics.get('circos_chromosomes_plotted', 'N/A')}/"
|
||
f"{metrics.get('circos_chromosomes_total', 'N/A')} plotted)\n"
|
||
)
|
||
f.write("\n")
|
||
else:
|
||
f.write("⚠️ **Circos绘图未生成**\n\n")
|
||
|
||
f.write("## 5. 产出文件\n")
|
||
for artifact_name, artifact_info in artifacts.items():
|
||
if isinstance(artifact_info, dict):
|
||
f.write(f"- **{artifact_name}**: {artifact_info.get('path', 'N/A')}\n")
|
||
f.write("\n")
|
||
|
||
f.write("## 6. 详细统计\n")
|
||
f.write("```json\n")
|
||
f.write(json.dumps(metrics, indent=2, ensure_ascii=False))
|
||
f.write("\n```\n")
|
||
|
||
return report_path
|
||
|
||
|
||
def write_manifest(
|
||
workspace: Path,
|
||
job_id: str,
|
||
run_id: str,
|
||
prefix: str,
|
||
start_time: datetime,
|
||
end_time: datetime,
|
||
artifacts: Dict[str, Dict],
|
||
metrics: Dict[str, Any],
|
||
parameters: Dict[str, Any],
|
||
tool_versions: Dict[str, str],
|
||
cmd_history: List[str],
|
||
status: str = "success"
|
||
) -> Path:
|
||
"""写入符合schema v1的manifest文件"""
|
||
elapsed_sec = (end_time - start_time).total_seconds()
|
||
rebased_parameters = rebase_manifest_payload_paths(workspace, parameters)
|
||
rebased_artifacts = rebase_manifest_payload_paths(workspace, artifacts)
|
||
rebased_metrics = rebase_manifest_payload_paths(workspace, metrics)
|
||
|
||
manifest_data = {
|
||
"manifest_version": "1.0",
|
||
"job_id": job_id,
|
||
"stage": "synteny",
|
||
"run_id": run_id,
|
||
"prefix": prefix,
|
||
|
||
"timestamp": end_time.isoformat(),
|
||
"status": status,
|
||
"start_time": start_time.isoformat(),
|
||
"end_time": end_time.isoformat(),
|
||
"elapsed_sec": round(elapsed_sec, 2),
|
||
|
||
"cmd": rebase_manifest_payload_paths(workspace, cmd_history),
|
||
"tool_versions": tool_versions,
|
||
|
||
"workspace_root": str(workspace.parent.absolute()),
|
||
"workspace": str((workspace / "final").absolute()),
|
||
"input_dir": str(current_input_dir(workspace).absolute()),
|
||
|
||
"inputs": [
|
||
{"type": "protein_fasta", "path": rebased_parameters.get("protein_fasta", "")},
|
||
{"type": "gff3", "path": rebased_parameters.get("gff3", "")},
|
||
{"type": "genome", "path": rebased_parameters.get("genome", "")}
|
||
],
|
||
|
||
"artifacts": rebased_artifacts,
|
||
|
||
"parameters": rebased_parameters,
|
||
"metrics": rebased_metrics,
|
||
|
||
"provenance": {
|
||
"derived_from": f"{prefix}.identify.manifest.json",
|
||
"stage_sequence": ["identify", "phylogeny", "synteny"]
|
||
}
|
||
}
|
||
for key in (
|
||
"scope_id",
|
||
"scope_mode",
|
||
"scope_file",
|
||
"analysis_route",
|
||
"member_set_id",
|
||
"target_species",
|
||
"reference_species",
|
||
"comparative_species",
|
||
"selected_species",
|
||
"family",
|
||
):
|
||
if key in rebased_parameters:
|
||
manifest_data[key] = rebased_parameters[key]
|
||
|
||
manifest_path = workspace / "final" / f"{prefix}.synteny.manifest.json"
|
||
manifest_path.parent.mkdir(parents=True, exist_ok=True)
|
||
|
||
print(f"生成Manifest文件: {manifest_path}")
|
||
|
||
with open(manifest_path, "w", encoding='utf-8') as f:
|
||
json.dump(manifest_data, f, indent=2, ensure_ascii=False)
|
||
|
||
if parameters.get("scope_id"):
|
||
scoped_manifest_path = workspace / "final" / f"{prefix}.{sanitize_token(parameters.get('scope_id', ''))}.synteny.manifest.json"
|
||
scoped_manifest_data = dict(manifest_data)
|
||
scoped_manifest_data["artifacts"] = materialize_scoped_artifacts(
|
||
artifacts=rebased_artifacts,
|
||
workspace=workspace,
|
||
prefix=prefix,
|
||
scope_id=parameters.get("scope_id", ""),
|
||
)
|
||
with open(scoped_manifest_path, "w", encoding='utf-8') as f:
|
||
json.dump(scoped_manifest_data, f, indent=2, ensure_ascii=False)
|
||
|
||
return manifest_path
|
||
|
||
|
||
def main():
|
||
"""主函数：执行Synteny分析流程"""
|
||
parser = argparse.ArgumentParser(
|
||
description="Synteny阶段：种内共线性分析",
|
||
formatter_class=argparse.RawDescriptionHelpFormatter,
|
||
epilog="""
|
||
示例：
|
||
python run_synteny.py --prefix Ft.SPL --workspace /path/to/workspace --cpu 4
|
||
|
||
输入要求：
|
||
- Identify阶段必须已成功运行
|
||
- 需要GFF3注释文件和基因组文件
|
||
|
||
输出产物：
|
||
- final/{prefix}.blast (BLAST比对结果)
|
||
- final/{prefix}.gff (过滤后的GFF)
|
||
- final/{prefix}.collinearity (MCScanX共线性结果)
|
||
- final/{prefix}.synteny_report.md (分析报告)
|
||
- final/{prefix}.synteny.manifest.json (机器清单)
|
||
"""
|
||
)
|
||
|
||
parser.add_argument("--prefix", required=True,
|
||
help="分析前缀，格式：物种.家族（如 Ft.SPL）")
|
||
parser.add_argument("--workspace", required=True,
|
||
help="工作空间目录（workspace/<job_id>/)")
|
||
parser.add_argument("--input-dir",
|
||
help="输入目录（默认：workspace/<job_id>/../input/<job_id>）")
|
||
parser.add_argument("--cpu", type=int, default=8,
|
||
help="CPU核心数（默认：8）")
|
||
parser.add_argument("--gff3",
|
||
help="GFF3注释文件路径")
|
||
parser.add_argument("--genome",
|
||
help="基因组FASTA文件路径")
|
||
parser.add_argument("--evalue", type=float, default=1e-10,
|
||
help="BLAST E-value阈值（默认：1e-10）")
|
||
parser.add_argument("--max-target-seqs", type=int, default=5,
|
||
help="最大目标序列数（默认：5）")
|
||
parser.add_argument("--use-blastp", action="store_true",
|
||
help="使用NCBI blastp代替diamond")
|
||
parser.add_argument("--log-file",
|
||
help="日志文件路径")
|
||
parser.add_argument("--gene-id-file",
|
||
help="基因ID列表文件路径（可选，用于Circos绘图）")
|
||
parser.add_argument("--scope-file",
|
||
help="Gene-family scope JSON from resolve_scope.py")
|
||
parser.add_argument("--analysis-route",
|
||
help="Scoped analysis route recorded in manifest, e.g. single_species")
|
||
parser.add_argument("--member-set-id",
|
||
help="Scoped member set identity recorded in manifest")
|
||
args = parser.parse_args()
|
||
|
||
# 记录开始时间
|
||
start_time = datetime.now()
|
||
run_id = f"synteny_{start_time.strftime('%Y%m%d_%H%M%S_%f')[:-3]}"
|
||
|
||
# 设置工作空间
|
||
workspace = Path(args.workspace)
|
||
job_id = workspace.name
|
||
|
||
# 设置输入目录
|
||
if args.input_dir:
|
||
input_dir = Path(args.input_dir)
|
||
else:
|
||
project_root = workspace.parent.parent
|
||
input_dir = project_root / "input" / job_id
|
||
|
||
scope_data: Dict[str, Any] = {}
|
||
scope_protein: Optional[Path] = None
|
||
scope_gff3: Optional[Path] = None
|
||
scope_genome: Optional[Path] = None
|
||
analysis_route = str(args.analysis_route or "").strip()
|
||
member_set_id = str(args.member_set_id or "").strip()
|
||
|
||
print("=" * 60)
|
||
print(f"🚀 启动共线性分析")
|
||
print("=" * 60)
|
||
print(f"分析前缀: {args.prefix}")
|
||
print(f"工作空间: {workspace}")
|
||
print(f"任务ID: {job_id}")
|
||
print(f"运行ID: {run_id}")
|
||
print(f"CPU核心: {args.cpu}")
|
||
|
||
cmd_history = []
|
||
|
||
try:
|
||
scope_file: Optional[Path] = None
|
||
if args.scope_file:
|
||
scope_file = Path(args.scope_file)
|
||
if not scope_file.exists():
|
||
raise FileNotFoundError(f"Scope file not found: {args.scope_file}")
|
||
scope_data = read_scope(scope_file)
|
||
scope_data, scope_protein, scope_gff3, scope_genome = resolve_synteny_scope_inputs(workspace, scope_file)
|
||
scope_targets = [str(x).strip() for x in (scope_data.get("target_species") or []) if str(x).strip()]
|
||
scope_target_species = scope_targets[0] if scope_targets else ""
|
||
scope_protein, scope_protein_source = prefer_representative_target_file(
|
||
input_dir, scope_target_species, "pep", scope_protein
|
||
)
|
||
scope_gff3, scope_gff_source = prefer_representative_target_file(
|
||
input_dir, scope_target_species, "gff", scope_gff3
|
||
)
|
||
scope_genome, scope_genome_source = prefer_representative_target_file(
|
||
input_dir, scope_target_species, "genome", scope_genome
|
||
)
|
||
if scope_protein_source == "representative":
|
||
print(f" Scope代表转录本PEP: {scope_protein}")
|
||
elif scope_protein_source == "scope_raw_fallback":
|
||
print(f" ⚠️ 未找到 {scope_target_species} 代表转录本PEP，回退使用scope原始PEP: {scope_protein}")
|
||
if scope_gff_source == "representative":
|
||
print(f" Scope代表转录本GFF: {scope_gff3}")
|
||
elif scope_gff_source == "scope_raw_fallback":
|
||
print(f" ⚠️ 未找到 {scope_target_species} 代表转录本GFF，回退使用scope原始GFF: {scope_gff3}")
|
||
if scope_genome_source == "representative":
|
||
print(f" Scope代表基因组FASTA: {scope_genome}")
|
||
elif scope_genome_source == "scope_raw_fallback":
|
||
print(f" ⚠️ 未找到 {scope_target_species} 代表基因组FASTA，回退使用scope原始genome: {scope_genome}")
|
||
if not analysis_route:
|
||
analysis_route = "single_species"
|
||
if not member_set_id and scope_data.get("scope_id") and analysis_route:
|
||
member_set_id = f"{scope_data.get('scope_id')}:{analysis_route}:all_members"
|
||
|
||
# 1. 准备目录结构
|
||
print("\n📁 准备目录结构...")
|
||
paths = prepare_directories(workspace)
|
||
|
||
# 2. 获取工具版本
|
||
print("\n🔧 检查工具版本...")
|
||
tool_versions = get_tool_versions()
|
||
for tool, version in tool_versions.items():
|
||
print(f" {tool}: {version}")
|
||
|
||
# 3. 验证输入文件
|
||
print("\n🔍 验证输入文件...")
|
||
inputs = validate_inputs(
|
||
workspace=workspace,
|
||
prefix=args.prefix,
|
||
input_dir=input_dir,
|
||
gff3=args.gff3 or scope_gff3,
|
||
genome=args.genome or scope_genome,
|
||
protein_fasta=scope_protein,
|
||
)
|
||
if not inputs.get("gene_id_file") and scope_data:
|
||
scoped_gene_id_file = find_scope_gene_id_file(workspace, args.prefix, scope_data)
|
||
if scoped_gene_id_file:
|
||
inputs["gene_id_file"] = scoped_gene_id_file
|
||
print(f" Scope基因ID文件: {scoped_gene_id_file}")
|
||
original_protein_fasta = inputs["protein_fasta"]
|
||
safe_protein_fasta, safe_stats = write_blast_safe_protein_fasta(
|
||
Path(original_protein_fasta),
|
||
paths["analysis_dir"] / f"{args.prefix}.blast_safe.pep.fa",
|
||
)
|
||
inputs["protein_fasta"] = safe_protein_fasta
|
||
print(
|
||
" BLAST-safe protein FASTA: "
|
||
f"{safe_protein_fasta} "
|
||
f"(records={safe_stats['records']}, removed_chars={safe_stats['removed_chars']})"
|
||
)
|
||
|
||
# 4. 运行BLAST比对
|
||
print("\n🔄 运行BLAST比对...")
|
||
blast_file = paths["analysis_dir"] / f"{args.prefix}.blast"
|
||
|
||
if args.use_blastp:
|
||
blast_result = run_blastp(
|
||
input_fasta=inputs["protein_fasta"],
|
||
output_blast=blast_file,
|
||
cpu=args.cpu,
|
||
evalue=args.evalue,
|
||
max_alignments=args.max_target_seqs
|
||
)
|
||
else:
|
||
# 优先尝试diamond
|
||
blast_result = run_diamond_blastp(
|
||
input_fasta=inputs["protein_fasta"],
|
||
output_blast=blast_file,
|
||
cpu=args.cpu,
|
||
evalue=args.evalue,
|
||
max_target_seqs=args.max_target_seqs
|
||
)
|
||
|
||
# 如果diamond失败，尝试blastp
|
||
if not blast_result.get("success", False):
|
||
print(" Diamond失败，尝试使用NCBI blastp...")
|
||
blast_result = run_blastp(
|
||
input_fasta=inputs["protein_fasta"],
|
||
output_blast=blast_file,
|
||
cpu=args.cpu,
|
||
evalue=args.evalue,
|
||
max_alignments=args.max_target_seqs
|
||
)
|
||
|
||
if not blast_result.get("success", False):
|
||
print("❌ BLAST比对失败")
|
||
raise RuntimeError(f"BLAST失败: {blast_result.get('stderr', '未知错误')}")
|
||
|
||
cmd_history.append(blast_result.get("command", ""))
|
||
|
||
# 5. 复制blast文件到工作目录
|
||
work_blast = paths["analysis_dir"] / f"{args.prefix}.blast"
|
||
import shutil
|
||
# 只有在源和目标不同时才复制
|
||
if blast_file != work_blast:
|
||
shutil.copy(blast_file, work_blast)
|
||
# 6. 过滤GFF3
|
||
print("\n🔍 过滤GFF3文件...")
|
||
gff_file = paths["analysis_dir"] / f"{args.prefix}.gff"
|
||
preferred_gff_ids = read_fasta_ids_for_matching(inputs["protein_fasta"])
|
||
gff_filter_result = filter_gff3(
|
||
gff3_file=inputs["gff3"],
|
||
output_gff=gff_file,
|
||
preferred_ids=preferred_gff_ids,
|
||
)
|
||
|
||
if not gff_filter_result.get("success", False):
|
||
print("❌ GFF3过滤失败")
|
||
raise RuntimeError(f"GFF3过滤失败")
|
||
|
||
work_gff = paths["analysis_dir"] / f"{args.prefix}.gff"
|
||
# 只有在源和目标不同时才复制
|
||
if gff_file != work_gff:
|
||
shutil.copy(gff_file, work_gff)
|
||
|
||
# 7. 运行duplicate_gene_classifier
|
||
print("\n🔍 运行复制基因分类...")
|
||
|
||
# 对于 MCScanX 和 duplicate_gene_classifier，使用物种前缀（如 Ft）而非完整前缀（如 Ft.SPL）
|
||
# 因为这些工具在处理带点的前缀时会有问题
|
||
species_prefix = args.prefix.split('.')[0] if '.' in args.prefix else args.prefix
|
||
|
||
# 复制文件并重命名为物种前缀
|
||
import shutil
|
||
work_blast_species = paths["analysis_dir"] / f"{species_prefix}.blast"
|
||
work_gff_species = paths["analysis_dir"] / f"{species_prefix}.gff"
|
||
if work_blast != work_blast_species:
|
||
shutil.copy(work_blast, work_blast_species)
|
||
if work_gff != work_gff_species:
|
||
shutil.copy(work_gff, work_gff_species)
|
||
|
||
# 使用物种前缀运行 duplicate_gene_classifier
|
||
classifier_result = run_duplicate_classifier(
|
||
work_dir=paths["analysis_dir"],
|
||
prefix=species_prefix,
|
||
blast_file=work_blast_species,
|
||
gff_file=work_gff_species
|
||
)
|
||
# 检查是否成功 - 只要有返回结果就算成功
|
||
if classifier_result and (classifier_result.get("returncode") == 0 or classifier_result.get("log_file")):
|
||
|
||
print(" 复制基因分类完成")
|
||
|
||
# 8. 运行MCScanX
|
||
print("\n🔍 运行MCScanX共线性分析...")
|
||
mcscanx_result = run_mcscanx(
|
||
work_dir=paths["analysis_dir"],
|
||
prefix=species_prefix
|
||
)
|
||
|
||
if mcscanx_result.get("success", False):
|
||
print(" 共线性分析完成")
|
||
else:
|
||
print(" ⚠️ MCScanX可能存在问题")
|
||
|
||
# 9. 准备Circos绘图数据（可选）
|
||
circos_data = {}
|
||
if mcscanx_result.get("success", False) and inputs.get("gene_id_file"):
|
||
print("\n🔍 准备Circos绘图数据...")
|
||
circos_data = run_circos_prepare(
|
||
gff_file=gff_file,
|
||
collinearity_file=mcscanx_result.get("collinearity_file", paths["analysis_dir"] / f"{args.prefix}.collinearity"),
|
||
gene_id_file=inputs["gene_id_file"],
|
||
output_prefix=paths["analysis_dir"] / f"{args.prefix}.circos",
|
||
genome_file=inputs["genome"]
|
||
)
|
||
|
||
# 10. 计算统计信息
|
||
print("\n📊 计算统计信息...")
|
||
metrics = {}
|
||
|
||
# BLAST统计
|
||
if blast_file.exists():
|
||
blast_stats = compute_blast_stats(blast_file)
|
||
metrics.update(blast_stats)
|
||
|
||
# GFF统计
|
||
metrics["gene_count"] = gff_filter_result.get("gene_count", 0)
|
||
|
||
# 共线性统计 - 使用 species_prefix
|
||
collinearity_file = paths["analysis_dir"] / f"{species_prefix}.collinearity"
|
||
if collinearity_file.exists():
|
||
collinearity_stats = compute_collinearity_stats(collinearity_file)
|
||
metrics.update(collinearity_stats)
|
||
|
||
for key in (
|
||
"circos_chromosomes_total",
|
||
"circos_chromosomes_plotted",
|
||
"circos_chromosomes_filtered",
|
||
"circos_chromosomes_filter_mode",
|
||
"circos_chromosomes_filtered_names",
|
||
):
|
||
if key in circos_data:
|
||
metrics[key] = circos_data[key]
|
||
|
||
# 10.5 Circos不可用或未产图时，使用MCScanX结果生成标准染色体-link后备图
|
||
fallback_plot = {}
|
||
circos_has_image = bool(
|
||
circos_data.get("success", False)
|
||
and (
|
||
(circos_data.get("svg_file") and Path(circos_data["svg_file"]).exists())
|
||
or (circos_data.get("png_file") and Path(circos_data["png_file"]).exists())
|
||
)
|
||
)
|
||
if collinearity_file.exists() and not circos_has_image:
|
||
print("\n🎨 Circos未产出图片，生成MCScanX染色体-link后备图...")
|
||
fallback_plot = generate_synteny_fallback_plot(
|
||
gff_file=gff_file,
|
||
collinearity_file=collinearity_file,
|
||
output_prefix=paths["analysis_dir"] / f"{args.prefix}.fallback",
|
||
gene_id_file=inputs.get("gene_id_file"),
|
||
)
|
||
if fallback_plot.get("success"):
|
||
print(
|
||
" 后备图完成: "
|
||
f"highlighted_pairs={fallback_plot.get('highlighted_pairs_plotted', 0)}, "
|
||
f"family_genes={fallback_plot.get('family_genes_plotted', 0)}"
|
||
)
|
||
else:
|
||
print(f" ⚠️ 后备图未生成: {fallback_plot.get('reason', 'unknown')}")
|
||
|
||
# 11. 准备artifacts
|
||
artifacts = {}
|
||
|
||
# blast文件
|
||
if blast_file.exists():
|
||
artifacts["blast"] = {
|
||
"artifact": "blast",
|
||
"path": f"synteny/analysis/{args.prefix}.blast",
|
||
"full_path": str(blast_file.absolute()),
|
||
"type": "blast",
|
||
"size": blast_file.stat().st_size,
|
||
"sha256": compute_file_hash(blast_file)
|
||
}
|
||
|
||
# gff文件
|
||
if gff_file.exists():
|
||
artifacts["gff"] = {
|
||
"artifact": "gff",
|
||
"path": f"synteny/analysis/{args.prefix}.gff",
|
||
"full_path": str(gff_file.absolute()),
|
||
"type": "gff",
|
||
"size": gff_file.stat().st_size,
|
||
"sha256": compute_file_hash(gff_file)
|
||
}
|
||
|
||
# 共线性文件
|
||
if collinearity_file.exists():
|
||
artifacts["collinearity"] = {
|
||
"artifact": "collinearity",
|
||
"path": f"synteny/analysis/{species_prefix}.collinearity",
|
||
"full_path": str(collinearity_file.absolute()),
|
||
"type": "text",
|
||
"size": collinearity_file.stat().st_size,
|
||
"sha256": compute_file_hash(collinearity_file)
|
||
}
|
||
|
||
# 复制基因分类日志 - 使用 species_prefix
|
||
dup_log = paths["analysis_dir"] / f"{species_prefix}.dup_class.log"
|
||
if not dup_log.exists():
|
||
# 也检查 args.prefix 版本
|
||
dup_log = paths["analysis_dir"] / f"{args.prefix}.dup_class.log"
|
||
|
||
if dup_log.exists():
|
||
artifacts["dup_class_log"] = {
|
||
"artifact": "dup_class_log",
|
||
"path": f"synteny/analysis/{args.prefix}.dup_class.log",
|
||
"full_path": str(dup_log.absolute()),
|
||
"type": "text",
|
||
"size": dup_log.stat().st_size,
|
||
"sha256": compute_file_hash(dup_log)
|
||
}
|
||
|
||
# Circos绘图文件
|
||
if circos_data.get("success", False):
|
||
# SVG文件 - 复制到final目录
|
||
if circos_data.get("svg_file") and Path(circos_data["svg_file"]).exists():
|
||
svg_file = Path(circos_data["svg_file"])
|
||
final_svg = paths["final_dir"] / f"{args.prefix}.circos.svg"
|
||
shutil.copy(svg_file, final_svg)
|
||
artifacts["circos_svg"] = {
|
||
"artifact": "circos_svg",
|
||
"path": f"final/{args.prefix}.circos.svg",
|
||
"full_path": str(final_svg.absolute()),
|
||
"type": "image",
|
||
"format": "svg",
|
||
"size": final_svg.stat().st_size,
|
||
"sha256": compute_file_hash(final_svg)
|
||
}
|
||
|
||
# PNG文件 - 复制到final目录
|
||
if circos_data.get("png_file") and Path(circos_data["png_file"]).exists():
|
||
png_file = Path(circos_data["png_file"])
|
||
final_png = paths["final_dir"] / f"{args.prefix}.circos.png"
|
||
shutil.copy(png_file, final_png)
|
||
artifacts["circos_png"] = {
|
||
"artifact": "circos_png",
|
||
"path": f"final/{args.prefix}.circos.png",
|
||
"full_path": str(final_png.absolute()),
|
||
"type": "image",
|
||
"format": "png",
|
||
"size": final_png.stat().st_size,
|
||
"sha256": compute_file_hash(final_png)
|
||
}
|
||
|
||
# Links文件
|
||
links_file = paths["analysis_dir"] / f"{args.prefix}.circos.links.txt"
|
||
if links_file.exists():
|
||
artifacts["circos_links"] = {
|
||
"artifact": "circos_links",
|
||
"path": f"synteny/analysis/{args.prefix}.circos.links.txt",
|
||
"full_path": str(links_file.absolute()),
|
||
"type": "text",
|
||
"size": links_file.stat().st_size,
|
||
"sha256": compute_file_hash(links_file)
|
||
}
|
||
|
||
if fallback_plot.get("success"):
|
||
for key, fmt in (("png_file", "png"), ("pdf_file", "pdf")):
|
||
src = Path(fallback_plot[key])
|
||
final_plot = paths["final_dir"] / f"{args.prefix}.synteny.{fmt}"
|
||
shutil.copy(src, final_plot)
|
||
artifacts[f"synteny_{fmt}"] = {
|
||
"artifact": f"synteny_{fmt}",
|
||
"path": f"final/{args.prefix}.synteny.{fmt}",
|
||
"full_path": str(final_plot.absolute()),
|
||
"type": "image" if fmt == "png" else "pdf",
|
||
"format": fmt,
|
||
"size": final_plot.stat().st_size,
|
||
"sha256": compute_file_hash(final_plot),
|
||
"source": "MCScanX fallback chromosome-link plot",
|
||
}
|
||
artifacts["synteny_plot_metadata"] = {
|
||
"artifact": "synteny_plot_metadata",
|
||
"path": "",
|
||
"type": "metadata",
|
||
"fallback_reason": fallback_plot.get("fallback_reason"),
|
||
"total_pairs_plotted_source": fallback_plot.get("total_pairs_plotted_source", 0),
|
||
"background_pairs_plotted": fallback_plot.get("background_pairs_plotted", 0),
|
||
"highlighted_pairs_plotted": fallback_plot.get("highlighted_pairs_plotted", 0),
|
||
"family_genes_plotted": fallback_plot.get("family_genes_plotted", 0),
|
||
}
|
||
|
||
# 12. 确定状态
|
||
status = "success"
|
||
if not blast_result.get("success", False):
|
||
status = "failed"
|
||
|
||
# 13. 准备参数
|
||
parameters = {
|
||
"cpu": args.cpu,
|
||
"prefix": args.prefix,
|
||
"evalue": args.evalue,
|
||
"max_target_seqs": args.max_target_seqs,
|
||
"use_blastp": args.use_blastp,
|
||
"stage": "synteny",
|
||
"job_id": job_id,
|
||
"run_id": run_id,
|
||
"protein_fasta": str(inputs["protein_fasta"]),
|
||
"gff3": str(inputs["gff3"]),
|
||
"genome": str(inputs["genome"]),
|
||
"tool_versions": tool_versions
|
||
}
|
||
if scope_data:
|
||
parameters.update({
|
||
"synteny_pep_source": locals().get("scope_protein_source", ""),
|
||
"synteny_gff_source": locals().get("scope_gff_source", ""),
|
||
"synteny_genome_source": locals().get("scope_genome_source", ""),
|
||
})
|
||
if scope_data:
|
||
parameters.update({
|
||
"scope_file": str(Path(args.scope_file).absolute()) if args.scope_file else "",
|
||
"scope_id": scope_data.get("scope_id", ""),
|
||
"scope_mode": scope_data.get("scope_mode", ""),
|
||
"analysis_route": analysis_route,
|
||
"member_set_id": member_set_id,
|
||
"target_species": scope_data.get("target_species", []),
|
||
"reference_species": scope_data.get("reference_species", []),
|
||
"comparative_species": scope_data.get("comparative_species", []),
|
||
"selected_species": scope_data.get("selected_species", []),
|
||
"family": scope_data.get("family", ""),
|
||
})
|
||
|
||
# 14. 写入报告
|
||
end_time = datetime.now()
|
||
|
||
print("\n📝 生成分析报告...")
|
||
report_path = write_report(
|
||
workspace=workspace,
|
||
prefix=args.prefix,
|
||
metrics=metrics,
|
||
parameters=parameters,
|
||
tool_versions=tool_versions,
|
||
diamond_result=blast_result,
|
||
gff_filter_result=gff_filter_result,
|
||
classifier_result=classifier_result,
|
||
mcscanx_result=mcscanx_result,
|
||
artifacts=artifacts
|
||
)
|
||
|
||
artifacts["report"] = {
|
||
"artifact": "report",
|
||
"path": f"final/{args.prefix}.synteny_report.md",
|
||
"full_path": str(report_path.absolute()),
|
||
"type": "markdown",
|
||
"size": report_path.stat().st_size,
|
||
"sha256": compute_file_hash(report_path)
|
||
}
|
||
|
||
# 15. 写入Manifest
|
||
print("\n📋 生成Manifest文件...")
|
||
manifest_path = write_manifest(
|
||
workspace=workspace,
|
||
job_id=job_id,
|
||
run_id=run_id,
|
||
prefix=args.prefix,
|
||
start_time=start_time,
|
||
end_time=end_time,
|
||
artifacts=artifacts,
|
||
metrics=metrics,
|
||
parameters=parameters,
|
||
tool_versions=tool_versions,
|
||
cmd_history=cmd_history,
|
||
status=status
|
||
)
|
||
|
||
artifacts["manifest"] = {
|
||
"artifact": "manifest",
|
||
"path": f"final/{args.prefix}.synteny.manifest.json",
|
||
"full_path": str(manifest_path.absolute()),
|
||
"type": "json",
|
||
"size": manifest_path.stat().st_size,
|
||
"sha256": compute_file_hash(manifest_path)
|
||
}
|
||
|
||
print("\n" + "=" * 60)
|
||
print(f"✅ 共线性分析完成！")
|
||
print(f"📊 基因数量: {metrics.get('gene_count', 'N/A')}")
|
||
print(f"🔗 共线性块数: {metrics.get('synteny_blocks', 'N/A')}")
|
||
print(f"📁 结果目录: {workspace}")
|
||
print("=" * 60)
|
||
|
||
return 0
|
||
|
||
except Exception as e:
|
||
print(f"\n❌ 共线性分析失败: {e}")
|
||
import traceback
|
||
traceback.print_exc()
|
||
|
||
# 写入失败manifest
|
||
end_time = datetime.now()
|
||
write_manifest(
|
||
workspace=workspace,
|
||
job_id=job_id,
|
||
run_id=run_id,
|
||
prefix=args.prefix,
|
||
start_time=start_time,
|
||
end_time=end_time,
|
||
artifacts={},
|
||
metrics={},
|
||
parameters={
|
||
"prefix": args.prefix,
|
||
"stage": "synteny",
|
||
"scope_file": str(Path(args.scope_file).absolute()) if args.scope_file else "",
|
||
"scope_id": scope_data.get("scope_id", ""),
|
||
"scope_mode": scope_data.get("scope_mode", ""),
|
||
"analysis_route": analysis_route,
|
||
"member_set_id": member_set_id,
|
||
"target_species": scope_data.get("target_species", []),
|
||
"reference_species": scope_data.get("reference_species", []),
|
||
"comparative_species": scope_data.get("comparative_species", []),
|
||
"selected_species": scope_data.get("selected_species", []),
|
||
"family": scope_data.get("family", ""),
|
||
},
|
||
tool_versions={},
|
||
cmd_history=cmd_history,
|
||
status="failed"
|
||
)
|
||
|
||
return 1
|
||
|
||
|
||
if __name__ == "__main__":
|
||
sys.exit(main())
|