diff --git a/.pre-commit-config.yaml b/.pre-commit-config.yaml
index ad56192..0294ad3 100644
--- a/.pre-commit-config.yaml
+++ b/.pre-commit-config.yaml
@@ -13,14 +13,14 @@ repos:
     rev: v2.4.1
     hooks:
       - id: codespell
-        additional_dependencies: ["tomli>=2.2.1"]
+        additional_dependencies: ["tomli>=2.4"]
   - repo: https://github.com/tox-dev/tox-ini-fmt
     rev: "1.7.1"
     hooks:
       - id: tox-ini-fmt
         args: ["-p", "fix"]
   - repo: https://github.com/tox-dev/pyproject-fmt
-    rev: "v2.15.0"
+    rev: "v2.15.2"
     hooks:
       - id: pyproject-fmt
   - repo: https://github.com/astral-sh/ruff-pre-commit
@@ -34,7 +34,7 @@ repos:
     hooks:
       - id: prettier
         additional_dependencies:
-          - prettier@3.6.2
+          - prettier@3.8.1
           - "@prettier/plugin-xml@3.4.2"
   - repo: meta
     hooks:
diff --git a/README.md b/README.md
index 73df965..3336a5a 100644
--- a/README.md
+++ b/README.md
@@ -6,7 +6,7 @@
 [![Downloads](https://static.pepy.tech/badge/pytest-env/month)](https://pepy.tech/project/pytest-env)
 
 A `pytest` plugin that sets environment variables from `pyproject.toml`, `pytest.toml`, `.pytest.toml`, or `pytest.ini`
-configuration files.
+configuration files. It can also load variables from `.env` files.
 
 ## Installation
 
@@ -104,6 +104,43 @@ Running `pytest tests_integration/` uses `DB_HOST = "test-db"` from the subdirec
 
 If no TOML file with a `pytest_env` section is found, the plugin falls back to the INI-style `env` key.
 
+### Loading `.env` files
+
+Use `env_files` to load variables from `.env` files. Files are loaded before inline `env` entries, so inline config
+takes precedence. Missing files are silently skipped. Paths are relative to the project root.
+
+```toml
+# pyproject.toml
+[tool.pytest_env]
+env_files = [".env", ".env.test"]
+API_KEY = "override_value"
+```
+
+```toml
+# pytest.toml or .pytest.toml
+[pytest_env]
+env_files = [".env"]
+```
+
+```ini
+# pytest.ini
+[pytest]
+env_files =
+    .env
+    .env.test
+```
+
+Files are parsed by [python-dotenv](https://github.com/theskumar/python-dotenv), supporting `KEY=VALUE` lines, `#`
+comments, `export` prefix, quoted values (with escape sequences in double quotes), and `${VAR:-default}` expansion:
+
+```shell
+# .env
+DATABASE_URL=postgres://localhost/mydb
+export SECRET_KEY='my-secret-key'
+DEBUG="true"
+MESSAGE="hello\nworld"
+```
+
 ### Examples
 
 **Expanding environment variables** — reference existing variables using `{VAR}` syntax:
diff --git a/pyproject.toml b/pyproject.toml
index 5e7acbb..db7be9e 100644
--- a/pyproject.toml
+++ b/pyproject.toml
@@ -2,7 +2,7 @@
 build-backend = "hatchling.build"
 requires = [
   "hatch-vcs>=0.5",
-  "hatchling>=1.27",
+  "hatchling>=1.28",
 ]
 
 [project]
@@ -36,12 +36,13 @@ dynamic = [
   "version",
 ]
 dependencies = [
-  "pytest>=9",
-  "tomli>=2.2.1; python_version<'3.11'",
+  "pytest>=9.0.2",
+  "python-dotenv>=1.2.1",
+  "tomli>=2.4; python_version<'3.11'",
 ]
 optional-dependencies.testing = [
   "covdefaults>=2.3",
-  "coverage>=7.10.7",
+  "coverage>=7.13.4",
   "pytest-mock>=3.15.1",
 ]
 urls.Homepage = "https://github.com/pytest-dev/pytest-env"
diff --git a/src/pytest_env/plugin.py b/src/pytest_env/plugin.py
index b66c683..2f38904 100644
--- a/src/pytest_env/plugin.py
+++ b/src/pytest_env/plugin.py
@@ -9,9 +9,11 @@
 from typing import TYPE_CHECKING, Any
 
 import pytest
+from dotenv import dotenv_values
 
 if TYPE_CHECKING:
     from collections.abc import Generator, Iterator
+    from pathlib import Path
 
 if sys.version_info >= (3, 11):  # pragma: >=3.11 cover
     import tomllib
@@ -23,6 +25,7 @@ def pytest_addoption(parser: pytest.Parser) -> None:
     """Add section to configuration files."""
     help_msg = "a line separated list of environment variables of the form (FLAG:)NAME=VALUE"
     parser.addini("env", type="linelist", help=help_msg, default=[])
+    parser.addini("env_files", type="linelist", help="a line separated list of .env files to load", default=[])
 
 
 @dataclass
@@ -43,6 +46,10 @@ def pytest_load_initial_conftests(
     parser: pytest.Parser,  # noqa: ARG001
 ) -> None:
     """Load environment variables from configuration files."""
+    for env_file in _load_env_files(early_config):
+        for key, value in dotenv_values(env_file).items():
+            if value is not None:
+                os.environ[key] = value
     for entry in _load_values(early_config):
         if entry.unset:
             os.environ.pop(entry.key, None)
@@ -53,8 +60,41 @@ def pytest_load_initial_conftests(
             os.environ[entry.key] = entry.value.format(**os.environ) if entry.transform else entry.value
 
 
+def _env_files_from_toml(early_config: pytest.Config) -> list[str]:
+    for path in chain.from_iterable([[early_config.rootpath], early_config.rootpath.parents]):
+        for pytest_toml_name in ("pytest.toml", ".pytest.toml", "pyproject.toml"):
+            pytest_toml_file = path / pytest_toml_name
+            if not pytest_toml_file.exists():
+                continue
+            with pytest_toml_file.open("rb") as file_handler:
+                try:
+                    config = tomllib.load(file_handler)
+                except tomllib.TOMLDecodeError:
+                    return []
+                if pytest_toml_name == "pyproject.toml":
+                    config = config.get("tool", {})
+                if (
+                    (pytest_env := config.get("pytest_env"))
+                    and isinstance(pytest_env, dict)
+                    and (raw := pytest_env.get("env_files"))
+                ):
+                    return [str(f) for f in (raw if isinstance(raw, list) else [raw])]
+                return []
+    return []
+
+
+def _load_env_files(early_config: pytest.Config) -> Generator[Path, None, None]:
+    if not (env_files := _env_files_from_toml(early_config)):
+        env_files = list(early_config.getini("env_files"))
+    for env_file_str in env_files:
+        if (resolved := early_config.rootpath / env_file_str).is_file():
+            yield resolved
+
+
 def _parse_toml_config(config: dict[str, Any]) -> Generator[Entry, None, None]:
     for key, entry in config.items():
+        if key == "env_files" and isinstance(entry, list):
+            continue
         if isinstance(entry, dict):
             unset = bool(entry.get("unset"))
             value = str(entry.get("value", "")) if not unset else ""
diff --git a/tests/test_env.py b/tests/test_env.py
index 1715494..3351fb3 100644
--- a/tests/test_env.py
+++ b/tests/test_env.py
@@ -3,10 +3,16 @@
 import os
 import re
 from pathlib import Path
+from typing import TYPE_CHECKING
 from unittest import mock
 
 import pytest
 
+from pytest_env.plugin import _env_files_from_toml  # noqa: PLC2701
+
+if TYPE_CHECKING:
+    import pytest_mock
+
 
 @pytest.mark.parametrize(
     ("env", "ini", "expected_env"),
@@ -318,6 +324,176 @@ def test_env_via_toml(  # noqa: PLR0913, PLR0917
     result.assert_outcomes(passed=1)
 
 
+@pytest.mark.parametrize(
+    ("env", "env_file_content", "config", "expected_env", "config_type"),
+    [
+        pytest.param(
+            {},
+            "MAGIC=alpha\nSORCERY=beta",
+            '[tool.pytest_env]\nenv_files = [".env"]',
+            {"MAGIC": "alpha", "SORCERY": "beta"},
+            "pyproject",
+            id="basic env file via pyproject toml",
+        ),
+        pytest.param(
+            {},
+            "MAGIC=alpha\nSORCERY=beta",
+            '[pytest_env]\nenv_files = [".env"]',
+            {"MAGIC": "alpha", "SORCERY": "beta"},
+            "pytest.toml",
+            id="basic env file via pytest toml",
+        ),
+        pytest.param(
+            {},
+            "MAGIC=alpha\nSORCERY=beta",
+            "[pytest]\nenv_files = .env",
+            {"MAGIC": "alpha", "SORCERY": "beta"},
+            "ini",
+            id="basic env file via ini",
+        ),
+        pytest.param(
+            {},
+            "# comment line\n\nMAGIC=alpha\n  # indented comment\n",
+            '[tool.pytest_env]\nenv_files = [".env"]',
+            {"MAGIC": "alpha"},
+            "pyproject",
+            id="comments and blank lines",
+        ),
+        pytest.param(
+            {},
+            "SINGLE='hello world'\nDOUBLE=\"hello world\"",
+            '[tool.pytest_env]\nenv_files = [".env"]',
+            {"SINGLE": "hello world", "DOUBLE": "hello world"},
+            "pyproject",
+            id="quoted values",
+        ),
+        pytest.param(
+            {},
+            "MAGIC=alpha",
+            '[tool.pytest_env]\nenv_files = [".env"]\nMAGIC = "beta"',
+            {"MAGIC": "beta"},
+            "pyproject",
+            id="inline overrides env file",
+        ),
+        pytest.param(
+            {},
+            "",
+            '[tool.pytest_env]\nenv_files = ["missing.env"]',
+            {},
+            "pyproject",
+            id="missing env file is skipped",
+        ),
+        pytest.param(
+            {},
+            "KEY_ONLY\nVALID=yes",
+            '[tool.pytest_env]\nenv_files = [".env"]',
+            {"VALID": "yes"},
+            "pyproject",
+            id="line without equals is skipped",
+        ),
+        pytest.param(
+            {},
+            "MAGIC=has=equals",
+            '[tool.pytest_env]\nenv_files = [".env"]',
+            {"MAGIC": "has=equals"},
+            "pyproject",
+            id="value with equals sign",
+        ),
+        pytest.param(
+            {},
+            "  MAGIC  =  alpha  ",
+            '[tool.pytest_env]\nenv_files = [".env"]',
+            {"MAGIC": "alpha"},
+            "pyproject",
+            id="whitespace around key and value",
+        ),
+        pytest.param(
+            {"MAGIC": "original"},
+            "MAGIC=from_file",
+            '[tool.pytest_env]\nenv_files = [".env"]\nMAGIC = {value = "from_file", skip_if_set = true}',
+            {"MAGIC": "from_file"},
+            "pyproject",
+            id="skip if set respects env file",
+        ),
+        pytest.param(
+            {},
+            "=no_key\nVALID=yes",
+            '[tool.pytest_env]\nenv_files = [".env"]',
+            {"VALID": "yes"},
+            "pyproject",
+            id="empty key is skipped",
+        ),
+        pytest.param(
+            {},
+            "",
+            '[tool.pytest_env]\nenv_files = "some_value"',
+            {"env_files": "some_value"},
+            "pyproject",
+            id="env_files as env var when string",
+        ),
+        pytest.param(
+            {},
+            "export MAGIC=alpha",
+            '[tool.pytest_env]\nenv_files = [".env"]',
+            {"MAGIC": "alpha"},
+            "pyproject",
+            id="export prefix",
+        ),
+        pytest.param(
+            {},
+            'MAGIC="hello\\nworld"',
+            '[tool.pytest_env]\nenv_files = [".env"]',
+            {"MAGIC": "hello\nworld"},
+            "pyproject",
+            id="escape sequences in double quotes",
+        ),
+        pytest.param(
+            {},
+            "MAGIC=alpha #comment",
+            '[tool.pytest_env]\nenv_files = [".env"]',
+            {"MAGIC": "alpha"},
+            "pyproject",
+            id="inline comment",
+        ),
+    ],
+)
+def test_env_via_env_file(  # noqa: PLR0913, PLR0917
+    testdir: pytest.Testdir,
+    env: dict[str, str],
+    env_file_content: str,
+    config: str,
+    expected_env: dict[str, str | None],
+    config_type: str,
+    request: pytest.FixtureRequest,
+) -> None:
+    tmp_dir = Path(str(testdir.tmpdir))
+    test_name = re.sub(r"\W|^(?=\d)", "_", request.node.callspec.id).lower()
+    Path(str(tmp_dir / f"test_{test_name}.py")).symlink_to(Path(__file__).parent / "template.py")
+    if env_file_content:
+        (tmp_dir / ".env").write_text(env_file_content, encoding="utf-8")
+    config_file_names = {"pyproject": "pyproject.toml", "pytest.toml": "pytest.toml", "ini": "pytest.ini"}
+    (tmp_dir / config_file_names[config_type]).write_text(config, encoding="utf-8")
+
+    new_env = {
+        **env,
+        "_TEST_ENV": repr(expected_env),
+        "PYTEST_DISABLE_PLUGIN_AUTOLOAD": "1",
+        "PYTEST_PLUGINS": "pytest_env.plugin",
+    }
+
+    with mock.patch.dict(os.environ, new_env, clear=True):
+        result = testdir.runpytest()
+
+    result.assert_outcomes(passed=1)
+
+
+def test_env_files_from_toml_bad_toml(tmp_path: Path, mocker: pytest_mock.MockerFixture) -> None:
+    (tmp_path / "pyproject.toml").write_text("bad toml", encoding="utf-8")
+    config = mocker.MagicMock()
+    config.rootpath = tmp_path
+    assert _env_files_from_toml(config) == []
+
+
 @pytest.mark.parametrize("toml_name", ["pytest.toml", ".pytest.toml", "pyproject.toml"])
 def test_env_via_pyproject_toml_bad(testdir: pytest.Testdir, toml_name: str) -> None:
     toml_file = Path(str(testdir.tmpdir)) / toml_name
diff --git a/tox.ini b/tox.ini
index 6987410..b10d44f 100644
--- a/tox.ini
+++ b/tox.ini
@@ -1,7 +1,7 @@
 [tox]
 requires =
-    tox>=4.30.3
-    tox-uv>=1.28
+    tox>=4.34.1
+    tox-uv>=1.29
 env_list =
     fix
     3.14
@@ -37,7 +37,7 @@ commands =
 description = run static analysis and style check using flake8
 skip_install = true
 deps =
-    pre-commit-uv>=4.1.5
+    pre-commit-uv>=4.2
 pass_env =
     HOMEPATH
     PROGRAMDATA
@@ -47,7 +47,7 @@ commands =
 [testenv:type]
 description = run type check on code base
 deps =
-    mypy==1.18.2
+    mypy==1.19.1
 commands =
     mypy --strict src
     mypy --strict tests
@@ -58,7 +58,7 @@ skip_install = true
 deps =
     check-wheel-contents>=0.6.3
     twine>=6.2
-    uv>=0.8.22
+    uv>=0.10.2
 commands =
     uv build --sdist --wheel --out-dir {env_tmp_dir} .
     twine check {env_tmp_dir}{/}*