diff --git a/src/_pytest/assertion/rewrite.py b/src/_pytest/assertion/rewrite.py
index 805d4c8b3..a01be76b4 100644
--- a/src/_pytest/assertion/rewrite.py
+++ b/src/_pytest/assertion/rewrite.py
@@ -27,8 +27,6 @@ from typing import Tuple
 from typing import TYPE_CHECKING
 from typing import Union
 
-import py
-
 from _pytest._io.saferepr import saferepr
 from _pytest._version import version
 from _pytest.assertion import util
@@ -37,6 +35,7 @@ from _pytest.assertion.util import (  # noqa: F401
 )
 from _pytest.config import Config
 from _pytest.main import Session
+from _pytest.pathlib import absolutepath
 from _pytest.pathlib import fnmatch_ex
 from _pytest.store import StoreKey
 
@@ -215,7 +214,7 @@ class AssertionRewritingHook(importlib.abc.MetaPathFinder, importlib.abc.Loader)
             return True
 
         if self.session is not None:
-            if self.session.isinitpath(py.path.local(fn)):
+            if self.session.isinitpath(absolutepath(fn)):
                 state.trace(f"matched test file (was specified on cmdline): {fn!r}")
                 return True
 
diff --git a/src/_pytest/config/argparsing.py b/src/_pytest/config/argparsing.py
index 9a4819655..5a09ea781 100644
--- a/src/_pytest/config/argparsing.py
+++ b/src/_pytest/config/argparsing.py
@@ -1,4 +1,5 @@
 import argparse
+import os
 import sys
 import warnings
 from gettext import gettext
@@ -14,8 +15,6 @@ from typing import Tuple
 from typing import TYPE_CHECKING
 from typing import Union
 
-import py
-
 import _pytest._io
 from _pytest.compat import final
 from _pytest.config.exceptions import UsageError
@@ -97,14 +96,14 @@ class Parser:
 
     def parse(
         self,
-        args: Sequence[Union[str, py.path.local]],
+        args: Sequence[Union[str, "os.PathLike[str]"]],
         namespace: Optional[argparse.Namespace] = None,
     ) -> argparse.Namespace:
         from _pytest._argcomplete import try_argcomplete
 
         self.optparser = self._getparser()
         try_argcomplete(self.optparser)
-        strargs = [str(x) if isinstance(x, py.path.local) else x for x in args]
+        strargs = [os.fspath(x) for x in args]
         return self.optparser.parse_args(strargs, namespace=namespace)
 
     def _getparser(self) -> "MyOptionParser":
@@ -128,7 +127,7 @@ class Parser:
 
     def parse_setoption(
         self,
-        args: Sequence[Union[str, py.path.local]],
+        args: Sequence[Union[str, "os.PathLike[str]"]],
         option: argparse.Namespace,
         namespace: Optional[argparse.Namespace] = None,
     ) -> List[str]:
@@ -139,7 +138,7 @@ class Parser:
 
     def parse_known_args(
         self,
-        args: Sequence[Union[str, py.path.local]],
+        args: Sequence[Union[str, "os.PathLike[str]"]],
         namespace: Optional[argparse.Namespace] = None,
     ) -> argparse.Namespace:
         """Parse and return a namespace object with known arguments at this point."""
@@ -147,13 +146,13 @@ class Parser:
 
     def parse_known_and_unknown_args(
         self,
-        args: Sequence[Union[str, py.path.local]],
+        args: Sequence[Union[str, "os.PathLike[str]"]],
         namespace: Optional[argparse.Namespace] = None,
     ) -> Tuple[argparse.Namespace, List[str]]:
         """Parse and return a namespace object with known arguments, and
         the remaining arguments unknown at this point."""
         optparser = self._getparser()
-        strargs = [str(x) if isinstance(x, py.path.local) else x for x in args]
+        strargs = [os.fspath(x) for x in args]
         return optparser.parse_known_args(strargs, namespace=namespace)
 
     def addini(
diff --git a/src/_pytest/fixtures.py b/src/_pytest/fixtures.py
index 273bcafd3..c24ab7069 100644
--- a/src/_pytest/fixtures.py
+++ b/src/_pytest/fixtures.py
@@ -648,12 +648,13 @@ class FixtureRequest:
             if has_params:
                 frame = inspect.stack()[3]
                 frameinfo = inspect.getframeinfo(frame[0])
-                source_path = py.path.local(frameinfo.filename)
+                source_path = absolutepath(frameinfo.filename)
                 source_lineno = frameinfo.lineno
-                rel_source_path = source_path.relto(funcitem.config.rootdir)
-                if rel_source_path:
-                    source_path_str = rel_source_path
-                else:
+                try:
+                    source_path_str = str(
+                        source_path.relative_to(funcitem.config.rootpath)
+                    )
+                except ValueError:
                     source_path_str = str(source_path)
                 msg = (
                     "The requested fixture has no parameter defined for test:\n"
@@ -876,7 +877,7 @@ class FixtureLookupError(LookupError):
 class FixtureLookupErrorRepr(TerminalRepr):
     def __init__(
         self,
-        filename: Union[str, py.path.local],
+        filename: Union[str, "os.PathLike[str]"],
         firstlineno: int,
         tblines: Sequence[str],
         errorstring: str,
@@ -903,7 +904,7 @@ class FixtureLookupErrorRepr(TerminalRepr):
                     f"{FormattedExcinfo.flow_marker}       {line.strip()}", red=True,
                 )
         tw.line()
-        tw.line("%s:%d" % (self.filename, self.firstlineno + 1))
+        tw.line("%s:%d" % (os.fspath(self.filename), self.firstlineno + 1))
 
 
 def fail_fixturefunc(fixturefunc, msg: str) -> "NoReturn":
diff --git a/src/_pytest/main.py b/src/_pytest/main.py
index eab3c9afd..d536f9d80 100644
--- a/src/_pytest/main.py
+++ b/src/_pytest/main.py
@@ -467,7 +467,7 @@ class Session(nodes.FSCollector):
         self.shouldfail: Union[bool, str] = False
         self.trace = config.trace.root.get("collection")
         self.startdir = config.invocation_dir
-        self._initialpaths: FrozenSet[py.path.local] = frozenset()
+        self._initialpaths: FrozenSet[Path] = frozenset()
 
         self._bestrelpathcache: Dict[Path, str] = _bestrelpath_cache(config.rootpath)
 
@@ -510,8 +510,8 @@ class Session(nodes.FSCollector):
 
     pytest_collectreport = pytest_runtest_logreport
 
-    def isinitpath(self, path: py.path.local) -> bool:
-        return path in self._initialpaths
+    def isinitpath(self, path: Union[str, "os.PathLike[str]"]) -> bool:
+        return Path(path) in self._initialpaths
 
     def gethookproxy(self, fspath: "os.PathLike[str]"):
         # Check if we have the common case of running
@@ -601,14 +601,14 @@ class Session(nodes.FSCollector):
         self.trace.root.indent += 1
 
         self._notfound: List[Tuple[str, Sequence[nodes.Collector]]] = []
-        self._initial_parts: List[Tuple[py.path.local, List[str]]] = []
+        self._initial_parts: List[Tuple[Path, List[str]]] = []
         self.items: List[nodes.Item] = []
 
         hook = self.config.hook
 
         items: Sequence[Union[nodes.Item, nodes.Collector]] = self.items
         try:
-            initialpaths: List[py.path.local] = []
+            initialpaths: List[Path] = []
             for arg in args:
                 fspath, parts = resolve_collection_argument(
                     self.config.invocation_params.dir,
@@ -669,13 +669,13 @@ class Session(nodes.FSCollector):
             # No point in finding packages when collecting doctests.
             if not self.config.getoption("doctestmodules", False):
                 pm = self.config.pluginmanager
-                confcutdir = py.path.local(pm._confcutdir) if pm._confcutdir else None
-                for parent in reversed(argpath.parts()):
-                    if confcutdir and confcutdir.relto(parent):
+                confcutdir = pm._confcutdir
+                for parent in (argpath, *argpath.parents):
+                    if confcutdir and parent in confcutdir.parents:
                         break
 
-                    if parent.isdir():
-                        pkginit = parent.join("__init__.py")
+                    if parent.is_dir():
+                        pkginit = py.path.local(parent / "__init__.py")
                         if pkginit.isfile() and pkginit not in node_cache1:
                             col = self._collectfile(pkginit, handle_dupes=False)
                             if col:
@@ -685,7 +685,7 @@ class Session(nodes.FSCollector):
 
             # If it's a directory argument, recurse and look for any Subpackages.
             # Let the Package collector deal with subnodes, don't collect here.
-            if argpath.check(dir=1):
+            if argpath.is_dir():
                 assert not names, "invalid arg {!r}".format((argpath, names))
 
                 seen_dirs: Set[py.path.local] = set()
@@ -717,15 +717,16 @@ class Session(nodes.FSCollector):
                             node_cache2[key] = x
                             yield x
             else:
-                assert argpath.check(file=1)
+                assert argpath.is_file()
 
-                if argpath in node_cache1:
-                    col = node_cache1[argpath]
+                argpath_ = py.path.local(argpath)
+                if argpath_ in node_cache1:
+                    col = node_cache1[argpath_]
                 else:
-                    collect_root = pkg_roots.get(argpath.dirname, self)
-                    col = collect_root._collectfile(argpath, handle_dupes=False)
+                    collect_root = pkg_roots.get(argpath_.dirname, self)
+                    col = collect_root._collectfile(argpath_, handle_dupes=False)
                     if col:
-                        node_cache1[argpath] = col
+                        node_cache1[argpath_] = col
 
                 matching = []
                 work: List[
@@ -782,9 +783,7 @@ class Session(nodes.FSCollector):
                 # first yielded item will be the __init__ Module itself, so
                 # just use that. If this special case isn't taken, then all the
                 # files in the package will be yielded.
-                if argpath.basename == "__init__.py" and isinstance(
-                    matching[0], Package
-                ):
+                if argpath.name == "__init__.py" and isinstance(matching[0], Package):
                     try:
                         yield next(iter(matching[0].collect()))
                     except StopIteration:
@@ -833,7 +832,7 @@ def search_pypath(module_name: str) -> str:
 
 def resolve_collection_argument(
     invocation_path: Path, arg: str, *, as_pypath: bool = False
-) -> Tuple[py.path.local, List[str]]:
+) -> Tuple[Path, List[str]]:
     """Parse path arguments optionally containing selection parts and return (fspath, names).
 
     Command-line arguments can point to files and/or directories, and optionally contain
@@ -875,4 +874,4 @@ def resolve_collection_argument(
             else "directory argument cannot contain :: selection parts: {arg}"
         )
         raise UsageError(msg.format(arg=arg))
-    return py.path.local(str(fspath)), parts
+    return fspath, parts
diff --git a/src/_pytest/monkeypatch.py b/src/_pytest/monkeypatch.py
index a052f693a..d012b8a53 100644
--- a/src/_pytest/monkeypatch.py
+++ b/src/_pytest/monkeypatch.py
@@ -4,7 +4,6 @@ import re
 import sys
 import warnings
 from contextlib import contextmanager
-from pathlib import Path
 from typing import Any
 from typing import Generator
 from typing import List
@@ -325,20 +324,14 @@ class MonkeyPatch:
 
         invalidate_caches()
 
-    def chdir(self, path) -> None:
+    def chdir(self, path: Union[str, "os.PathLike[str]"]) -> None:
         """Change the current working directory to the specified path.
 
-        Path can be a string or a py.path.local object.
+        Path can be a string or a path object.
         """
         if self._cwd is None:
             self._cwd = os.getcwd()
-        if hasattr(path, "chdir"):
-            path.chdir()
-        elif isinstance(path, Path):
-            # Modern python uses the fspath protocol here LEGACY
-            os.chdir(str(path))
-        else:
-            os.chdir(path)
+        os.chdir(path)
 
     def undo(self) -> None:
         """Undo previous changes.
diff --git a/src/_pytest/nodes.py b/src/_pytest/nodes.py
index 98bd581b9..fee0770eb 100644
--- a/src/_pytest/nodes.py
+++ b/src/_pytest/nodes.py
@@ -480,10 +480,14 @@ class Collector(Node):
             excinfo.traceback = ntraceback.filter()
 
 
-def _check_initialpaths_for_relpath(session, fspath):
+def _check_initialpaths_for_relpath(
+    session: "Session", fspath: py.path.local
+) -> Optional[str]:
     for initial_path in session._initialpaths:
-        if fspath.common(initial_path) == initial_path:
-            return fspath.relto(initial_path)
+        initial_path_ = py.path.local(initial_path)
+        if fspath.common(initial_path_) == initial_path_:
+            return fspath.relto(initial_path_)
+    return None
 
 
 class FSCollector(Collector):
diff --git a/src/_pytest/pathlib.py b/src/_pytest/pathlib.py
index 8875a28f8..2e452eb1c 100644
--- a/src/_pytest/pathlib.py
+++ b/src/_pytest/pathlib.py
@@ -30,8 +30,6 @@ from typing import Set
 from typing import TypeVar
 from typing import Union
 
-import py
-
 from _pytest.compat import assert_never
 from _pytest.outcomes import skip
 from _pytest.warning_types import PytestWarning
@@ -456,7 +454,7 @@ class ImportPathMismatchError(ImportError):
 
 
 def import_path(
-    p: Union[str, py.path.local, Path],
+    p: Union[str, "os.PathLike[str]"],
     *,
     mode: Union[str, ImportMode] = ImportMode.prepend,
 ) -> ModuleType:
@@ -482,7 +480,7 @@ def import_path(
     """
     mode = ImportMode(mode)
 
-    path = Path(str(p))
+    path = Path(p)
 
     if not path.exists():
         raise ImportError(path)
diff --git a/testing/test_assertrewrite.py b/testing/test_assertrewrite.py
index 84d5276e7..ffe18260f 100644
--- a/testing/test_assertrewrite.py
+++ b/testing/test_assertrewrite.py
@@ -17,8 +17,6 @@ from typing import Mapping
 from typing import Optional
 from typing import Set
 
-import py
-
 import _pytest._code
 import pytest
 from _pytest.assertion import util
@@ -1311,7 +1309,7 @@ class TestEarlyRewriteBailout:
         import importlib.machinery
 
         self.find_spec_calls: List[str] = []
-        self.initial_paths: Set[py.path.local] = set()
+        self.initial_paths: Set[Path] = set()
 
         class StubSession:
             _initialpaths = self.initial_paths
@@ -1346,7 +1344,7 @@ class TestEarlyRewriteBailout:
         pytester.makepyfile(test_foo="def test_foo(): pass")
         pytester.makepyfile(bar="def bar(): pass")
         foobar_path = pytester.makepyfile(foobar="def foobar(): pass")
-        self.initial_paths.add(py.path.local(foobar_path))
+        self.initial_paths.add(foobar_path)
 
         # conftest files should always be rewritten
         assert hook.find_spec("conftest") is not None
diff --git a/testing/test_main.py b/testing/test_main.py
index 3e94668e8..f45607abc 100644
--- a/testing/test_main.py
+++ b/testing/test_main.py
@@ -4,13 +4,12 @@ import re
 from pathlib import Path
 from typing import Optional
 
-import py.path
-
 import pytest
 from _pytest.config import ExitCode
 from _pytest.config import UsageError
 from _pytest.main import resolve_collection_argument
 from _pytest.main import validate_basetemp
+from _pytest.pytester import Pytester
 from _pytest.pytester import Testdir
 
 
@@ -109,40 +108,37 @@ def test_validate_basetemp_integration(testdir):
 
 class TestResolveCollectionArgument:
     @pytest.fixture
-    def invocation_dir(self, testdir: Testdir) -> py.path.local:
-        testdir.syspathinsert(str(testdir.tmpdir / "src"))
-        testdir.chdir()
+    def invocation_path(self, pytester: Pytester) -> Path:
+        pytester.syspathinsert(pytester.path / "src")
+        pytester.chdir()
 
-        pkg = testdir.tmpdir.join("src/pkg").ensure_dir()
-        pkg.join("__init__.py").ensure()
-        pkg.join("test.py").ensure()
-        return testdir.tmpdir
+        pkg = pytester.path.joinpath("src/pkg")
+        pkg.mkdir(parents=True)
+        pkg.joinpath("__init__.py").touch()
+        pkg.joinpath("test.py").touch()
+        return pytester.path
 
-    @pytest.fixture
-    def invocation_path(self, invocation_dir: py.path.local) -> Path:
-        return Path(str(invocation_dir))
-
-    def test_file(self, invocation_dir: py.path.local, invocation_path: Path) -> None:
+    def test_file(self, invocation_path: Path) -> None:
         """File and parts."""
         assert resolve_collection_argument(invocation_path, "src/pkg/test.py") == (
-            invocation_dir / "src/pkg/test.py",
+            invocation_path / "src/pkg/test.py",
             [],
         )
         assert resolve_collection_argument(invocation_path, "src/pkg/test.py::") == (
-            invocation_dir / "src/pkg/test.py",
+            invocation_path / "src/pkg/test.py",
             [""],
         )
         assert resolve_collection_argument(
             invocation_path, "src/pkg/test.py::foo::bar"
-        ) == (invocation_dir / "src/pkg/test.py", ["foo", "bar"])
+        ) == (invocation_path / "src/pkg/test.py", ["foo", "bar"])
         assert resolve_collection_argument(
             invocation_path, "src/pkg/test.py::foo::bar::"
-        ) == (invocation_dir / "src/pkg/test.py", ["foo", "bar", ""])
+        ) == (invocation_path / "src/pkg/test.py", ["foo", "bar", ""])
 
-    def test_dir(self, invocation_dir: py.path.local, invocation_path: Path) -> None:
+    def test_dir(self, invocation_path: Path) -> None:
         """Directory and parts."""
         assert resolve_collection_argument(invocation_path, "src/pkg") == (
-            invocation_dir / "src/pkg",
+            invocation_path / "src/pkg",
             [],
         )
 
@@ -156,16 +152,16 @@ class TestResolveCollectionArgument:
         ):
             resolve_collection_argument(invocation_path, "src/pkg::foo::bar")
 
-    def test_pypath(self, invocation_dir: py.path.local, invocation_path: Path) -> None:
+    def test_pypath(self, invocation_path: Path) -> None:
         """Dotted name and parts."""
         assert resolve_collection_argument(
             invocation_path, "pkg.test", as_pypath=True
-        ) == (invocation_dir / "src/pkg/test.py", [])
+        ) == (invocation_path / "src/pkg/test.py", [])
         assert resolve_collection_argument(
             invocation_path, "pkg.test::foo::bar", as_pypath=True
-        ) == (invocation_dir / "src/pkg/test.py", ["foo", "bar"])
+        ) == (invocation_path / "src/pkg/test.py", ["foo", "bar"])
         assert resolve_collection_argument(invocation_path, "pkg", as_pypath=True) == (
-            invocation_dir / "src/pkg",
+            invocation_path / "src/pkg",
             [],
         )
 
@@ -191,13 +187,11 @@ class TestResolveCollectionArgument:
         ):
             resolve_collection_argument(invocation_path, "foobar", as_pypath=True)
 
-    def test_absolute_paths_are_resolved_correctly(
-        self, invocation_dir: py.path.local, invocation_path: Path
-    ) -> None:
+    def test_absolute_paths_are_resolved_correctly(self, invocation_path: Path) -> None:
         """Absolute paths resolve back to absolute paths."""
-        full_path = str(invocation_dir / "src")
+        full_path = str(invocation_path / "src")
         assert resolve_collection_argument(invocation_path, full_path) == (
-            py.path.local(os.path.abspath("src")),
+            Path(os.path.abspath("src")),
             [],
         )
 
@@ -206,7 +200,7 @@ class TestResolveCollectionArgument:
         drive, full_path_without_drive = os.path.splitdrive(full_path)
         assert resolve_collection_argument(
             invocation_path, full_path_without_drive
-        ) == (py.path.local(os.path.abspath("src")), [])
+        ) == (Path(os.path.abspath("src")), [])
 
 
 def test_module_full_path_without_drive(testdir):
diff --git a/testing/test_nodes.py b/testing/test_nodes.py
index f3824c570..bae31f0a3 100644
--- a/testing/test_nodes.py
+++ b/testing/test_nodes.py
@@ -1,3 +1,4 @@
+from typing import cast
 from typing import List
 from typing import Type
 
@@ -73,17 +74,21 @@ def test__check_initialpaths_for_relpath() -> None:
     class FakeSession1:
         _initialpaths = [cwd]
 
-    assert nodes._check_initialpaths_for_relpath(FakeSession1, cwd) == ""
+    session = cast(pytest.Session, FakeSession1)
+
+    assert nodes._check_initialpaths_for_relpath(session, cwd) == ""
 
     sub = cwd.join("file")
 
     class FakeSession2:
         _initialpaths = [cwd]
 
-    assert nodes._check_initialpaths_for_relpath(FakeSession2, sub) == "file"
+    session = cast(pytest.Session, FakeSession2)
+
+    assert nodes._check_initialpaths_for_relpath(session, sub) == "file"
 
     outside = py.path.local("/outside")
-    assert nodes._check_initialpaths_for_relpath(FakeSession2, outside) is None
+    assert nodes._check_initialpaths_for_relpath(session, outside) is None
 
 
 def test_failure_with_changed_cwd(pytester: Pytester) -> None: