diff --git a/productmd/composeinfo.py b/productmd/composeinfo.py
index bf853cc..a5d0a38 100644
--- a/productmd/composeinfo.py
+++ b/productmd/composeinfo.py
@@ -838,9 +838,14 @@ def _serialize_v2(self, data):
                 field = getattr(self, name)
                 value = field.get(arch, None)
                 if value:
-                    # Use stored Location if available (round-trip)
-                    loc = self._locations.get(name, {}).get(arch, None)
-                    if loc is not None:
+                    # Check if value is already a Location object (direct assignment)
+                    if isinstance(value, Location):
+                        # User assigned a Location directly - use it and store for round-trip
+                        self._locations.setdefault(name, {})[arch] = value
+                        paths.setdefault(name, {})[arch] = value.serialize()
+                    # Use stored Location if available (round-trip from deserialization)
+                    elif name in self._locations and arch in self._locations[name]:
+                        loc = self._locations[name][arch]
                         paths.setdefault(name, {})[arch] = loc.serialize()
                     else:
                         # Synthesize Location from path string
diff --git a/tests/test_composeinfo_v2.py b/tests/test_composeinfo_v2.py
index 42336c3..967c422 100644
--- a/tests/test_composeinfo_v2.py
+++ b/tests/test_composeinfo_v2.py
@@ -413,3 +413,73 @@ def test_no_paths_variant(self):
             data = {}
             ci.serialize(data, force_version=version)
             assert "Server" in data["payload"]["variants"]
+
+
+class TestVariantPathsLocationAssignment:
+    """Tests for issue #227: assigning Location objects to variant paths."""
+
+    def test_assign_location_object_to_variant_path(self):
+        """Test that assigning a Location object directly works as expected."""
+        ci = _create_composeinfo()
+        _add_server_variant(ci)
+
+        # Create a Location with different url and local_path
+        loc = Location(
+            url="https://pulp.example.com/pulp/content/Server/x86_64",
+            local_path="Server/x86_64/os",
+            size=1234567,
+            checksum="sha256:" + "a" * 64,
+        )
+
+        # This should work - directly assign Location to repository path
+        ci.variants["Server"].paths.repository["x86_64"] = loc
+
+        # Verify the assignment worked correctly
+        # Getting back should return the Location object
+        stored_loc = ci.variants["Server"].paths.repository["x86_64"]
+        assert isinstance(stored_loc, Location)
+        assert stored_loc.url == "https://pulp.example.com/pulp/content/Server/x86_64"
+        assert stored_loc.local_path == "Server/x86_64/os"
+        assert stored_loc.size == 1234567
+        assert stored_loc.checksum == "sha256:" + "a" * 64
+
+    def test_assign_string_to_variant_path_backward_compatible(self):
+        """Test that assigning a string still works (backward compatibility)."""
+        ci = _create_composeinfo()
+        _add_server_variant(ci)
+
+        # Assign a string (backward compatible behavior)
+        ci.variants["Server"].paths.repository["x86_64"] = "Server/x86_64/os"
+
+        # Verify the path is set
+        assert ci.variants["Server"].paths.repository["x86_64"] == "Server/x86_64/os"
+
+        # No Location should be stored (v1.x behavior)
+        paths = ci.variants["Server"].paths
+        assert "repository" not in paths._locations or "x86_64" not in paths._locations.get("repository", {})
+
+    def test_location_assignment_serializes_correctly(self):
+        """Test that assigned Location objects serialize correctly in v2.0."""
+        ci = _create_composeinfo()
+        _add_server_variant(ci)
+
+        # Assign Location using direct assignment
+        loc = Location(
+            url="https://cdn.example.com/Server/x86_64/os",
+            local_path="Server/x86_64/os",
+            size=5555,
+            checksum="sha256:" + "c" * 64,
+        )
+        ci.variants["Server"].paths.repository["x86_64"] = loc
+
+        # Serialize as v2.0
+        data = {}
+        ci.serialize(data, force_version=VERSION_2_0)
+
+        # Check serialized data
+        repo = data["payload"]["variants"]["Server"]["paths"]["repository"]["x86_64"]
+        assert isinstance(repo, dict)
+        assert repo["url"] == "https://cdn.example.com/Server/x86_64/os"
+        assert repo["local_path"] == "Server/x86_64/os"
+        assert repo["size"] == 5555
+        assert repo["checksum"] == "sha256:" + "c" * 64