diff --git a/tests/test_metadata_serialization.py b/tests/test_metadata_serialization.py
index c7457482cc..c10798f611 100644
--- a/tests/test_metadata_serialization.py
+++ b/tests/test_metadata_serialization.py
@@ -490,6 +490,11 @@ def test_invalid_succinct_roles_serialization(self, test_data: str) -> None:
                 {"keyids": ["keyid1"], "name": "b", "terminating": true, "paths": ["fn1"], "threshold": 3}, \
                 {"keyids": ["keyid2"], "name": "root", "terminating": true, "paths": ["fn2"], "threshold": 4} ] \
             }',
+        "using top-level role name in a different case": '{"keys": { \
+                "keyid1" : {"keytype": "rsa", "scheme": "rsassa-pss-sha256", "keyval": {"public": "foo"}}}, \
+            "roles": [ \
+                {"keyids": ["keyid1"], "name": "Root", "terminating": true, "paths": ["fn1"], "threshold": 3}] \
+            }',
         "roles and succinct_roles set": '{"keys": { \
                 "keyid1" : {"keytype": "rsa", "scheme": "rsassa-pss-sha256", "keyval": {"public": "foo"}}, \
                 "keyid2" : {"keytype": "ed25519", "scheme": "ed25519", "keyval": {"public": "bar"}}}, \
diff --git a/tuf/api/_payload.py b/tuf/api/_payload.py
index 7b63cf5e01..82e2d402b5 100644
--- a/tuf/api/_payload.py
+++ b/tuf/api/_payload.py
@@ -1428,7 +1428,11 @@ def __init__(
 
         if roles is not None:
             for role in roles:
-                if not role or role in TOP_LEVEL_ROLE_NAMES:
+                # Compare case-insensitively: the client stores delegated
+                # metadata as "<quoted rolename>.json", and quoting leaves
+                # ASCII letters alone, so "Root" and "root" are the same file
+                # on a case-insensitive filesystem.
+                if not role or role.lower() in TOP_LEVEL_ROLE_NAMES:
                     raise ValueError(
                         "Delegated roles cannot be empty or use top-level "
                         "role names"