diff --git a/pandas/io/excel/_base.py b/pandas/io/excel/_base.py
index 0b284fd4e9750..5e0ef4d6193f2 100644
--- a/pandas/io/excel/_base.py
+++ b/pandas/io/excel/_base.py
@@ -1592,9 +1592,9 @@ def __init__(
 
             xlrd_version = Version(get_version(xlrd))
 
-        ext = None
         if engine is None:
             # Only determine ext if it is needed
+            ext: str | None
             if xlrd_version is not None and isinstance(path_or_buffer, xlrd.Book):
                 ext = "xls"
             else:
@@ -1611,32 +1611,6 @@ def __init__(
             if engine == "auto":
                 engine = get_default_engine(ext, mode="reader")
 
-        if engine == "xlrd" and xlrd_version is not None:
-            if ext is None:
-                # Need ext to determine ext in order to raise/warn
-                if isinstance(path_or_buffer, xlrd.Book):
-                    ext = "xls"
-                else:
-                    ext = inspect_excel_format(
-                        path_or_buffer, storage_options=storage_options
-                    )
-
-            # Pass through if ext is None, otherwise check if ext valid for xlrd
-            if ext and ext != "xls" and xlrd_version >= Version("2"):
-                raise ValueError(
-                    f"Your version of xlrd is {xlrd_version}. In xlrd >= 2.0, "
-                    f"only the xls format is supported. Install openpyxl instead."
-                )
-            elif ext and ext != "xls":
-                stacklevel = find_stack_level()
-                warnings.warn(
-                    f"Your version of xlrd is {xlrd_version}. In xlrd >= 2.0, "
-                    f"only the xls format is supported. Install "
-                    f"openpyxl instead.",
-                    FutureWarning,
-                    stacklevel=stacklevel,
-                )
-
         assert engine is not None
         self.engine = engine
         self.storage_options = storage_options
diff --git a/pandas/io/excel/_xlrd.py b/pandas/io/excel/_xlrd.py
index 171705dee6e59..c556e4c68c6c0 100644
--- a/pandas/io/excel/_xlrd.py
+++ b/pandas/io/excel/_xlrd.py
@@ -30,7 +30,7 @@ def __init__(
             Object to be parsed.
         {storage_options}
         """
-        err_msg = "Install xlrd >= 1.0.0 for Excel support"
+        err_msg = "Install xlrd >= 2.0.1 for xls Excel support"
         import_optional_dependency("xlrd", extra=err_msg)
         super().__init__(filepath_or_buffer, storage_options=storage_options)
 
diff --git a/pandas/tests/io/excel/__init__.py b/pandas/tests/io/excel/__init__.py
index 9136153101e23..419761cbe1d6d 100644
--- a/pandas/tests/io/excel/__init__.py
+++ b/pandas/tests/io/excel/__init__.py
@@ -9,8 +9,4 @@
     pytest.mark.filterwarnings(
         "ignore:This method will be removed in future versions:DeprecationWarning"
     ),
-    # GH 38571
-    pytest.mark.filterwarnings(
-        "ignore:.*In xlrd >= 2.0, only the xls format is supported:FutureWarning"
-    ),
 ]
diff --git a/pandas/tests/io/excel/test_xlrd.py b/pandas/tests/io/excel/test_xlrd.py
index 30dddbd7de50b..1f8fb4b801356 100644
--- a/pandas/tests/io/excel/test_xlrd.py
+++ b/pandas/tests/io/excel/test_xlrd.py
@@ -2,8 +2,6 @@
 
 import pytest
 
-from pandas.compat._optional import import_optional_dependency
-
 import pandas as pd
 import pandas._testing as tm
 
@@ -39,35 +37,13 @@ def test_read_xlrd_book(read_ext_xlrd, datapath):
     tm.assert_frame_equal(result, expected)
 
 
-def test_excel_file_warning_with_xlsx_file(datapath):
-    # GH 29375
-    path = datapath("io", "data", "excel", "test1.xlsx")
-    has_openpyxl = import_optional_dependency("openpyxl", errors="ignore") is not None
-    if not has_openpyxl:
-        with tm.assert_produces_warning(
-            FutureWarning,
-            raise_on_extra_warnings=False,
-            match="The xlrd engine is no longer maintained",
-        ):
-            ExcelFile(path, engine=None)
-    else:
-        with tm.assert_produces_warning(None):
-            pd.read_excel(path, "Sheet1", engine=None)
-
-
-def test_read_excel_warning_with_xlsx_file(datapath):
+def test_read_xlsx_fails(datapath):
     # GH 29375
+    from xlrd.biffh import XLRDError
+
     path = datapath("io", "data", "excel", "test1.xlsx")
-    has_openpyxl = import_optional_dependency("openpyxl", errors="ignore") is not None
-    if not has_openpyxl:
-        with pytest.raises(
-            ValueError,
-            match="Your version of xlrd is ",
-        ):
-            pd.read_excel(path, "Sheet1", engine=None)
-    else:
-        with tm.assert_produces_warning(None):
-            pd.read_excel(path, "Sheet1", engine=None)
+    with pytest.raises(XLRDError, match="Excel xlsx file; not supported"):
+        pd.read_excel(path, engine="xlrd")
 
 
 @pytest.mark.parametrize(