diff --git a/src/core/scene/scene-process/service/editors/prefab-editor.ts b/src/core/scene/scene-process/service/editors/prefab-editor.ts
index 62bc63e93..c1d8702b6 100644
--- a/src/core/scene/scene-process/service/editors/prefab-editor.ts
+++ b/src/core/scene/scene-process/service/editors/prefab-editor.ts
@@ -33,6 +33,7 @@ export class PrefabEditor extends BaseEditor {
 
         // 实例化预制体
         const instance = instantiate(prefabAsset);
+        editorPrefabUtils.preparePrefabRootForEditing(instance);
         this.virtualScene.addChild(instance);
         await this.ensurePreviewCanvasForUI(instance);
 
@@ -84,6 +85,7 @@ export class PrefabEditor extends BaseEditor {
         if (!instance) {
             throw new Error(`reload 失败，找不到预制体根节点: ${prefabName}`);
         }
+        editorPrefabUtils.preparePrefabRootForEditing(instance);
         this.entity.instance = instance;
         Prefab._utils.applyTargetOverrides(this.entity.instance);
         await this.ensurePreviewCanvasForUI(this.entity.instance);
diff --git a/src/core/scene/scene-process/service/prefab/prefab-editor-utils.ts b/src/core/scene/scene-process/service/prefab/prefab-editor-utils.ts
index c45fd1585..afa579694 100644
--- a/src/core/scene/scene-process/service/prefab/prefab-editor-utils.ts
+++ b/src/core/scene/scene-process/service/prefab/prefab-editor-utils.ts
@@ -4,6 +4,13 @@ import { prefabUtils } from './utils';
 type UUIDMap = Map<string, string | UUIDMap>;
 
 class EditorPrefabUtils {
+    preparePrefabRootForEditing(node: Node) {
+        const prefabInfo = node['_prefab'];
+        if (prefabInfo) {
+            prefabInfo.instance = undefined;
+        }
+    }
+
     serialize(node: Node) {
         // 校验数据
         prefabUtils.checkMountedRootData(node, true);
diff --git a/src/core/scene/test/prefab-editor-preview-canvas.test.ts b/src/core/scene/test/prefab-editor-preview-canvas.test.ts
index 173963d51..368547d16 100644
--- a/src/core/scene/test/prefab-editor-preview-canvas.test.ts
+++ b/src/core/scene/test/prefab-editor-preview-canvas.test.ts
@@ -49,42 +49,65 @@ jest.mock('../scene-process/service/prefab/prefab-editor-utils', () => ({
         restorePrefabUUID: jest.fn(),
         generateSceneAsset: jest.fn(),
         removePrefabInstanceRoots: jest.fn(),
+        preparePrefabRootForEditing: jest.fn((node: any) => {
+            if (node?._prefab) {
+                node._prefab.instance = undefined;
+            }
+        }),
     },
 }));
 
-import { instantiate } from 'cc';
+import { find, instantiate } from 'cc';
 import { sceneUtils } from '../scene-process/service/scene/utils';
 import { PrefabEditor } from '../scene-process/service/editors/prefab-editor';
 
-async function openPrefabWith(prefabRoot: any, scene = new MockScene('virtual-scene')): Promise<MockScene> {
+function createPrefabRoot(name: string, options?: { hasCanvas?: boolean; hasUI?: boolean; nestedChild?: any }) {
+    return {
+        name,
+        uuid: `${name}-uuid`,
+        parent: null,
+        children: options?.nestedChild ? [options.nestedChild] : [],
+        _prefab: { fileId: `${name}-file-id`, instance: { fileId: `${name}-instance-file-id` } },
+        getComponentInChildren: jest.fn((type: unknown) => type === MockCanvas && options?.hasCanvas ? {} : null),
+        getComponentsInChildren: jest.fn((type: unknown) => type === MockUITransform && options?.hasUI !== false ? [{}] : []),
+    };
+}
+
+async function openPrefabWith(prefabRoot: any, scene = new MockScene('virtual-scene')): Promise<{ editor: PrefabEditor; scene: MockScene; }> {
     (sceneUtils.runScene as jest.Mock).mockResolvedValue(scene);
     (sceneUtils.loadAny as jest.Mock).mockResolvedValue({});
     (instantiate as unknown as jest.Mock).mockReturnValue(prefabRoot);
+    const editor = new PrefabEditor();
 
-    await new PrefabEditor().open({
+    await editor.open({
         uuid: 'prefab-uuid',
         name: 'LabelPrefab',
         type: 'prefab',
         url: 'db://assets/LabelPrefab.prefab',
     } as never);
 
-    return scene;
+    return { editor, scene };
 }
 
 describe('PrefabEditor preview Canvas', () => {
     beforeEach(() => {
         jest.clearAllMocks();
+        (globalThis as any).EditorExtends = {
+            serialize: jest.fn(() => ({ json: 'scene-asset' })),
+            Node: {
+                getNode: jest.fn(),
+            },
+        };
+    });
+
+    afterEach(() => {
+        delete (globalThis as any).EditorExtends;
     });
 
     it('hosts a UI prefab without its own Canvas under an editor-only Canvas when opened', async () => {
         const scene = new MockScene('virtual-scene');
         const previewCanvas = { name: 'should_hide_in_hierarchy' };
-        const prefabRoot = {
-            name: 'LabelPrefab',
-            parent: null,
-            getComponentInChildren: jest.fn((type: unknown) => type === MockCanvas ? null : null),
-            getComponentsInChildren: jest.fn((type: unknown) => type === MockUITransform ? [{}] : []),
-        };
+        const prefabRoot = createPrefabRoot('LabelPrefab');
 
         createShouldHideInHierarchyCanvasNode.mockResolvedValue(previewCanvas);
 
@@ -95,30 +118,48 @@ describe('PrefabEditor preview Canvas', () => {
     });
 
     it('does not create a preview Canvas when the prefab already owns one', async () => {
-        const prefabRoot = {
-            name: 'CanvasPrefab',
-            parent: null,
-            getComponentInChildren: jest.fn((type: unknown) => type === MockCanvas ? {} : null),
-            getComponentsInChildren: jest.fn((type: unknown) => type === MockUITransform ? [{}] : []),
-        };
+        const prefabRoot = createPrefabRoot('CanvasPrefab', { hasCanvas: true });
 
-        const scene = await openPrefabWith(prefabRoot);
+        const { scene } = await openPrefabWith(prefabRoot);
 
         expect(createShouldHideInHierarchyCanvasNode).not.toHaveBeenCalled();
         expect(prefabRoot.parent).toBe(scene);
     });
 
     it('does not create a preview Canvas for prefabs without UI components', async () => {
-        const prefabRoot = {
-            name: 'MeshPrefab',
-            parent: null,
-            getComponentInChildren: jest.fn(() => null),
-            getComponentsInChildren: jest.fn(() => []),
-        };
+        const prefabRoot = createPrefabRoot('MeshPrefab', { hasUI: false });
 
-        const scene = await openPrefabWith(prefabRoot);
+        const { scene } = await openPrefabWith(prefabRoot);
 
         expect(createShouldHideInHierarchyCanvasNode).not.toHaveBeenCalled();
         expect(prefabRoot.parent).toBe(scene);
     });
+
+    it('clears root prefab instance but keeps nested child instance when opened', async () => {
+        const nestedChild = createPrefabRoot('NestedChild');
+        const prefabRoot = createPrefabRoot('RootPrefab', { nestedChild });
+
+        await openPrefabWith(prefabRoot);
+
+        expect(prefabRoot._prefab.instance).toBeUndefined();
+        expect(nestedChild._prefab.instance).toEqual({ fileId: 'NestedChild-instance-file-id' });
+    });
+
+    it('clears reloaded root prefab instance but keeps nested child instance after reload', async () => {
+        const nestedChild = createPrefabRoot('NestedChild');
+        const openedRoot = createPrefabRoot('RootPrefab', { nestedChild });
+        const { editor } = await openPrefabWith(openedRoot);
+        const reloadedNestedChild = createPrefabRoot('ReloadedNestedChild');
+        const reloadedRoot = createPrefabRoot('RootPrefab', { nestedChild: reloadedNestedChild });
+        const reloadedScene = new MockScene('reloaded-scene');
+
+        (sceneUtils.runSceneImmediateByJson as jest.Mock).mockResolvedValue(reloadedScene);
+        ((globalThis as any).EditorExtends.Node.getNode as jest.Mock).mockReturnValue(reloadedRoot);
+        (find as unknown as jest.Mock).mockReturnValue(null);
+
+        await editor.reload();
+
+        expect(reloadedRoot._prefab.instance).toBeUndefined();
+        expect(reloadedNestedChild._prefab.instance).toEqual({ fileId: 'ReloadedNestedChild-instance-file-id' });
+    });
 });