The invention discloses a guiding and locking device for a vertical rotating type anti-flooding protective door. The device specifically comprises a guiding and locking stand column unit, a hydraulicdrive locking unit and a hydraulic drive locking unit mounting support, wherein the guiding and locking stand column unit is installed at the bottom of the cantilever end of a door body, the hydraulicdrive locking unit is arranged on the hydraulic drive locking unit mounting support, and the hydraulic drive locking unit mounting support is fixed on the ground according to the limit position afterthe door body is opened. When the door body is opened to the limit position, a hydraulic cylinder in the hydraulic drive locking unit drives two sets of spring bolt units to hold tightly a locking stand column on the guiding and locking stand column unit so as to complete locking of the door body, and therefore, the door body is fixed; and the device has the advantages of being simple in structure, small in occupied space, high in locking speed, large in locking torque and stable and reliable in operation.