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In this paper,we present a method for localization of a rail autonomous pesticide spraying and sampling robot working in greenhouse using an absolute localization system.Design and implementation of the localization system comes from the usage of beacon systems each of which is composed of an RF single receiver and an ultrasonic transmitter.The RF single receiver gets the synchronization signal from the mobile robot,and the ultrasonic transmitter sends ultrasonic signal,thus the distance from the beacon to the ultrasonic receiver can be measured.The position of a beacon in coordinate system of robot can be calculated according to distance information from the beacons to two ultrasonic receivers which are mounted on the robot.Based on the coordinate transformation,the position of a mobile robot can be calculated from the beacons absolute position information in the global coordinate system.Experiments demonstrate the effectiveness of the proposed method in real world applications.