In several ways of introduction of foreign gene into plants, laser microbeam puncture is a potential and available method for transformation of foreign genes directly into plant tissue. The purpose of this research is uptaking the laser microbeam puncture technique to introduce pBI121 plasmid DNA containing GUS gene into Lotus. The laser microbeam irradiation source was a third harmonic Nd∶YAG laser. Before irradiation, the cells of cotyledon must be pretreated in high osmotic buffer. Then cultured the punctured tissues on screening media containing kanamycin to select green transgenic plantlets. After the seedings were selected on screening medium (km 100 mg/l) for three generations, we got four transgenic plants. Two of them were tested by PCR with GUS special primer. The results were positive, and proved that the GUS gene had been introduced into Lotus. We have established a laser microbeam puncture transformation system of Lotus, and proved that this system was easy to operate and replicate with relative high transformation frequency, thus providing a new way for high plant genetic transformation.