Phosphate, a key substrate in biological activities, is one of the least available macronutrients restricting crop production in many ecosystems. Genetic improvement of plant phosphate uptake is an important issue for sustainable agriculture. A phytase gene (MtPHY1) and a purple acid phosphatase gene (MtPAP1), both isolated from the model legume Medicago truncatula, were transgenically expressed in white clover (Trifolium repens L.) under the control of the constitutive CaMV35S promoter and the root-specific MtPT1 promoter. Transcripts were detected in roots of the transgenic plants. Phytase or acid phosphatase (APase) activities in root apoplasts of the transgenic plants were increased up to three fold compared to the wild type control. When the plants were grown 80 days in sand pots with organic P (phytate, Po) as the sole P supply, shoot dry weight of two transgenic plants almost doubled that of the control, and one of them showed no significant difference from its counterpart grown with inorganic P (Pi) supply. The relative biomass production of the transgenics in the Po treatment were over 90% and 80% of those in the Pi treatment in hydroponic culture (40 days) and sand pot (80 days), respectively; whereas biomass of the wild type control in the Po treatment was only about 50% of the Pi treatment in both hydroponic and sand cultures. In addition, shoot P concentrations in transgenic plants were significantly increased compared to the control. Transgenic plants accumulated much higher amount of total P (up to 2.6 fold after 80 days of growth) than the control in sand pots supplied with Po. The results showed that transgenic expression of MtPHY1 or MtPAP1 in white clover plants increased their abilities of utilizing organic phosphorus in response to P deficiency.