A low-cost and facile preparation of water-soluble photoluminescent carbon nanodots (CDs) with a quantum yield of approximately 12.4% by hydrothermal method utilizing the leaves of Olea Europaea, a large number of planted trees in southwest of China, as a carbon source is developed for the first time. The prepared photoluminescent CDs not only show favorable photoluminescent properties, but also possess intrinsic peroxidase-like activity for colorimetric and UV-Vis absorption detection of hydrogen oxide (H2O2) and glucose. This sensing system exhibits excellent sensitivity toward H2O2 and glucose with the limit of detection as low as 0.6 microM and 5.2 microM. The practical use of this system for glucose determination in serum samples is also demonstrated successfully. The stability and low cost of photoluminescent CDs make them a powerful tool for a wide range of potential applications in biochemical analysis.