ABSTRACT: Pinus cultivation without fertilization is a common practice in southern Brazil, which can induce a decline in the availability of phosphorus (P) in the soil. The purpose of this study was to evaluate the changes in phosphorus fractions in a Humic Cambisol subjected to continuous Pinus taeda L. cultivation without fertilization. Two forest stands were evaluated, after 16 years of Pinus cultivation (1st crop) and 49 years (3rd crop), when soil samples were collected (layers 0-10, 10-20, 20-40, 40-60, and 60-80cm) from six trenches per forest. In soil samples, the P contained in organic (Po) and inorganic (Pi) forms was determined by sequential chemical fractionation. Labile inorganic P fractions remained unchanged after the different cultivation periods. However, the labile organic fractions declined from the first to the third cycle (from 70.6 to 39.8mg dm-3 in the 0-10cm layer), indicating that these forms influence the buffering capacity of labile Pi. The moderately labile organic P acted as a P drain, increasing its percentage of the total, from 34.7 to 56.3%, from the first to the third crop. Soil cultivation for 49 years with Pinus taeda resulted in a reduction of the organic P content, indicating that for this soil use, this P form should be used to diagnose P availability and fertilization requirements.