To evaluate the relationships between soil inorganic phosphorus P (Pi) fractions, the soil P test and plant parameters such as plant P uptake, dry matter yield, tissue P concentration and relative yield, glasshouse experiments and chemical analyses were conducted on 13 calcareous soils from six agricultural and seven adjacent bushland (virgin soil) sites. Four rates of P (0, 15, 30, 60 mg/kg soil) were applied as reagent-grade KH2PO4 to the soils in a randomised complete block design with three replications. Perennial ryegrass (Lolium perenne cv. Roper) was grown and forage was harvested five times over a period of 210 days. Successive harvesting resulted in the depletion of plant available P as measured by NaHCO3-extractable P, which coincided with the decrease in the plant dry matter yield and P uptake. After five harvests, the order of reduction in Pi fractions induced by cropping without added P was Ca10-P>Al-P>Ca2-P>Ca8-P>occluded-P>Fe-P for the virgin soils and Ca2-P>Al-P>Ca10-P>Ca8-P>Fe-P>occluded-P for the agricultural soils. The order of abundance of Pi fractions for P treated-soils was non-occluded Al and Fe phosphate (Al-P+Fe-P)>secondary Ca-bound P (Ca2-P+Ca8-P)>acid-extractable P (Ca10- P)>occluded-P for both virgin and agricultural soils. Although a marked proportion of added P was transformed into less soluble Al and Fe phosphates, successive harvesting had depleted considerable percentages of these fractions. Highly significant (p<0.001) relationships were found for P uptake vs. Olsen-P, P uptake vs. Pi fractions (Ca2-P, Ca10-P, Al-P, Ca8-P, Fe-P) and Olsen-P vs. Pi fractions. NaHCO3-extractable P seems to be adequate for evaluating plant available P in calcareous soils. However, the closer relationship for the Fe-P fraction vs plant P uptake than for Olsen-P versus plant P uptake indicates that NaHCO3 may not provide the best estimate of plant available P for calcareous soils. Using stepwise regression analysis, it was found that the Ca2-P fraction was most predictive of P uptake (60%), total dry matter (68%), relative yield (74%) and Olsen-P (69%), followed by the Fe-P fraction.