Biochar has the potential to store carbon (C) in soils on a millennial time scale and hence it is proposed as a tool to aid in the mitigation of climate change. However, the presence of biochar in soil can induce either a positive or negative priming effect on native soil C, or the converse, which may either reduce or enhance the C storage potential of biochar. Thus far, priming effects between soil and biochar have been predominately assessed in the exclusion of plants. Therefore, this study set out with the aim to assess the priming effect of plants, i.e., rhizosphere priming effect (RPE) in the presence and absence of biochar and within different soil types. Three soils (Arenosol, Cambisol and Ferralsol) were used in full factorial combination with or without soybean plants and with or without 2% blue mallee biochar that was produced at 500 °C by slow pyrolysis. Plants were labelled with an isotopically depleted δ13C signature to that of the soil and biochar to allow the separation of plant-derived CO2–C from the total CO2–C. Carbon dioxide was trapped three times over a period of 13 days. Subsequent titration of the CO2 trap samples followed by IRMS analysis was used to quantify the CO2–C captured and its source. Biochar was found to have no effect on plant or microbial biomass. Plant treatments had significantly higher overall respiration rates than those without plants. Plants induced a negative priming in the Arenosol which was similar in the absence and presence of biochar. In the Cambisol, biochar induced a significant negative RPE in comparison to the positive RPE in the control. The RPE in the Ferralsol was positive and substantially decreased in the presence of biochar. Our results suggest that blue mallee biochar amendments may partially offset the positive RPE, or reduce it further where it is already negative.