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It is not clear how the number of 15 N-injection needles affects the estimated results of gross N transformation rates in the presence of plants. To evaluate this, a 15 N tracing study of sandy loam soil and a silty loam soil in the presence of maize (Zea mays L., cv. "Zhengdan 958") was conducted and the Ntraceplant tool was used to quantify gross N transformation rates. Our results showed that the number of 15 N-injection needles could significantly influence the estimated results of gross N transformation rates in the studied soil–plant systems. There was no discernible difference in the gross rates of N mineralization (Min), the oxidation of NH4+ to NO3− (ONH4), the oxidation of recalcitrant organic-N to NO3− (ONrec), the NH4+ and NO3− plant uptake between 4-needle and 6-needle injections for both studied soils. However, for the 1-needle and 2-needle injections in the silty loam soil, Min was significantly lower than the 4-needle and 6-needle injections. While, in sandy loam soil, Min measured with 1-needle injection was significantly higher than the 4-needle and 6-needle injections. The ONH4 and ONrec also varied for the 1-needle and 2-needle injections, compared to the 4-needle and 6-needle injections. The total plant N uptake rate declined with the increase in the number of injection needles. Based on these results and considering the simplicity of the experimental procedure, we suggest that the 4-needle injection could be used in the 15 N tracing studies conducted in pot (diameter = 6.4 cm in this study) experiments with plants, i.e., one needle for about 8 cm2. [ABSTRACT FROM AUTHOR]