Purpose: To analyze the relationship between functional threshold power (FTP) and the lactate threshold (LT). Methods: A total of 20 male cyclists performed an incremental test in which LT was determined. At least 48 h later, they performed a 20-min time trial, and 95% of the mean power output was defined as FTP. Participants were divided into recreational (peak power output < 4.5 W·kg⁻¹; n = 11) or trained cyclists (peak power output > 4.5 W·kg⁻¹; n = 9) according to their fitness status. Results: The FTP (240 [35] W) was overall not significantly different (effect size = 0.20; limits of agreement = −2.4% [11.5%]) from the LT (246 [24] W), and both markers were strongly correlated (r = .95; P < .0001). Accounting for the participants’ fitness status, no significant differences were found between FTP and LT (effect size = 0.22; limits of agreement =2.1% [7.8%]) in trained cyclists, but FTP was significantly lower than the LT (P = .0004, effect size = 0.81; limits of agreement =−6.5% [8.3%]) in recreational cyclists. A significant relationship was found between relative peak power output and the bias between FTP and the LT markers (r = .77; P < .0001). Conclusions: FTP is a valid field test-based marker for the assessment of endurance fitness. However, caution should be taken when using FTP interchangeably with LT, as the bias between markers seems to depend on the athlete’s fitness status. Whereas FTP provides a good estimate of LT in trained cyclists, in recreational cyclists, it may underestimate LT.