Tolerant wheat cultivars yield well when sown in fields infested with the root‐lesion nematode Pratylenchus thornei, which is present in 67% of fields in the subtropical grain region of eastern Australia. Wheat breeding programmes require accurate phenotyping to select germplasm with superior tolerance to P. thornei. This study investigated normalised difference vegetation index (NDVI) as a phenotypic tool to predict the tolerance of wheat cultivars on low and high P. thornei population densities. Three, 2‐year field experiments used a resistant and a susceptible wheat cultivar in the first year to develop low and high P. thornei populations. In the second year, 36 wheat cultivars were sown on these plots. A NTech Greenseeker was used to determine the NDVI of each plot at regular times during the season and grain yield was measured at crop maturity. There was an inverse relationship between P. thornei population densities and the NDVI for intolerant wheat cultivars. Regression analysis showed a highly predictive response between the yield tolerance index and NDVI with R2 ranging from 0.85 (n = 36) to 0.93 (n = 36) for the three experiments. The area under the disease progress curve with respect to NDVI was highly predictive of yield tolerance (R2 = 0.92; n = 36) when there were high populations (9,091 P. thornei/kg), but not when populations were low (578 P. thornei/kg). Tolerant cultivars can be identified by NDVI when sown on soil containing high populations (>2,500 P. thornei/kg) by measurement at approximately 1,000 degree days after sowing. Greenseeker is a valuable tool for wheat breeders to select germplasm with tolerance of P. thornei.