Monitoring vertical profile of leaf water content (LWC) within wheat canopies after head emergence is vital significant for increasing crop yield. However, the estimation of vertical distribution of LWC from remote sensing data is still challenging due to the effects of wheat spikes and the efficacy of sensor measurement from the nadir direction. Using two-year field experiments with different growth stages after head emergence, N rates, wheat cultivars, we investigated the vertical distribution of LWC within canopies, the changes of canopy reflectance after spikes removal, the relationship between spectral indices and LWC in the upper-, middle- and bottom-layer. The interrelationship among vertical LWC were constructed, and four ratio of reflectance difference (RRD) type of indices were proposed based on the published WI and NDWSI indices to determine vertical distribution of LWC. The results indicated a bell shape distribution of LWC in wheat plants with the highest value appeared at the middle layer, and significant linear correlations between middle-LWC vs. upper-LWC and middle-LWC vs. bottom-LWC (r ≥ 0.92) were identified. The effects of wheat spikes on spectral reflectance mainly occurred in near infrared to shortwave infrared regions, which then decreased the accuracy of LWC estimation. Spectral indices at the middle layer outperformed the other two layers in LWC assessment and were less susceptible to wheat spikes effects, in particular, the newly proposed narrow-band WI-4 and NDWSI-4 indices exhibited great potential in tracking the changes of middle-LWC (R2
= 0.82 and 0.84, respectively). By taking into account the effects of wheat spikes and the interrelationship of vertical LWC within canopies, an indirect induction strategy was developed for modeling the upper-LWC and bottom-LWC. It was found that the indirect induction models based on the WI-4 and NDWSI-4 indices were more effective than the models obtained from conventional direct estimation method, with R2
of 0.78 and 0.81 for the upper-LWC estimation, and 0.75 and 0.74 for the bottom-LWC estimation, respectively.
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