Leaf-rolling in maize crops: from leaf scoring to canopy-level measurements for phenotyping.

Leaf rolling in maize crops is one of the main plant reactions to water stress that can be visually scored in the field. However, leaf-scoring techniques do not meet the high-throughput requirements needed by breeders for efficient phenotyping. Consequently, this study investigated the relationship between leaf-rolling scores and changes in canopy structure that can be determined by high-throughput remote-sensing techniques. Experiments were conducted in 2015 and 2016 on maize genotypes subjected to water stress. Leaf-rolling was scored visually over the whole day around the flowering stage. Concurrent digital hemispherical photographs were taken to evaluate the impact of leaf-rolling on canopy structure using the computed fraction of intercepted diffuse photosynthetically active radiation, FIPARdif. The results showed that leaves started to roll due to water stress around 09:00 h and leaf-rolling reached its maximum around 15:00 h (the photoperiod was about 05:00-20:00 h). In contrast, plants maintained under well-watered conditions did not show any significant rolling during the same day. A canopy-level index of rolling (CLIR) is proposed to quantify the diurnal changes in canopy structure induced by leaf-rolling. It normalizes for the differences in FIPARdif between genotypes observed in the early morning when leaves are unrolled, as well as for yearly effects linked to environmental conditions. Leaf-level rolling score was very strongly correlated with changes in canopy structure as described by the CLIR (r2=0.86, n=370). The daily time course of rolling was characterized using the amplitude of variation, and the rate and the timing of development computed at both the leaf and canopy levels. Results obtained from eight genotypes common between the two years of experiments showed that the amplitude of variation of the CLIR was the more repeatable trait (Spearman coefficient ρ=0.62) as compared to the rate (ρ=0.29) and the timing of development (ρ=0.33). The potential of these findings for the development of a high-throughput method for determining leaf-rolling based on aerial drone observations are considered.