Sorghum genotypes vary in their response to higher sowing density, but the traits explaining these variations are unknown. In the present study, a 3D-imaging based approach identified the phenotypic traits responsible for the genetic variation in sorghum's response to high sowing density. Twenty sorghum genotypes, some varying in their response to density, were grown and 3D-images were collected weekly between weeks 4–6. From these images, 80 phenotypic traits, including 33 architectural and 47 multispectral, were extracted. The within-genotype means of these 80 traits, and two indicators of the sowing density response (Biomass ratio (Br) and Transpiration ratio (Tr)), measured in a previous study with 13 common genotypes, were used in a Spearman correlation analysis. Seventeen and four traits were strongly correlated with Br and Tr, respectively. The majority of these traits, predominantly architectural, strongly suggest that, under high sowing density, a fuller light interception, having more leaf area in the lower canopy, lead to a larger Br, while more vertically aligned leaves favour larger Tr values, which related to higher water use efficiency in another study. Furthermore, a Principal Component Analysis (PCA) indicated traits contributing to better photosynthesis could be used to estimate Br. Similarly, a combination of traits relating to leaf angle were good indicators of the genetic variation in Tr values. These results provide insights about the strategies some sorghum genotypes have developed to thrive under higher sowing density and that could be used as biomarkers for the breeding of density-resistant cultivars.

In semi-arid tropical areas, sorghum is sown at very low planting densities. Hence, increasing plant density represents an opportunity to improve productivity. However, assessing the expected increase in water needs is critical prior to testing higher densities under rainfed conditions. This was tested with a panel of elite cultivars in field and lysimiter experiments, and testing the effects of two density treatment, high (HD, 22 plants.m(-)(2)) and low (LD, 11 plants.m(-)(2)), on grain and biomass yield and on water use and water use efficiency (WUE). Doubling the conventional sowing density significantly increased biomass and grain yield, with a genotypic variability in the biomass response. No link was found between the response to density and the maintenance of the tillering capacity, whereas the response to density was somewhat explained by a differential increase in the leaf area index under high density (r=0.43 P<0.05). Lysimeter experiments showed that, compared with the conventional density, the high-density treatment had 62% increase in biomass vs a 38% increase in water use, resulting in a 17% higher WUE on average of the genotypes tested. There was an appreciable genotypic variability in this degree of WUE increase under high density. The most striking result was the very tight positive link between the biomass response to density and the differential increase in WUE in the dry season (r=0.91 P<0.0001), whereas in the wet season this link was negative (r=-0.48 P<0.02). This work shows that intensifying sorghum production by increasing sowing density is possible, in the short term using cultivars that show the largest WUE increase under high density, in the longer term by breeding high-density adapted cultivars, targeting plant traits that explain the tight link between higher WUE and higher yield under high density.

Abstract Water scarcity is already set to be one of the main issues of the 21st century, because of competing needs between civil, industrial, and agricultural use. Agriculture is currently the largest user of water, but its share is bound to decrease as societies develop and clearly it needs to become more water efficient. Improving water use efficiency (WUE) at the plant level is important, but translating this at the farm/landscape level presents considerable challenges. As we move up from the scale of cells, organs, and plants to more integrated scales such as plots, fields, farm systems, and landscapes, other factors such as trade-offs need to be considered to try to improve WUE. These include choices of crop variety/species, farm management practices, landscape design, infrastructure development, and ecosystem functions, where human decisions matter. This review is a cross-disciplinary attempt to analyse approaches to addressing WUE at these different scales, including definitions of the metrics of analysis and consideration of trade-offs. The equations we present in this perspectives paper use similar metrics across scales to make them easier to connect and are developed to highlight which levers, at different scales, can improve WUE. We also refer to models operating at these different scales to assess WUE. While our entry point is plants and crops, we scale up the analysis of WUE to farm systems and landscapes.

Introduction:Pearlmillet is themain subsistence crop for smallholder farmers systemswhere it is grown at low plant density. Intensifying pearl millet cultivation could boost productivity although it may have trade-offs. Increasing planting density would indeed increase the leaf area and the related water budget, whereas a denser canopy could create a more favorable canopymicroclimate to the benefit of the water use efficiency (WUE) of the crops. The first aim of this work was to test the yield response of popular pearlmillet varieties to an increased density and to assess possible genotypic variation in this response. The second aim was to measure the water use and the WUE of the crop in different densities.Method:To this end we designed several field and lysimetric experiments To increase the robustness of the results, these trials were carried out in India and Senegal, using two independent sets of genotypes adapted to both sites.Results:In the field, the higher sowing density significantly increased yield in all genotypes when trials were carried out in high evaporative demand conditions. There was no genotype x density interaction in these trials, suggesting no genotypic variation in the response to density increase. The high-density treatment also decreased the vapor pressure deficit (VPD) in the canopies, both in the field and in the lysimeter experiments. In the lysimeter trials, although the higher density treatment increased water use, the resulting increase in biomass was proportionally higher, hence increasingWUE of the crops in all genotypes under high density. The increase in yield under high density was closely related to the increase in WUE, although this link was more tight in the high- than in the low evaporative demand seasons. This confirmed a strong environmental effect on the response to density of all genotypes tested.Discussion:Although they did not open a scope for breeding density tolerant cultivars, these results highlight the possibility to improve pearl millet yield by increasing the density, targeting specifically areas facing high evaporative demand.

Indoor experiments with individual plants often show that transpiration rate is restricted under high vapor pressure deficit (VPD), resulting in a plateau of transpiration that increases water use efficiency (WUE) of some genotypes. We tested this hypothesis outdoors during dry or rainy seasons of India and Senegal, based on the response of the transpiration of canopy-grown sorghum plants to the reference evapotranspiration that takes both light and VPD into account. This response showed no plateau at high evaporative demand in 47 genotypes, but a large genetic variability was observed for the slope of the relationship over the whole range of evaporative demand. Unexpectedly, this slope was genetically correlated with WUE in two experiments with high evaporative demand: genotypes that most transpired had the highest WUE. Conversely, a negative correlation was observed under low evaporative demand. Genotypes with high WUE and response to evaporative demand were also those allowing maximum light penetration into the canopy. We suggest that this caused the observed high WUE of these genotypes because leaves within the canopy had sufficient light for photosynthesis whereas we observed a lower VPD in the canopy than in open air when leaf area index reached 2.5-3, thereby decreasing transpiration.Highlights The transpiration response to evaporative demand was genetically variable and correlated to WUE: genotypes that most transpired had highest light penetration towards leaves subjected to lower VPD than in air.### Competing Interest StatementThe authors have declared no competing interest.

Improving crop water use efficiency, the amount of carbon assimilated as biomass per unit of water used by a plant, is of major importance as water for agriculture becomes scarcer. In rice, the genetic bases of transpiration efficiency, the derivation of water use efficiency at the whole-plant scale, and its putative component trait transpiration restriction under high evaporative demand remain unknown. These traits were measured in 2019 in a panel of 147 African rice (Oryza glaberrima) genotypes known to be potential sources of tolerance genes to biotic and abiotic stresses. Our results reveal that higher transpiration efficiency is associated with transpiration restriction in African rice. Detailed measurements in a subset of highly contrasted genotypes in terms of biomass accumulation and transpiration confirmed these associations and suggested that root to shoot ratio played an important role in transpiration restriction. Genome wide association studies identified marker-trait associations for transpiration response to evaporative demand, transpiration efficiency, and its residuals, with links to genes involved in water transport and cell wall patterning. Our data suggest that root-shoot partitioning is an important component of transpiration restriction that has a positive effect on transpiration efficiency in African rice. Both traits are heritable and define targets for breeding rice with improved water use strategies.

Because water availability is the most important environmental factor limiting crop production, improving water use efficiency, the amount of carbon fixed per water used, is a major target for crop improvement. In rice, the genetic bases of transpiration efficiency, the derivation of water use efficiency at the whole-plant scale, and its putative component trait transpiration restriction under high evaporative demand, remain unknown. These traits were measured in a panel of 147 African rice Oryza glaberrima genotypes, known as potential sources of tolerance genes to biotic and abiotic stresses. Our results reveal that higher transpiration efficiency is associated with transpiration restriction in African rice. Detailed measurements in a subset of highly differentiated genotypes confirmed these associations and suggested that the root to shoot ratio played an important role in transpiration restriction. Genome wide association studies identified marker-trait associations for transpiration response to evaporative demand, transpiration efficiency and its residuals, that links to genes involved in water transport and cell wall patterning. Our data suggest that root shoot partitioning is an important component of transpiration restriction that has a positive effect on transpiration efficiency in African rice. Both traits are heritable and define targets for breeding rice with improved water use strategies.