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Following the recent development of high-throughput phenotyping platforms for plant research, the number of individual plants grown together in a same experiment has raised, sometimes at the expense of pot size. However, root restriction in excessively small pots affects plant growth and carbon partitioning, andmayinteract with other stresses targeted in these experiments. In work reported here, we investigated the interactive effects of pot size and soil water deficit on multiple growth-related traits from the cellular to the whole-plant scale in oilseed rape (Brassica napus L.). The effects of pot size on responses to water deficit and allometric relationships revealed strong, multilevel interactions between pot size and watering regime. Notably, water deficit increased the root : shoot ratio in large pots, but not in small pots. At the cellular scale, water deficit decreased epidermal leaf cell area in large pots, but not in small pots. These results were consistent with changes in the level of endoreduplication factor in leaf cells. Our study illustrates the disturbing interaction of pot size with water deficit and raises the need to carefully consider this factor in the frame of the current development of high-throughput phenotyping experiments. [ABSTRACT FROM AUTHOR]

Background: Effects of abiotic and biotic stresses on plant photosynthetic performance lead to fitness and yield decrease. The maximum quantum efficiency of photosystem II (Fv/Fm) is a parameter of chlorophyll fluorescence (ChlF) classically used to track changes in photosynthetic performance. Despite recent technical and methodological advances in ChlF imaging, the spatio-temporal heterogeneity of Fv/Fm still awaits for standardized and accurate quantification. Results: We developed a method to quantify the dynamics of spatial heterogeneity of photosynthetic efficiency through the distribution-based analysis of Fv/Fm values. The method was applied to Arabidopsis thaliana grown under well-watered and severe water deficit (survival rate of 40%). First, whole-plant Fv/Fm shifted from unimodal to bimodal distributions during plant development despite a constant mean Fv/Fm under well-watered conditions. The establishment of a bimodal distribution of Fv/Fm reflects the occurrence of two types of leaf regions with contrasted photosynthetic efficiency. The distance between the two modes (called S) quantified the whole-plant photosynthetic heterogeneity. The weighted contribution of the most efficient/healthiest leaf regions to whole-plant performance (called Wmax) quantified the spatial efficiency of a photosynthetically heterogeneous plant. Plant survival to water deficit was associated to high S values, as well as with strong and fast recovery of Wmax following soil rewatering. Hence, during stress surviving plants had higher, but more efficient photosynthetic heterogeneity compared to perishing plants. Importantly, S allowed the discrimination between surviving and perishing plants four days earlier than the mean Fv/Fm. A sensitivity analysis from simulated dynamics of Fv/Fm showed that parameters indicative of plant tolerance and/or stress intensity caused identifiable changes in S and Wmax. Finally, an independent comparison of six Arabidopsis accessions grown under well-watered conditions indicated that S and Wmax are related to the genetic variability of growth. Conclusions: The distribution-based analysis of ChlF provides an efficient tool for quantifying photosynthetic heterogeneity and performance. S and Wmax are good indicators to estimate plant survival under water stress. Our results suggest that the dynamics of photosynthetic heterogeneity are key components of plant growth and tolerance to stress. [ABSTRACT FROM AUTHOR]

Mutualistic bacteria can alter plant phenotypes and confer new abilities to plants. Some plant growth-promoting rhizobacteria (PGPR) are known to improve both plant growth and tolerance to multiple stresses, including drought, but reports on their effects on plant survival under severe water deficits are scarce. We investigated the effect of Phyllobacterium brassicacearum STM196 strain, a PGPR isolated from the rhizosphere of oilseed rape, on survival, growth and physiological responses of Arabidopsis thaliana to severe water deficits combining destructive and non-destructive high-throughput phenotyping. Soil inoculation with STM196 greatly increased the survival rate of A. thaliana under several scenarios of severe water deficit. Photosystem II efficiency, assessed at the whole-plant level by high-throughput fluorescence imaging (Fv/Fm), was related to the probability of survival and revealed that STM196 delayed plant mortality. Inoculated surviving plants tolerated more damages to the photosynthetic tissues through a delayed dehydration and a better tolerance to low water status. Importantly, STM196 allowed a better recovery of plant growth after rewatering and stressed plants reached a similar biomass at flowering than non-stressed plants. Our results highlight the importance of plant-bacteria interactions in plant responses to severe drought and provide a new avenue of investigations to improve drought tolerance in agriculture. [ABSTRACT FROM AUTHOR]

• The high-throughput phenotypic analysis of Arabidopsis thaliana collections requires methodological progress and automation. Methods to impose stable and reproducible soil water deficits are presented and were used to analyse plant responses to water stress. • Several potential complications and methodological difficulties were identified, including the spatial and temporal variability of micrometeorological conditions within a growth chamber, the difference in soil water depletion rates between accessions and the differences in developmental stage of accessions the same time after sowing. Solutions were found. • Nine accessions were grown in four experiments in a rigorously controlled growth-chamber equipped with an automated system to control soil water content and take pictures of individual plants. One accession, An1, was unaffected by water deficit in terms of leaf number, leaf area, root growth and transpiration rate per unit leaf area. • Methods developed here will help identify quantitative trait loci and genes involved in plant tolerance to water deficit. [ABSTRACT FROM AUTHOR]