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Transpiration efficiency (TE), the biomass produced per unit of water transpired, is a key trait for crop performance under limited water. As water becomes scarce, increasing TE would contribute to increase crop drought tolerance. This study is a first step to explore pearl millet genotypic variability for TE on a large and representative diversity panel. We analyzed TE on 537 pearl millet genotypes, including inbred lines, test-cross hybrids, and hybrids bred for different agroecological zones. Three lysimeter trials were conducted in 2012, 2013 and 2015, to assess TE both under well-watered and terminal-water stress conditions. We recorded grain yield to assess its relationship with TE. Up to two-fold variation for TE was observed over the accessions used. Mean TE varied between inbred and testcross hybrids, across years and was slightly higher under water stress. TE also differed among hybrids developed for three agroecological zones, being higher in hybrids bred for the wetter zone, underlining the importance of selecting germplasm according to the target area. Environmental conditions triggered large Genotype x Environment (GxE) interactions, although TE showed some high heritability. Transpiration efficiency was the second contributor to grain yield after harvest index, highlighting the importance of integrating it into pearl millet breeding programs. Future research on TE in pearl millet should focus (i) on investigating the causes of its plasticity i.e. the GxE interaction (ii) on studying its genetic basis and its association with other important physiological traits., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Grégoire et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Stomatal conductance ( g s ) is a crucial component of plant physiology, as it links plant productivity and water loss through transpiration. Estimating g s indirectly through leaf temperature ( T l ) measurement is common for reducing the high labor cost associated with direct g s measurement. However, the relationship between observed T l and g s can be notably affected by local environmental conditions, canopy structure, measurement scale, sample size, and g s itself. To better understand and quantify the variation in the relationship between T l measurements to g s , this study analyzed the sensitivity of T l to g s using a high-resolution three-dimensional model that resolves interactions between microclimate and canopy structure. The model was used to simulate the sensitivity of T l to g s across different environmental conditions, aggregation scales (point measurement, infrared thermometer, and thermographic image), and sample sizes. Results showed that leaf-level sensitivity of T l to g s was highest under conditions of high net radiation flux, high vapor pressure deficit, and low boundary layer conductance. The study findings also highlighted the trade-off between measurement scale and sample size to maximize sensitivity. Smaller scale measurements (e.g., thermocouple) provided maximal sensitivity because they allow for exclusion of shaded leaves and the ground, which have low sensitivity. However, large sample sizes (up to 50 to 75) may be needed to differentiate genotypes. Larger-scale measurements (e.g., thermal camera) reduced sample size requirements but include low-sensitivity elements in the measurement. This work provides a means of estimating leaf-level sensitivity and offers quantitative guidance for balancing scale and sample size issues., Competing Interests: Competing interests: The authors declare that they have no competing interests., (Copyright © 2024 Ismael K. Mayanja et al.)

Seedling root traits impact plant establishment under challenging environments. Pearl millet is one of the most heat and drought tolerant cereal crops that provides a vital food source across the sub-Saharan Sahel region. Pearl millet's early root system features a single fast-growing primary root which we hypothesize is an adaptation to the Sahelian climate. Using crop modeling, we demonstrate that early drought stress is an important constraint in agrosystems in the Sahel where pearl millet was domesticated. Furthermore, we show that increased pearl millet primary root growth is correlated with increased early water stress tolerance in field conditions. Genetics including genome-wide association study and quantitative trait loci (QTL) approaches identify genomic regions controlling this key root trait. Combining gene expression data, re-sequencing and re-annotation of one of these genomic regions identified a glutaredoxin-encoding gene PgGRXC9 as the candidate stress resilience root growth regulator. Functional characterization of its closest Arabidopsis homolog AtROXY19 revealed a novel role for this glutaredoxin (GRX) gene clade in regulating cell elongation. In summary, our study suggests a conserved function for GRX genes in conferring root cell elongation and enhancing resilience of pearl millet to its Sahelian environment., Competing Interests: Cd, AG, BS, MD, CB, AH, JA, SP, MS, JO, CT, JB, MS, CM, HD, BP, JD, AC, AP, CB, MP, PN, PG, PG, YG, VV, JR, MB, NK, SG, DW, YV, LL No competing interests declared, TM is affiliated with Be More Specific. The author has no financial interests to declare, (© 2023, de la Fuente, Grondin et al.)

Peanuts ( Arachis hypogaea L.) are important high-protein and oil-containing legume crops adapted to arid to semi-arid regions. The yield and quality of peanuts are complex quantitative traits that show high environmental influence. In this study, a recombinant inbred line population (RIL) (Valencia-C × JUG-03) was developed and phenotyped for nine traits under two environments. A genetic map was constructed using 1323 SNP markers spanning a map distance of 2003.13 cM. Quantitative trait loci (QTL) analysis using this genetic map and phenotyping data identified seventeen QTLs for nine traits. Intriguingly, a total of four QTLs, two each for 100-seed weight (HSW) and shelling percentage (SP), showed major and consistent effects, explaining 10.98% to 14.65% phenotypic variation. The major QTLs for HSW and SP harbored genes associated with seed and pod development such as the seed maturation protein-encoding gene, serine-threonine phosphatase gene, TIR-NBS-LRR gene, protein kinase superfamily gene, bHLH transcription factor -encoding gene, isopentyl transferase gene, ethylene-responsive transcription factor -encoding gene and cytochrome P450 superfamily gene. Additionally, the identification of 76 major epistatic QTLs, with PVE ranging from 11.63% to 72.61%, highlighted their significant role in determining the yield- and quality-related traits. The significant G × E interaction revealed the existence of the major role of the environment in determining the phenotype of yield-attributing traits. Notably, the seed maturation protein-coding gene in the vicinity of major QTLs for HSW can be further investigated to develop a diagnostic marker for HSW in peanut breeding. This study provides understanding of the genetic factor governing peanut traits and valuable insights for future breeding efforts aimed at improving yield and quality.

Sorghum production system in the semi-arid region of Africa is characterized by low yields which are generally attributed to high rainfall variability, poor soil fertility, and biotic factors. Production constraints must be well understood and quantified to design effective sorghum-system improvements. This study uses the state-of-the-art in silico methods and focuses on characterizing the sorghum production regions in Mali for drought occurrence and its effects on sorghum productivity. For this purpose, we adapted the APSIM-sorghum module to reproduce two cultivated photoperiod-sensitive sorghum types across a latitude of major sorghum production regions in Western Africa. We used the simulation outputs to characterize drought stress scenarios. We identified three main drought scenarios: (i) no-stress; (ii) early pre-flowering drought stress; and (iii) drought stress onset around flowering. The frequency of drought stress scenarios experienced by the two sorghum types across rainfall zones and soil types differed. As expected, the early pre-flowering and flowering drought stress occurred more frequently in isohyets < 600 mm, for the photoperiod-sensitive, late-flowering sorghum type. In isohyets above 600 mm, the frequency of drought stress was very low for both cultivars. We quantified the consequences of these drought scenarios on grain and biomass productivity. The yields of the highly-photoperiod-sensitive sorghum type were quite stable across the higher rainfall zones > 600 mm, but was affected by the drought stress in the lower rainfall zones < 600 mm. Comparatively, the less photoperiod-sensitive cultivar had notable yield gain in the driest regions < 600 mm. The results suggest that, at least for the tested crop types, drought stress might not be the major constraint to sorghum production in isohyets > 600 mm. The findings from this study provide the entry point for further quantitative testing of the Genotype × Environment × Management options required to optimize sorghum production in Mali., Supplementary Information: The online version contains supplementary material available at 10.1007/s13593-023-00909-5., Competing Interests: Conflict of interestThe authors declare no conflict of interest., (© The Author(s) 2024.)

Water will be a major limitation to food production in the 21st century, and drought issues already prevail in many parts of the world. Finding solutions to ensure that farmers harvest profitable crops, and secure food supplies for families and feed for animals that will provide for them through to the next season are urgent necessities. The Interdrought community has been addressing this issue for almost 30 years in a series of international conferences, characterized by a multi-disciplinary approach across the domains of molecular biology, physiology, genetics, agronomy, breeding, environmental and social sciences, policy, and systems modeling. This special issue presents papers from the 7th edition of the conference, the first to be held in Africa, that paid special attention to drought in a smallholder context, adding a 'system' dimension to the crop focus from the previous Interdrought events (Varshney et al., 2018; Hammer et al., 2021)., (© The Author(s) 2023. Published by Oxford University Press on behalf of the Society for Experimental Biology. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

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., (© The Author(s) 2023. Published by Oxford University Press on behalf of the Society for Experimental Biology.)

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., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Pilloni, Faye, Kakkera, Kholova, Badji, Faye and Vadez.)

Drought stress is one of the major constraints for crop production in the Sahel region of Africa. Here, we explore the potential to use natural genetic variation to build on the inherent drought tolerance of an elite sorghum cultivar, Teshale, that has been bred for Ethiopian conditions including chronic drought. We evaluated a backcross nested-association mapping population using 12 diverse founder lines crossed with Teshale under three drought-prone environments in Ethiopia. All 12 populations averaged higher head exsertion and lower leaf senescence than the recurrent parent in the two most stressful environments, reflecting new drought resilience mechanisms from the donors. A total of 154 quantitative trait loci (QTLs) were detected for eight drought-responsive traits, and their validity was supported by the fact that 113 (73.4%) overlapped with QTLs previously detected for the same traits, concentrated in regions previously associated with 'stay-green' traits. Allele effects showed that some favourable alleles are already present in the Ethiopian cultivar; however, the exotic donors offer rich scope for increasing drought resilience. Using model-selected SNPs associated with the eight traits identified in this study and three in a companion study, phenotypic prediction accuracies for grain yield were equivalent to genome-wide SNPs and were significantly better than random SNPs, indicating that the selected traits are predictive of sorghum grain yield., (© The Author(s) 2022. Published by Oxford University Press on behalf of the Society for Experimental Biology. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

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., (© The Author(s) 2022. Published by Oxford University Press on behalf of the Society for Experimental Biology. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Plant function arises from a complex network of structural and physiological traits. Explicit representation of these traits, as well as their connections with other biophysical processes, is required to advance our understanding of plant-soil-climate interactions. We used the Terrestrial Regional Ecosystem Exchange Simulator (TREES) to evaluate physiological trait networks in maize. Net primary productivity (NPP) and grain yield were simulated across five contrasting climate scenarios. Simulations achieving high NPP and grain yield in high precipitation environments featured trait networks conferring high water use strategies: deep roots, high stomatal conductance at low water potential ("risky" stomatal regulation), high xylem hydraulic conductivity and high maximal leaf area index. In contrast, high NPP and grain yield was achieved in dry environments with low late-season precipitation via water conserving trait networks: deep roots, high embolism resistance and low stomatal conductance at low leaf water potential ("conservative" stomatal regulation). We suggest that our approach, which allows for the simultaneous evaluation of physiological traits, soil characteristics and their interactions (i.e., networks), has potential to improve our understanding of crop performance in different environments. In contrast, evaluating single traits in isolation of other coordinated traits does not appear to be an effective strategy for predicting plant performance., (© 2022 John Wiley & Sons Ltd.)

Selection criteria that co-optimize water use efficiency and yield are needed to promote plant productivity in increasingly challenging and variable drought scenarios, particularly dryland cereals in the semi-arid tropics. Optimizing water use efficiency and yield fundamentally involves transpiration dynamics, where restriction of maximum transpiration rate helps to avoid early crop failure, while maximizing grain filling. Transpiration restriction can be regulated by multiple mechanisms and involves cross-organ coordination. This coordination involves complex feedbacks and feedforwards over time scales ranging from minutes to weeks, and from spatial scales ranging from cell membrane to crop canopy. Aquaporins have direct effect but various compensation and coordination pathways involve phenology, relative root and shoot growth, shoot architecture, root length distribution profile, as well as other architectural and anatomical aspects of plant form and function. We propose gravimetric phenotyping as an integrative, cross-scale solution to understand the dynamic, interwoven, and context-dependent coordination of transpiration regulation. The most fruitful breeding strategy is likely to be that which maintains focus on the phene of interest, namely, daily and season level transpiration dynamics. This direct selection approach is more precise than yield-based selection but sufficiently integrative to capture attenuating and complementary factors., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Burridge, Grondin and Vadez.)

Biosilica accumulation in plant tissues is related to the transpiration stream, which in turn depends on water availability. Nevertheless, the debate on whether genetically and environmentally controlled mechanisms of biosilica deposition are directly connected to water availability is still open. We aim at clarifying the system which leads to the deposition of biosilica in Sorghum bicolor , Pennisetum glaucum , and Eleusine coracana , expanding our understanding of the physiological role of silicon in crops well-adapted to arid environments, and simultaneously advancing the research in archaeological and paleoenvironmental studies. We cultivated ten traditional landraces for each crop in lysimeters, simulating irrigated and rain-fed scenarios in arid contexts. The percentage of biosilica accumulated in leaves indicates that both well-watered millet species deposited more biosilica than the water-stressed ones. By contrast, sorghum accumulated more biosilica with respect to the other two species, and biosilica accumulation was independent of the water regime. The water treatment alone did not explain either the variability of the assemblage or the differences in the biosilica accumulation. Hence, we hypothesize that genetics influence the variability substantially. These results demonstrate that biosilica accumulation differs among and within C4 species and that water availability is not the only driver in this process.

Pearl millet [ Pennisetum glaucum (L) R. Br.] is an important cereal crop of the semiarid tropics, which can withstand prolonged drought and heat stress. Considering an active involvement of the aquaporin (AQP) genes in water transport and desiccation tolerance besides several basic functions, their potential role in abiotic stress tolerance was systematically characterized and functionally validated. A total of 34 AQP genes from P. glaucum were identified and categorized into four subfamilies, viz., plasma membrane intrinsic proteins (PIPs), tonoplast intrinsic proteins (TIPs), nodulin-26-like intrinsic proteins (NIPs), and small basic intrinsic proteins (SIPs). Sequence analysis revealed that PgAQPs have conserved characters of AQP genes with a closer relationship to sorghum. The PgAQPs were expressed differentially under high vapor pressure deficit (VPD) and progressive drought stresses where the PgPIP2;6 gene showed significant expression under high VPD and drought stress. Transgenic tobacco plants were developed by heterologous expression of the PgPIP2;6 gene and functionally characterized under different abiotic stresses to further unravel their role. Transgenic tobacco plants in the T 2 generations displayed restricted transpiration and low root exudation rates in low- and high-VPD conditions. Under progressive drought stress, wild-type (WT) plants showed a quick or faster decline of soil moisture than transgenics. While under heat stress, PgPIP2;6 transgenics showed better adaptation to heat (40°C) with high canopy temperature depression (CTD) and low transpiration; under low-temperature stress, they displayed lower transpiration than their non-transgenic counterparts. Cumulatively, lower transpiration rate (Tr), low root exudation rate, declined transpiration, elevated CTD, and lower transpiration indicate that PgPIP2;6 plays a role under abiotic stress tolerance. Since the PgPIP2;6 transgenic plants exhibited better adaptation against major abiotic stresses such as drought, high VPD, heat, and cold stresses by virtue of enhanced transpiration efficiency, it has the potential to engineer abiotic stress tolerance for sustained growth and productivity of crops., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Reddy, Dhaware, Sivasakthi, Divya, Nagaraju, Sri Cindhuri, Kavi Kishor, Bhatnagar-Mathur, Vadez and Sharma.)

Late embryogenesis abundant (LEA) genes display distinct functions in response to abiotic stresses in plants. In pearl millet (Pennisetum glaucum L.), a total of 21 PgLEA genes were identified and classified into six groups including LEA1, LEA2, LEA3, LEA5, LEA7, and dehydrins (DHN). Open reading frames (ORFs) of PgLEAs range from 291 bp (PgLEA1-1) to 945 bp (PgLEA2-11) and distributed randomly among the seven chromosomes. Phylogenetic analysis revealed that all PgLEA proteins are closely related to sorghum LEA proteins. The PgLEAs were found to be expressed differentially under high progressive vapor pressure deficit (VPD), PgLEA7 was significantly expressed under high VPD and was selected for functional validation. In silico analysis of the PgLEA promoter regions revealed abiotic stress-specific cis-acting elements such as ABRE, CCAAT, MYBS, and LTRE. Based on the type of motifs, PgLEAPC promoter (758 bp), its deletion 1 (PgLpd1, 349 bp) and deletion 2 (PgLpd2, 125 bp) were cloned into the plant expression vector pMDC164 having the promoter-less uidA gene. All the three plant expression vectors were introduced into tobacco through Agrobacterium tumefaciens-mediated transformation to obtain T 1 and T 2 generations of transgenic plants. Based on expression of the uidA gene, tissue-specific expression was observed in mature stems, roots and seedlings of PgLEAPC and PgLpd1 carrying transgenics only. While the transgenic PgLEAPC plants displayed significantly higher uidA expression in the stem and root tissues under salt, drought, heat, and cold stresses, very low or no expression was observed in PgLpd1 and PgLpd2 transgenics under the tested stress conditions. The results of this study indicate that the complete promoter of PgLEAPC plays a role in developing abiotic stress tolerance in plants., (© 2021 Scandinavian Plant Physiology Society.)

We have previously reported that there is a tight link between high transpiration efficiency (TE; shoot biomass per unit water transpired) and restriction of transpiration under high vapor pressure deficit (VPD). In this study, we examine other factors affecting TE among major C4 cereals, namely species' differences, soil type, and source-sink relationships. We found that TE in maize (10 genotypes) was higher overall than in pearl millet (10 genotypes), and somewhat higher than in sorghum (16 genotypes). Overall, transpiration efficiency was higher in high-clay than in sandy soil under high VPD, but the effect was species-dependent with maize showing large variations in TE and yield across different soil types whilst pearl millet showed no variation in TE. This suggested that species fitness was specific to soil type. Removal of cobs drastically decreased TE in maize under high VPD, but removal of panicles did not have the same effect in pearl millet, suggesting that source-sink balance also drove variations in TE. We interpret the differences in TE between species as being accounted for by differences in the capacity to restrict transpiration under high VPD, with breeding history possibly having favored the source-sink balance in maize. This suggests that there is also scope to increase TE in pearl millet and sorghum through breeding. With regards to soil conditions, our results indicate that it appears to be critical to consider hydraulic characteristics and the root system together in order to better understand stomatal regulation and restriction of transpiration under high VPD. Finally, our results highlight the importance of sink strength in regulating transpiration/photosynthesis, and hence in influencing TE., (© The Author(s) 2021. Published by Oxford University Press on behalf of the Society for Experimental Biology.)

A deep understanding of the genetic control of drought tolerance and iron deficiency tolerance is essential to hasten the process of developing improved varieties with higher tolerance through genomics-assisted breeding. In this context, an improved genetic map with 1205 loci was developed spanning 2598.3 cM with an average 2.2 cM distance between loci in the recombinant inbred line (TAG 24 × ICGV 86031) population using high-density 58K single nucleotide polymorphism (SNP) "Axiom&#95; Arachis " array. Quantitative trait locus (QTL) analysis was performed using extensive phenotyping data generated for 20 drought tolerance- and two iron deficiency tolerance-related traits from eight seasons (2004-2015) at two locations in India, one in Niger, and one in Senegal. The genome-wide QTL discovery analysis identified 19 major main-effect QTLs with 10.0-33.9% phenotypic variation explained (PVE) for drought tolerance- and iron deficiency tolerance- related traits. Major main-effect QTLs were detected for haulm weight (20.1% PVE), SCMR (soil plant analytical development (SPAD) chlorophyll meter reading, 22.4% PVE), and visual chlorosis rate (33.9% PVE). Several important candidate genes encoding glycosyl hydrolases; malate dehydrogenases; microtubule-associated proteins; and transcription factors such as MADS-box, basic helix-loop-helix (bHLH), NAM, ATAF, and CUC (NAC), and myeloblastosis (MYB) were identified underlying these QTL regions. The putative function of these genes indicated their possible involvement in plant growth, development of seed and pod, and photosynthesis under drought or iron deficiency conditions in groundnut. These genomic regions and candidate genes, after validation, may be useful to develop molecular markers for deploying genomics-assisted breeding for enhancing groundnut yield under drought stress and iron-deficient soil conditions.

The rapid development of phenotyping technologies over the last years gave the opportunity to study plant development over time. The treatment of the massive amount of data collected by high-throughput phenotyping (HTP) platforms is however an important challenge for the plant science community. An important issue is to accurately estimate, over time, the genotypic component of plant phenotype. In outdoor and field-based HTP platforms, phenotype measurements can be substantially affected by data-generation inaccuracies or failures, leading to erroneous or missing data. To solve that problem, we developed an analytical pipeline composed of three modules: detection of outliers, imputation of missing values, and mixed-model genotype adjusted means computation with spatial adjustment. The pipeline was tested on three different traits (3D leaf area, projected leaf area, and plant height), in two crops (chickpea, sorghum), measured during two seasons. Using real-data analyses and simulations, we showed that the sequential application of the three pipeline steps was particularly useful to estimate smooth genotype growth curves from raw data containing a large amount of noise, a situation that is potentially frequent in data generated on outdoor HTP platforms. The procedure we propose can handle up to 50% of missing values. It is also robust to data contamination rates between 20 and 30% of the data. The pipeline was further extended to model the genotype time series data. A change-point analysis allowed the determination of growth phases and the optimal timing where genotypic differences were the largest. The estimated genotypic values were used to cluster the genotypes during the optimal growth phase. Through a two-way analysis of variance (ANOVA), clusters were found to be consistently defined throughout the growth duration. Therefore, we could show, on a wide range of scenarios, that the pipeline facilitated efficient extraction of useful information from outdoor HTP platform data. High-quality plant growth time series data is also provided to support breeding decisions. The R code of the pipeline is available at https://github.com/ICRISAT-GEMS/SpaTemHTP., (Copyright © 2020 Kar, Garin, Kholová, Vadez, Durbha, Tanaka, Iwata, Urban and Adinarayana.)

Background: Restricting transpiration under high vapor pressure deficit (VPD) is a promising water-saving trait for drought adaptation. However, it is often measured under controlled conditions and at very low throughput, unsuitable for breeding. A few high-throughput phenotyping (HTP) studies exist, and have considered only maximum transpiration rate in analyzing genotypic differences in this trait. Further, no study has precisely identified the VPD breakpoints where genotypes restrict transpiration under natural conditions. Therefore, outdoors HTP data (15 min frequency) of a chickpea population were used to automate the generation of smooth transpiration profiles, extract informative features of the transpiration response to VPD for optimal genotypic discretization, identify VPD breakpoints, and compare genotypes., Results: Fifteen biologically relevant features were extracted from the transpiration rate profiles derived from load cells data. Genotypes were clustered (C1, C2, C3) and 6 most important features (with heritability > 0.5) were selected using unsupervised Random Forest. All the wild relatives were found in C1, while C2 and C3 mostly comprised high TE and low TE lines, respectively. Assessment of the distinct p-value groups within each selected feature revealed highest genotypic variation for the feature representing transpiration response to high VPD condition. Sensitivity analysis on a multi-output neural network model (with R of 0.931, 0.944, 0.953 for C1, C2, C3, respectively) found C1 with the highest water saving ability, that restricted transpiration at relatively low VPD levels, 56% (i.e. 3.52 kPa) or 62% (i.e. 3.90 kPa), depending whether the influence of other environmental variables was minimum or maximum. Also, VPD appeared to have the most striking influence on the transpiration response independently of other environment variable, whereas light, temperature, and relative humidity alone had little/no effect., Conclusion: Through this study, we present a novel approach to identifying genotypes with drought-tolerance potential, which overcomes the challenges in HTP of the water-saving trait. The six selected features served as proxy phenotypes for reliable genotypic discretization. The wild chickpeas were found to limit water-loss faster than the water-profligate cultivated ones. Such an analytic approach can be directly used for prescriptive breeding applications, applied to other traits, and help expedite maximized information extraction from HTP data., Competing Interests: Competing interestsThe authors declare that they have no competing interests., (© The Author(s) 2020.)

Pearl millet is a key cereal for food security in arid and semi-arid regions but its yield is increasingly threatened by water stress. Physiological mechanisms relating to conservation of soil water or increased water use efficiency can alleviate that stress. Aquaporins (AQP) are water channels that mediate root water transport, thereby influencing plant hydraulics, transpiration and soil water conservation. However, AQP remain largely uncharacterized in pearl millet. Here, we studied AQP function in root water transport in two pearl millet lines contrasting for water use efficiency (WUE). We observed that these lines also contrasted for root hydraulic conductivity (Lpr) and AQP contribution to Lpr. The line with lower WUE showed significantly higher AQP contribution to Lpr. To investigate AQP isoforms contributing to Lpr, we developed genomic approaches to first identify the entire AQP family in pearl millet and secondly, characterize the plasma membrane intrinsic proteins (PIP) gene expression profile. We identified and annotated 33 AQP genes in pearl millet, among which ten encoded PIP isoforms. PgPIP1-3 and PgPIP1-4 were significantly more expressed in the line showing lower WUE, higher Lpr and higher AQP contribution to Lpr. Overall, our study suggests that the PIP1 AQP family are the main regulators of Lpr in pearl millet and may possibly be associated with mechanisms associated to whole plant water use. This study paves the way for further investigations on AQP functions in pearl millet hydraulics and adaptation to environmental stresses., Competing Interests: The authors have declared that no competing interests exist.

Terminal drought substantially reduces chickpea yield. Reducing water use at vegetative stage by reducing transpiration under high vapor pressure deficit (VPD), i.e. under dry/hot conditions, contributes to drought adaptation. We hypothesized that this trait could relate to differences in a genotype's dependence on root water transport pathways and hydraulics. Transpiration rate responses in conservative and profligate chickpea genotypes were evaluated under increasing VPD in the presence/absence of apoplastic and cell-to-cell transport inhibitors. Conservative genotypes ICC 4958 and ICC 8058 restricted transpiration under high VPD compared to the profligate genotypes ICC 14799 and ICC 867. Profligate genotypes were more affected by aquaporin inhibition of the cell-to-cell pathway than conservative genotypes, as measured by the root hydraulic conductance and transpiration under high VPD. Aquaporin inhibitor treatment also led to a larger reduction in root hydraulic conductivity in profligate than in conservative genotypes. In contrast, blockage of the apoplastic pathway in roots decreased transpiration more in conservative than in profligate genotypes. Interestingly, conservative genotypes had high early vigour, whereas profligate genotypes had low early vigour. In conclusion, profligate genotypes depend more on the cell-to-cell pathway, which might explain their higher root hydraulic conductivity, whereas water-saving by restricting transpiration led to higher dependence on the apoplastic pathway. This opens the possibility to screen for conservative or profligate chickpea phenotypes using inhibitors, itself opening to the search of the genetic basis of these differences., (© 2020 German Society for Plant Sciences and The Royal Botanical Society of the Netherlands.)

In the present study, the promoter region of the pearl millet heat shock protein 10 (PgHsp10) gene was cloned and characterized. The PgHsp10 promoter (PgHsp10pro) sequence region has all the cis-motifs required for tissue and abiotic stress inducibility. The complete PgHsp10pro (PgHsp10PC) region and a series of 5' truncations of PgHsp10 (PgHsp10D1 and PgHsp10D2) and an antisense form of PgHsp10pro (PgHsp10AS) were cloned into a plant expression vector (pMDC164) through gateway cloning. All four constructs were separately transformed into tobacco through Agrobacterium-mediated genetic transformation, and PCR-confirmed transgenic plants progressed to T 1 and T 2 generations. The T 2 transgenic tobacco plants comprising all PgHsp10pro fragments were used for GUS histochemical and qRT-PCR assays in different tissues under control and abiotic stresses. The PgHsp10PC pro expression was specific to stem and seedlings under control conditions. Under different abiotic stresses, particularly heat stress, PgHsp10PCpro had relatively higher activity than PgHsp10D1pro, PgHsp10D2pro and PgHsp10ASpro. PgHsp10pro from a stress resilient crop like pearl millet responds positively to a range of abiotic stresses, in particular heat, when expressed in heterologous plant systems such as tobacco. Hence, PgHsp10pro appears to be a potential promoter candidate for developing heat and drought stress-tolerant crop plants., Competing Interests: Declaration of competing interest The authors declare that they have no conflict of interest., (Copyright © 2020. Published by Elsevier B.V.)

This study compared maize, sorghum and pearl-millet, leading C 4 cereals, for the transpiration rate (TR) response to increasing atmospheric and soil water stress. The TR response to transiently increasing VPD (0.9-4.1 kPa) and the transpiration and leaf area expansion response to progressive soil drying were measured in controlled conditions at early vegetative stage in 10-16 genotypes of each species grown in moderate or high vapor pressure deficit (VPD) conditions. Maize grown under moderate VPD conditions restricted TR under high VPD, but not sorghum and pearl millet. By contrast, when grown under high VPD, all species increased TR upon increasing VPD, suggesting a loss of TR responsiveness. Sorghum and pearl-millet grown under high VPD reduced leaf area, but not maize. Upon progressive soil drying, maize reduced transpiration at higher soil moisture than sorghum and pearl millet, especially under high VPD, and leaf area expansion declined at similar or lower soil moisture than transpiration in maize and sorghum. It is concluded that maize conserves water by restricting transpiration upon increasing VPD and under higher soil moisture than sorghum and millet, giving maize significantly higher TE, whereas sorghum and pearl millet rely mostly on reduced leaf area and somewhat on transpiration restriction., Competing Interests: Declaration of Competing Interest The authors declare that there are no conflicts of interest., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Genetically engineered plants have varied applications in agriculture for enhancing the values of food and feed. Genetic engineering aims to introduce selected genetic regions with desirable traits into target plants for both spatial and temporal expressions. Promoters are the key elements responsible for regulating gene expressions by modulating the transcription factors (TFs) through recognition of RNA polymerases. Based on their recognition and expression, RNA polymerases were categorized into RNA pol II and pol III promoters. Promoter activity and specificity are the two prime parameters in regulating the transgene expression. Since the use of constitutive promoters like Cauliflower mosaic virus (CaMV) 35S may lead to adverse effects on nontarget organisms or ecosystem, inducible/tissue specific promoters and/or the RNA pol III promoters provide myriad opportunities for gene expressions with controlled regulation and with minimum adverse effects. Besides their role in transgene expression, their influence in synthetic biology and genome editing are also discussed. This review provides an update on the importance, current prospects, and insight into the advantages and disadvantages of promoters reported thus far would help to utilize them in the endeavour to develop nutritionally and agronomically improved transgenic crops for commercialization.

Pearl millet is a C 4 cereal crop that grows in arid and semi-arid climatic conditions with the remarkable abiotic stress tolerance. It contributed to the understanding of stress tolerance not only at the physiological level but also at the genetic level. In the present study, we functionally cloned and characterized three abiotic stress-inducible promoters namely cytoplasmic Apx1 (Ascorbate peroxidase), Dhn (Dehydrin), and Hsc70 (Heat shock cognate) from pearl millet. Sequence analysis revealed that all three promoters have several cis-acting elements specific for temporal and spatial expression. PgApx pro, PgDhn pro and PgHsc70 pro were fused with uidA gene in Gateway-based plant transformation pMDC164 vector and transferred into tobacco through leaf-disc method. While PgApx pro and PgDhn pro were active in seedling stages, PgHsc70 pro was active in stem and root tissues of the T 2 transgenic tobacco plants under control conditions. Higher activity was observed under high temperature and drought, and less in salt and cold stress conditions. Further, all three promoters displayed higher GUS gene expression in the stem, moderate expression in roots, and less expression in leaves under similar conditions. While RT-qPCR data showed that PgApx pro and PgDhn pro were expressed highly in high temperature, salt and drought, PgHsc70 pro was fairly expressed during high temperature stress only. Histochemical and RT-qPCR assays showed that all three promoters are inducible under abiotic stress conditions. Thus, these promoters appear to be immediate candidates for developing abiotic stress tolerant crops as these promoter-driven transgenics confer high degree of tolerance in comparison with the wild-type (WT) plants.

"Stay-green" crop phenotypes have been shown to impact drought tolerance and nutritional content of several crops. We aimed to genetically describe and functionally dissect the particular stay-green phenomenon found in chickpeas with a green cotyledon color of mature dry seed and investigate its potential use for improvement of chickpea environmental adaptations and nutritional value. We examined 40 stay-green accessions and a set of 29 BC2F4-5 stay-green introgression lines using a stay-green donor parent ICC 16340 and two Indian elite cultivars (KAK2, JGK1) as recurrent parents. Genetic studies of segregating populations indicated that the green cotyledon trait is controlled by a single recessive gene that is invariantly associated with the delayed degreening (extended chlorophyll retention). We found that the chickpea ortholog of Mendel's I locus of garden pea, encoding a SGR protein as very likely to underlie the persistently green cotyledon color phenotype of chickpea. Further sequence characterization of this chickpea ortholog CaStGR1 (CaStGR1, for carietinum stay-green gene 1) revealed the presence of five different molecular variants (alleles), each of which is likely a loss-of-function of the chickpea protein (CaStGR1) involved in chlorophyll catabolism. We tested the wild type and green cotyledon lines for components of adaptations to dry environments and traits linked to agronomic performance in different experimental systems and different levels of water availability. We found that the plant processes linked to disrupted CaStGR1 gene did not functionality affect transpiration efficiency or water usage. Photosynthetic pigments in grains, including provitaminogenic carotenoids important for human nutrition, were 2-3-fold higher in the stay-green type. Agronomic performance did not appear to be correlated with the presence/absence of the stay-green allele. We conclude that allelic variation in chickpea CaStGR1 does not compromise traits linked to environmental adaptation and agronomic performance, and is a promising genetic technology for biofortification of provitaminogenic carotenoids in chickpea.

Pearl millet plays a major role in food security in arid and semi-arid areas of Africa and India. However, it lags behind the other cereal crops in terms of genetic improvement. The recent sequencing of its genome opens the way to the use of modern genomic tools for breeding. Our study aimed at identifying genetic components involved in early drought stress tolerance as a first step toward the development of improved pearl millet varieties or hybrids. A panel of 188 inbred lines from West Africa was phenotyped under early drought stress and well-irrigated conditions. We found a strong impact of drought stress on yield components. This impact was variable between inbred lines. We then performed an association analysis with a total of 392,493 SNPs identified using Genotyping-by-Sequencing (GBS). Correcting for genetic relatedness, genome wide association study identified QTLs for biomass production in early drought stress conditions and for stay-green trait. In particular, genes involved in the sirohaem and wax biosynthesis pathways were found to co-locate with two of these QTLs. Our results might contribute to breed pearl millet lines with improved yield under drought stress., Competing Interests: The authors have declared that no competing interests exist.

Key Message: Four genetic regions associated with water use traits, measured at different levels of plant organization, and with agronomic traits were identified within a previously reported region for terminal water deficit adaptation on linkage group 2. Close linkages between these traits showed the value of phenotyping both for agronomic and secondary traits to better understand plant productive processes. Water saving traits are critical for water stress adaptation of pearl millet, whereas maximizing water use is key to the absence of stress. This research aimed at demonstrating the close relationship between traits measured at different levels of plant organization, some putatively involved in water stress adaptation, and those responsible for agronomic performance. A fine-mapping population of pearl millet, segregating for a previously identified quantitative trait locus (QTL) for adaptation to terminal drought stress on LG02, was phenotyped for traits at different levels of plant organization in different experimental environments (pot culture, high-throughput phenotyping platform, lysimeters, and field). The linkages among traits across the experimental systems were analysed using principal component analysis and QTL co-localization approach. Four regions within the LG02-QTL were found and revealed substantial co-mapping of water use and agronomic traits. These regions, identified across experimental systems, provided genetic evidence of the tight linkages between traits phenotyped at a lower level of plant organization and agronomic traits assessed in the field, therefore deepening our understanding of complex traits and then benefiting both geneticists and breeders. In short: (1) under no/mild stress conditions, increasing biomass and tiller production increased water use and eventually yield; (2) under severe stress conditions, water savings at vegetative stage, from lower plant vigour and fewer tillers in that population, led to more water available during grain filling, expression of stay-green phenotypes, and higher yield.

Grain legumes form an important component of the human diet, provide feed for livestock, and replenish soil fertility through biological nitrogen fixation. Globally, the demand for food legumes is increasing as they complement cereals in protein requirements and possess a high percentage of digestible protein. Climate change has enhanced the frequency and intensity of drought stress, posing serious production constraints, especially in rainfed regions where most legumes are produced. Genetic improvement of legumes, like other crops, is mostly based on pedigree and performance-based selection over the past half century. To achieve faster genetic gains in legumes in rainfed conditions, this review proposes the integration of modern genomics approaches, high throughput phenomics, and simulation modelling in support of crop improvement that leads to improved varieties that perform with appropriate agronomy. Selection intensity, generation interval, and improved operational efficiencies in breeding are expected to further enhance the genetic gain in experimental plots. Improved seed access to farmers, combined with appropriate agronomic packages in farmers' fields, will deliver higher genetic gains. Enhanced genetic gains, including not only productivity but also nutritional and market traits, will increase the profitability of farming and the availability of affordable nutritious food especially in developing countries.

Genotypic differences in transpiration rate responses to high vapour pressure deficit (VPD) was earlier reported. Here we tested the hypothesis that this limitation could relate to different degrees of dependence on the apoplastic (spaces between cells), and symplastic water transport pathways (through cells via aquaporin-facilitated transport), which are known to have different hydraulic conductivities. The low transpiration rate (Tr) genotype PRLT 2/89/33 either restricted its transpiration under high VPD, or was more sensitive to VPD than H77/833-2, when grown hydroponically or in soil. The slope of the transpiration response to an ascending series of VPD was lower in whole plants than in de-rooted shoots. In addition, the transpiration response of detached leaves to moderately high VPD (2.67kPa), normalised against leaves exposed to constant VPD (1.27kPa), was similar in low and high Tr genotypes. This suggested that roots hydraulics were a substantial limitation to water flow in pearl millet, especially under high VPD. The dependence on the apoplastic and symplastic water transport pathways was investigated by assessing the transpiration response of plants treated with inhibitors specific to the AQP-mediated symplastic pathway (AgNO3 and H2O2) and to the apoplastic pathway (precipitates of Cu(Fe(CN)6) or Cu(CuFe(CN)6)). When CuSO4 alone was used, Cu ions caused an inhibition of transpiration in both genotypes and more so in H77/833-2. The transpiration of high Tr H77/833-2 was decreased more by AQP inhibitors under low VPD (1.8kPa) than in PRLT 2/89/33, whereas under high VPD (4.2kPa), the transpiration of PRLT 2/89/33 was decreased more by AQP inhibitors than in H77/833-2. The transpiration rate of detached leaves from H77/833-2 when treated with AgNO3 decreased more than in PRLT 2/89/33. Although the root hydraulic conductivity of both genotypes was similar, it decreased more upon the application of a symplastic inhibitor in H77/833-2. The transpiration of low Tr PRLT 2/89/33 was decreased more by apoplastic inhibitors under both low and high VPD. Then the hydraulic conductivity decreased more upon the application of an apoplastic inhibitor in PRLT 2/89/33. In conclusion, both pathways contributed to water transport, and their contribution varied with environmental conditions and genotypes. Roots were a main source of hydraulic limitation in these genotypes of pearl millet, although a leaf limitation was not excluded. The similarity between genotypes in root hydraulic conductivity under normal conditions also suggests changes in this conductivity upon changes in the evaporative demand. The low Tr genotype depended more on the apoplastic pathway for water transport, whereas the high Tr genotype depended on both pathway, may be by 'tuning-up' the symplastic pathway under high transpiration demand, very likely via the involvement of aquaporins.

Domesticated species are impacted in unintended ways during domestication and breeding. Changes in the nature and intensity of selection impart genetic drift, reduce diversity, and increase the frequency of deleterious alleles. Such outcomes constrain our ability to expand the cultivation of crops into environments that differ from those under which domestication occurred. We address this need in chickpea, an important pulse legume, by harnessing the diversity of wild crop relatives. We document an extreme domestication-related genetic bottleneck and decipher the genetic history of wild populations. We provide evidence of ancestral adaptations for seed coat color crypsis, estimate the impact of environment on genetic structure and trait values, and demonstrate variation between wild and cultivated accessions for agronomic properties. A resource of genotyped, association mapping progeny functionally links the wild and cultivated gene pools and is an essential resource chickpea for improvement, while our methods inform collection of other wild crop progenitor species.

Background: Terminal drought stress leads to substantial annual yield losses in chickpea (Cicer arietinum L.). Adaptation to water limitation is a matter of matching water supply to water demand by the crop. Therefore, harnessing the genetics of traits contributing to plant water use, i.e. transpiration rate and canopy development dynamics, is important to design crop ideotypes suited to a varying range of water limited environments. With an aim of identifying genomic regions for plant vigour (growth and canopy size) and canopy conductance traits, 232 recombinant inbred lines derived from a cross between ICC 4958 and ICC 1882, were phenotyped at vegetative stage under well-watered conditions using a high throughput phenotyping platform (LeasyScan)., Results: Twenty one major quantitative trait loci (M-QTLs) were identified for plant vigour and canopy conductance traits using an ultra-high density bin map. Plant vigour traits had 13 M-QTLs on CaLG04, with favourable alleles from high vigour parent ICC 4958. Most of them co-mapped with a previously fine mapped major drought tolerance "QTL-hotspot" region on CaLG04. One M-QTL was found for canopy conductance on CaLG03 with the ultra-high density bin map. Comparative analysis of the QTLs found across different density genetic maps revealed that QTL size reduced considerably and % of phenotypic variation increased as marker density increased., Conclusion: Earlier reported drought tolerance hotspot is a vigour locus. The fact that canopy conductance traits, i.e. the other important determinant of plant water use, mapped on CaLG03 provides an opportunity to manipulate these loci to tailor recombinants having low/high transpiration rate and plant vigour, fitted to specific drought stress scenarios in chickpea.

Pearl millet is a crop of the semi-arid tropics having high degree of genetic diversity and variable tolerance to drought stress. To investigate drought tolerance mechanism that possibly accounts for differences in drought tolerance, four recombinant inbred lines from a high resolution cross (HRC) were selected for variability in their transpiration rate (Tr) response to vapour pressure deficit (VPD) conditions. The differential Tr response of the genotypes to increased VPD conditions was used to classify the genotypes as sensitive or insensitive to high VPD. Aquaporin (AQP) genes PgPIP1;1, PgPIP1;2, PgPIP2;1, PgPIP2;3, PgPIP2;6, PgTIP1;1 and PgTIP2;2 were cloned. Phylogenetic analysis revealed that the cloned PgAQPs were evolutionarily closer to maize AQPs than to rice. PgAQP genes, including PgPIP1;1 and PgPIP2;6 in root tissue showed a significant expression pattern with higher expression in VPD-insensitive genotypes than VPD-sensitive genotypes under low VPD conditions (1.2kPa) i.e when there is no high evaporative demand from the atmosphere. PgAQP genes (PgPIP2;1 in leaf and root tissues; PgPIP1;2 and PgTIP2;2 in leaf and PgPIP2;6 in root) followed a diurnal rhythm in leaves and roots that have either higher or lower expression levels at different time intervals. Under high VPD conditions (4.21kPa), PgPIP2;3 showed higher transcript abundance in VPD-insensitive genotypes, and PgPIP2;1 in VPD-sensitive genotypes, while rest of the PgAQPs showed differential expression. Our current hypothesis is that these differences in the expression of AQP genes under different VPDs suggests a role of the AQPs in tuning the water transport pathways with variation between genotypes., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Under conditions of high vapor pressure deficit (VPD) and soil drying, restricting transpiration is an important avenue to gain efficiency in water use. The question we raise in this article is whether breeding for agro-ecological environments that differ for the rainfall have selected for traits that control plant water use. These are measured in pearl millet materials bred for zones varying in rainfall (8 combinations of parent and F 1 -hybrids, 18 F 1 -hybrids and then 40 F 1 -hybrids). In all cases, we found an agro-ecological variation in the slope of the transpiration response to increasing VPD, and parental line variation in the transpiration response to soil drying within hybrids/parent combinations. The hybrids adapted to lower rainfall had higher transpiration response curves than those from the highest rainfall zones, but showed no variation in how transpiration responded to soil drying. The genotypes bred for lower rainfall zones showed lower leaf area, dry matter, thicker leaves, root development, and exudation, than the ones bred for high rainfall zone when grown in the low VPD environment of the greenhouse, but there was no difference in their root length neither on the root/shoot index in these genotypes. By contrast, when grown under high VPD conditions outdoors, the lower rainfall hybrids had the highest leaf, tiller, and biomass development. Finally, under soil drying the genotypes from the lower rainfall accumulated less biomass than the ones from higher rainfall zone, and so did the parental lines compared to the hybrids. These differences in the transpiration response and growth clearly showed that breeding for different agro-ecological zones also bred for different genotype strategies in relation to traits related to plant water use. Highlights : • Variation in transpiration response reflected breeding for agro-ecological zones • Different growth strategies depended on the environmental conditions • Different ideotypes reflected rainfall levels in specific agro-ecological zones.

Developing a sustainable agricultural model is one of the great challenges of the coming years. The agricultural practices inherited from the Green Revolution of the 1960s show their limits today, and new paradigms need to be explored to counter rising issues such as the multiplication of climate-change related drought episodes. Two such new paradigms are the use of functional-structural plant models to complement and rationalize breeding approaches and a renewed focus on root systems as untapped sources of plant amelioration. Since the late 1980s, numerous functional and structural models of root systems were developed and used to investigate the properties of root systems in soil or lab-conditions. In this review, we focus on the conception and use of such root models in the broader context of research on root-driven drought tolerance, on the basis of root system architecture (RSA) phenotyping. Such models result from the integration of architectural, physiological and environmental data. Here, we consider the different phenotyping techniques allowing for root architectural and physiological study and their limits. We discuss how QTL and breeding studies support the manipulation of RSA as a way to improve drought resistance. We then go over the integration of the generated data within architectural models, how those architectural models can be coupled with functional hydraulic models, and how functional parameters can be measured to feed those models. We then consider the assessment and validation of those hydraulic models through confrontation of simulations to experimentations. Finally, we discuss the up and coming challenges facing root systems functional-structural modeling approaches in the context of breeding.

Lower plant transpiration rate (TR) under high vapor pressure deficit (VPD) conditions and early plant vigor are proposed as major traits influencing the rate of crop water use and possibly the fitness of chickpea lines to specific terminal drought conditions-this being the major constraint limiting chickpea productivity. The physiological mechanisms underlying difference in TR under high VPD and vigor are still unresolved, and so is the link between vigor and TR. Lower TR is hypothesized to relate to hydraulic conductance differences. Experiments were conducted in both soil (Vertisol) and hydroponic culture. The assessment of the TR response to increasing VPD showed that high vigor genotypes had TR restriction under high VPD, and this was confirmed in the early vigor parent and progeny genotype (ICC 4958 and RIL 211) having lower TR than the late vigor parent and progeny genotype (ICC 1882 and RIL 022). Inhibition of water transport pathways [apoplast and symplast (aquaporins)] in intact plants led to a lower transpiration inhibition in the early vigor/low TR genotypes than in the late vigor/high TR genotypes. De-rooted shoot treatment with an aquaporin inhibitor led to a lower transpiration inhibition in the early vigor/low TR genotypes than in the late vigor/high TR genotypes. Early vigor genotypes had lower root hydraulic conductivity than late vigor/high TR genotypes. Under inhibited conditions (apoplast, symplast), root hydraulic conductivity was reduced more in the late vigor/high TR genotypes than in the early vigor/low TR genotypes. We interpret that early vigor/low TR genotypes have a lower involvement of aquaporins in water transport pathways and may also have a smaller apoplastic pathway than high TR genotypes, which could explain the transpiration restriction under high VPD and would be helpful to conserve soil water under high evaporative demand. These findings open an opportunity for breeding to tailor genotypes with different "dosage" of these traits toward adaptation to varying drought-prone environments.

Water deficit under nearly all field conditions is the major constraint on plant yields. Other than empirical observations, very little progress has been made in developing crop plants in which specific physiological traits for drought are expressed. As a consequence, there was little known about under what conditions and to what extent drought impacts crop yield. However, there has been rapid progress in recent years in understanding and developing a limited-transpiration trait under elevated atmospheric vapor pressure deficit to increase plant growth and yield under water-deficit conditions. This review paper examines the physiological basis for the limited-transpiration trait as result of low plant hydraulic conductivity, which appears to be related to aquaporin activity. Methodology was developed based on aquaporin involvement to identify candidate genotypes for drought tolerance of several major crop species. Cultivars of maize and soybean are now being marketed specifically for arid conditions. Understanding the mechanism of the limited-transpiration trait has allowed a geospatial analyses to define the environments in which increased yield responses can be expected. This review highlights the challenges and approaches to finally develop physiological traits contributing directly to plant improvement for water-limited environments., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Groundnut production is limited in Sub-Saharan Africa and water deficit or "drought," is often considered as the main yield-limiting factor. However, no comprehensive study has assessed the extent and intensity of "drought"-related yield decreases, nor has it explored avenues to enhance productivity. Hence, crop simulation modeling with SSM (Simple Simulation Modeling) was used to address these issues. To palliate the lack of reliable weather data as input to the model, the validity of weather data generated by Marksim, a weather generator, was tested. Marksim provided good weather representation across a large gradient of rainfall, representative of the region, and although rainfall generated by Marksim was above observations, run-off from Marksim data was also higher, and consequently simulations using observed or Marksim weather agreed closely across this gradient of weather conditions (root mean square of error = 99 g m -2 ; R 2 = 0.81 for pod yield). More importantly, simulation of yield changes upon agronomic or genetic alterations in the model were equally predicted with Marksim weather. A 1° × 1° grid of weather data was generated. "Drought"-related yield reduction were limited to latitudes above 12-13° North in West Central Africa (WCA) and to the Eastern fringes of Tanzania and Mozambique in East South Africa (ESA). Simulation and experimental trials also showed that doubling the sowing density of Spanish cultivars from 20 to 40 plants m -2 would increase yield dramatically in both WCA and ESA. However, increasing density would require growers to invest in more seeds and likely additional labor. If these trade-offs cannot be alleviated, genetic improvement would then need to re-focus on a plant type that is adapted to the current low sowing density, like a runner rather than a bush plant type, which currently receives most of the genetic attention. Genetic improvement targeting "drought" adaptation should also be restricted to areas where water is indeed an issue, i.e., above 12-13°N latitude in WCA and the Eastern fringes of Tanzania and Mozambique.

Chickpeas are often grown under receding soil moisture and suffer ~50% yield losses due to drought stress. The timing of soil water use is considered critical for the efficient use of water under drought and to reduce yield losses. Therefore the root growth and the soil water uptake of 12 chickpea genotypes known for contrasts in drought and rooting response were monitored throughout the growth period both under drought and optimal irrigation. Root distribution reduced in the surface and increased in the deep soil layers below 30cm in response to drought. Soil water uptake was the maximum at 45-60cm soil depth under drought whereas it was the maximum at shallower (15-30 and 30-45cm) soil depths when irrigated. The total water uptake under drought was 1-fold less than optimal irrigation. The amount of water left unused remained the same across watering regimes. All the drought sensitive chickpea genotypes were inferior in root distribution and soil water uptake but the timing of water uptake varied among drought tolerant genotypes. Superiority in water uptake in most stages and the total water use determined the best adaptation. The water use at 15-30cm soil depth ensured greater uptake from lower depths and the soil water use from 90-120cm soil was critical for best drought adaptation. Root length density and the soil water uptake across soil depths were closely associated except at the surface or the ultimate soil depths of root presence.

Chickpea, the second most important legume crop, suffers major yield losses by terminal drought stress (DS). Stronger root system is known to enhance drought yields but this understanding remains controversial. To understand precisely the root traits contribution towards yield, 12 chickpea genotypes with well-known drought response were field evaluated under drought and optimal irrigation. Root traits, such as root length density (RLD), total root dry weight (RDW), deep root dry weight (deep RDW) and root:shoot ratio (RSR), were measured periodically by soil coring up to 1.2 m soil depth across drought treatments. Large variations were observed for RLD, RDW, deep RDW and RSR in both the drought treatments. DS increased RLD below 30 cm soil depth, deep RDW, RSR but decreased the root diameter. DS increased the genetic variation in RDW more at the penultimate soil depths. Genetic variation under drought was the widest for RLD ∼50 DAS, for deep RDW ∼50-75 DAS and for RSR at 35 DAS. Genotypes ICC 4958, ICC 8261, Annigeri, ICC 14799, ICC 283 and ICC 867 at vegetative stage and genotypes ICC 14778, ICCV 10, ICC 3325, ICC 14799 and ICC 1882 at the reproductive phase produced greater RLD. Path- and correlation coefficients revealed strong positive contributions of RLD after 45 DAS, deep RDW at vicinity of maturity and RSR at early podfill stages to yield under drought. Breeding for the best combination of profuse RLD at surface soil depths, and RDW at deeper soil layers, was proposed to be the best selection strategy, for an efficient water use and an enhanced terminal drought tolerance in chickpea.

As the incidence of water deficit and heat stress increases in many production regions there is an increasing requirement for crops adapted to these stresses. Thus it is essential to match water supply and demand, particularly during grain-filling. Here we integrate Grime's ecological strategies approach with traditional drought resistance/yield component frameworks describing plant responses to water deficit. We demonstrate that water use is a function of both short and longer term trade-offs between competing demands for carbon. Agricultural crop adaptation is based on escape. Rapid growth rates and high reproductive investment maximize yield, and stress is avoided through a closely regulated, climate-appropriate annual phenology. Crops have neither the resources nor morphological capacity to withstand long periods of intense water deficit. Thus, under terminal drought, yield potential is traded off against drought escape, such that drought postponing and/or tolerance traits which extend the growing season and/or divert source from reproductive sinks are maladaptive. However, these traits do play a supporting role against transient water deficits, allowing longer season cultivars to survive by mining water through deeper roots, or restricting transpiration. Recognizing these trade-offs made within escape-strategy limits will allow breeders to integrate complementary adaptive traits to transient and terminal water deficits., (Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.)

Rapid leaf area development may be attractive under a number of cropping conditions to enhance the vigor of crop establishment and allow rapid canopy closure for maximizing light interception and shading of weed competitors. This study was undertaken to determine (1) if parameters describing leaf area development varied among ten peanut ( Arachis hypogeae L.) genotypes grown in field and pot experiments, (2) if these parameters were affected by the planting density, and (3) if these parameters varied between Spanish and Virginia genotypes. Leaf area development was described by two steps: prediction of main stem number of nodes based on phyllochron development and plant leaf area dependent based on main stem node number. There was no genetic variation in the phyllochron measured in the field. However, the phyllochron was much longer for plants grown in pots as compared to the field-grown plants. These results indicated a negative aspect of growing peanut plants in the pots used in this experiment. In contrast to phyllochron, there was no difference in the relationship between plant leaf area and main stem node number between the pot and field experiments. However, there was genetic variation in both the pot and field experiments in the exponential coefficient (PLAPOW) of the power function used to describe leaf area development from node number. This genetic variation was confirmed in another experiment with a larger number of genotypes, although possible G × E interaction for the PLAPOW was found. Sowing density did not affect the power function relating leaf area to main stem node number. There was also no difference in the power function coefficient between Spanish and Virginia genotypes. SSM (Simple Simulation model) reliably predicted leaf canopy development in groundnut. Indeed the leaf area showed a close agreement between predicted and observed values up to 60000 cm 2  m -2 . The slightly higher prediction in India and slightly lower prediction in Niger reflected GxE interactions. Until more understanding is obtained on the possible GxE interaction effects on the canopy development, a generic PLAPOW value of 2.71, no correction for sowing density, and a phyllochron on 53 °C could be used to model canopy development in peanut.

Chickpea is the second most important legume crop largely grown under semi-arid tropics where terminal drought is one of the major constraints for its productivity. A trait-based selection had been considered more beneficial in drought tolerance breeding to overcome the environmental influence on drought yields. Large number of traits had been suggested in literature, with less indication on their importance and priority, for use in such breeding programs resulting in poor utilization of critical traits in drought tolerance breeding. To identify the most critical traits that contribute to grain yield under drought, 12 chickpea genotypes, with well-defined drought response, were field evaluated by sampling at regular intervals during the cropping period. Large range of variation was observed for shoot biomass productivity, specific leaf area (SLA) and leaf area index (LAI) at different days after sowings (DAS), canopy temperature depression (CTD) at mid-reproductive stages, growth duration and both morphological and analytical yield components. Grain yield under drought was closely associated with the rate of partitioning (p), crop growth rate (C), CTD, phenology, LAI at mid-pod fill stage, pod number m -2 at maturity, shoot biomass at reproductive growth stages and SLA at physiological maturity. The shoot trait(s) were prioritized based on their significance and contribution to drought tolerance. The trait(s) that conferred tolerance varied across genotypes. The order of traits/plant functions identified as important and critical for the drought tolerance were p, C, CTD, growth duration and other related traits. Relatively less important traits were LAI, SLA at the mid reproductive stage and pod number per unit area at maturity. The traits Dr, seeds pod -1 and 100-seed weight were found to be least important. Breeding for the best combination of p and C with the right phenology was proposed to be the best selection strategy to enhance terminal drought tolerance in chickpea.

West Africa (WA) is among the most food insecure regions. Rapid human population growth and stagnating crop yields greatly contribute to this fact. Poor soil fertility, especially low plant available phosphorus (P) is constraining food production in the region. P-fertilizer use in WA is among the lowest in the world due to inaccessibility and high prices, often unaffordable to resource-poor subsistence farmers. This article provides an overview of soil P-deficiency in WA and opportunities to overcome it by exploiting sorghum and pearl millet genetic diversity. The topic is examined from the perspectives of plant breeding, soil science, plant physiology, plant nutrition, and agronomy, thereby referring to recent results obtained in a joint interdisciplinary research project, and reported literature. Specific objectives are to summarize: (1) The global problem of P scarcity and how it will affect WA farmers; (2) Soil P dynamics in WA soils; (3) Plant responses to P deficiency; (4) Opportunities to breed for improved crop adaptation to P-limited conditions; (5) Challenges and trade-offs for improving sorghum and pearl millet adaptation to low-P conditions in WA; and (6) Systems approaches to address soil P-deficiency in WA. Sorghum and pearl millet in WA exhibit highly significant genetic variation for P-uptake efficiency, P-utilization efficiency, and grain yield under P-limited conditions indicating the possibility of breeding P-efficient varieties. Direct selection under P-limited conditions was more efficient than indirect selection under high-P conditions. Combining P-uptake and P-utilization efficiency is recommendable for WA to avoid further soil mining. Genomic regions responsible for P-uptake, P-utilization efficiency, and grain yield under low-P have been identified in WA sorghum and pearl millet, and marker-assisted selection could be possible once these genomic regions are validated. Developing P-efficient genotypes may not, however, be a sustainable solution in itself in the long-term without replenishing the P removed from the system in harvested produce. We therefore propose the use of integrated soil fertility management and systems-oriented management such as enhanced crop-tree-livestock integration in combination with P-use-efficiency-improved varieties. Recycling P from animal bones, human excreta and urine are also possible approaches toward a partially closed and efficient P cycle in WA.

Heat shock proteins (Hsp10) belong to the ubiquitous family of heat-shock molecular chaperones found in the organelles of both prokaryotes and eukaryotes. Chaperonins assist the folding of nascent and stress-destabilized proteins. A cDNA clone encoding a 10 kDa Hsp was isolated from pearl millet, Pennisetum glaucum (L.) by screening a heat stress cDNA library. The fulllength PgHsp10 cDNA consisted of 297 bp open reading frame (ORF) encoding a 98 amino acid polypeptide with a predicted molecular mass of 10.61 kDa and an estimated isoelectric point (pI) of 7.95. PgHsp10 shares 70-98 % sequence identity with other plant homologs. Phylogenetic analysis revealed that PgHsp10 is evolutionarily close to the maize Hsp10 homolog. The predicted 3D model confirmed a conserved eight-stranded ß-barrel with active site between the ß-barrel comprising of eight-strands, with conserved domain VLLPEYGG sandwiched between two ß-sheets. The gene consisted of 3 exons and 2 introns, while the position and phasing of these introns were conserved similar to other plant Hsp10 family genes. In silico analysis of the promoter region of PgHsp10 presented several distinct set of cis-elements and transcription factor binding sites. Quantitative RT-PCR analysis showed that PgHsp10 gene was differentially expressed in response to abiotic stresses with the highest level of expression under heat stress conditions. Results of this study provide useful information regarding the role of chaperonins in stress regulation and generated leads for further elucidation of their function in plant stress tolerance.

Water limitation is a major yield limiting factor in groundnut and transpiration efficiency (TE) is considered the main target for improvement, but TE being difficult to measure it has mostly been screened with surrogates. The paper re-explore the contribution of TE to grain yield in peanut by using a novel experimental approach in which TE is measured gravimetrically throughout the crop life cycle, in addition to measurement of TE surrogates. Experimentation was carried out with the groundnut reference collection (n = 288), across seasons varying for the evaporative demand (vapor pressure deficit, VPD) and across both fully irrigated and intermittent water stress conditions. There was large genotypic variation for TE under water stress in both low and high VPD season but the range was larger (5-fold) in the high- than in the low-VPD season (2-fold). Under water stress in both seasons, yield was closely related to the harvest index (HI) while TE related directly to yield only in the high VPD season. After discounting the direct HI effect on yield, TE explained a large portion of the remaining yield variations in both seasons, although marginally in the low VPD season. By contrast, the total water extracted from the soil profile, which varied between genotypes, did not relate directly to pod yield and neither to the yield residuals unexplained by HI. Surrogates for TE (specific leaf area, SLA, and SPAD chlorophyll meter readings, SCMR) never showed any significant correlation to TE measurements. Therefore, TE is an important factor explaining yield differences in groundnut under high VPD environments, suggesting that stomatal regulation under high VPD, rather than high photosynthetic rate as proposed earlier, may have a key role to play in the large TE differences found, which open new opportunities to breed improved groundnut for high VPD.

Traits influencing plant water use eventually define the fitness of genotypes for specific rainfall environments. We assessed the response of several water use traits to vapour pressure deficit (VPD) in pearl millet (Pennisetum glaucum (L.) R.Br.) genotypes known to differ in drought adaptation mechanisms: PRLT 2/89-33 (terminal drought-adapted parent), H 77/833-2 (terminal drought-sensitive parent) and four near-isogenic lines introgressed with a terminal drought tolerance quantitative trait locus (QTL) from PRLT 2/89-33 (ICMR01029, ICMR01031, ICMR02042, and ICMR02044). Plant water use traits at various levels of plant organisation were evaluated in seven experiments in plants exposed either transiently or over the long term to different VPD regimes: biomass components, transpiration (water usage per time unit) and transpiration rate (TR) upon transient VPD increase (gH2Ocm-2h-1)), transpiration efficiency (g dry biomass per kg H2O transpired), leaf expansion rate (cm per thermal time unit) and root anatomy (endodermis dimensions)). High VPD decreased biomass accumulation by reducing tillering, the leaf expansion rate and the duration of leaf expansion; decreased root endodermis cell size; and increased TR and the rate of TR increase upon gradual short-term VPD increases. Such changes may allow plants to increase their water transport capacity in a high VPD environment and are genotype-specific. Some variation in water use components was associated with terminal drought adaptation QTL. Knowledge of water use traits' plasticity in growth environments that varied in evaporative demand, and on their genetic determinacy, is necessary to develop trait-based breeding approaches to complex constraints.

Accurate and reliable gene expression data from qPCR depends on stable reference gene expression for potential gene functional analyses. In this study, 15 reference genes were selected and analyzed in various sample sets including abiotic stress treatments (salt, cold, water stress, heat, and abscisic acid) and tissues (leaves, roots, seedlings, panicle, and mature seeds). Statistical tools, including geNorm, NormFinder and RefFinder, were utilized to assess the suitability of reference genes based on their stability rankings for various sample groups. For abiotic stress, PP2A and CYP were identified as the most stable genes. In contrast, EIF4α was the most stable in the tissue sample set, followed by PP2A; PP2A was the most stable in all the sample set, followed by EIF4α. GAPDH, and UBC1 were the least stably expressed in the tissue and all the sample sets. These results also indicated that the use of two candidate reference genes would be sufficient for the optimization of normalization studies. To further verify the suitability of these genes for use as reference genes, SbHSF5 and SbHSF13 gene expression levels were normalized using the most and least stable sorghum reference genes in root and water stressed-leaf tissues of five sorghum varieties. This is the first systematic study of the selection of the most stable reference genes for qPCR-related assays in Sorghum bicolor that will potentially benefit future gene expression studies in sorghum and other closely related species.

Aquaporin activity and root anatomy may affect root hydraulic properties under drought stress. To better understand the function of aquaporins in rice root water fluxes under drought, we studied the root hydraulic conductivity (Lpr) and root sap exudation rate (Sr) in the presence or absence of an aquaporin inhibitor (azide) under well-watered conditions and following drought stress in six diverse rice varieties. Varieties varied in Lpr and Sr under both conditions. The contribution of aquaporins to Lpr was generally high (up to 79% under well-watered conditions and 85% under drought stress) and differentially regulated under drought. Aquaporin contribution to Sr increased in most varieties after drought, suggesting a crucial role for aquaporins in osmotic water fluxes during drought and recovery. Furthermore, root plasma membrane aquaporin (PIP) expression and root anatomical properties were correlated with hydraulic traits. Three chromosome regions highly correlated with hydraulic traits of the OryzaSNP panel were identified, but did not co-locate with known aquaporins. These results therefore highlight the importance of aquaporins in the rice root radial water pathway, but emphasize the complex range of additional mechanisms related to root water fluxes and drought response., (© 2015 John Wiley & Sons Ltd.)

This brief communication contains a description of the 2002-2010 annual panel collected by the Tsimane' Amazonian Panel Study team. The study took place among the Tsimane', a native Amazonian society of forager-horticulturalists. The team tracked a wide range of socio-economic and anthropometric variables from all residents (633 adults ≥16 years; 820 children) in 13 villages along the Maniqui River, Department of Beni. The panel is ideally suited to examine how market exposure and modernization affect the well-being of a highly autarkic population and to examine human growth in a non-Western rural setting., (Copyright © 2015 Elsevier B.V. All rights reserved.)