Leaf relative water content at 50% stomatal conductance measured by noninvasive NMR is linked to climate of origin in nine species of eucalypt.

Stomata are the gatekeepers of plant water use and must quickly respond to changes in plant water status to ensure plant survival under fluctuating environmental conditions. The mechanism for their closure is highly sensitive to disturbances in leaf water status, which makes isolating their response to declining water content difficult to characterise and to compare responses among species. Using a small-scale non-destructive nuclear magnetic resonance spectrometer as a leaf water content sensor, we measure the stomatal response to rapid induction of water deficit in the leaves of nine species of eucalypt from contrasting climates. We found a strong linear correlation between relative water content at 50% stomatal conductance (RWCgs50 ) and mean annual temperature at the climate of origin of each species. We also show evidence for stomata to maintain control over water loss well below turgor loss point in species adapted to warmer climates and secondary increases in stomatal conductance despite declining water content. We propose that RWCgs50 is a promising trait to guide future investigations comparing stomatal responses to water deficit. It may provide a useful phenotyping trait to delineate tolerance and adaption to hot temperatures and high leaf-to-air vapour pressure deficits.

The Metabolic Profile of Young, Watered Chickpea Plants Can Be Used as a Biomarker to Predict Seed Number under Terminal Drought.

Chickpea is the second-most-cultivated legume globally, with India and Australia being the two largest producers. In both of these locations, the crop is sown on residual summer soil moisture and left to grow on progressively depleting water content, finally maturing under terminal drought conditions. The metabolic profile of plants is commonly, correlatively associated with performance or stress responses, e.g., the accumulation of osmoprotective metabolites during cold stress. In animals and humans, metabolites are also prognostically used to predict the likelihood of an event (usually a disease) before it occurs, e.g., blood cholesterol and heart disease. We sought to discover metabolic biomarkers in chickpea that could be used to predict grain yield traits under terminal drought, from the leaf tissue of young, watered, healthy plants. The metabolic profile (GC-MS and enzyme assays) of field-grown chickpea leaves was analysed over two growing seasons, and then predictive modelling was applied to associate the most strongly correlated metabolites with the final seed number plant-1. Pinitol (negatively), sucrose (negatively) and GABA (positively) were significantly correlated with seed number in both years of study. The feature selection algorithm of the model selected a larger range of metabolites including carbohydrates, sugar alcohols and GABA. The correlation between the predicted seed number and actual seed number was R2 adj = 0.62, demonstrating that the metabolic profile could be used to predict a complex trait with a high degree of accuracy. A previously unknown association between D-pinitol and hundred-kernel weight was also discovered and may provide a single metabolic marker with which to predict large seeded chickpea varieties from new crosses. The use of metabolic biomarkers could be used by breeders to identify superior-performing genotypes before maturity is reached.

Metabolic, physiological and anatomical responses of soybean plants under water deficit and high temperature condition.

Water deficit (WD) combined with high temperature (HT) is the major factor limiting agriculture worldwide, and it is predicted to become worse according to the current climate change scenario. It is thus important to understand how current cultivated crops respond to these stress conditions. Here we investigated how four soybean cultivars respond to WD and HT isolated or in combination at metabolic, physiological, and anatomical levels. The WD + HT increased the level of stress in soybean plants when compared to plants under well-watered (WW), WD, or HT conditions. WD + HT exacerbates the increases in ascorbate peroxidase activity, which was associated with the greater photosynthetic rate in two cultivars under WD + HT. The metabolic responses to WD + HT diverge substantially from plants under WW, WD, or HT conditions. Myo-inositol and maltose were identified as WD + HT biomarkers and were connected to subnetworks composed of catalase, amino acids, and both root and leaf osmotic potentials. Correlation-based network analyses highlight that the network heterogeneity increased and a higher integration among metabolic, physiological, and morphological nodes is observed under stress conditions. Beyond unveiling biochemical and metabolic WD + HT biomarkers, our results collectively highlight that the mechanisms behind the acclimation to WD + HT cannot be understood by investigating WD or HT stress separately.

Carbon isotope and soluble metabolites reflect physiological status among contrasting faba bean genotypes in response to water deficit.

Identification and validation of biomarkers and bioindicators to select genotypes with superior tolerance to water deficit (WD) under field conditions are paramount to plant breeding programs. However, the co-occurrence of different abiotic stresses such as WD, heat, and radiation makes it difficult to develop generalized protocols to monitor the physiological health of the plant system. The study assessed the most abundant carbohydrates and sugar alcohols in five faba bean (Vicia faba) genotypes under field conditions and the abundance of naturally occurring carbon isotopes in bulk leaf material to predict water use efficiency (WUE). Plant water status and biomass accumulation were also assessed. Among the accumulated sugars, inter-specific variation in glucose was most prevalent and was found at a higher concentration (8.52 mg g-1 leaf) in rainfed trial. myo-Inositol concentrations followed that of glucose accumulation in that the rainfed trial had higher amounts compared to the irrigated trial. WUE calculated from carbon isotope abundance was consistently offset with measured WUE from measurements of leaf gas exchange. All genotypes demonstrated significant relationships between predicted and measured WUE (p < 0.05) apart from control variety PBA Warda. Thus, bulk leaf-level carbon isotope abundance can be used to calculate WUE and used as an effective selection criterion for improving WUE in faba bean breeding programs under field conditions.

Temporal imaging of drug dynamics in live cells using stimulated Raman scattering microscopy and a perfusion cell culture system.

Stimulated Raman scattering (SRS) microscopy is a powerful technique for visualising the cellular uptake and distribution of drugs and small molecules in live cells under biocompatible imaging conditions. The use of bio-orthogonal groups within the drug molecule, including alkynes and nitriles, has enabled the direct detection of a plethora of bioactive molecules in a minimally perturbative fashion. Limited progress has been made towards real-time detection of drug uptake and distribution into live cells under physiological conditions, despite the accordant potential it presents for preclinical drug development. SRS microscopy has been applied to the study of cellular dynamics of the drug 7RH, which is a potent inhibitor of dicoidin domain receptor 1 (DDR1) and prevents cellular adhesion, proliferation and migration in vitro. The uptake of 7RH into a variety of mammalian cell models was shown to be independent of DDR1 expression. Using a perfusion chamber, the recurrent treatment of live cancer cells was achieved, enabling 7RH uptake to be visualised in real-time using SRS microscopy, after which the viability of the same cellular population was assessed using commercially available fluorescent markers in a multimodal imaging experiment. The effect of 7RH treatment in combination with the chemotherapeutic, cisplatin was investigated using sequential perfusion and time-lapse imaging in the same live cell population, to demonstrate the application of the approach. SRS microscopy also identified potent inhibition of cellular adhesion and migration in breast cancer cell models with increasing 7RH treatment concentrations, thus representing a novel read-out methodology for phenotypic assays of this kind. The direct assessment of drug-cell interactions under physiological conditions offers significant potential for the preclinical drug development process.

Effect of Drought and Low P on Yield and Nutritional Content in Common Bean.

Common bean (Phaseolus vulgaris L.) production in the tropics typically occurs in rainfed systems on marginal lands where yields are low, primarily as a consequence of drought and low phosphorus (P) availability in soil. This study aimed to investigate the physiological and chemical responses of 12 bush bean genotypes for adaptation to individual and combined stress factors of drought and low P availability. Water stress and P deficiency, both individually and combined, decreased seed weight and aboveground biomass by ∼80%. Water deficit and P deficiency decreased photosynthesis and stomatal conductance during plant development. Maximum rates of carboxylation, electron transport, and triose phosphate utilization were superior for two common bean genotypes (SEF60 and NCB226) that are better adapted to combined stress conditions of water deficit and low P compared to the commercial check (DOR390). In response to water deficit treatment, carbon isotope fractionation in the leaf tissue decreased at all developmental stages. Within the soluble leaf fraction, combined water deficit and low P, led to significant changes in the concentration of key nutrients and amino acids, whereas no impact was detected in the seed. Our results suggest that common bean genotypes have a degree of resilience in yield development, expressed in traits such as pod harvest index, and conservation of nutritional content in the seed. Further exploration of the chemical and physiological traits identified here will enhance the resilience of common bean production systems in the tropics.

In situ pod growth rate reveals contrasting diurnal sensitivity to water deficit in Phaseolus vulgaris.

The development of reproductive tissues determines plant fecundity and yield. Loading of resources into the developing reproductive tissue is thought to be under the co-limiting effects of source and sink strength. The dynamics of this co-limitation are unknown, largely due to an inability to measure the flux of resources into a developing sink. Here we use nuclear magnetic resonance (NMR) sensors to measure sink strength by quantifying rates of pod dry matter accumulation (pod loading) in Phaseolus vulgaris at 13-min intervals across the diel period. Rates of pod loading showed contrasting variation across light and dark periods during the onset of water deficit. In addition, rates of pod loading appeared decoupled from net photosynthetic rates when adjusted to the plant scale. Combined, these observations illustrate that the rate of pod development varies under water limitation and that continuous, non-invasive methodologies to measure sink strength provide insight into the governing processes that determine the development of reproductive tissues.

Investigating Nutrient Supply Effects on Plant Growth and Seed Nutrient Content in Common Bean.

Low soil fertility commonly limits growth and yield production of common bean (Phaseolus vulgaris L.) in tropical regions. Impacts of nutrient limitations on production volume are well studied and are a major factor in reducing crop yields. This study characterised the impact of reduced nutrient supply on carbon assimilation and nutrient content of leaf, phloem sap and reproductive tissues of common bean grown in a controlled environment in order to detect chemical markers for changes in nutritional content. Leaf gas exchange measurements were undertaken over plant development to characterise changes to carbon assimilation under reduced nutrient supply. Samples of leaf, phloem sap and pod tissue of common bean were analysed for carbon isotope discrimination, mineral nutrient content, and amino acid concentration. Despite declines in nutrient availability leading to decreased carbon assimilation and reductions in yield, amino acid concentration was maintained in the pod tissue. Common bean can maintain the nutritional content of individual pods under varying nutrient availabilities demonstrating the resilience of processes determining the viability of reproductive tissues.

Phloem sap metabolites vary according to the interactive effects of nutrient supply and seasonal conditions in Eucalyptus globulus (Labill).

Improving the efficiency of fertilizer application is paramount to both the sustainability and profitability of forest plantations. Therefore, developing reliable, cost-effective tools to assess tree nutritional status is of great interest. This investigation sought to assess the use of phloem sap-derived metabolites as an indicator of nutritional status on a background of seasonal water availability of Eucalyptus globulus (Labill) trees grown under field conditions. Phloem is a central conduit for long-distance transport and signaling in plants and offers great promise in reflecting plant-scale resource limitations. Changes in the abundance of solutes and isotopes in phloem sap are sensitive to environmental cues. With a focus on both water and nutrient availability, we characterize patterns in phloem sugars, amino acids and the abundance of carbon isotopes in phloem sap obtained from E. globulus among different seasons and fertilizer treatments. Phloem-derived total amino acid concentration was found to increase with an increasing nitrogen (N) supply; however, this response was lost with the concurrent addition of phosphorus and at the highest level of N supply. Significant seasonal variation in all measured parameters was also detected, highlighting the need for caution in making quantitative relationships with growth. Broader implications of the interactive effects of both water supply and multi-nutrient additions and relationships with growth are discussed.

Chemical composition and reproductive functionality of contrasting faba bean genotypes in response to water deficit.

Water deficit (WD), a major contributor to yield reductions in faba bean (Vicia faba), is a complex phenomenon that varies across daily to seasonal cycles. Several studies have identified various morphological and physiological indicators of WD tolerance, which generally show limited water use during WD. Limited information is available on the impact of WD on nutrient content and reproductive biology of the faba bean. We studied carbohydrates, amino acids, mineral nutrients and the abundance of naturally occurring carbon isotopes (δ13 C) in leaf and grain tissues of faba bean genotypes grown under well-watered (WW) and WD conditions. δ13 C of leaf tissues were found to indicate changes in water use due to WD but this was not reflected in grain tissues. Nutrient concentrations with regard to amino acids and minerals were not influenced by WD. However, carbohydrate accumulation was found to be significant for WD, specifically through the presence of a higher concentration of myo-inositol in WD leaf tissues. Alternatively, sucrose concentration in grain tissues was reduced under WD treatment. WD hampered reproductive functionality by reducing pollen viability and germination with the severity and duration of stress and this reduction was less prominent in the drought-tolerant genotype (AC0805#4912) compared to the sensitive one (11NF010c-4). It was also demonstrated that WD caused developmental impairment in the stamen and pistil, where the pistil appeared more sensitive than stamen. These findings suggest that WD impairs pollen viability and pistil function reducing yield volume, but the nutrient content of the resulting yield is not significantly affected.

Wide variation in the suboptimal distribution of photosynthetic capacity in relation to light across genotypes of wheat.

Suboptimal distribution of photosynthetic capacity in relation to light among leaves reduces potential whole-canopy photosynthesis. We quantified the degree of suboptimality in 160 genotypes of wheat by directly measuring photosynthetic capacity and daily irradiance in flag and penultimate leaves. Capacity per unit daily irradiance was systematically lower in flag than penultimate leaves in most genotypes, but the ratio (γ) of capacity per unit irradiance between flag and penultimate leaves varied widely across genotypes, from less than 0.5 to over 1.2. Variation in γ was most strongly associated with differences in photosynthetic capacity in penultimate leaves, rather than with flag leaf photosynthesis or canopy light penetration. Preliminary genome-wide association analysis identified nine strong marker-trait associations with this trait, which should be validated in future work in other environments and/or materials. Our modelling suggests canopy photosynthesis could be increased by up to 5 % under sunny conditions by harnessing this variation through selective breeding for increased γ.

The preceding root system drives the composition and function of the rhizosphere microbiome.

BACKGROUND: The soil environment is responsible for sustaining most terrestrial plant life, yet we know surprisingly little about the important functions carried out by diverse microbial communities in soil. Soil microbes that inhabit the channels of decaying root systems, the detritusphere, are likely to be essential for plant growth and health, as these channels are the preferred locations of new root growth. Understanding the microbial metagenome of the detritusphere, and how it responds to agricultural management such as crop rotations and soil tillage, is vital for improving global food production. RESULTS: This study establishes an in-depth soil microbial gene catalogue based on the living-decaying rhizosphere niches in a cropping soil. The detritusphere microbiome regulates the composition and function of the rhizosphere microbiome to a greater extent than plant type: rhizosphere microbiomes of wheat and chickpea were homogenous (65-87% similarity) in the presence of decaying root (DR) systems but were heterogeneous (3-24% similarity) where DR was disrupted by tillage. When the microbiomes of the rhizosphere and the detritusphere interact in the presence of DR, there is significant degradation of plant root exudates by the rhizosphere microbiome, and genes associated with membrane transporters, carbohydrate and amino acid metabolism are enriched. CONCLUSIONS: The study describes the diversity and functional capacity of a high-quality soil microbial metagenome. The results demonstrate the contribution of the detritusphere microbiome in determining the metagenome of developing root systems. Modifications in root microbial function through soil management can ultimately govern plant health, productivity and food security.

Biochemical and physiological impacts of zinc sulphate, potassium phosphite and hydrogen sulphide in mitigating stress conditions in soybean.

Soybean is the most widely grown oilseed in the world. It is an important source of protein and oil which are derived from its seeds. Drought stress is a major constraint to soybean yields. Finding alternative methods to mitigate the water stress for soybean is useful to maintain adequate crop yields. The aim of this study was to evaluate the morpho-physiological, biochemical and metabolic changes in soybean plants in two ontogenetic stages, under exposure to water deficit and treatment with zinc sulphate (ZS), potassium phosphite (PP) or hydrogen sulphide (HS). We carried out two independent experiments in the V4 and R1 development stages consisting of the following treatments: well-watered control (WW, 100% maximum water holding capacity, MWHC), water deficit (WD, 50% MWHC), PP + WW, PP + WD, HS + WW, HS + WD, ZS + WW and ZS + WD. The experimental design consisted of randomized blocks with eight treatments with five replicates. Morphological, physiological and metabolic analyses were performed 8 days after the start of the treatments for both experiments. We identified two tolerance mechanisms acting in response to compound application during water stress: the first involved the upregulation of antioxidant enzyme activity and the second involved the accumulation of soluble sugars, free amino acids and proline to facilitate osmotic adjustment. Both mechanisms are related to the maintenance of the photosynthetic parameters and cell membrane integrity. This report suggests the potential agricultural use of these compounds to mitigate drought effects in soybean plants.

Field drought conditions impact yield but not nutritional quality of the seed in common bean (Phaseolus vulgaris L.).

Drought substantially limits seed yield of common bean (Phaseolus vulgaris L.) in the tropics. Understanding the interaction of drought on yield and the nutrient concentration of the seed is vital in order to supply nutrition to the millions of consumers who rely on common bean as a staple crop. Nevertheless, the impact of drought on common bean for both yield and nutrient concentration has not yet been concurrently investigated in a field environment. Using 10 bred lines developed by CIAT and its partners for their improved adaptation to drought and phosphorus deficiency, this study characterised the impact of drought on yield and nutrient concentration for leaf and seed tissue of common bean grown in the field. Drought significantly reduced leaf area (by ~50%), harvest index (by ~60%), yield (by ~70%), seed weight (by ~25%) and enriched carbon isotope abundance (δ13C) in the seed. Within the soluble leaf fraction, drought significantly decreased the concentration of mineral nutrients and amino acids, whereas no negative effect on the concentration of nutrients and amino acids was detected within the seed. Genotypic variation in nutrient concentration in both the leaf and seed tissue was identified and should be explored further to identify traits that may confer tolerance to abiotic stress.

Water Deficit Elicits a Transcriptional Response of Genes Governing d-pinitol Biosynthesis in Soybean (Glycine max).

d-pinitol is the most commonly accumulated sugar alcohol in the Leguminosae family and has been observed to increase significantly in response to abiotic stress. While previous studies have identified genes involved in d-pinitol synthesis, no study has investigated transcript expression in planta. The present study quantified the expression of several genes involved in d-pinitol synthesis in different plant tissues and investigated the accumulation of d-pinitol, myo-inositol and other metabolites in response to a progressive soil drought in soybean (Glycine max). Expression of myo-inositol 1-phosphate synthase (INPS), the gene responsible for the conversion of glucose-6-phosphate to myo-inositol-1-phosphate, was significantly up regulated in response to a water deficit for the first two sampling weeks. Expression of myo-inositol O-methyl transferase (IMT1), the gene responsible for the conversion of myo-inositol into d-ononitol was only up regulated in stems at sampling week 3. Assessment of metabolites showed significant changes in their concentration in leaves and stems. d-Pinitol concentration was significantly higher in all organs sampled from water deficit plants for all three sampling weeks. In contrast, myo-inositol, had significantly lower concentrations in leaf samples despite up regulation of INPS suggesting the transcriptionally regulated flux of carbon through the myo-inositol pool is important during water deficit.

PARbars: Cheap, Easy to Build Ceptometers for Continuous Measurement of Light Interception in Plant Canopies.

Ceptometry is a technique used to measure the transmittance of photosynthetically active radiation through a plant canopy using multiple light sensors connected in parallel on a long bar. Ceptometry is often used to infer properties of canopy structure and light interception, notably leaf area index (LAI) and effective plant area index (PAIeff). Due to the high cost of commercially available ceptometers, the number of measurements that can be taken is often limited in space and time. This limits the usefulness of ceptometry for studying genetic variability in light interception, and precludes thorough analysis of, and correction for, biases that can skew measurements depending on the time of day. We developed continuously logging ceptometers (called PARbars) that can be produced for USD $75 each and yield high quality data comparable to commercially available alternatives. Here we provide detailed instruction on how to build and calibrate PARbars, how to deploy them in the field and how to estimate PAI from collected transmittance data. We provide representative results from wheat canopies and discuss further considerations that should be made when using PARbars.

Corrigendum: Root Exudation of Primary Metabolites: Mechanisms and Their Roles in Plant Responses to Environmental Stimuli.

[This corrects the article DOI: 10.3389/fpls.2019.00157.].

Rate of photosynthetic induction in fluctuating light varies widely among genotypes of wheat.

Crop photosynthesis and yield are limited by slow photosynthetic induction in sunflecks. We quantified variation in induction kinetics across diverse genotypes of wheat for the first time. Following a preliminary study that hinted at wide variation in induction kinetics across 58 genotypes, we grew 10 genotypes with contrasting responses in a controlled environment and quantified induction kinetics of carboxylation capacity (Vcmax) from dynamic A versus ci curves after a shift from low to high light (from 50 µmol m-2 s-1 to 1500 µmol m-2 s-1), in five flag leaves per genotype. Within-genotype median time for 95% induction (t95) of Vcmax varied 1.8-fold, from 5.2 min to 9.5 min. Our simulations suggest that non-instantaneous induction reduces daily net carbon gain by up to 15%, and that breeding to speed up Vcmax induction in the slowest of our 10 genotypes to match that in the fastest genotype could increase daily net carbon gain by up to 3.4%, particularly for leaves in mid-canopy positions (cumulative leaf area index ≤1.5 m2 m-2), those that experience predominantly short-duration sunflecks, and those with high photosynthetic capacities.

Root Exudation of Primary Metabolites: Mechanisms and Their Roles in Plant Responses to Environmental Stimuli.

Root exudation is an important process determining plant interactions with the soil environment. Many studies have linked this process to soil nutrient mobilization. Yet, it remains unresolved how exudation is controlled and how exactly and under what circumstances plants benefit from exudation. The majority of root exudates including primary metabolites (sugars, amino acids, and organic acids) are believed to be passively lost from the root and used by rhizosphere-dwelling microbes. In this review, we synthetize recent advances in ecology and plant biology to explain and propose mechanisms by which root exudation of primary metabolites is controlled, and what role their exudation plays in plant nutrient acquisition strategies. Specifically, we propose a novel conceptual framework for root exudates. This framework is built upon two main concepts: (1) root exudation of primary metabolites is driven by diffusion, with plants and microbes both modulating concentration gradients and therefore diffusion rates to soil depending on their nutritional status; (2) exuded metabolite concentrations can be sensed at the root tip and signals are translated to modify root architecture. The flux of primary metabolites through root exudation is mostly located at the root tip, where the lack of cell differentiation favors diffusion of metabolites to the soil. We show examples of how the root tip senses concentration changes of exuded metabolites and translates that into signals to modify root growth. Plants can modify the concentration of metabolites either by controlling source/sink processes or by expressing and regulating efflux carriers, therefore challenging the idea of root exudation as a purely unregulated passive process. Through root exudate flux, plants can locally enhance concentrations of many common metabolites, which can serve as sensors and integrators of the plant nutritional status and of the nutrient availability in the surrounding environment. Plant-associated micro-organisms also constitute a strong sink for plant carbon, thereby increasing concentration gradients of metabolites and affecting root exudation. Understanding the mechanisms of and the effects that environmental stimuli have on the magnitude and type of root exudation will ultimately improve our knowledge of processes determining soil CO2 emissions, ecosystem functioning, and how to improve the sustainability of agricultural production.