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Aims: Root system architecture (RSA) plays an important role in the plant’s ability to sustain yield under abiotic stresses such as drought. Preceding crops (precrops) can affect the yield of the proceeding crop, partially by affecting the RSA. This experiment aims to explore the interactions between precrop identity, crop genotype and drought at early growth stages. Methods: Rhizotrons, sized 60 × 80 × 3.5 cm, were used to assess the early root growth of two winter wheat (Triticum aestivum L.) genotypes, using precroptreated soil around the seedlings and differing water regimes. The rhizotrons were automatically imaged 3 times a week to track root development. Results: Precrop-treated soil affected the RSA and changes caused by the reduced water treatment (RWT) were different depending on the precrop. Largest of these was the 36% reduction in root depth after wheat, but 44% after OSR. This indicates that effects caused by the precrop can be simulated, at least partially, by transferring precrop-treated soils to controlled environments. The genotypes had differential RSA and reacted differently to the RWT, with Julius maintaining an 8.8-13.1% deeper root system compared to Brons in the RWT. In addition, the combined environmental treatment affected the genotypes differently. Conclusion: Our results could help explain discrepancies found from using precrops to enhance yield as they indicate differences in the preceding crop effect when experiencing drought stress. Further, these differences are affected by genotypic interactions, which can be used to select and adapt crop genotypes for specific crop rotations, depending on the year. Additionally, we have shown a viable method of stimulating a partial precrop effect at the seedling stage in a controlled greenhouse setting using field soil around the germinated seed. [ABSTRACT FROM AUTHOR]

Suboptimal temperatures at sowing and emergence affect the early development of maize, with potentially irreversible effects later in the growing season. We studied recovery from cold stress of an inbred line (B73) and 13 Doubled Haploid lines derived from a European flint maize landrace. After a cold treatment (20–12 °C, day–night) from sowing to seedling establishment, seedlings were transplanted and grown in the greenhouse until the V8 stage (eight leaves fully developed), when we measured agronomically relevant plant traits and spectral indices of mature leaves. Survival rates of transplanted seedlings after cold treatment ranged from 10% to 100%. After a strong delay in early development due to cold, the surviving plants were able to compensate for this delay at later stages of recovery. They reached the V8 stage after only five more growing degree days than plants grown under the control treatment (25–18 °C, day–night). Plants from the most cold-tolerant genotypes (PE0401 and PE0100) accumulated more root and shoot biomass at the end of the recovery phase compared with the same genotypes exposed to the control treatment. The genotypes with the most plastic leaf morphological traits (PE0161 and PE0072) had little reduction in leaf biomass at the end of the recovery phase in comparison with less responsive genotypes such as PE0171. We conclude that genotypes that survived cold treatment with minimal cold damage of seedling leaves can be candidates for further cold recovery studies and breeding. Nevertheless, such studies must take trait acclimation for other suboptimal conditions into consideration. [ABSTRACT FROM AUTHOR]

Automated high-throughput plant phenotyping (HTPP) enables non-invasive, fast and standardized evaluations of a large number of plants for size, development, and certain physiological variables. Many research groups recognize the potential of HTPP and have made significant investments in HTPP infrastructure, or are considering doing so. To make optimal use of limited resources, it is important to plan and use these facilities prudently and to interpret the results carefully. Here we present a number of points that users should consider before purchasing, building or utilizing such equipment. They relate to (1) the financial and time investment for acquisition, operation, and maintenance, (2) the constraints associated with such machines in terms of flexibility and growth conditions, (3) the pros and cons of frequent nondestructive measurements, (4) the level of information provided by proxy traits, and (5) the utilization of calibration curves. Using data from an Arabidopsis experiment, we demonstrate how diurnal changes in leaf angle can impact plant size estimates from top-view cameras, causing deviations of more than 20% over the day. Growth analysis data from another rosette species showed that there was a curvilinear relationship between total and projected leaf area. Neglecting this curvilinearity resulted in linear calibration curves that, although having a high r² (> 0.92), also exhibited large relative errors. Another important consideration we discussed is the frequency at which calibration curves need to be generated and whether different treatments, seasons, or genotypes require distinct calibration curves. In conclusion, HTPP systems have become a valuable addition to the toolbox of plant biologists, provided that these systems are tailored to the research questions of interest, and users are aware of both the possible pitfalls and potential involved. [ABSTRACT FROM AUTHOR]

Summary: In natural environments, plants are exposed to rapidly changing light. Maintaining photosynthetic efficiency while avoiding photodamage requires equally rapid regulation of photoprotective mechanisms. We asked what the operation frequency range of regulation is in which plants can efficiently respond to varying light.Chlorophyll fluorescence, P700, plastocyanin, and ferredoxin responses of wild‐types Arabidopsis thaliana were measured in oscillating light of various frequencies. We also investigated the npq1 mutant lacking violaxanthin de‐epoxidase, the npq4 mutant lacking PsbS protein, and the mutants crr2‐2, and pgrl1ab impaired in different pathways of the cyclic electron transport.The fastest was the PsbS‐regulation responding to oscillation periods longer than 10 s. Processes involving violaxanthin de‐epoxidase dampened changes in chlorophyll fluorescence in oscillation periods of 2 min or longer. Knocking out the PGR5/PGRL1 pathway strongly reduced variations of all monitored parameters, probably due to congestion in the electron transport. Incapacitating the NDH‐like pathway only slightly changed the photosynthetic dynamics.Our observations are consistent with the hypothesis that nonphotochemical quenching in slow light oscillations involves violaxanthin de‐epoxidase to produce, presumably, a largely stationary level of zeaxanthin. We interpret the observed dynamics of photosystem I components as being formed in slow light oscillations partially by thylakoid remodeling that modulates the redox rates. [ABSTRACT FROM AUTHOR]

In recent years, quinoa (Chenopodium quinoa Willd.), an ancient Andean region crop, has received increased research attention because it is an excellent source of nutrients and also of bioactive phenolic compounds, which are potentially beneficial for human health. However, variation in the content and type of these metabolites in quinoa genetic resources remains, to a large extent, unexplored. We evaluated the composition of free and bound phenolic forms in the seeds of 111 Chilean quinoa accessions by using LC-DAD-MS/MS. The relative phenolic content ranged from 35.51 mg/100 g to 93.23 mg/100 g of seed dry weight. The free phenolic fraction accounted for 72% of the total phenolic content, while the bound fraction represented the remaining 28% of the total phenolic content. Our study also revealed a significant degree of variation in terms of individual phenolic compounds such as rutin, vanillic acid, quercetin, and their derivatives, which can have important implications for quinoa's nutritional and functional properties. We conclude that our data reveal a significant phenotypic variation of bioactive phenolic content in the examined germplasm, which could be exploited in current and future genetic improvement programs in quinoa. [ABSTRACT FROM AUTHOR]

This study aims to highlight the major effects of biochar incorporation into potting soil substrate on plant growth and performance in early growth stages of five elite Italian varieties of durum wheat (Triticum durum). The biochars used were obtained from two contrasting feedstocks, namely wood chips and wheat straw, by gasification under high temperature conditions, and were applied in a greenhouse experiment either as pure or as nutrient-activated biochar obtained by incubation with digestate. The results of the experiment showed that specific genotypes as well as different treatments with biochar have significant effects on plant response when looking at shoot traits related to growth. The evaluated genotypes could be clustered in two main distinct groups presenting, respectively, significantly increasing (Duilio, Iride, and Saragolla varieties) and decreasing (Marco Aurelio and Grecale varieties) values of projected shoot system area (PSSA), fresh weight (FW), dry weight (DW), and plant water loss by evapotranspiration (ET). All these traits were correlated with Pearson correlation coefficients ranging from 0.74 to 0.98. Concerning the treatment effect, a significant alteration of the mentioned plant traits was observed when applying biochar from wheat straw, characterized by very high electrical conductivity (EC), resulting in a reduction of 34.6% PSSA, 43.2% FW, 66.9% DW, and 36.0% ET, when compared to the control. Interestingly, the application of the same biochar after nutrient spiking with digestate determined about a 15–30% relief from the abovementioned reduction induced by the application of the sole pure wheat straw biochar. Our results reinforce the current basic knowledge available on biological soil amendments as biochar and digestate. [ABSTRACT FROM AUTHOR]

Chlorophyll fluorescence (ChlF) is a powerful non‐invasive technique for probing photosynthesis. Although proposed as a method for drought tolerance screening, ChlF has not yet been fully adopted in physiological breeding, mainly due to limitations in high‐throughput field phenotyping capabilities. The light‐induced fluorescence transient (LIFT) sensor has recently been shown to reliably provide active ChlF data for rapid and remote characterisation of plant photosynthetic performance. We used the LIFT sensor to quantify photosynthesis traits across time in a large panel of durum wheat genotypes subjected to a progressive drought in replicated field trials over two growing seasons. The photosynthetic performance was measured at the canopy level by means of the operating efficiency of Photosystem II (Fq′/Fm′) and the kinetics of electron transport measured by reoxidation rates (Fr1′ and Fr2′). Short‐ and long‐term changes in ChlF traits were found in response to soil water availability and due to interactions with weather fluctuations. In mild drought, Fq′/Fm′ and Fr2′ were little affected, while Fr1′ was consistently accelerated in water‐limited compared to well‐watered plants, increasingly so with rising vapour pressure deficit. This high‐throughput approach allowed assessment of the native genetic diversity in ChlF traits while considering the diurnal dynamics of photosynthesis. Photosynthetic performance of a large durum wheat panel under progressive drought in the field was quantified using the light‐induced fluorescence transient (LIFT) sensor for high‐throughput phenotyping. Photosynthesis traits showed high genetic variability, aiding physiological crop breeding. [ABSTRACT FROM AUTHOR]

Plant growth depends on the diurnal regulation of cellular processes, but it is not well understood if and how transcriptional regulation controls diurnal fluctuations at the protein level. Here, we report a high‐resolution Arabidopsis thaliana (Arabidopsis) leaf rosette proteome acquired over a 12 hr light:12 hr dark diurnal cycle and the phosphoproteome immediately before and after the light‐to‐dark and dark‐to‐light transitions. We quantified nearly 5,000 proteins and 800 phosphoproteins, of which 288 fluctuated in their abundance and 226 fluctuated in their phosphorylation status. Of the phosphoproteins, 60% were quantified for changes in protein abundance. This revealed six proteins involved in nitrogen and hormone metabolism that had concurrent changes in both protein abundance and phosphorylation status. The diurnal proteome and phosphoproteome changes involve proteins in key cellular processes, including protein translation, light perception, photosynthesis, metabolism and transport. The phosphoproteome at the light–dark transitions revealed the dynamics at phosphorylation sites in either anticipation of or response to a change in light regime. Phosphorylation site motif analyses implicate casein kinase II and calcium/calmodulin‐dependent kinases among the primary light–dark transition kinases. The comparative analysis of the diurnal proteome and diurnal and circadian transcriptome established how mRNA and protein accumulation intersect in leaves during the diurnal cycle of the plant. The manuscript provides quantitative information of diurnal changes in the accumulation and phosphorylation of proteins in Arabidopsis thaliana rosettes grown in a 12 hr photoperiod. The highly resolved time‐scale of the datasets offers new proteome‐level insights for future targeted studies. [ABSTRACT FROM AUTHOR]

In legumes, pod shattering occurs when mature pods dehisce along the sutures, and detachment of the valves promotes seed dispersal. In Phaseolus vulgaris (L). the major locus qPD5.1-Pv for pod indehiscence was identified recently. We developed a BC4/F4 introgression line population and narrowed the major locus down to a 22.5 kb region. Here, gene expression and a parallel histological analysis of dehiscent and indehiscent pods identified an AtMYB26 orthologue as the best candidate for loss of pod shattering, on a genomic region ~11 kb downstream of the highest associated peak. Based on mapping and expression data, we propose early and fine up-regulation of PvMYB26 in dehiscent pods. Detailed histological analysis establishes that pod indehiscence is associated with the lack of a functional abscission layer in the ventral sheath, and that the key anatomical modifications associated with pod shattering in common bean occur early during pod development. We finally propose that loss of pod shattering in legumes resulted from histological convergent evolution and that it is the result of selection at orthologous loci. [ABSTRACT FROM AUTHOR]

Water stress (WS) during spike development strongly affects final grain yield and grain quality in cereals. Proline, an osmoprotectant amino-acid, may contribute to alleviating the effects of cell and tissue dehydration. We studied five spring barley genotypes contrasting in their drought response, including two introgression lines, S42IL-143 and S42IL-141 , harboring a Pyrroline-5-carboxylate synthase1- P5cs1 allele originating from the wild barley accession ISR42-8. We tested the hypothesis that barley genotypes harboring a wild allele at P5cs1 locus are comparatively more drought-tolerant at the reproductive stage by inducing proline accumulation in their immature spikes. At the booting stage, we subjected plants to well-watered and WS treatments until physiological maturity. Several morpho-physiological traits had significant genotype by treatment interaction and reduction under WS. Varying levels of genotypic proline accumulation and differences in WS tolerance were observed. Spike proline accumulation was higher than leaf proline accumulation for all genotypes under WS. Also, introgression lines carrying a wild allele at P5cs1 locus had a markedly higher spike and leaf proline content compared with the other genotypes. These introgression lines showed milder drought symptoms compared with elite genotypes, remained photosynthetically active under WS, and maintained their intrinsic water use efficiency. These combined responses contributed to the achievement of higher final seed productivity. Magnetic resonance imaging (MRI) of whole spikes at the soft dough stage showed an increase in seed abortion among the elite genotypes compared with the introgression lines 15 days after WS treatment. Our results suggest that proline accumulation at the reproductive stage contributes to the maintenance of grain formation under water shortage. [ABSTRACT FROM AUTHOR]

Plant phenotyping enables noninvasive quantification of plant structure and function and interactions with environments. High-capacity phenotyping reaches hitherto inaccessible phenotypic characteristics. Diverse, challenging, and valuable applications of phenotyping have originated among scientists, prebreeders, and breeders as they study the phenotypic diversity of genetic resources and apply increasingly complex traits to crop improvement. Noninvasive technologies are used to analyze experimental and breeding populations. We cover the most recent research in controlled-environment and field phenotyping for seed, shoot, and root traits. Select field phenotyping technologies have become state of the art and show promise for speeding up the breeding process in early generations. We highlight the technologies behind the rapid advances in proximal and remote sensing of plants in fields. We conclude by discussing the new disciplines working with the phenotyping community: data science, to address the challenge of generating FAIR (findable, accessible, interoperable, and reusable) data, and robotics, to apply phenotyping directly on farms. [ABSTRACT FROM AUTHOR]

Glutathione is considered a key metabolite for stress defense and elevated levels have frequently been proposed to positively influence stress tolerance. To investigate whether glutathione affects plant performance and the drought tolerance of plants, wild-type Arabidopsis plants and an allelic series of five mutants (rax1, pad2, cad2, nrc1, and zir1) with reduced glutathione contents between 21 and 63% compared to wild-type glutathione content were phenotypically characterized for their shoot growth under control and water-limiting conditions using a shoot phenotyping platform. Under non-stress conditions the zir1 mutant with only 21% glutathione showed a pronounced dwarf phenotype. All other mutants with intermediate glutathione contents up to 62% in contrast showed consistently slightly smaller shoots than the wild-type. Moderate drought stress imposed through water withdrawal until shoot growth ceased showed that wild-type plants and all mutants responded similarly in terms of chlorophyll fluorescence and growth retardation. These results lead to the conclusion that glutathione is important for general plant performance but that the glutathione content does not affect tolerance to moderate drought conditions typically experienced by crops in the field. [ABSTRACT FROM AUTHOR]

Our phenotyping and hormonal study has characterized the role of cytokinins (CK) in the drought and recovery responses of Arabidopsis thaliana. CK down-regulation was achieved by overexpression of the gene for CK deactivating enzyme cytokinin oxidase/dehydrogenase (CKX): constitutive (35S:CKX) or at the stress onset using a dexamethasone-inducible pOp/LhGR promoter (DEX:CKX). The 35S:CKX plants exhibited slow ontogenesis and higher expression levels of stress-associated genes, e.g., AtP5CS1, already at well-watered conditions. CK down-regulation resulted during drought in higher stress tolerance (indicated by relatively low up-regulation of the expression of drought stress marker gene AtRD29B) accompanied with lower leaf water loss. Nevertheless, these plants exhibited slow and delayed recovery after re-watering. CK levels were increased at the stress onset by stimulation of the expression of CK biosynthetic gene isopentenyl transferase (ipt) (DEX:IPT) or by application of exogenous CK meta-topolin. After water withdrawal, long-term CK elevation resulted in higher water loss in comparison with CKX transformants as well as with plants overexpressing ipt driven by senescence-inducible SAG12 promoter (SAG:IPT), which gradually enhanced CKs during the stress progression. In all cases, CK up-regulation resulted in fast andmore vigorous recovery. All drought-stressed plants exhibited growth suppression associated with elevation of abscisic acid and decrease of auxins and active CKs (with the exception of SAG:IPT plants). Apart from the ipt overexpressers, also increase of jasmonic and salicylic acid was found. [ABSTRACT FROM AUTHOR]

Background. Soil moisture deficiency causes yield reduction and instability in faba bean (Vicia faba L.) production. The extent of sensitivity to drought stress varies across accessions originating from diverse moisture regimes of the world. Hence, we conducted successive greenhouse experiments in pots and rhizotrons to explore diversity in root responses to soil water deficit. Methods. A set of 89 accessions from wet and dry growing regions of the world was defined according to the Focused Identification of Germplasm Strategy and screened in a perlite-sand medium under well watered conditions in a greenhouse experiment. Stomatal conductance, canopy temperature, chlorophyll concentration, and root and shoot dry weights were recorded during the fifth week of growth. Eight accessions representing the range of responses were selected for further investigation. Starting five days after germination, they were subjected to a root phenotyping experiment using the automated phenotyping platform GROWSCREEN-Rhizo. The rhizotrons were filled with peat-soil under well watered and water limited conditions. Root architectural traits were recorded five, 12, and 19 days after the treatment (DAT) began. Results. In the germplasm survey, accessions from dry regions showed significantly higher values of chlorophyll concentration, shoot and root dry weights than those from wet regions. Root and shoot dry weight as well as seed weight, and chlorophyll concentration were positively correlated with each other. Accession DS70622 combined higher values of root and shoot dry weight than the rest. The experiment in GROWSCREEN-Rhizo showed large differences in root response to water deficit. The accession by treatment interactions in taproot and second order lateral root lengths were significant at 12 and 19 DAT, and the taproot length was reduced up to 57% by drought. The longest and deepest root systems under both treatment conditions were recorded by DS70622 and DS11320, and total root length of DS70622 was three times longer than that of WS99501, the shortest rooted accession. The maximum horizontal distribution of a root system and root surface coverage were positively correlated with taproot and total root lengths and root system depth. DS70622 and WS99501 combined maximum and minimum values of these traits, respectively. Thus, roots of DS70622 and DS11320, from dry regions, showed drought-avoidance characteristics whereas those of WS99501 and Mèlodie/2, from wet regions, showed the opposite. Discussion. The combination of the germplasm survey and use of GROWSCREEN-Rhizo allowed exploring of adaptive traits and detection of root phenotypic markers for potential drought avoidance. The greater root system depth and root surface coverage, exemplified by DS70622 and DS11320, can now be tested as new sources of drought tolerance. [ABSTRACT FROM AUTHOR]

Volume carving is a well established method for visual hull reconstruction and has been successfully applied in plant phenotyping, especially for 3d reconstruction of small plants and seeds. When imaging larger plants at still relatively high spatial resolution (≤1 mm), well known implementations become slow or have prohibitively large memory needs. Here we present and evaluate a computationally efficient algorithm for volume carving, allowing e.g., 3D reconstruction of plant shoots. It combines a well-known multi-grid representation called "Octree" with an efficient image region integration scheme called "Integral image." Speedup with respect to less efficient octree implementations is about 2 orders of magnitude, due to the introduced refinement strategy "Mark and refine." Speedup is about a factor 1.6 compared to a highly optimized GPU implementation using equidistant voxel grids, even without using any parallelization. We demonstrate the application of this method for trait derivation of banana and maize plants. [ABSTRACT FROM AUTHOR]

The productivity of many important crops is significantly threatened by water shortage, and the elevated atmospheric CO2 can significantly interact with physiological processes and crop responses to drought. We examined the effects of three different CO2 concentrations (historical ~300 ppm, ambient ~400 ppm and elevated ~700 ppm) on physiological traits of oilseed rape ( Brassica napus L.) seedlings subjected to well-watered and reduced water availability. Our data show (1) that, as expected, increasing CO2 level positively modulates leaf photosynthetic traits, leaf water-use efficiency and growth under non-stressed conditions, although a pronounced acclimation of photosynthesis to elevated CO2 occurred; (2) that the predicted elevated CO2 concentration does not reduce total evapotranspiration under drought when compared with present (400 ppm) and historical (300 ppm) concentrations because of a larger leaf area that does not buffer transpiration; and (3) that accordingly, the physiological traits analysed decreased similarly under stress for all CO2 concentrations. Our data support the hypothesis that increasing CO2 concentrations may not significantly counteract the negative effect of increasing drought intensity on Brassica napus performance. [ABSTRACT FROM AUTHOR]