Characterizing stay-green in barley across diverse environments: unveiling novel haplotypes.

KEY MESSAGE: There is variation in stay-green within barley breeding germplasm, influenced by multiple haplotypes and environmental conditions. The positive genetic correlation between stay-green and yield across multiple environments highlights the potential as a future breeding target. Barley is considered one of the most naturally resilient crops making it an excellent candidate to dissect the genetics of drought adaptive component traits. Stay-green, is thought to contribute to drought adaptation, in which the photosynthetic machinery is maintained for a longer period post-anthesis increasing the photosynthetic duration of the plant. In other cereal crops, including wheat, stay-green has been linked to increased yield under water-limited conditions. Utilizing a panel of diverse barley breeding lines from a commercial breeding program we aimed to characterize stay-green in four environments across two years. Spatiotemporal modeling was used to accurately model senescence patterns from flowering to maturity characterizing the variation for stay-green in barley for the first time. Environmental effects were identified, and multi-environment trait analysis was performed for stay-green characteristics during grain filling. A consistently positive genetic correlation was found between yield and stay-green. Twenty-two chromosomal regions with large effect haplotypes were identified across and within environment types, with ten being identified in multiple environments. In silico stacking of multiple desirable haplotypes showed an opportunity to improve the stay-green phenotype through targeted breeding. This study is the first of its kind to model barley stay-green in a large breeding panel and has detected novel, stable and environment specific haplotypes. This provides a platform for breeders to develop Australian barley with custom senescence profiles for improved drought adaptation.

Fusarium wilt constrains mungbean yield due to reduction in source availability.

Mungbean is an important source of plant protein for consumers and a high-value export crop for growers across Asia, Australia and Africa. However, many commercial cultivars are highly vulnerable to biotic stresses, which rapidly reduce yield within the season. Fusarium oxysporum is a soil-borne pathogen that is a growing concern for mungbean growers globally. This pathogen causes Fusarium wilt by infecting the root system of the plant resulting in devastating yield reductions. To understand the impact of Fusarium on mungbean development and productivity and to identify tolerant genotypes, a panel of 23 diverse accessions was studied. Field trials conducted in 2016 and 2021 in Warwick, Queensland, Australia under rainfed conditions investigated the variation in phenology, canopy and yield component traits under disease and disease-free conditions. Analyses revealed a high degree of genetic variation for all traits. By comparing the performance of these traits across these two environments, we identified key traits that underpin yield under disease and disease-free conditions. Aboveground biomass components at 50 % flowering were identified as significant drivers of yield development under disease-free conditions and when impacted by Fusarium resulted in up to 96 % yield reduction. Additionally, eight genotypes were identified to be tolerant to Fusarium. These genotypes were found to display differing phenological and morphological behaviours, thereby demonstrating the potential to breed tolerant lines with a range of diverse trait variations. The identification of tolerant genotypes that sustain yield under disease pressure may be exploited in crop improvement programs.

Coherent terahertz laser feedback interferometry for hydration sensing in leaves.

The response of terahertz to the presence of water content makes it an ideal analytical tool for hydration monitoring in agricultural applications. This study reports on the feasibility of terahertz sensing for monitoring the hydration level of freshly harvested leaves of Celtis sinensis by employing a imaging platform based on quantum cascade lasers and laser feedback interferometry. The imaging platform produces wide angle high resolution terahertz amplitude and phase images of the leaves at high frame rates allowing monitoring of dynamic water transport and other changes across the whole leaf. The complementary information in the resulting images was fed to a machine learning model aiming to predict relative water content from a single frame. The model was used to predict the change in hydration level over time. Results of the study suggest that the technique could have substantial potential in agricultural applications.

Characterization of 'QTL-hotspot' introgression lines reveals physiological mechanisms and candidate genes associated with drought adaptation in chickpea.

'QTL-hotspot' is a genomic region on linkage group 04 (CaLG04) in chickpea (Cicer arietinum) that harbours major-effect quantitative trait loci (QTLs) for multiple drought-adaptive traits, and it therefore represents a promising target for improving drought adaptation. To investigate the mechanisms underpinning the positive effects of 'QTL-hotspot' on seed yield under drought, we introgressed this region from the ICC 4958 genotype into five elite chickpea cultivars. The resulting introgression lines (ILs) and their parents were evaluated in multi-location field trials and semi-controlled conditions. The results showed that the 'QTL-hotspot' region improved seed yield under rainfed conditions by increasing seed weight, reducing the time to flowering, regulating traits related to canopy growth and early vigour, and enhancing transpiration efficiency. Whole-genome sequencing data analysis of the ILs and parents revealed four genes underlying the 'QTL-hotspot' region associated with drought adaptation. We validated diagnostic KASP markers closely linked to these genes using the ILs and their parents for future deployment in chickpea breeding programs. The CaTIFY4b-H2 haplotype of a potential candidate gene CaTIFY4b was identified as the superior haplotype for 100-seed weight. The candidate genes and superior haplotypes identified in this study have the potential to serve as direct targets for genetic manipulation and selection for chickpea improvement.

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.

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.

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.

An evaluation of inkjet printed amino acid fingerprint test targets for ninhydrin process monitoring - and some observations.

Ninhydrin was implemented as the primary police method of developing latent fingermarks on paper, cardboard and some other porous surfaces from the late 1960s. Some researchers have used individual amino acids, or mixtures of amino acids, as a method of testing the effectiveness of reagent formulations. It was not however known whether simple mixtures of amino acids could effectively emulate latent fingermarks in reactions with reagents such as ninhydrin. The first part of this study compared the effects of ninhydrin fingermark treatments used internationally in various police laboratories on test targets created by inkjet printing graduated concentrations of a representative mixture of amino acids in a series of blocks on paper. Variations in intensity of development were observed between laboratories which used various formulations and heat and humidity post treatment protocols. In a further trial in 2015 several participants in the International Fingerprint Research Group (IFRG) meeting processed test targets in their own laboratories and submitted them for measurement. Again, variation in developed intensity was observed. The depletion of the activity of ninhydrin solutions during use was also investigated in early evaluations of the test targets. An established fingerprint laboratory then processed a number of samples from a batch of targets to examine batch consistency. This was followed by designing a new test target which enabled comparisons between the developed intensity of printed test target blocks alongside depletion series of split, natural donor fingermarks. A panel of 20 donors provided depletion fingermarks and four ninhydrin formulations and treatment protocols were used. The developed test target blocks were scanned, intensity of development measured, and the results compared with the fingermark development which was evaluated by three assessors using two types of scale. Good correlation between the intensity of the developed test targets and latent fingermark quality and intensity scores was observed with the four ninhydrin treatment protocols, including some which used deliberately downgraded ninhydrin concentrations. This type of evaluation was carried out a second time to investigate modified heat and humidity protocols. The use of such test targets for routine reagent quality control and process verification would appear to be far more accurate and reliable than the use of small numbers of donor fingermarks. It is not clear why the different ninhydrin formulations investigated in the latter part of the work have very different optimum post treatment heating regimes.

Need for speed: manipulating plant growth to accelerate breeding cycles.

To develop more productive and resilient crops that are capable of feeding 10 billion people by 2050, we must accelerate the rate of genetic improvement in plant breeding programs. Speed breeding manipulates the growing environment by regulating light and temperature for the purpose of rapid generation advance. Protocols are now available for a range of short-day and long-day species and the approach is highly compatible with other cutting-edge breeding tools such as genomic selection. Here, we highlight how speed breeding hijacks biological processes for applied plant breeding outcomes and provide a case study examining wheat growth and development under speed breeding conditions. The establishment of speed breeding facilities worldwide is expected to provide benefits for capacity building, discovery research, pre-breeding, and plant breeding to accelerate the development of productive and robust crops.

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.

Limitations to using phloem sap to assess tree water and nutrient status.

Rapid, reliable tools are needed to infer physiological and nutritional health for managing forest systems. Understanding the processes governing tree health is central to the development of these tools. Non-foliar approaches such as the collection of phloem sap reflect processes governing both the use and acquisition of plant water and nutrients at a wide range of temporal (diurnal to seasonal) and spatial (canopy) scales. Despite this, phloem sap is not commonly employed due to an incomplete understanding of transport and post-photosynthetic processes and their effects on chemical concentrations and carbon isotope discrimination. We highlight the need to characterize the influences of storage, remobilization and transport on the concentrations of metabolites to address the time and spatial decoupling of phloem contents to that of environmental stimuli. A conceptual framework is suggested to focus research on key phenomena regarding metabolite transport and highlight significant advantages, misconceptions and limitations to its application.

Chemical and isotopic markers detect water deficit and its influence on nutrient allocation in Phaseolus vulgaris.

The impact of drought on plant growth and yield has been widely studied and is considered a major limitation to crops reaching yield potential. Less known is the impact of water deficit on the nutritional quality of the resulting yield. This study characterised the impact of water deficit on carbon assimilation, modelled water use efficiency from carbon isotope discrimination and analysed the concentration of mineral nutrients, amino acids and sugars in leaf, phloem and pod pools collected from Phaseolus vulgaris L. (common bean) grown in a controlled environment. Water deficit led to an isohydric response, impacting on carbon isotope abundance in all tissues though not translating to any significant treatment differences in water use efficiency or nutrient content in tissues over the course of plant development. The results obtained in this study demonstrate that nutrient content of P. vulgaris yield was not impacted by the availability of water. The absence of significant changes in the nutrient content of individual seeds highlights the plasticity of developing reproductive tissue to changes in whole plant water availability.

Source-Sink Relationships in Crop Plants and Their Influence on Yield Development and Nutritional Quality.

For seed crops, yield is the cumulative result of both source and sink strength for photoassimilates and nutrients over the course of seed development. Source strength for photoassimilates is dictated by both net photosynthetic rate and the rate of photoassimilate remobilisation from source tissues. This review focuses on the current understanding of how the source-sink relationship in crop plants influences rates of yield development and the resilience of yield and nutritional quality. We present the limitations of current approaches to accurately measure sink strength and emphasize differences in coordination between photosynthesis and yield under varying environmental conditions. We highlight the potential to exploit source-sink dynamics, in order to improve yields and emphasize the importance of resilience in yield and nutritional quality with implications for plant breeding strategies.

Carbon Isotope Composition of Carbohydrates and Polyols in Leaf and Phloem Sap of Phaseolus vulgaris L. Influences Predictions of Plant Water Use Efficiency.

The use of carbon isotope abundance (δ(13)C) to assess plant carbon acquisition and water use has significant potential for use in crop management and plant improvement programs. Utilizing Phaseolus vulgaris L. as a model system, this study demonstrates the occurrence and sensitivity of carbon isotope fractionation during the onset of abiotic stresses between leaf and phloem carbon pools. In addition to gas exchange data, compound-specific measures of carbon isotope abundance and concentrations of soluble components of phloem sap were compared with major carbohydrate and sugar alcohol pools in leaf tissue. Differences in both δ(13)C and concentration of metabolites were found in leaf and phloem tissues, the magnitude of which responded to changing environmental conditions. These changes have inplications for the modeling of leaf-level gas exchange based upon δ(13)C natural abundance. Estimates of δ(13)C of low molecular weight carbohydrates and polyols increased the precision of predictions of water use efficiency compared with those based on bulk soluble carbon. The use of this technique requires consideration of the dynamics of the δ(13)C pool under investigation. Understanding the dynamics of changes in δ(13)C during movement and incorporation into heterotrophic tissues is vital for the continued development of tools that provide information on plant physiological performance relating to water use.

Toward designed singlet fission: solution photophysics of two indirectly coupled covalent dimers of 1,3-diphenylisobenzofuran.

In order to identify optimal conditions for singlet fission, we are examining the photophysics of 1,3-diphenylisobenzofuran (1) dimers covalently coupled in various ways. In the two dimers studied presently, the coupling is weak. The subunits are linked via the para position of one of the phenyl substituents, in one case (2) through a CH2 linker and in the other (3) directly, but with methyl substituents in ortho positions forcing a nearly perpendicular twist between the two joint phenyl rings. The measurements are accompanied with density functional theory (DFT) and time-dependent DFT (TD-DFT) calculations. Although in neat solid state, 1 undergoes singlet fission with a rate constant higher than 10(11) s(-1); in nonpolar solutions of 2 and 3, the triplet formation rate constant is less than 10(6) s(-1) and fluorescence is the only significant event following electronic excitation. In polar solvents, fluorescence is weaker because the initial excited singlet state S1 equilibrates by sub-nanosecond charge transfer with a nonemissive dipolar species in which a radical cation of 1 is attached to a radical anion of 1. Most of this charge transfer species decays to S0, and some is converted into triplet T1 with a rate constant near 10(8) s(-1). Experimental uncertainties prevent an accurate determination of the number of T1 excitations that result when a single S1 excitation changes into triplet excitation. It would be one if the charge-transfer species undergoes ordinary intersystem crossing and two if it undergoes the second step of two-step singlet fission. The triplet yield maximizes below room temperature to a value of roughly 9% for 3 and 4% for 2. Above ∼360 K, some of the S1 molecules of 3 are converted into an isomeric charge-transfer species with a shorter lifetime, possibly with a twisted intramolecular charge transfer (TICT) structure. This is not observed in 2.

Recent advances in singlet fission.

A survey is provided of recent progress in the understanding of singlet fission, a spin-allowed process in which a singlet excited molecule shares its energy with a ground-state neighbor to produce two triplet excited molecules. It has been observed to occur in single-crystal, polycrystalline, and amorphous solids, on timescales from 80 fs to 25 ps, producing triplet yields as high as 200%. Photovoltaic devices using the effect have shown external quantum efficiencies in excess of 100%. Almost all the efficient materials are alternant hydrocarbons of the acene series or their simple derivatives, and it is argued that a wider structural variety would be desirable. The current state of the development of molecular structure design rules, based on first-principles theoretical considerations, is described along with initial examples of implementation.

Singlet exciton fission for solar cell applications: energy aspects of interchromophore coupling.

Singlet exciton fission, a process that converts one singlet exciton to a pair of triplet excitons, has the potential to enhance the efficiency of both bulk heterojunction and dye-sensitized solar cells and is understood in crystals but not well understood in molecules. Previous studies have identified promising building blocks for singlet fission in molecular systems, but little work has investigated how these individual chromophores should be combined to maximize triplet yield. We consider the effects of chemically connecting two chromophores to create a coupled chromophore pair and compute how various structural choices alter the thermodynamic and kinetic parameters likely to control singlet fission yield. We use density functional theory to compute the electron transfer matrix element and the thermodynamics of fission for several promising chromophore pairs and find a trade-off between the desire to maximize this element and the desire to keep the singlet fission process exoergic. We identify promising molecular systems for singlet fission and suggest future experiments.

Toward designed singlet fission: electronic states and photophysics of 1,3-diphenylisobenzofuran.

Single crystal molecular structure and solution photophysical properties are reported for 1,3-diphenylisobenzofuran (1), of interest as a model compound in studies of singlet fission. For the ground state of 1 and of its radical cation (1(+*)) and anion (1(-*)), we report the UV-visible absorption spectra, and for neutral 1, also the magnetic circular dichroism (MCD) and the decomposition of the absorption spectrum into purely polarized components, deduced from fluorescence polarization. These results were used to identify a series of singlet excited states. For the first excited singlet and triplet states of 1, the transient visible absorption spectra, S(1) --> S(x) and sensitized T(1) --> T(x), and single exponential lifetimes, tau(F) = approximately 5.3 ns and tau(T) = approximately 200 micros, are reported. The spectra and lifetimes of S(1) --> S(0) fluorescence and sensitized T(1) --> T(x) absorption of 1 were obtained in a series of solvents, as was the fluorescence quantum yield, Phi(F) = 0.95-0.99. No phosphorescence has been detected. The first triplet excitation energy of solid 1 (11,400 cm(-1)) was obtained by electron energy loss spectroscopy, in agreement with previously reported solution values. The fluorescence excitation spectrum suggests an onset of a nonradiative channel at approximately 37,000 cm(-1). Excitation energies and relative transition intensities are in agreement with those of ab initio (CC2) calculations after an empirical 3000 cm(-1) adjustment of the initial state energy to correct differentially for a better quality description of the initial relative to the terminal state of an absorption transition. The interpretation of the MCD spectrum used the semiempirical PPP method, whose results for the S(0) --> S(x) spectrum require no empirical adjustment and are otherwise nearly identical with the CC2 results in all respects including the detailed nature of the electronic excitation. The ground state geometry of 1 was also calculated by the MP2, B3LYP, and CAS methods. The calculations provided a prediction of changes of molecular geometry upon excitation or ionization and permitted an interpretation of the spectra in terms of molecular orbitals involved. Computations suggest that 1 can exist as two nearly isoenergetic conformers of C(2) or C(s) symmetry. Linear dichroism measurements in stretched polyethylene provide evidence for their existence and show that they orient to different degrees, permitting a separation of their spectra in the region of the purely polarized first absorption band. Their excitation energies are nearly identical, but the Franck-Condon envelopes of their first transition differ to a surprising degree.

Crystal structure and the paraelectric-to-ferroelectric phase transition of nanoscale BaTiO3.

We have investigated the paraelectric-to-ferroelectric phase transition of various sizes of nanocrystalline barium titanate (BaTiO3) by using temperature-dependent Raman spectroscopy and powder X-ray diffraction (XRD). Synchrotron X-ray scattering has been used to elucidate the room temperature structures of particles of different sizes by using both Rietveld refinement and pair distribution function (PDF) analysis. We observe the ferroelectric tetragonal phase even for the smallest particles at 26 nm. By using temperature-dependent Raman spectroscopy and XRD, we find that the phase transition is diffuse in temperature for the smaller particles, in contrast to the sharp transition that is found for the bulk sample. However, the actual transition temperature is almost unchanged. Rietveld and PDF analyses suggest increased distortions with decreasing particle size, albeit in conjunction with a tendency to a cubic average structure. These results suggest that although structural distortions are robust to changes in particle size, what is affected is the coherency of the distortions, which is decreased in the smaller particles.