Natural hybridization occurs in sites where Pinus halepensis Mill. and Pinus brutia Ten. overlap geographically. Studies have shown that these hybrids demonstrate vigorous growth. Currently, there is no efficient method for hybrid identification. In the present study, a transcriptome database that was established from P. halepensis and P. brutia provided 111,388 potential SNP markers for hybrid identification. We selected 295 SNPs that were verified in silico using the Integrative Genomics Viewer (IGV) program. Forty-three SNPs were confirmed in vitro using several methods, including CAPS, HRM, TaqMan, KASP, and direct sequencing. Seven SNPs that were used for genotyping P. brutia trees in three planted sites along the climatic gradient of Israel demonstrated hybrid incidences of 2.5, 6.3, and 9.4%. The trunk volume of the hybrids in these sites was 2.9, 2.4, and 1.2 times larger than that of their P. brutia neighboring trees. Differences among the sites in the hybrid vigorous growth extent might have resulted from several factors, including genetic source, stand age, thinning history, and site conditions. However, the overall vigorous growth of the hybrids that was more pronounced in the arid site highlights the potential of interspecific hybridization as a means for improving the drought resistance of forest trees. This work provides a reliable SNP database for the identification of P. brutia x P. halepensis hybrids for the potential utilization of these hybrids to confront changes in climate.

The KNOTTED1-LIKE HOMEOBOX PROTEIN1 (KD1) gene is highly expressed in flower and leaf abscission zones (AZs), and KD1 was reported to regulate tomato flower pedicel abscission via alteration of the auxin gradient and response in the flower AZ (FAZ). The present work was aimed to further examine how KD1 regulates signaling factors and regulatory genes involved in pedicel abscission, by using silenced KD1 lines and performing a large-scale transcriptome profiling of the FAZ before and after flower removal, using a customized AZ-specific microarray. The results highlighted a differential expression of regulatory genes in the FAZ of KD1-silenced plants compared to the wild-type. In the TAPG4::antisense KD1-silenced plants, KD1 gene expression decreased before flower removal, resulting in altered expression of regulatory genes, such as epigenetic modifiers, transcription factors, posttranslational regulators, and antioxidative defense factors occurring at zero time and before affecting auxin levels in the FAZ detected at 4 h after flower removal. The expression of additional regulatory genes was altered in the FAZ of KD1-silenced plants at 4-20 h after flower removal, thereby leading to an inhibited abscission phenotype, and downregulation of genes involved in abscission execution and defense processes. Our data suggest that KD1 is a master regulator of the abscission process, which promotes abscission of tomato flower pedicels. This suggestion is based on the inhibitory effect of KD1 silencing on flower pedicel abscission that operates via alteration of various regulatory pathways, which delay the competence acquisition of the FAZ cells to respond to ethylene signaling.

Tef (Eragrostis tef (Zucc.) Trotter) is an annual small grain, panicle bearing, C-4 cereal crop native to Ethiopia, where it is a major staple food. The objectives of the present study were to characterize the responses of two tef genotypes to escalating phosphorus (P) and potassium (K) levels and to determine an optimum range for P and K at which tef performance is maximized. Two experiments were carried out in the Gilat Research Station, each testing two different genotypes of tef (405B and 406W), one experiment in pots in controlled conditions, and the other in the field. In both experiments, the highest grain yield increased until 6 mg L-1 P, and declined at 12 mg L-1 P. The decline was precipitous and significant in the pot experiment, and gradual and statistically insignificant in the field experiment. In the pots experiment, the grain yield increased until 40 mg L-1 K, with no significant decrease thereafter. The effect of K concentration was only seen in the grain yield and not in the size of the other plant organs. In the field experiment, grain yield was highest at 80 mg L-1 K, but it was not statistically different from 40 mg L-1. The effect of K on growth was only apparent at maturity and not at flowering.

The exposure of plants to pharmaceuticals via treated wastewater irrigation and biosolid application presents an important route of chronic exposure of crops to a wide variety of bioactive pollutants. This paper presents a novel approach which aims to improve our understanding of the interactions of bioactive pollutants with plants through the concept of plant pharmacology and two main sub-divisions: (i) plant pharmacokinetics which describes the fate of exogenous xenobiotics in the plant based on the processes of absorption, distribution, metabolism and accumulation (ADMA), processes that are analogous to pharmacokinetics in animals; and (ii) plant pharmacodynamics that proposes that exogenous xenobiotics interact with plant enzymes and biochemical pathways, establishing a relationship with pharmacological concepts and emphasizing the importance of exposure-response interactions. The concept of plant pharmacology and its two subdivisions provide a foundation for the development of in-depth knowledge regarding the fate of xenobiotics in plants and establishing plant pharmacokinetic-pharmacodynamic models that include both the ADMA processes and time-dependent response of the plant to these compounds. This concept provides a new perspective on pharmacovigilance, focusing on plant-xenobiotic compound interactions, and a conceptual framework for understanding the fate and interactions of these bioactive molecules in agricultural systems, to enable more accurate risks assessments of environmental and human health.

Plants are subjected to fluctuations in light intensity, and this might cause unbalanced photosynthetic electron fluxes and overproduction of reactive oxygen species (ROS). Electrons needed for ROS detoxification are drawn, at least partially, from the cellular glutathione (GSH) pool via the ascorbate-glutathione cycle. Here, we explore the dynamics of the chloroplastic glutathione redox potential (chl-E-GSH) using high-temporal-resolution monitoring of Arabidopsis (Arabidopsis thaliana) lines expressing the reduction-oxidation sensitive green fluorescent protein 2 (roGFP2) in chloroplasts. This was carried out over several days under dynamic environmental conditions and in correlation with PSII operating efficiency. Peaks in chl-E-GSH oxidation during dark-to-light and light-to-dark transitions were observed. Increasing light intensities triggered a binary oxidation response, with a threshold around the light saturating point, suggesting two regulated oxidative states of the chl-E-GSH. These patterns were not affected in npq1 plants, which are impaired in non-photochemical quenching. Oscillations between the two oxidation states were observed under fluctuating light in WT and npq1 plants, but not in pgr5 plants, suggesting a role for PSI photoinhibition in regulating the chl-E-GSH dynamics. Remarkably, pgr5 plants showed an increase in chl-E-GSH oxidation during the nights following light stresses, linking daytime photoinhibition and nighttime GSH metabolism. This work provides a systematic view of the dynamics of the in vivo chloroplastic glutathione redox state during varying light conditions.

Alternative water sources for agriculture are in high demand in a world with diminishing fresh water (FW) availability. Treated wastewater (TWW) offers a reliable alternative, but increasing evidence is pointing to damage to TWW irrigated orchards planted in clay soils related to soil hypoxia. The mechanisms responsible for this hypoxia have not been extensively studied to date. The aim of this study was to elucidate meaningful insights into the mechanisms responsible for the hypoxia in TWW irrigated orchards planted in clay soils using a novel approach whereby parameters describing the soil oxygen and water temporal dynamics are analyzed. To that end, soil oxygen and soil water tension (SWT) measurements from a two year field experiment comparing TWW to FW irrigation in an avocado orchard planted in a clay soil (60 % clay) were used. The deterioration in oxygen levels occurred as the irrigation season progressed, and the oxygen availability decreased with depth (10-35 cm depth). During August-September, when the lowest oxygen concentrations were measured, the water content at which oxygen supply matched oxygen consumption at 35-cm depth did not differ between treatments (similar to 50 mbar), but the TWW irrigated soil experienced similar to 47 % more time at wetter conditions. Lower oxygen decline rates were observed in the TWW irrigated plots which countered the previous concept that TWW leads to increased soil oxygen consumption. The findings point towards the rate of soil drying as the prime cause of differences - TWW irrigated plots dried in a rate which is nearly 4-times smaller than that in FW irrigated plots during the dark and light hours, reflecting slower drainage and water uptake respectively. It is suggested that soil hypoxia induced by the low soil drainage in TWW irrigated clay soils impairs tree water uptake, which further hinders the soil oxygen levels. Based on these results management tools are suggested to allow sustainable irrigation with TWW in the future. Furthermore, the work demonstrates how analysis of parameters describing the oxygen hourly changes can be utilized to gain mechanistic insights unto processes affecting the oxygen regime in the soil.

Floral guides are patterned cues that direct the pollinator to the plant reproductive organs. The spatial distribution of showy visual and olfactory traits allows efficient plant-pollinator interactions. Data on the mechanisms underlying floral volatile patterns or their interactions with pollinators are lacking. Here we characterize the spatial emission patterns of volatiles from the corolla of the model plant Petunia x hybrida and reveal the ability of honeybees to distinguish these patterns. Along the adaxial epidermis, in correlation with cell density, the petal base adjacent to reproductive organs emitted significantly higher levels of volatiles than the distal petal rim. Volatile emission could also be differentiated between the two epidermal surfaces: emission from the adaxial side was significantly higher than that from the abaxial side. Similar emission patterns were also observed in other petunias, Dianthus caryophyllus (carnation) and Argyranthemum frutescens (Marguerite daisy). Analyses of transcripts involved in volatile production/emission revealed lower levels of the plasma-membrane transporter ABCG1 in the abaxial versus adaxial epidermis. Transient overexpression of ABCG1 enhanced emission from the abaxial epidermis to the level of the adaxial epidermis, suggesting its involvement in spatial emission patterns in the epidermal layers. Proboscis extension response experiments showed that differences in emission levels along the adaxial epidermis, that is, petal base versus rim, detected by GC-MS are also discernible by honeybees.

This review deals with modification of montmorillonite and other clay-minerals and clays by interacting them with organic cations, for producing slow release formulations of herbicides, and efficient removal of pollutants from water by filtration. Elaboration is on incorporating initially the organic cations in micelles and liposomes, then producing complexes denoted micelle- or liposome-clay nano-particles. The material characteristics (XRD, Freeze-fracture electron microscopy, adsorption) of the micelle- or liposome-clay complexes are different from those of a complex of the same composition (organo-clay), which is formed by interaction of monomers of the surfactant with the clay-mineral, or clay. The resulting complexes have a large surface area per weight; they include large hydrophobic parts and (in many cases) have excess of a positive charge. The organo-clays formed by preadsorbing organic cations with long alkyl chains were also addressed for adsorption and slow release of herbicides. Another examined approach includes ``adsorptive'' clays modified by small quaternary cations, in which the adsorbed organic cation may open the clay layers, and consequently yield a high exposure of the siloxane surface for adsorption of organic compounds. Small scale and field experiments demonstrated that slow release formulations of herbicides prepared by the new complexes enabled reduced contamination of ground water due to leaching, and exhibited enhanced herbicidal activity. Pollutants removed efficiently from water by the new complexes include (i) hydrophobic and anionic organic molecules, such as herbicides, dissolved organic matter; pharmaceuticals, such as antibiotics and non-steroidal drugs; (ii) inorganic anions, e.g., perchlorate and (iii) microorganisms, such as bacteria, including cyanobacteria (and their toxins). Model calculations of adsorption and kinetics of filtration, and estimation of capacities accompany the survey of results and their discussion.

Journal of Experimental Botany, Volume 71, Issue 21, 2 December 2020, Pages 6830–6843, https://doi.org/10.1093/jxb/eraa258In the original publication of this article, in the penultimate sentence of the legend to Fig 3, there were errors in identifying coloured lines. These should read: “Direct polarization (blue line) samples all the Si atoms in the sample, while 1H-cross-polarization (black line) samples Si atoms in proximity to protons.” instead of “: “Direct polarization (black line) samples all the Si atoms in the sample, while 1H-cross-polarization (blue line) samples Si atoms in proximity to protons.” This error has now been corrected online.

Environmental stresses are among the major factors that limit crop productivity and plant growth. Various nondestructive approaches for monitoring plant stress states have been developed. However, early sensing of the initial biochemical events during stress responses remains a significant challenge. In this work, we established whole-plant redox imaging using potato (Solanum tuberosum) plants expressing a chloroplast-targeted redox-sensitive green fluorescence protein 2 (roGFP2), which reports the glutathione redox potential (E-GSH). Ratiometric imaging analysis demonstrated the probe response to redox perturbations induced by H2O2, DTT, or a GSH biosynthesis inhibitor. We mapped alterations in the chloroplast E-GSH under several stress conditions including, high-light (HL), cold, and drought. An extremely high increase in chloroplast E-GSH was observed under the combination of HL and low temperatures, conditions that specifically induce PSI photoinhibition. Intriguingly, we noted a higher reduced state in newly developed compared with mature leaves under steady-state and stress conditions, suggesting a graded stress sensitivity as part of the plant strategies for coping with stress. The presented observations suggest that whole-plant redox imaging can serve as a powerful tool for the basic understanding of plant stress responses and applied agricultural research, such as toward improving phenotyping capabilities in breeding programs and early detection of stress responses in the field.

Reclaimed wastewater is increasingly used to irrigate agriculture crops. We have previously shown that carbamazepine (CBZ), an anticonvulsant human medication, not entirely discarded during wastewater purification, induces a stress response in tomatoes grown with roots bathing in CBZ-containing water. Induction of stress-related osmoprotectants (sugars, amino acids, proteins) was conspicuous in CBZ-treated seedlings. Here, tomato seedlings were grown in pots watered with increasing concentrations of CBZ. Soluble sugars effectively reacted to CBZ in both leaves and roots. However, the induction of stress-related amino acids and proteins was relevant in roots, but insignificant in leaves. Therefore, roots may be the site where CBZ stress is exerted. Moreover, roots may protect the whole plant from the pharmaceutical. Tomato crops endure biotic stresses, caused by whitefly (Bemisia tabaci) infestation and by the tomato yellow leaf curl begomovirus (TYLCV) they vector. The interplay between CBZ, TYLCV infection and B. tabaci was studied in tomato. Whiteflies preferred CBZ-treated plantlets than control plants, which may be due to increased amount of sugars in leaves. The increased amount of viruliferous whiteflies on CBZ-treated plants is expected to be accompanied by a rise in virus amounts. In fact, CBZ caused a reduction of TYLCV amounts. CBZ-dependent activation of autophagy degradation may explain this decrease in virus amounts. TYLCV infection mitigates the activation of stress markers associated with CBZ treatment. Altogether, CBZ in the water used to irrigate tomatoes grown in pots causes a relatively weak plant stress response, but is definitively sensed by insect and by virus.

Aim Genetic diversity is a major determinant for the capacity of species to persist and adapt to their environments. Unravelling the factors affecting genetic differentiation is crucial to understand how genetic diversity is shaped and species may react to changing environments. We investigated the drivers of genetic differentiation and their interplay with the evolutionary history in a wild pea to test how those may have affected the distribution of genetic diversity. Location Mediterranean basin, western Asia. Taxon Pisum sativum (Fabaceae). Methods We employed restriction site associated DNA (RAD) sequencing of a collection of 81 wild pea samples. The influence of environmental factors on genetic differentiation was tested by estimating the association of these factors and genetic variation using gradient forest analysis. Stairwayplots were used to estimate past effective population sizes and clines of allele frequencies were analysed to detect past expansion patterns. Association of single nucleotide polymorphisms with environmental variables was tested to find signatures of local adaptation. Results The effect of the tested factors influencing genetic differentiation was variable among genetic clusters. Climate predictors were most important in all clusters. Land cover was more important in clusters from areas strongly influenced by human land use. We found statistically significant associations of 3623 SNPs with environmental variables. Most of these SNPs were correlated with latitude followed by temperature. Wild peas went through a genetic bottleneck during the last glacial period followed by population recovery. The detected range expansion patterns suggested an eastward range expansion of the European cluster to Turkey and thereof southwards and eastwards. Main conclusion Drivers of genetic differentiation do not act in a uniform manner within a species. The distribution of genetic variation and the processes affecting this distribution have to be viewed in the context of the hierarchical structure of genetic variation and the environment of its entities to understand how genetic diversity was shaped and may change in the future.

The variability in leaf form in nature is immense. Leaf patterning occurs by differential growth, taking place during a limited window of morphogenetic activity at the leaf marginal meristem. While many regulators have been implicated in the designation of the morphogenetic window and in leaf patterning, how these effectors interact to generate a particular form is still not well understood. We investigated the interaction among different effectors of tomato (Solanum lycopersicum) compound-leaf development, using genetic and molecular analyses. Mutations in the tomato auxin response factor SlARF5/SlMP, which normally promotes leaflet formation, suppressed the increased leaf complexity of mutants with extended morphogenetic window. Impaired activity of the NAC/CUC transcription factor GOBLET (GOB), which specifies leaflet boundaries, also reduced leaf complexity in these backgrounds. Analysis of genetic interactions showed that the patterning factors SlMP, GOB and the MYB transcription factor LYRATE (LYR) coordinately regulate leaf patterning by modulating in parallel different aspects of leaflet formation and shaping. This work places an array of developmental regulators in a morphogenetic context. It reveals how organ-level differentiation rate and local growth are coordinated to sculpture an organ. These concepts are applicable to the coordination of pattering and differentiation in other species and developmental processes.

Abstract Ecological selection is a major driver of community assembly. Selection is classified as stabilizing when species with intermediate trait values gain the highest reproductive success, whereas selection is considered directional when fitness is highest for species with extreme trait values. Previous studies have investigated the effects of different selection types on trait distribution, but the effects of selection on species diversity have remained unclear. Here, we propose a framework for inferring the type and strength of selection by studying species diversity and trait distribution together against null expectations. We use a simulation model to confirm our prediction that directional selection should lead to lower species diversity than stabilizing selection despite a similar effect on trait community-weighted variance. We apply the framework to a mesocosm system of annual plants to test whether differences in species diversity between two habitats that vary in productivity are related to differences in selection on seed mass. We show that, in both habitats, species diversity was lower than the null expectation, but that species diversity was lower in the more productive habitat. We attribute this difference to strong directional selection for large-seeded species in the productive habitat as indicated by trait community-weighted mean being higher and community-weighted variance being lower than the null expectations. In the less productive habitat, we found that community-weighted variance was higher than expected by chance, suggesting that seed mass could be a driver of niche partitioning under such conditions. Altogether, our results suggest that viewing species diversity and trait distribution as interrelated patterns driven by the same process, ecological selection, is helpful in understanding community assembly.

Abstract Grapevine leafroll disease (GLD) is a globally spreading viral infection that causes major economic losses by reducing crop yield, plant longevity, and berry quality, with no effective treatment. Grapevine leafroll associated virus-3 (GLRaV-3) is the most severe, prevalent GLD strain affecting wine production. Here, the ability of RNA interference (RNAi), a non-GMO gene-silencing pathway, to treat GLRaV-3 in infected Cabernet Sauvignon grapevines is evaluated. Lipid-modified polyethylenimine (lmPEI) is synthesized as the carrier for long double-stranded RNA (dsRNA, 250-bp-long) that targets RNA polymerase and coat protein is a gene target that are conserved in the GLRaV-3 genome. Self-assembled dsRNA–lmPEI particles, 220 nm in diameter, display inner ordered domains spaced 7.3 ± 2 nm from one another, correlating to lmPEI wrapping spirally around the dsRNA. The particles effectively protect RNA from degradation by ribonucleases and show to increase uptake rate into plant cells as a result of the lipid component comprising the RNA carrier. In three field experiments, a single dose of foliar sprayed treatment of the RNA-particles knocks down GLRaV-3 titer, and multiple doses of the treatment keep the viral titer at baseline and trigger recovery of the vine and berries. This study demonstrates RNAi as a promising platform for treating viral diseases in agriculture.

Summary Steroidal glycoalkaloids (SGAs) are protective metabolites constitutively produced by Solanaceae species. Genes and enzymes generating the vast structural diversity of SGAs have been largely identified. Yet, mechanisms of hormone pathways coordinating defence (jasmonate; JA) and growth (gibberellin; GA) controlling SGAs metabolism remain unclear. We used tomato to decipher the hormonal regulation of SGAs metabolism during growth vs defence tradeoff. This was performed by genetic and biochemical characterisation of different JA and GA pathways components, coupled with in vitro experiments to elucidate the crosstalk between these hormone pathways mediating SGAs metabolism. We discovered that reduced active JA results in decreased SGA production, while low levels of GA or its receptor led to elevated SGA accumulation. We showed that MYC1 and MYC2 transcription factors mediate the JA/GA crosstalk by transcriptional activation of SGA biosynthesis and GA catabolism genes. Furthermore, MYC1 and MYC2 transcriptionally regulate the GA signalling suppressor DELLA that by itself interferes in JA-mediated SGA control by modulating MYC activity through protein–protein interaction. Chemical and fungal pathogen treatments reinforced the concept of JA/GA crosstalk during SGA metabolism. These findings revealed the mechanism of JA/GA interplay in SGA biosynthesis to balance the cost of chemical defence with growth.

Drought intensity as experienced by plants depends upon soil moisture status and atmospheric variables such as temperature, radiation, and air vapour pressure deficit (VPD). Although the role of shoot architecture with these edaphic and atmospheric factors is well-characterized, the extent to which shoot and root dynamic interactions as a continuum are controlled by genotypic variation is less known. Here, we targeted these interactions using a wild emmer introgression line (IL20) with a distinct drought-induced shift in the shoot-to-root ratio and its drought-sensitive recurrent parent Svevo. Using a gravimetric platform, we show that IL20 maintained higher root water influx and gas exchange under drought stress, which supported a greater growth. Interestingly, the advantage of IL20 in root water influx and transpiration was expressed earlier during the daily diurnal cycle under lower VPD and therefore supported higher transpiration efficiency. Application of structural equation model indicates that under drought, VPD and radiation are antagonistic to transpiration rate, whereas the root water influx operates as feedback for the higher atmospheric responsiveness of leaves. Collectively, our results suggest that a drought-induced shift in root-to-shoot ratio can improve plant water uptake potential in a short preferable time window determined by both water and atmospheric parameters.

Hydrated silica (SiO2·nH2O) aggregates in the root endodermis of grasses. Application of soluble silicates (Si) to roots is associated with variations in the balance of reactive oxygen species (ROS), increased tolerance to a broad range of stresses affecting ROS levels, and early lignin deposition. In sorghum (Sorghum bicolor L.), silica aggregation is patterned in an active silicification zone (ASZ) by a special type of aromatic material with a signature typical to lignin. Since lignin polymerization is mediated by ROS, we studied the formation of root lignin and silica under varied conditions of ROS by modulating hydrogen peroxide (H2O2) concentration in the growth medium. Sorghum seedlings were grown hydroponically and supplemented with Si, H2O2, and KI, a salt that catalyzes H2O2 decomposition. Lignin and silica deposits in the endodermis were studied by histology, scanning electron and Raman microscopies. Cell wall composition was quantified by thermal gravimetric analysis. The endodermal H2O2 concentration correlated to the extent of the aromatic polymer deposition along the root, but did not affect its patterning in spots. Our results show that the ASZ spots were necessary for root silica aggregation, and suggest that silicification is intensified under oxidative stress as a result of increased ASZ lignin-like deposition.

Abstract Polyvinyl alcohol (PVA)-based hydrogels are promising biomaterials for tissue engineering printing applications. However, one of their main disadvantages is their inability to support cell attachment, which is a critical feature for the preparation of biological scaffolds. The goal of this study was to develop a printable, cell-supportive PVA-based bioink with tunable mechanical properties, without using animal-derived polymers which potentially harbor human pathogens. An ultraviolet light (UV) curable PVA-methacrylate (PVA-MA) polymer mixed with Cys-Arg-Gly-Asp (CRGD) peptide was developed. This peptide holds the integrin receptor binding sequence – RGD, that can enhance cell attachment. The additional cysteine was designed to enable its thiol binding under UV to methacrylate groups of the UV curable PVA-MA. Vero cell, as an adherent cell model was used to assess the hydrogel's cell adhesion. It was found that the PVA-MA-CRGD formula enables the preparation of hydrogels with excellent cell attachment and had even shown superior cell attachment properties relative to added gelatin. Adding hyaluronic acid (HA) as a rheologic modulator enabled the printing of this new formula. Our overall data demonstrates the applicability of this mixture as a bioink for soft tissue engineering such as skin, adipose, liver or kidney tissue.

Unrevealing the genetic makeup of crop morpho-agronomic traits is essential for improving yield quality and sustainability. Sesame (Sesamum indicum L.) is one of the oldest oil-crops in the world. Despite its economic and agricultural importance, it is an ‘orphan crop-plant’ that has undergone limited modern selection, and, as a consequence preserved wide genetic diversity. Here we established a new sesame panel (SCHUJI) that contains 184 genotypes representing wide phenotypic variation and is geographically distributed. We harnessed the natural variation of this panel to perform genome-wide association studies for morpho-agronomic traits under the Mediterranean climate conditions.