Targeting editing of tomato SPEECHLESS cis-regulatory regions generates plants with altered stomatal density in response to changing climate conditions.

Flexible developmental programs enable plants to customize their organ size and cellular composition. In leaves of eudicots, the stomatal lineage produces two essential cell types, stomata and pavement cells, but the total numbers and ratio of these cell types can vary. Central to this flexibility is the stomatal lineage initiating transcription factor, SPEECHLESS (SPCH). Here we show, by multiplex CRISPR/Cas9 editing of SlSPCH cis-regulatory sequences in tomato, that we can identify variants with altered stomatal development responses to light and temperature cues. Analysis of tomato leaf development across different conditions, aided by newly-created tools for live-cell imaging and translational reporters of SlSPCH and its paralogues SlMUTE and SlFAMA, revealed the series of cellular events that lead to the environmental change-driven responses in leaf form. Plants bearing the novel SlSPCH variants generated in this study are powerful resources for fundamental and applied studies of tomato resilience in response to climate change.

Phosphate Additives for Aging Inhibition of Impregnated Activated Carbon against Hazardous Gases.

Impregnated activated carbons (IACs) used in air filtration gradually lose their efficacy for the chemisorption of noxious gases when exposed to humidity due to impregnated metal deactivation. In order to stabilize IACs against aging, and to prolong the filters' shelf life, inorganic phosphate compounds (phosphoric acid and its three salts, NaHPO4, Na2HPO4, and Na3PO4) were used as anti-aging additives for two different chromium-free IACs impregnated with copper, zinc, molybdenum, and triethylenediamine (TEDA). Phosphoric acid, monosodium, and disodium phosphate were found to be very efficient in inhibiting the aging of IACs over long periods against cyanogen chloride (the test agent) chemisorption, with the latter being the most efficient. However, the efficiency of phosphate as an anti-aging additive was not well correlated with its ability to inhibit the migration of metal impregnants, especially copper, from the interior to the external surface of carbon granules. Unlike organic additives, the inorganic phosphate additives did not decrease the surface area of the IAC or its physical adsorption capacity for toluene. Using a phosphate additive in IAC used in collective protection and personal filters can improve the safety of the user and the environment and dramatically reduce the need to replace these filters after exposure to humid environments. This has safety, economic, logistical, and environmental advantages.

PlantACT! - how to tackle the climate crisis.

Greenhouse gas (GHG) emissions have created a global climate crisis which requires immediate interventions to mitigate the negative effects on all aspects of life on this planet. As current agriculture and land use contributes up to 25% of total GHG emissions, plant scientists take center stage in finding possible solutions for a transition to sustainable agriculture and land use. In this article, the PlantACT! (Plants for climate ACTion!) initiative of plant scientists lays out a road map of how and in which areas plant scientists can contribute to finding immediate, mid-term, and long-term solutions, and what changes are necessary to implement these solutions at the personal, institutional, and funding levels.

Evolution of polarity protein BASL and the capacity for stomatal lineage asymmetric divisions.

Asymmetric and oriented stem cell divisions enable the continued production of patterned tissues. The molecules that guide these divisions include several "polarity proteins" that are localized to discrete plasma membrane domains, are differentially inherited during asymmetric divisions, and whose scaffolding activities can guide division plane orientation and subsequent cell fates. In the stomatal lineages on the surfaces of plant leaves, asymmetric and oriented divisions create distinct cell types in physiologically optimized patterns. The polarity protein BREAKING OF ASYMMETRY IN THE STOMATAL LINEAGE (BASL) is a major regulator of stomatal lineage division and cell fate asymmetries in Arabidopsis, but its role in the stomatal lineages of other plants is unclear. Here, using phylogenetic and functional assays, we demonstrate that BASL is a eudicot-specific polarity protein. Dicot BASL orthologs can polarize in heterologous systems and rescue the Arabidopsis BASL mutant. The more widely distributed BASL-like proteins, although they share BASL's conserved C-terminal domain, are neither polarized nor do they function in asymmetric divisions of the stomatal lineage. Comparison of BASL protein localization and loss of function BASL phenotypes in Arabidopsis and tomato revealed previously unappreciated differences in how asymmetric cell divisions are employed for pattern formation in different species. This multi-species analysis therefore provides insight into the evolution of a unique polarity regulator and into the developmental choices available to cells as they build and pattern tissues.

Studying the Physical and Chemical Properties of Polydimethylsiloxane Matrix Reinforced by Nanostructured TiO2 Supported on Mesoporous Silica.

In this study, a reactive adsorbent filler was integrated into a polymeric matrix as a novel reactive protective barrier without undermining its mechanical, thermal, and chemical properties. For this purpose, newly synthesized TiO2/MCM/polydimethylsiloxane (PDMS) composites were prepared, and their various properties were thoroughly studied. The filler, TiO2/MCM, is based on a (45 wt%) TiO2 nanoparticle catalyst inside the pores of ordered mesoporous silica, MCM-41, which combines a high adsorption capacity and catalytic capability. This study shows that the incorporation of TiO2/MCM significantly enhances the composite's Young's modulus in terms of tensile strength, as an optimal measurement of 1.6 MPa was obtained, compared with that of 0.8 MPa of pristine PDMS. The composites also showed a higher thermal stability, a reduction in the coefficient of thermal expansion (from 290 to 110 ppm/°C), a 25% reduction in the change in the normalized specific heat capacity, and an increase in the thermal degradation temperatures. The chemical stability in organic environments was improved, as toluene swelling decreased by 40% and the contact angle increased by ~15°. The enhanced properties of the novel synthesized TiO2/MCM/PDMS composite can be used in various applications where a high adsorption capacity and catalytic/photocatalytic activity are required, such as in protective equipment, microfluidic applications, and chemical sensor devices.

Explosive vapour/particles detection using SERS substrates and a hand-held Raman detector.

We developed and optimized surface-enhanced Raman spectrometry (SERS) methods for trace analysis of explosive vapour and particles using a hand-held Raman spectrometer in the field. At first, limits of detection (LODs) using SERS methods based on a colloidal suspension of gold nanoparticles were measured under alkaline conditions and are as follows: pentaerythritol tetranitrate (PETN) (1.5 × 10-6 M, 6.9 ng), 1,3,5,7-tetranitro-1,3,5,7-tetrazoctane (HMX), 8.1 × 10-6 M, 35 ng; urea nitrate (UN), 9.2 × 10-4 M, 165 ng; 2,4,6-trinitrotoluene (TNT), 1.1 × 10-7 M, 0.35 ng. We developed SERS substrates that demonstrate the wide applicability of this technique for use in the field for explosive vapour and particles adsorbed on a surface based on Au nanoparticles that were optimal for the detection of the target materials in solution. Au nanoparticles were modified onto quartz fibres or a polyurethane sponge for vapour/particles detection. SERS detection of vapours of 2,4-dinitrotoluene (2,4-DNT) and 1,3-dinitrobenzene (1,3-DNB) was shown by sampling vapours onto Au-modified quartz fibres followed by hand-held Raman analysis with estimated minimum detection levels of 3.6 ng and 54 ng, respectively. The detection of 2,4-DNT using sponge-based SERS decorated with Au nanoparticles was also demonstrated; however, the sensitivity was lower than that observed using quartz fibres. The detection of TNT on a surface was performed by utilizing quartz-fibres precoated with alumina and modified with Au nanoparticles, and the detection of 10 µg (0.53 µg cm-2) of TNT was demonstrated.

Characterization of the cytokinin sensor TCSv2 in arabidopsis and tomato.

BACKGROUND: Hormones are crucial to plant life and development. Being able to follow the plants hormonal response to various stimuli and throughout developmental processes is an important and increasingly widespread tool. The phytohormone cytokinin (CK) has crucial roles in the regulation of plant growth and development. RESULTS: Here we describe a version of the CK sensor Two Component signaling Sensor (TCS), referred to as TCSv2. TCSv2 has a different arrangement of binding motifs when compared to previous TCS versions, resulting in increased sensitivity in some examined tissues. Here, we examine the CK responsiveness and distribution pattern of TCSv2 in arabidopsis and tomato. CONCLUSIONS: The increased sensitivity and reported expression pattern of TCSv2 make it an ideal TCS version to study CK response in particular hosts, such as tomato, and particular tissues, such as leaves and flowers.

Colloidal-like aggregation of a functional amyloid protein.

Functional amyloid proteins are self-secreted by microbial cells that aggregate into extracellular networks and provide microbial colonies with mechanical stability and resistance to antibiotic treatment. In order to understand the formation mechanism of functional amyloid networks, their aggregation has been studied in vitro under different physical conditions, such as temperature, salt concentration, and pH. Typical aggregates' morphologies include fibers or plaques, the latter resembling amyloid aggregates in neurodegenerated brains. Here, we studied the pH-reduction-induced aggregation of TasA, an extracellular functional amyloid appearing as fibers in biofilms of the soil bacterium, Bacillus subtilis. We used turbidity and zeta potential measurements, electron microscopy, atomic force microscopy, and static light scattering measurements, to characterize the aggregates of TasA and to compare them with colloidal aggregates. We further studied the aggregation of TasA in the presence of negatively charged nanoparticles and showed that nanoparticles co-aggregated with TasA, and that the co-aggregation was hindered sterically. Based on these studies, we concluded that, similarly to colloidal aggregation, TasA aggregation occurs due to surface potential modulations and that the aggregation is followed by a rearrangement process. Shedding light on the aggregation mechanism of TasA, our results can be used for the design of TasA aggregation inhibitors and promoters.

Mutations in the tomato gibberellin receptors suppress xylem proliferation and reduce water loss under water-deficit conditions.

Low gibberellin (GA) activity in tomato (Solanum lycopersicum) inhibits leaf expansion and reduces stomatal conductance. This leads to lower transpiration and improved water status under transient drought conditions. Tomato has three GIBBERELLIN-INSENSITIVE DWARF1 (GID1) GA receptors with overlapping activities and high redundancy. We tested whether mutation in a single GID1 reduces transpiration without affecting growth and productivity. CRISPR-Cas9 gid1 mutants were able to maintain higher leaf water content under water-deficit conditions. Moreover, while gid1a exhibited normal growth, it showed reduced whole-plant transpiration and better recovery from dehydration. Mutation in GID1a inhibited xylem vessel proliferation, which led to lower hydraulic conductance. In stronger GA mutants, we also found reduced xylem vessel expansion. These results suggest that low GA activity affects transpiration by multiple mechanisms: it reduces leaf area, promotes stomatal closure, and reduces xylem proliferation and expansion, and as a result, xylem hydraulic conductance. We further examined if gid1a performs better than the control M82 in the field. Under these conditions, the high redundancy of GID1s was lost and gid1a plants were semi-dwarf, but their productivity was not affected. Although gid1a did not perform better under drought conditions in the field, it exhibited a higher harvest index.

The Tomato DELLA Protein PROCERA Acts in Guard Cells to Promote Stomatal Closure.

Plants employ stomatal closure and reduced growth to avoid water deficiency damage. Reduced levels of the growth-promoting hormone gibberellin (GA) lead to increased tolerance to water deficit, but the underlying mechanism is unknown. Here, we show that the tomato (Solanum lycopersicum) DELLA protein PROCERA (PRO), a negative regulator of GA signaling, acts in guard cells to promote stomatal closure and reduce water loss in response to water deficiency by increasing abscisic acid (ABA) sensitivity. The loss-of-function pro mutant exhibited increased stomatal conductance and rapid wilting under water deficit stress. Transgenic tomato overexpressing constitutively active stable DELLA proteins (S-della) displayed the opposite phenotype. The effects of S-della on stomatal aperture and water loss were strongly suppressed in the ABA-deficient mutant sitiens, indicating that these effects of S-della are ABA dependent. While DELLA had no effect on ABA levels, guard cell ABA responsiveness was increased in S-della and reduced in pro plants compared with the wild type. Expressing S-della under the control of a guard-cell-specific promoter was sufficient to increase stomatal sensitivity to ABA and to reduce water loss under water deficit stress but had no effect on leaf size. This result indicates that DELLA promotes stomatal closure independently of its effect on growth.

Uncovering DELLA-Independent Gibberellin Responses by Characterizing New Tomato procera Mutants.

Gibberellin (GA) regulates plant development primarily by triggering the degradation/deactivation of the DELLA proteins. However, it remains unclear whether all GA responses are regulated by DELLAs. Tomato (Solanum lycopersicum) has a single DELLA gene named PROCERA (PRO), and its recessive pro allele exhibits constitutive GA activity but retains responsiveness to external GA. In the loss-of-function mutant pro(ΔGRAS), all examined GA developmental responses were considerably enhanced relative to pro and a defect in seed desiccation tolerance was uncovered. As pro, but not pro(ΔGRAS), elongation was promoted by GA treatment, pro may retain residual DELLA activity. In agreement with homeostatic feedback regulation of the GA biosynthetic pathway, we found that GA20oxidase1 expression was suppressed in pro(ΔGRAS) and was not affected by exogenous GA3. In contrast, expression of GA2oxidase4 was not affected by the elevated GA signaling in pro(ΔGRAS) but was strongly induced by exogenous GA3. Since a similar response was found in Arabidopsis thaliana plants with impaired activity of all five DELLA genes, we suggest that homeostatic GA responses are regulated by both DELLA-dependent and -independent pathways. Transcriptome analysis of GA-treated pro(ΔGRAS) leaves suggests that 5% of all GA-regulated genes in tomato are DELLA independent.

The effects of aging on the dynamic adsorption of hazardous organic vapors on impregnated activated carbon.

The effects of an eight-year natural aging of ASC impregnated activated carbon on the adsorption capacity and breakthrough times of model organic vapors and of the nerve agent sarin were investigated. Aging delayed methanol breakthrough from dry air on pre-dried carbon, but shortened the breakthrough time of both methanol and hexane under relative humidity (RH) of 30-85% on pre-humidified carbon. Aging also shortened the breakthrough time of the less volatile model compound 2-methoxyethanol, especially under RH of 60-85%. Aging significantly reduced the protection capacity against sarin at RH of 85%. The effects of aging on physisorption are attributed to enhanced hydrogen-bonding capability and strength of the interaction between water and adsorption sites on the carbon surface.

Decontamination of adsorbed chemical warfare agents on activated carbon using hydrogen peroxide solutions.

Mild treatment with hydrogen peroxide solutions (3-30%) efficiently decomposes adsorbed chemical warfare agents (CWAs) on microporous activated carbons used in protective garments and air filters. Better than 95% decomposition of adsorbed sulfur mustard (HD), sarin, and VX was achieved at ambient temperatures within 1-24 h, depending on the H2O2 concentration. HD was oxidized to the nontoxic HD-sulfoxide. The nerve agents were perhydrolyzed to the respective nontoxic methylphosphonic acids. The relative rapidity of the oxidation and perhydrolysis under these conditions is attributed to the microenvironment of the micropores. Apparently, the reactions are favored due to basic sites on the carbon surface. Our findings suggest a potential environmentally friendly route for decontamination of adsorbed CWAs, using H2O2 without the need of cosolvents or activators.

The Arabidopsis gibberellin methyl transferase 1 suppresses gibberellin activity, reduces whole-plant transpiration and promotes drought tolerance in transgenic tomato.

Previous studies have shown that reduced gibberellin (GA) level or signal promotes plant tolerance to environmental stresses, including drought, but the underlying mechanism is not yet clear. Here we studied the effects of reduced levels of active GAs on tomato (Solanum lycopersicum) plant tolerance to drought as well as the mechanism responsible for these effects. To reduce the levels of active GAs, we generated transgenic tomato overexpressing the Arabidopsis thaliana GA METHYL TRANSFERASE 1 (AtGAMT1) gene. AtGAMT1 encodes an enzyme that catalyses the methylation of active GAs to generate inactive GA methyl esters. Tomato plants overexpressing AtGAMT1 exhibited typical GA-deficiency phenotypes and increased tolerance to drought stress. GA application to the transgenic plants restored normal growth and sensitivity to drought. The transgenic plants maintained high leaf water status under drought conditions, because of reduced whole-plant transpiration. The reduced transpiration can be attributed to reduced stomatal conductance. GAMT1 overexpression inhibited the expansion of leaf-epidermal cells, leading to the formation of smaller stomata with reduced stomatal pores. It is possible that under drought conditions, plants with reduced GA activity and therefore, reduced transpiration, will suffer less from leaf desiccation, thereby maintaining higher capabilities and recovery rates.

W/O microemulsions as dendrimer nanocarriers: an EPR study.

A complex system, based on a dendrimer solubilized in the aqueous core of water-in-oil microemulsion, may combine the advantages of both dendrimers and microemulsions to provide better control of drug release. We report for the first time the use of EPR technique to determine the effect of solubilized dendrimer on the structure of the microemulsion. The solubilized poly(propyleneimine) (PPI-G2) interacts with sodium bis(2-ethylhexyl) sulfosuccinate (AOT). EPR analysis provided information on polarity, microviscosity, and molecular order of the systems. Polarity and microviscosity increased from unloaded water-in-oil microemulsion to the system loaded with 0.2 wt % PPI-G2, but remained unchanged with higher PPI-G2 loads. The degree of order also increased with 0.2 wt % PPI-G2 with only minor additional increase with larger quantities (25 wt %) of PPI-G2. Variations in pH only slightly affected the structure of microemulsion in the absence and presence of the loaded dendrimers. Aliphatic oils with longer lipophilic chains enhanced the structural order of the microemulsion. On increasing water content, polarity and degree of order increased. PPI-G2 dendrimer in small loads is attracted by the negatively charged AOT and thus intercalates in the interface of the droplets. Yet, at higher PPI-G2 loads, the excess molecules are solubilized in the water core.

Type and location of interaction between hyperbranched polymers and liposomes. Relevance to design of a potentially advanced drug delivery nanosystem (aDDnS).

Advanced drug delivery nanosystems (aDDnSs) combining liposomal and dendritic materials have only recently appeared in the research field of drug delivery. The nature and localization of the interactions between the components of such systems are not yet fully described. In this study, liposomes are combined with hyperbranched polyesters for the development of new aDDnSs. The polymer-lipid interactions along with their dependence on the polyesters pseudogeneration number and the liposomal lipid composition have been examined. The results indicate that the interaction between the materials takes place in the headgroup region, where H-bonds between the polymers terminal hydroxyls and the phospholipids phosphate moiety are formed. Due to the polymers' compact imperfect structure, which varies with pseudogeneration number, no linear trends are observed with increasing pseudogeneration number. Moreover, it is shown that high percentages of cholesterol in the lipid bilayer affect the penetration of the polymers in the headgroup region.

Solubilization of a dendrimer into a microemulsion.

The present work investigates, for the first time, a system comprising a dendrimer incorporated into the water core of water-in-oil (W/O) microemulsion (ME). A second generation (G-2) poly(propyleneimine) dendrimer (PPI) was solubilized into W/O ME composed of AOT (sodium bis(2-ethylhexyl)sulfosuccinate), heptane, and water. Such a model system possessing the benefits of both dendrimers and ME, can potentially offer superior control of drug administration. The localization of PPI within the system, its specific interactions with the components of the carrier, and its effect on the ME structure was explored by SAXS, DSC, ATR-FTIR, and electrical conductivity measurements. Considerable water binding by PPI, accompanied by partial dehydration of AOT polar heads, was detected by ATR-FTIR and DSC analysis, suggesting that PPI acted as a "water pump". In addition, SAXS measurements showed periodicity increase and disordering of the droplets. Hence, localization of PPI within the core and interfacial regions of the droplets was assumed. Direct electrostatic interactions between PPI and the sulfonate group were not noticed, since the dendrimer molecules were mostly not protonated in the current basic environment at pH 12. However, slight hydrogen bonding between PPI and the S=O groups allowed the dendrimer to behave as a "spacer" between sodium and sulfonate ions. This affected the electrical conductivity behavior of the system, revealing that PPI favored the percolation process. Most likely, PPI decreased the rigidity of the interfacial layer, facilitating the diffusion of sodium ions through the channels. The characterized model system can be advantageously utilized to design specific delivery vehicles, allowing administration of dendrimers as a therapeutic agent from host MEs.

Decomposition of adsorbed VX on activated carbons studied by 31P MAS NMR.

The fate of the persistent OP nerve agent O-ethyl S-[2-(diisopropylamino)ethyl] methylphosphonothioate (VX) on granular activated carbons that are used for gas filtration was studied by means of 31P magic angle spinning (MAS) NMR spectroscopy. VX as vapor or liquid was adsorbed on carbon granules, and MAS NMR spectra were recorded periodically. The results show that at least 90% of the adsorbed VX decomposes within 20 days or less to the nontoxic ethyl methylphosphonic acid (EMPA) and bis(S-2-diisopropylaminoethane) {(DES)2}. Decomposition occurred irrespective of the phase from which VX was loaded, the presence of metal impregnation on the carbon surface, and the water content of the carbon. Theoretical and practical aspects of the degradation are discussed.