Controlling In Planta Gold Nanoparticle Synthesis and Size for Catalysis.

Gold nanoparticles (Au-NPs) are used as catalysts for a diverse range of industrial applications. Currently, Au-NPs are synthesized chemically, but studies have shown that plants fed Au deposit, this element naturally as NPs within their tissues. The resulting plant material can be used to make biomass-derived catalysts. In vitro studies have shown that the addition of specific, short (∼10 amino acid) peptide/s to solutions can be used to control the NP size and shape, factors that can be used to optimize catalysts for different processes. Introducing these peptides into the model plant species, Arabidopsis thaliana (Arabidopsis), allows us to regulate the diameter of nanoparticles within the plant itself, consequently influencing the catalytic performance in the resulting pyrolyzed biomass. Furthermore, we show that overexpressing the copper and gold COPPER TRANSPORTER 2 (COPT2) in Arabidopsis increases the uptake of these metals. Adding value to the Au-rich biomass offers the potential to make plant-based remediation and stabilization of mine wastes financially feasible. Thus, this study represents a significant step toward engineering plants for the sustainable recovery of finite and valuable elements from our environment.

Cyclic guanosine monophosphate improves salt tolerance in Solanum lycopersicum.

The cyclic nucleotide cyclic guanosine monophosphate (cGMP) is a powerful cell signaling molecule involved in biotic and abiotic stress perception and signal transduction. In the model plant Arabidopsis thaliana, salt and osmotic stress rapidly induce increase in cGMP which plays role by modulating the activity of monovalent cation transporters, possibly by direct binding to these proteins and by altering the expression of many abiotic stress responsive genes. In a recent study, a membrane permeable analogue of cGMP (8-bromo-cGMP) was found to have a promotive effect on soluble sugar, flavonoids and lignin content, and membrane integrity in Solanum lycopersicum seedlings under salt stress. However, it remains to be elucidated how salt stress affects the endogenous cGMP level in S. lycopersicum and if Br-cGMP-induced improvement in salt tolerance in S. lycopersicum involves altered cation fluxes. The current study was conducted to answer these questions. A rapid increase (within 30 s) in endogenous cGMP level was determined in S. lycopersicum roots after treatment with 100 mM NaCl. Addition of membrane permeable Br-cGMP in growth medium remarkably ameliorated the inhibitory effects of NaCl on seedlings' growth parameters, chlorophyll content and net photosynthesis rate. In salt stressed plants, Br-cGMP significantly decreased Na+ content by reducing its influx and increasing efflux while it improved plants K+ content by reducing its efflux and enhancing influx. Furthermore, supplementation with Br-cGMP improved plant's proline content and total antioxidant capacity, resulting in markedly decreased electrolyte leakage under salt stress. Br-cGMP increased the expression of Na+/H+ antiporter genes in roots and shoots of S. lycopersicum growing under salt stress, potentially enhancing plant's ability to sequester Na+ into the vacuole. The findings of this study provide insights into the mechanism of cGMP-induced salt stress tolerance in S. lycopersicum.

Induction of silicon defences in wheat landraces is local, not systemic, and driven by mobilization of soluble silicon to damaged leaves.

In response to herbivory, many grasses, including crops such as wheat, accumulate significant levels of silicon (Si) as an antiherbivore defence. Damage-induced increases in Si can be localized in damaged leaves or be more systemic, but the mechanisms leading to these differences in Si distribution remain untested. Ten genetically diverse wheat landraces (Triticum aestivum) were used to assess genotypic variation in Si induction in response to mechanical damage and how this was affected by exogenous Si supply. Total and soluble Si levels were measured in damaged and undamaged leaves as well as in the phloem to test how Si was allocated to different parts of the plant after damage. Localized, but not systemic, induction of Si defences occurred, and was more pronounced when plants had supplemental Si. Damaged plants had significant increases in Si concentration in their damaged leaves, while the Si concentration in undamaged leaves decreased, such that there was no difference in the average Si concentration of damaged and undamaged plants. The increased Si in damaged leaves was due to the redirection of soluble Si, present in the phloem, from undamaged to damaged plant parts, potentially a more cost-effective defence mechanism for plants than increased Si uptake.

The role of roots and rhizosphere in providing tolerance to toxic metals and metalloids.

Human activity and natural processes have led to the widespread dissemination of metals and metalloids, many of which are toxic and have a negative impact on plant growth and development. Roots, as the first point of contact, are essential in endowing plants with tolerance to excess metal(loid) in the soil. The most important root processes that contribute to tolerance are: adaptation of transport processes that affect uptake efflux and long-distance transport of metal(loid)s; metal(loid) detoxification within root cells via conjugation to thiol rich compounds and subsequent sequestration in the vacuole; plasticity in root architecture; the presence of bacteria and fungi in the rhizosphere that impact on metal(loid) bioavailability; the role of root exudates. In this review, we provide details on these processes and assess their relevance on the detoxification of arsenic, cadmium, mercury and zinc in crops. Furthermore, we assess which of these strategies have been tested in field conditions and whether they are effective in terms of improving crop metal(loid) tolerance.

Shoot potassium content provides a physiological marker to screen cotton genotypes for osmotic and salt tolerance.

Drought and salinity are considered two major abiotic stresses that diminish cotton production worldwide. Studying common morphological and physiological responses in cotton cultivars may help plant biologists to develop and apply standard screening criteria for either of these stresses and for their combination. Therefore, this research aimed to assess the suitability of several physiological parameters as diagnostic to report on osmotic and salinity tolerance in six elite cotton genotypes. Data for relative growth rate (RGR), RGR-reduction, potassium (K+) concentrations in roots, xylem sap and shoots, stomatal conductance (gs) and net photosynthesis rate (Pn) were assessed. Based on RGR and RGR-reduction, we observed an association between osmotic tolerance and salinity tolerance of cotton genotypes. Furthermore, this study found that tolerant cotton genotypes were better able to maintain high RGR, tissue K+, and gas exchange under both hyperosmotic and saline conditions. Shoot K+ levels showed high negative correlations with both osmotic and salinity stress and emerged as a convenient and suitable parameter to assess cotton tolerance to either stress.Novelty statementCotton (Gossypium hirsutum) is a leading fiber crop that is cultivated in more than 52 countries. Much of the land where cotton is grown faces co-occurring drought and salinity abiotic stress which negatively impacts cotton yield and fiber quality. In the present study, cotton genotypes were identified with tolerance to both hyperosmolarity and salinity. Furthermore, we show that shoot potassium content is a diagnostic trait that reports on both osmotic and salinity stress and hence a convenient tool for screening cotton germplasm.

A role for the OsHKT 2;1 sodium transporter in potassium use efficiency in rice.

Increasing the potassium use efficiency (KUE) of crops is important for agricultural sustainability. However, a greater understanding of this complex trait is required to develop new, high-KUE cultivars. To this end, a genome-wide association study (GWAS) was applied to diverse rice (Oryza sativa L.) genotypes grown under potassium-stressed and -replete conditions. Using high-stringency criteria, the genetic architecture of KUE was uncovered, together with the breadth of physiological responses to low-potassium stress. Specifically, three quantitative trait loci (QTLs) were identified, which contained >90 candidate genes. Of these, the sodium transporter gene OsHKT2;1 emerged as a key factor that impacts on KUE based on (i) the correlation between shoot Na+ and KUE, and (ii) higher levels of HKT2;1 expression in high-KUE lines.

Changes in Expression Level of OsHKT1;5 Alters Activity of Membrane Transporters Involved in K+ and Ca2+ Acquisition and Homeostasis in Salinized Rice Roots.

In rice, the OsHKT1;5 gene has been reported to be a critical determinant of salt tolerance. This gene is harbored by the SKC1 locus, and its role was attributed to Na+ unloading from the xylem. No direct evidence, however, was provided in previous studies. Also, the reported function of SKC1 on the loading and delivery of K+ to the shoot remains to be explained. In this work, we used an electrophysiological approach to compare the kinetics of Na+ uptake by root xylem parenchyma cells using wild type (WT) and NIL(SKC1) plants. Our data showed that Na+ reabsorption was observed in WT, but not NIL(SKC1) plants, thus questioning the functional role of HKT1;5 as a transporter operating in the direct Na+ removal from the xylem. Instead, changes in the expression level of HKT1;5 altered the activity of membrane transporters involved in K+ and Ca2+ acquisition and homeostasis in the rice epidermis and stele, explaining the observed phenotype. We conclude that the role of HKT1;5 in plant salinity tolerance cannot be attributed to merely reducing Na+ concentration in the xylem sap but triggers a complex feedback regulation of activities of other transporters involved in the maintenance of plant ionic homeostasis and signaling under stress conditions.

Genome-wide association studies to identify rice salt-tolerance markers.

Salinity is an ever increasing menace that affects agriculture worldwide. Crops such as rice are salt sensitive, but its degree of susceptibility varies widely between cultivars pointing to extensive genetic diversity that can be exploited to identify genes and proteins that are relevant in the response of rice to salt stress. We used a diversity panel of 306 rice accessions and collected phenotypic data after short (6 h), medium (7 d) and long (30 d) salinity treatment (50 mm NaCl). A genome-wide association study (GWAS) was subsequently performed, which identified around 1200 candidate genes from many functional categories, but this was treatment period dependent. Further analysis showed the presence of cation transporters and transcription factors with a known role in salinity tolerance and those that hitherto were not known to be involved in salt stress. Localization analysis of single nucleotide polymorphisms (SNPs) showed the presence of several hundred non-synonymous SNPs (nsSNPs) in coding regions and earmarked specific genomic regions with increased numbers of nsSNPs. It points to components of the ubiquitination pathway as important sources of genetic diversity that could underpin phenotypic variation in stress tolerance.

The Nodulin 26-like intrinsic membrane protein OsNIP3;2 is involved in arsenite uptake by lateral roots in rice.

Previous studies have shown that the Nodulin 26-like intrinsic membrane protein (NIP) Lsi1 (OsNIP2;1) is involved in arsenite [As(III)] uptake in rice (Oryza sativa). However, the role of other rice NIPs in As(III) accumulation in planta remains unknown. In the present study, we investigated the role OsNIP3;2 in As(III) uptake in rice. When expressed in Xenopus laevis oocytes, OsNIP3;2 showed a high transport activity for As(III). Quantitative real-time RT-PCR showed that the expression of OsNIP3;2 was suppressed by 5 µM As(III), but enhanced by 20 and 100 µM As(III). Transgenic rice plants expressing OsNIP3;2pro-GUS showed that the gene was predominantly expressed in the lateral roots and the stele region of the primary roots. Transient expression of OsNIP3;2:GFP fusion protein in rice protoplasts showed that the protein was localized in the plasma membrane. Knockout of OsNIP3;2 significantly decreased As concentration in the roots, but had little effect on shoot As concentration. Synchrotron microfocus X-ray fluorescence showed decreased As accumulation in the stele of the lateral roots in the mutants compared with wild-type. Our results indicate that OsNIP3;2 is involved in As(III) uptake by lateral roots, but its contribution to As accumulation in the shoots is limited.

Overexpression of the potassium channel TPKb in small vacuoles confers osmotic and drought tolerance to rice.

Potassium (K(+) ) is the most important cationic nutrient for all living organisms. Vacuolar two-pore K(+) (TPK) channels are important players in the regulation of cellular levels of K(+) but have not been characterised in rice. In order to assess the role of OsTPKb, a K(+) selective ion channel predominantly expressed in the tonoplast of small vacuoles, we generated overexpressing (OX) lines using a constitutive promoter and compared their phenotypes with control plants. Relative to control plants, OX lines showed better growth when exposed to low-K(+) or water stress conditions. K(+) uptake was greater in OX lines which may be driven by increased AKT1 and HAK1 activity. The enhanced K(+) uptake led to tissue K(+) levels that were raised in roots and shoots. Furthermore, energy dispersive X-ray (EDX) analyses showed a higher cytoplasm: vacuole K(+) ratio which is likely to contribute to the increased stress tolerance. In all, the data suggest that TPKb can alter the K(+) status of small vacuoles, which is important for general cellular K(+) homeostasis which, in turn, affects stress tolerance.

Overexpression of the rice AKT1 potassium channel affects potassium nutrition and rice drought tolerance.

Potassium (K(+)) is the most important cationic nutrient for all living organisms and has roles in most aspects of plant physiology. To assess the impact of one of the main K(+) uptake components, the K(+) inward rectifying channel AKT1, we characterized both loss of function and overexpression of OsAKT1 in rice. In many conditions, AKT1 expression correlated with K(+) uptake and tissue K(+) levels. No salinity-related growth phenotype was observed for either loss or gain of function mutants. However, a correlation between AKT1 expression and root Na(+) when the external Na/K ratio was high suggests that there may be a role for AKT1 in Na(+) uptake in such conditions. In contrast to findings with Arabidopsis thaliana, we did not detect any change in growth of AKT1 loss of function mutants in the presence of NH4 (+) Nevertheless, NH4 (+)-dependent inhibition was detected during K(+) uptake assays in loss of function and wild type plants, depending on pre-growth conditions. The most prominent result of OsAKT1 overexpression was a reduction in sensitivity to osmotic/drought stress in transgenic plants: the data suggest that AKT1 overexpression improved rice osmotic and drought stress tolerance by increasing tissue levels of K(+), especially in the root.

Plasma membrane cyclic nucleotide gated calcium channels control land plant thermal sensing and acquired thermotolerance.

Typically at dawn on a hot summer day, land plants need precise molecular thermometers to sense harmless increments in the ambient temperature to induce a timely heat shock response (HSR) and accumulate protective heat shock proteins in anticipation of harmful temperatures at mid-day. Here, we found that the cyclic nucleotide gated calcium channel (CNGC) CNGCb gene from Physcomitrella patens and its Arabidopsis thaliana ortholog CNGC2, encode a component of cyclic nucleotide gated Ca(2+) channels that act as the primary thermosensors of land plant cells. Disruption of CNGCb or CNGC2 produced a hyper-thermosensitive phenotype, giving rise to an HSR and acquired thermotolerance at significantly milder heat-priming treatments than in wild-type plants. In an aequorin-expressing moss, CNGCb loss-of-function caused a hyper-thermoresponsive Ca(2+) influx and altered Ca(2+) signaling. Patch clamp recordings on moss protoplasts showed the presence of three distinct thermoresponsive Ca(2+) channels in wild-type cells. Deletion of CNGCb led to a total absence of one and increased the open probability of the remaining two thermoresponsive Ca(2+) channels. Thus, CNGC2 and CNGCb are expected to form heteromeric Ca(2+) channels with other related CNGCs. These channels in the plasma membrane respond to increments in the ambient temperature by triggering an optimal HSR, leading to the onset of plant acquired thermotolerance.

Sodium in plants: perception, signalling, and regulation of sodium fluxes.

Although not essential for most plants, sodium (Na(+)) can be beneficial to plants in many conditions, particularly when potassium (K(+)) is deficient. As such it can be regarded a 'non-essential' or 'functional' nutrient. By contrast, the many salinized areas around the globe force plants to deal with toxicity from high levels of Na(+) in the environment and within tissues. Progress has been made in identifying the relevant membrane transporters involved in the uptake and distribution of Na(+). The latter is important in the context of mitigating salinity stress but also for the optimization of Na(+) as an abundantly available functional nutrient. In both cases plants are likely to require mechanism(s) to monitor Na(+) concentration, possibly in multiple compartments, to regulate gene expression and transport activities. Extremely little is known about whether such mechanisms are present and if so, how they operate, either at the cellular or the tissue level. This paper gives an overview of the regulatory and potential sensing mechanisms that pertain to Na(+), in both the context of salt stress and Na(+) as a nutrient.

Rice two-pore K+ channels are expressed in different types of vacuoles.

Potassium (K+) is a major nutrient for plant growth and development. Vacuolar K+ ion channels of the two-pore K+ (TPK) family play an important role in maintaining K+ homeostasis. Several TPK channels were previously shown to be expressed in the lytic vacuole (LV) tonoplast. Plants also contain smaller protein storage vacuoles (PSVs) that contain membrane transporters. However, the mechanisms that define how membrane proteins reach different vacuolar destinations are largely unknown. The Oryza sativa genome encodes two TPK isoforms (TPKa and TPKb) that have very similar sequences and are ubiquitously expressed. The electrophysiological properties of both TPKs were comparable, showing inward rectification and voltage independence. In spite of high levels of similarity in sequence and transport properties, the cellular localization of TPKa and TPKb channels was different, with TPKa localization predominantly at the large LV and TPKb primarily in smaller PSV-type compartments. Trafficking of TPKa was sensitive to brefeldin A, while that of TPKb was not. The use of TPKa:TPKb chimeras showed that C-terminal domains are crucial for the differential targeting of TPKa and TPKb. Site-directed mutagenesis of C-terminal residues that were different between TPKa and TPKb identified three amino acids that are important in determining ultimate vacuolar destination.

Arbuscular mycorrhizal fungi native from a Mediterranean saline area enhance maize tolerance to salinity through improved ion homeostasis.

Soil salinity restricts plant growth and productivity. Na(+) represents the major ion causing toxicity because it competes with K(+) for binding sites at the plasma membrane. Inoculation with arbuscular mycorrhizal fungi (AMF) can alleviate salt stress in the host plant through several mechanisms. These may include ion selection during the fungal uptake of nutrients from the soil or during transfer to the host plant. AM benefits could be enhanced when native AMF isolates are used. Thus, we investigated whether native AMF isolated from an area with problems of salinity and desertification can help maize plants to overcome the negative effects of salinity stress better than non-AM plants or plants inoculated with non-native AMF. Results showed that plants inoculated with two out the three native AMF had the highest shoot dry biomass at all salinity levels. Plants inoculated with the three native AMF showed significant increase of K(+) and reduced Na(+) accumulation as compared to non-mycorrhizal plants, concomitantly with higher K(+) /Na(+) ratios in their tissues. For the first time, these effects have been correlated with regulation of ZmAKT2, ZmSOS1 and ZmSKOR genes expression in the roots of maize, contributing to K(+) and Na(+) homeostasis in plants colonized by native AMF.

Exploring the potential: Can arsenic (As) resistant silicate-solubilizing bacteria manage the dual effects of silicon on As accumulation in rice?

Rice (Oryza sativa L.) cultivation in regions marked by elevated arsenic (As) concentrations poses significant health concerns due to As uptake by the plant and its subsequent entry into the human food chain. With rice serving as a staple crop for a substantial share of the global population, addressing this issue is critical for food security. In flooded paddy soils, where As availability is pronounced, innovative strategies to reduce As uptake and enhance agricultural sustainability are mandatory. Silicon (Si) and Si nanoparticles have emerged as potential candidates to mitigate As accumulation in rice. However, their effects on As uptake exhibit complexity, influenced by initial Si levels in the soil and the amount of Si introduced through fertilization. While low Si additions may inadvertently increase As uptake, higher Si concentrations may alleviate As uptake and toxicity. The interplay among existing Si and As availability, Si supplementation, and soil biogeochemistry collectively shapes the outcome. Adding water-soluble Si fertilizers (e.g., Na2SiO3 and K2SiO3) has demonstrated efficacy in mitigating As toxicity stress in rice. Nonetheless, the expense associated with these fertilizers underscores the necessity for low cost innovative solutions. Silicate-solubilizing bacteria (SSB) resilient to As hold promise by enhancing Si availability by accelerating mineral dissolution within the rhizosphere, thereby regulating the Si biogeochemical cycle in paddy soils. Promoting SSB could make cost-effective Si sources more soluble and, consequently, managing the intricate interplay of Si's dual effects on As accumulation in rice. This review paper offers a comprehensive exploration of Si's nuanced role in modulating As uptake by rice, emphasizing the potential synergy between As-resistant SSB and Si availability enhancement. By shedding light on this interplay, we aspire to shed light on an innovative attempt for reducing As accumulation in rice while advancing agricultural sustainability.

Development of a phenotyping protocol for combined drought and salinity stress at seedling stage in rice.

Introduction: The case of combined drought and salinity stress is increasingly becoming a constraint to rice production, especially in coastal areas and river deltas where low rainfall not only reduces soil moisture levels but also reduces the flow of river water, resulting in intrusion of saline sea-water. A standardized screening method is needed in order to systematically evaluate rice cultivars under combined drought+salinity at the same time because sequential stress of salinity followed by drought or vice-versa is not similar to simultaneous stress effects. Therefore, we aimed to develop a screening protocol for combined drought+salinity stress applied to soil-grown plants at seedling stage. Methods: The study system used 30-L soil-filled boxes, which allowed a comparison of plant growth under control conditions, individual drought and salinity stress, as well as combined drought+salinity. A set of salinity tolerant and drought tolerant cultivars were tested, together with several popular but salinity and drought-susceptible varieties that are grown in regions prone to combined drought+salinity. A range of treatments were tested including different timings of the drought and salinity application, and different severities of stress, in order to determine the most effective that resulted in visible distinction among cultivars. The challenges related to determining a protocol with repeatable seedling stage stress treatment effects while achieving a uniform plant stand are described here. Results: The optimized protocol simultaneously applied both stresses by planting into saline soil at 75% of field capacity which was then allowed to undergo progressive drydown. Meanwhile, physiological characterization revealed that chlorophyll fluorescence at seedling stage correlated well with grain yield when drought stress was applied to vegetative stage only. Discussion: The drought+salinity protocol developed here can be used for screening rice breeding populations as part of a pipeline to develop new rice varieties with improved adaptation to combined stresses.

Measurement of cellular cGMP in plant cells and tissues using the endogenous fluorescent reporter FlincG.

The cyclic nucleotide cGMP has been shown to play important roles in plant development and responses to abiotic and biotic stress. To date, the techniques that are available to measure cGMP in plants are limited by low spatial and temporal resolution. In addition, tissue destruction is necessary. To circumvent these drawbacks we have used the δ-FlincG fluorescent protein to create an endogenous cGMP sensor that can report cellular cGMP levels with high resolution in time and space in living plant cells. δ-FlincG in transient and stably expressing cells shows a dissociation constant for cGMP of around 200 nm giving it a dynamic range of around 20-2000 nm. Stimuli that were previously shown to alter cGMP in plant cells (nitric oxide and gibberrellic acid) evoked pronounced fluorescence signals in single cells and in root tissues, providing evidence that δ-FlincG reports changes in cellular cGMP in a physiologically relevant context.

Over-expression of an Na+-and K+-permeable HKT transporter in barley improves salt tolerance.

Soil salinity is an increasing menace that affects agriculture across the globe. Plant adaptation to high salt concentrations involves integrated functions, including control of Na+ uptake, translocation and compartmentalization. Na+ transporters belonging to the HKT family have been shown to be involved in tolerance to mild salt stress in glycophytes such as Arabidopsis, wheat and rice by contributing to Na+ exclusion from aerial tissues. Here, we have analysed the role of the HKT transporter HKT2;1, which is permeable to K+ and Na+, in barley, a relatively salt-tolerant crop that displays a salt-including behaviour. In Xenopus oocytes, HvHKT2;1 co-transports Na+ and K+ over a large range of concentrations, displaying low affinity for Na+, variable affinity for K+ depending on external Na+ concentration, and inhibition by K+ (K(i) approximately 5 mm). HvHKT2;1 is predominantly expressed in the root cortex. Transcript levels are up-regulated in both roots and shoots by low K+ growth conditions, and in shoots by high Na+ growth conditions. Over-expression of HvHKT2;1 led to enhanced Na+ uptake, higher Na+ concentrations in the xylem sap, and enhanced translocation of Na+ to leaves when plants were grown in the presence of 50 or 100 mm NaCl. Interestingly, these responses were correlated with increased barley salt tolerance. This suggests that one of the factors that limits barley salt tolerance is the capacity to translocate Na+ to the shoot rather than accumulation or compartmentalization of this cation in leaf tissues. Thus, over-expression of HvHKT2;1 leads to increased salt tolerance by reinforcing the salt-including behaviour of barley.