Absence Of Salt Research Articles

Synergistic Effect of Plant-Growth-Promoting Rhizobacteria Improves Strawberry Growth and Flowering with Soil Salinization and Increased Atmospheric CO2 Levels and Temperature Conditions

Biofertilization with plant-growth-promoting rhizobacteria (PGPR) can positively affect the growth and health of host plants and reinforce their tolerance of stressors. Here, we investigate the use of isolated PGPR consortia from halophytes to improve strawberry growth and flowering performance under saline and elevated CO2 and temperature conditions. Growth, flower bud production, and the photosynthetic apparatus response were determined in strawberry plants grown at 0 and 85 mmol L−1 NaCl and in two atmospheric CO2-temperature combinations (400/700 ppm and 25/+4 °C, respectively). Biofertilization improved strawberry plant growth and flower bud production, independently of salinity conditions, at ambient CO2 and 25 °C, while bacterial inoculation only had a positive effect on plant growth in the presence of salt in high CO2 and at +4 °C. Biofertilizers 1 and 3 generated the largest biomass of strawberries at 400 ppm CO2 and 0 and 85 mmol L−1 NaCl, respectively, while biofertilizer 1 did so in the presence of salt and in an atmosphere enriched with CO2 and at +4 °C. The effect of the consortia was mediated by bacterial strain PGP properties, rather than by an improvement in the photosynthetic rate of the plants. Furthermore, biofertilizers 1 and 2 increased the number of flower buds in the absence of salt, while biofertilizers 3 and 4 did so for salt-inoculated plants at 400 ppm CO2 and at 25 °C. There was no effect of inoculation on flower bud production of plants grown at high CO2 and at +4 °C. Finally, we concluded that the effect of bacterial inoculation on strawberry growth and flowering depended on the type of bacterial strain and growth conditions. This highlights the importance of developing studies considering stress interaction to assess the real potential of biofertilizers.

Detection of Biosignatures by Capillary Electrophoresis Mass Spectrometry in the Presence of Salts Relevant to Ocean Worlds Missions.

Capillary electrophoresis (CE) is a promising liquid-based technique for in situ chemical analysis on ocean worlds that allows the detection of a wide range of organic molecules relevant to the search for life. CE coupled with mass spectrometry (MS) is particularly valuable as it also enables the discovery of unknown compounds. Here we demonstrate that CE coupled to MS via electrospray ionization (ESI) can readily analyze samples containing up to half the saturation levels of salts relevant to ocean worlds when using 5 M acetic acid as the separation media. A mixture containing amino acids, peptides, nucleobases, and nucleosides was analyzed in the presence of two salts, NaCl and MgSO4, based on their relevance to Europa and Enceladus. We demonstrate here CE-MS limits of detection for these organics ranging from 0.05 to 1 μM (8 to 89 ppb) in the absence of salts. More importantly, we demonstrate here for the first time that organics in the low micromolar range (1-50 μM) are detected by CE-MS in the presence of 3 M NaCl without desalting, preconcentration, or derivatization. This demonstration highlights how CE-MS is uniquely suited for organic analysis on future missions to ocean worlds.

Heteroaggregation between particles modified by polyelectrolyte multilayers

The heteroaggregation of particles is interesting in many fields and particularly in the field of ceramic shaping. In general, heteroaggregation is obtained by mixing powders with opposite surface charges, which may require in some cases surface modifications of the particles. Here we propose to study the behavior of suspensions composed of a mixture of particles with positive and negative charges, created by the adsorption of polyelectrolyte multilayers on their surface. Particular attention is paid to the effect of the number of layers of adsorbed polyelectrolytes and of the ionic strength. This study shows that these surface modifications allow, whatever the number of layers, to have heteroaggregation in the absence of salt. Moreover, depending on the number of adsorbed layers, the addition of salt has a different effect on the suspension behavior. These phenomena can be explained by screening effects already mentioned in the literature for unmixed suspensions of particles modified by polyelectrolyte multilayers. As a consequence, this study highlights that the study of heteroaggregation in these systems is a very simple way to analyze this effect of screening which is difficult to observe on non-mixed suspensions.

From agricultural wastes to a resource: Kiwi Peels, as long-lasting, recyclable adsorbent, to remove emerging pollutants from water. The case of Ciprofloxacin removal

This work accounts for the first example of dry Kiwi Peels to remove emerging pollutants from water. Both the inner and outer sides of Kiwi Peels were characterized by using in synergy FTIR-ATR, TG, and SEM analyses before and after its use and re-use, proposing it as long-lasting, recyclable adsorbent material. Among the tested 14 emerging pollutants, 6 of them were successfully removed by Kiwi Peels, also if present in mixtures, calculating the Kiwi Peels maximum adsorption capacities, occurring in the range of 1–4 mg/g. To infer information about the behavior of Kiwi Peels during water treatments, Ciprofloxacin, a well-known and largely used antibiotic, was selected as a dangerous model contaminant. The roles of ionic strength, pH values, adsorbent/pollutant amounts, and temperature values during the adsorption process were assessed, giving physical and chemical information about the whole adsorption process. The thermodynamics, the adsorption isotherms, and kinetics were studied. The Freundlich model, with a good correlation, well described the obtained results, indicating the heterogenous character of the Ciprofloxacin adsorption with the formation of a multilayer of pollutant molecules onto the adsorbent surface. If, on the one hand, in the range of temperatures 283–303 K, the Ciprofloxacin adsorption was favored by increasing the temperature, on the other hand, the adsorption was hindered by further incrementing of temperature due to the pollutant desorption. Until the occurrence of desorption, the process occurred with ΔH° and ΔS° > 0 with a ΔG° < 0. Both the pseudo-first and pseudo-second-order kinetic equations seemed to describe the process with the applicability of the Weber-Morris model. The results suggested the main presence of electrostatic interaction between the pollutant and adsorbent by changing the pH values and ionic strength of Ciprofloxacin solutions. As a whole, from the obtained results, the best condition to remove Ciprofloxacin is neutral pH in the absence of salt at 303 K. The Kiwi Peels and Ciprofloxacin recycling was also investigated by performing desorption experiments using 0.1 M MgCl2 solution, increasing the Kiwi Peels maximum adsorption capacity at least from 4 to 40 mg/g for Ciprofloxacin. At least 10 cycles of adsorption/desorption were performed, desorbing almost 75% of adsorbed pollutants during each run. Preliminary information about the possibility of inducing the solid-state pollutant photodegradation by using Advanced Oxidation Processes was also explored, giving a possible alternative for pollutant desorption and adsorbent recycling.

Transcriptional Analyses of Genes Related to Fodder Qualities in Giant Leucaena Under Different Stress Environments.

Leucaena leucocephala subsp. glabrata (giant leucaena) is a tree legume, whose foliage is used as a fodder for animals because of its high protein content. In spite of being a highly nutritious fodder, giant leucaena foliage has two undesirable secondary metabolites, mimosine and tannin. The amounts of mimosine and tannin in giant leucaena foliage are known to vary under different environmental conditions. Giant leucaena was grown under different salinity, pH and nitrogen availability conditions. It produced the highest amounts of mimosine at pH 6.0–7.0, whereas, variation in soil pH did not affect tannin concentrations. Salinity stress had negative effects on both mimosine and tannin concentrations, while nitrogen abundance promoted both mimosine and tannin production. Seven genes for mimosine and tannin metabolism were isolated from a transcriptome library of giant leucaena. These were mimosine synthase, mimosinase, chalcone synthase, flavanone 3β-hydroxylase, dihydroflavonol reductase, leucoanthocyanidin reductase, and anthocyanidin reductase. The highest level of mimosine synthase activity was observed in the absence of salt in the soils. Mimosine synthase activities had strong positive correlation with mimosine concentrations in the foliage (R2 = 0.78) whereas mimosinase expression did not appear to have a direct relationship with salt concentrations. The expression of mimosine synthase was significantly higher in the leucaena foliage under nitrogen abundant condition than in nitrogen deficiency conditions, while mimosinase expression was significantly higher under nitrogen deficiency condition than in nitrogen abundance conditions. Mimosine concentrations in the foliage were positively correlated with the expression levels of mimosine synthase but not mimosinase. Similarly, the concentrations of tannin were positively correlated with expression levels of dihydroflavonol reductase, leucoanthocyanidin reductase, and anthocyanidin reductase. Understanding of the environmental conditions that promote or inhibit transcription of the genes for mimosine and tannin biosynthesis should help to design environmental conditions that inhibit transcription of these genes, resulting in reduced levels of these compounds in the leucaena foliage.

Temperature dependent single-chain structure of poly[3-(acrylamidopropyl-dimethyl-ammonium) propyl-1-sulfonate] via small-angle neutron scattering.

Responsive polyzwitterionic materials have become important for a range of applications such as environmental remediation and targeted drug delivery. Much is known about the macroscopic phase-behaviors of such materials, but how the smaller scale single-chain structures of polyzwitterions respond to external stimuli is not well understood, especially at temperatures close to their phase boundaries. Such chain conformation responses are important in directing larger-scale associative properties. Here, we study the temperature dependent single-chain structure of a model polysulfobetaine, poly[3-(acrylamidopropyl-dimethyl-ammonium) propyl-1-sulfonate], using small angle neutron scattering. In the absence of salt, we find that temperature has a large effect on solvent quality with a decreasing trend from good solvent conditions at 50 °C to poor solvent at 10 °C (a temperature just above the cloud point of 7.6 °C) and an estimated theta temperature of 39 °C. When 100 mM NaCl is present, the solvent quality is good with weak temperature dependence. Without salt present, the polymer chain appears to have a nearly Gaussian coil conformation and the backbone becomes slightly more rigid as the temperature is lowered to the cloud point as determined by the Debye-local rod model on a Kratky plot. The addition of salt has a notable effect on the intra-chain correlations where an increase in chain dimensions to a swollen coil conformation and an increase in chain rigidity is observed at 100 mM NaCl in D2O, however, with a negligible temperature dependence.

Assembly of aluminum oxide particles with lignin-acrylic acid polymer in saline systems

The flocculation process is widely used for separating particles from suspension systems. To make this process more environmentally friendly, a biobased flocculant was proposed to be used in this work. This study involves the experimental evaluation of a flocculation system of aluminum oxide particles (with two different sizes) and a kraft lignin-acrylic acid (KL-AA) polymer, as a biobased-flocculant, with advanced tools, such as focused beam reflectance measurement, vertical scanning analyzer, and gravitational scanning analyzer. A statistical framework was adapted to model the experimental results based on the mixture distribution theory, which led to a more in-depth analysis of the shape and size of flocs in the heterogeneous suspension system. The results revealed that the flocculation process of aluminum oxide suspension was altered by the salinity of the medium as well as the size of aluminum oxide particles. It was found that the flocs formed with KL-AA at a higher NaCl concentration had smaller sizes and exhibited higher shear resistance than the flocs formed in the absence of salt. Furthermore, by increasing the concentration of NaCl, the particles tended to possess more spherical shapes. The results also showed that the fractal dimension for flocculated particles increased by concentrating NaCl in the suspensions. Under gravity and centrifugal forces, the impact of KL-AA in flocculating particles was insignificant in the presence of salt.

Effect of biphosphate salt on dipalmitoylphosphatidylcholine bilayer deformation by Tat polypeptide.

Translocation of positively charged cell penetrating peptides (CPP) through cell membrane is important in drug delivery. Here we report all-atom molecular dynamics simulations to investigate how a biphosphate salt in a solvent affects the interaction of a CPP, HIV-1 Tat peptide with model dipalmitoylphosphatidylcholine (DPPC) lipid bilayer. Tat peptide has a large number of basic arginines and a couple of polar glutamines. We observe that in absence of salt, the basic residues of the polypeptide get localized in the vicinity of the membrane without altering the bilayer properties much; polypeptide induce local thinning of the bilayer membrane at the area of localization. In presence of biphosphate salt, the basic residues, dressed by the biphosphate ions, are repelled by the phosphate head groups of the lipid molecules. However, polar glutamine prefers to stay in the vicinity of the bilayer. This leads to larger local bilayer thickness at the contact point by the polar residue and non-uniform bilayer thickness profile. The thickness deformation of bilayer structure disappears upon mutating the polar residue, suggesting importance of the polar residue in bilayer deformation. Our studies point to control bilayer deformation by appropriate peptide sequence and solvent conditions.

Exogenous application of gibberellic acid and silicon to promote salinity tolerance in pea (Pisum sativum L.) through Na+ exclusion

Salinity is a worldwide problem limiting the plant growth and risking food security. This study was conducted to examine exogenous application of silicon (Si), gibberellic acid (GA3) upon the ion transport, growth, yield, and antioxidant enzymes activities of pea plant in saline conditions. Two pea varieties Meteor-FSD and Samrina Zard were pre-treated with GA3 (10-4 M) for 12 h. Plants were allowed to grow with or without silicon in washed silica sand. Ten days old seedlings were shifted in pots with 10 kg soil. Twenty-five days old plants were exposed to 0 and 5 dS m−1 sodium stress. Results showed that exogenous application of GA3 + Si was the best treatment for increasing plant biomass and yield in the presence and absence of NaCl. Furthermore, application of Si or GA3 enhanced chlorophyll content in the leaves, thereby increasing the net assimilation rate of pea varieties under NaCl stress by increasing the antioxidant enzyme activity. Treatment of Si alone or in combination with GA3 significantly reduced Na+ movement in both pea varieties. Results showed that Si has more prominent role than GA3 alone to build-up high plant biomass, yield, soluble protein content and reduction of Na+ transport. Samrina Zard variety showed higher yield, shoot and root dry weight as compared to Meteor-FSD variety in presence and absence of salt. It was concluded that Si can be used as a nutrient for pea under saline or non-saline conditions. Moreover, application of GA3 has a potential role for increasing salinity tolerance, mostly in sensitive pea varieties.

Effects of Chemical Priming on the Germination of the Ornamental Halophyte Lobularia maritima under NaCl Salinity

Lobularia maritima or sweet alyssum (Brassicaceae) is an annual littoral halophyte, naturally thriving on sandy beaches. In addition to its obvious interest as a naturally salt-tolerant plant, this species is mainly cultivated as an ornamental plant in many countries. Laboratory experiments were carried out to assess the effects of salinity on seed germination and on germination recovery from the effects of saline conditions after transfer to distilled water. Seed germination responses were determined at 0, 50, 100, 200 and 300 mM NaCl. Salt (NaCl) does not affect the germination of L. maritima if applied at a moderate dose of 50 mM. For higher concentrations of NaCl, there is a decrease in the germination rate at 100 and 200 mM NaCl or even a total inhibition of germination at 300 mM. Salt lowers or inhibits germination only through osmotic effects. To improve the germination of L. maritima under high salinity, seeds were pretreated with KNO3, thiourea, proline and salicylic acid. The germination of seeds is improved by KNO3 in the presence or absence of salt, while thiourea increases the final germination without affecting the germination rate. Salicylic acid amplifies the effect of salt, while proline delays germination without stopping it completely. These findings indicate that the application of KNO3 and thiourea may be used to improve seed germination of L. maritima, which is of great interest for cultivating this plant for landscaping purposes in saline soils.

Solution design to extend the pH range of the pH-responsive precipitation of a CspB fusion protein

Cell surface protein B (CspB) from Corynebacterium glutamicum has been developed as a reversible pH-responsive tag for protein purification. CspB fusion proteins precipitate at acidic pH, after that they completely dissolve at neutral pH. This property has been used in a non-chromatographic protein purification method named pH-responsive Precipitation-Redissolution of CspB tag Purification (pPRCP). However, it is difficult to apply pPRCP to proteins that are unstable under acidic conditions. In an effort to shift the precipitation pH to a milder range, we investigated the solution conditions of CspB-fused Teriparatide (CspB50TEV-Teriparatide) during the process of pH-responsive precipitation using pPRCP. The purified CspB50TEV-Teriparatide in buffer without additives precipitated at pH 5.3. By contrast, CspB50TEV-Teriparatide in buffer with 0.5 M Na2SO4 precipitated at pH 6.6 because of the kosmotropic effect. Interestingly, the pH at which precipitation occurred was independent of the protein concentration. The precipitated CspB50TEV-Teriparatide was fully redissolved at above pH 8.0 in the presence or absence of salt. The discovery that proteins can be precipitated at a mild pH will allow pPRCP to be applied to acid-sensitive proteins.

Heat and cold denaturation of yeast frataxin: The effect of pressure

Yfh1 is a yeast protein with the peculiar characteristic to undergo, in the absence of salt, cold denaturation at temperatures above the water freezing point. This feature makes the protein particularly interesting for studies aiming at understanding the rules that determine protein fold stability. Here, we present the phase diagram of Yfh1 unfolding as a function of pressure (0.1–500 MPa) and temperature 278–313 K (5–40°C) both in the absence and in the presence of stabilizers using Trp fluorescence as a monitor. The protein showed a remarkable sensitivity to pressure: at 293 K, pressures around 10 MPa are sufficient to cause 50% of unfolding. Higher pressures were required for the unfolding of the protein in the presence of stabilizers. The phase diagram on the pressure-temperature plane together with a critical comparison between our results and those found in the literature allowed us to draw conclusions on the mechanism of the unfolding process under different environmental conditions.

A short helix regulates conversion of dimeric and 24-meric ferritin architectures

The inter-subunit interaction at the protein interfaces plays a key role in protein self-assembly, through which enabling protein self-assembly controllable is of great importance for preparing the novel nanoscale protein materials with unexplored properties. Different from normal 24-meric ferritin, archaeal ferritin, Thermotoga maritima ferritin (TmFtn) naturally occurs as a dimer, which can assemble into a 24-mer nanocage induced by salts. However, the regulation mechanism of protein self-assembly underlying this phenomenon remains unclear. Here, a combination of the computational energy simulation and key interface reconstruction revealed that a short helix involved interactions at the C4 interface are mainly responsible for the existence of such dimer. Agreeing with this idea, deletion of such short helix of each subunit triggers it to be a stable dimer, which losses the ability to reassemble into 24-meric ferritin in the presence of salts in solution. Further support for this idea comes from the observation that grafting a small helix from human H ferritin onto archaeal subunit resulted in a stable 24-mer protein nanocage even in the absence of salts. Thus, these findings demonstrate that adjusting the interactions at the protein interfaces appears to be a facile, effective approach to control subunit assembly into different protein architectures.

Polyextremophilic Chitinolytic Activity by a Marine Strain (IG119) of Clonostachys&nbsp;rosea.

The investigation for novel unique extremozymes is a valuable business for which the marine environment has been overlooked. The marine fungus Clonostachys rosea IG119 was tested for growth and chitinolytic enzyme production at different combinations of salinity and pH using response surface methodology. RSM modelling predicted best growth in-between pH 3.0 and 9.0 and at salinity of 0–40‰, and maximum enzyme activity (411.137 IU/L) at pH 6.4 and salinity 0‰; however, quite high production (>390 IU/L) was still predicted at pH 4.5–8.5. The highest growth and activity were obtained, respectively, at pH 4.0 and 8.0, in absence of salt. The crude enzyme was tested at different salinities (0–120‰) and pHs (2.0–13.0). The best activity was achieved at pH 4.0, but it was still high (in-between 3.0 and 12.0) at pH 2.0 and 13.0. Salinity did not affect the activity in all tested conditions. Overall, C. rosea IG119 was able to grow and produce chitinolytic enzymes under polyextremophilic conditions, and its crude enzyme solution showed more evident polyextremophilic features. The promising chitinolytic activity of IG119 and the peculiar characteristics of its chitinolytic enzymes could be suitable for several biotechnological applications (i.e., degradation of salty chitin-rich materials and biocontrol of spoiling organisms, possibly solving some relevant environmental issues).

Review on the influence of NaCl on the quality parameters of soft wheat dough

The presence or absence of Sodium Chloride on the quality parameters of soft wheat dough samples was identified. 176 flour samples were analyzed for their protein content, Zeleny sedimentation value and by means of the Chopin Alveograph and Brabender Farinograph, with no salt and with 1.5% salt addition. Aubusson, Boleroand Blasco were studied by means of the Rapid Visco Analyzer at three levels of salt addition (0%, 1.5% and 3.0%). The results of the study confirmed the role of salt in strengthening the wheat gluten network and thus the gas retention of dough and in affecting yeast activity. Protein content, sedimentation value according to Zeleny, Alveograph and Farinograph parameters were analyzed. The protein content (% d.m.) were ranged from 8.2 to 15.0 covering the range of values that are commonly found in wheat commercialized for bakery products manufacturing and Zeleny sedimentation Index value ranges from 18 to 68 mL indicating that values below 20 mL are unsuitable for bread baking. Considering the Alveograph parameters obtained with salt ranges from 72 to 413 whereas values ranged from 42 to 342 in the absence of salt. T-test resulted in significant difference (p < 0.01) between all Farinograph and Alveograph parameters with or without salt. Two way analysis of variance showed that all parameters are influenced by the cultivar (p < 0.01), but did not confirm an interaction between the two factors (salt × cultivar). Differences between parameters with and without salt resulted to be of quite variable extent between samples.

Corrosion behaviour of Fe-25Cr alloy in wet CO2 gas at 650 °C: Effects of chloride deposits and Si+Mn alloying addition

A model ferritic Fe-25wt% Cr alloy exposed to an Ar-60CO2-20H2O gas mixture at 650 °C experienced rapid oxide scale growth and simultaneous internal carburization. When a deposit of NaCl-KCl was present on the alloy surface, it developed a more porous oxide scale and more extensive carburization. In the absence of salt, alloy additions of Si+Mn led to improved protection: slow oxide scaling and no carburization. In the presence of salt, this protection was much reduced. These effects are discussed in terms of oxide volatilisation and ferrite chromium concentration depletion by carburization.

Pulmonary tuberculosis and the Gerson's diet

Dr. B. Gettkant, director of the Schneberg tuberculosis sanatorium, also deals with pulmonary tuberculosis and the Gerson's diet (D. m. W. 1929, no. 43). Observing 20 severe pulmonary tbc patients, Gettkant came to the conclusion that in no case did the Gerson's diet affect the course of pulmonary TBK: 11 out of 20 people died, 8 remained inoperative, the process went ahead and the fate of 1 remained unknown; with the Gerson's diet there is undoubtedly an improvement in lupus, in bone and articular TBK, but not in pulmonary. As a great difficulty in carrying out a diet a. indicates the absence of salt: the TBC b-th already have a poor appetite, and desalted food, despite its best preparation, soon arouses disgust, and the b-th's appetite still falls.

Rate Constants and Rheological Properties of the Ultrasonic Degradation of Carboxymethyl Cellulose

To degrade the polysaccharide with high molecular weight the effectiveness of ultrasound for carboxymethyl cellulose (CMC) in aqueous solution has been studied at a time of 60 min and temperature of 25 ̊C and for polymer solution with concentration up 0.1 g/L. The Huggins equation was appropriate to apply to the intrinsic viscosity of CMC before sonication to understand the influence of salt on degradation, CMC solution was premixed with 0.1 M NaCl, before ultrasonication. Developed a kinetic model, successfully implement to predict and quantify rates of degradation and efficiency. The values of reaction rate constants and reaction orders were found related to the salt and the concentration used, suggesting that, the presence and absence of salts could increase or decrease the degradation by ultrasonic radiation through adjusting the molecular conformation of CMC

Local Ordering of Molten Salts at NiO Crystal Interfaces Promotes High-Index Faceting.

Given the strong influence of surface structure on the reactivity of heterogeneous catalysts, understanding the mechanisms that control crystal morphology is an important component of designing catalytic materials with targeted shape and functionality. Herein, we employ density functional theory to examine the impact of growth media on NiO crystal faceting in line with experimental findings, showing that molten‐salt synthesis in alkali chlorides (KCl, LiCl, and NaCl) imposes shape selectivity on NiO particles. We find that the production of NiO octahedra is attributed to the dissociative adsorption of H2O, whereas the formation of trapezohedral particles is associated with the control of the growth kinetics exerted by ordered salt structures on high‐index facets. To our knowledge, this is the first observation that growth inhibition of metal‐oxide facets occurs by a localized ordering of molten salts at the crystal–solvent interface. These findings provide new molecular‐level insight on kinetics and thermodynamics of molten‐salt synthesis as a predictive route to shape‐engineer metal‐oxide crystals.

Seven Years at High Salinity-Experimental Evolution of the Extremely Halotolerant Black Yeast Hortaea werneckii.

The experimental evolution of microorganisms exposed to extreme conditions can provide insight into cellular adaptation to stress. Typically, stress-sensitive species are exposed to stress over many generations and then examined for improvements in their stress tolerance. In contrast, when starting with an already stress-tolerant progenitor there may be less room for further improvement, it may still be able to tweak its cellular machinery to increase extremotolerance, perhaps at the cost of poorer performance under non-extreme conditions. To investigate these possibilities, a strain of extremely halotolerant black yeast Hortaea werneckii was grown for over seven years through at least 800 generations in a medium containing 4.3 M NaCl. Although this salinity is well above the optimum (0.8–1.7 M) for the species, the growth rate of the evolved H. werneckii did not change in the absence of salt or at high concentrations of NaCl, KCl, sorbitol, or glycerol. Other phenotypic traits did change during the course of the experimental evolution, including fewer multicellular chains in the evolved strains, significantly narrower cells, increased resistance to caspofungin, and altered melanisation. Whole-genome sequencing revealed the occurrence of multiple aneuploidies during the experimental evolution of the otherwise diploid H. werneckii. A significant overrepresentation of several gene groups was observed in aneuploid regions. Taken together, these changes suggest that long-term growth at extreme salinity led to alterations in cell wall and morphology, signalling pathways, and the pentose phosphate cycle. Although there is currently limited evidence for the adaptive value of these changes, they offer promising starting points for future studies of fungal halotolerance.