Carbon Starvation Research Articles

Modeling the mechanisms of conifer mortality under seawater exposure.

Relative sea level rise (SLR) increasingly impacts coastal ecosystems through the formation of ghost forests. To predict the future of coastal ecosystems under SLR and changing climate, it is important to understand the physiological mechanisms underlying coastal tree mortality and to integrate this knowledge into dynamic vegetation models. We incorporate the physiological effect of salinity and hypoxia in a dynamic vegetation model in the Earth system land model, and used the model to investigate the mechanisms of mortality of conifer forests on the west and east coast sites of USA, where trees experience different form of sea water exposure. Simulations suggest similar physiological mechanisms can result in different mortality patterns. At the east coast site that experienced severe increases in seawater exposure, trees loose photosynthetic capacity and roots rapidly, and both storage carbon and hydraulic conductance decrease significantly within a year. Over time, further consumption of storage carbon that leads to carbon starvation dominates mortality. At the west coast site that gradually exposed to seawater through SLR, hydraulic failure dominates mortality because root loss impacts on conductance are greater than the degree of storage carbon depletion. Measurements and modeling focused on understanding the physiological mechanisms of mortality is critical to reducing predictive uncertainty.

Understanding the effects of carbon and nitrogen starvation on the comparative secretomes of Ganoderma boninense and Ganoderma tornatum

Ganoderma boninense is the most pathogenic fungal species of oil palm trees. However, there is limited understanding of the host - pathogen interaction. Secreted proteins play a critical role during the early infection process which involves competition between the fungal and host populations for nutrient resources necessary for continual growth. To identify the secreted proteins that may play a role in pathogenicity during infection stage, we compared the secretome profiles of G. boninense versus G. tornatum under carbon and nitrogen starvation by using shotgun proteomics approach. In total, 250 non-redundant proteins were identified, and statistical data analysis indicated that 68 proteins were increased while 104 proteins decreased in abundance in T3 for G. boninense whereas in G. tornatum, 42 proteins were increased, and 27 proteins were decreased in abundance in T3. Further investigation via bioinformatic tools helped us to classify the carbohydrate-active enzymes (CAZymes) proteins into four classes: oxidoreductases, transferases, hydrolases, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Interestingly, oxidoreductase enzymes were identified only in G. boninense. These putative proteins might contribute to the different levels of pathogenicity of G. boninense and G. tornatum toward oil palm. The newly discovered protein information may help researchers better understand the secretory mechanisms of G. boninense and the process by which it invades oil palm root tissues. The discovered seretome proteins, along with their subclassification and functional annotation also serve as a database for Ganoderma species that is easy to access and retrieve.

Carbon starvation considerably accelerated nickel corrosion by Desulfovibrio vulgaris

Carbon starvation can affect the activity of microbes, thereby affecting the metabolism and the extracellular electron transfer (EET) process of biofilm. In the present work, the microbiologically influenced corrosion (MIC) behavior of nickel (Ni) was investigated under organic carbon starvation by Desulfovibrio vulgaris. Starved D. vulgaris biofilm was more aggressive. Extreme carbon starvation (0% CS level) reduced weight loss due to the severe weakening of biofilm. The corrosion rate of Ni (based on weight loss) was sequenced as 10% CS level > 50% CS level > 100 CS level > 0% CS level. Moderate carbon starvation (10% CS level) caused the deepest pit of Ni in all the carbon starvation treatments, with a maximal pit depth of 18.8 μm and a weight loss of 2.8 mg·cm−2 (0.164 mm·y−1). The corrosion current density (icorr) of Ni for the 10% CS level was as high as 1.62 × 10−5 A·cm−2, which was approximately 2.9-fold greater than the full-strength medium (5.45 × 10−6 A·cm−2). The electrochemical data corresponded to the corrosion trend revealed by weight loss. The various experimental data rather convincingly pointed to the Ni MIC of D. vulgaris following the EET-MIC mechanism despite a theoretically low Ecell value (+33 mV).

Rhodopsin-mediated nutrient uptake by cultivated photoheterotrophic Verrucomicrobiota

Rhodopsin photosystems convert light energy into electrochemical gradients used by the cell to produce ATP, or for other energy-demanding processes. While these photosystems are widespread in the ocean and have been identified in diverse microbial taxonomic groups, their physiological role in vivo has only been studied in few marine bacterial strains. Recent metagenomic studies revealed the presence of rhodopsin genes in the understudied Verrucomicrobiota phylum, yet their distribution within different Verrucomicrobiota lineages, their diversity, and function remain unknown. In this study, we show that more than 7% of Verrucomicrobiota genomes (n = 2916) harbor rhodopsins of different types. Furthermore, we describe the first two cultivated rhodopsin-containing strains, one harboring a proteorhodopsin gene and the other a xanthorhodopsin gene, allowing us to characterize their physiology under laboratory-controlled conditions. The strains were isolated in a previous study from the Eastern Mediterranean Sea and read mapping of 16S rRNA gene amplicons showed the highest abundances of these strains at the deep chlorophyll maximum (source of their inoculum) in winter and spring, with a substantial decrease in summer. Genomic analysis of the isolates suggests that motility and degradation of organic material, both energy demanding functions, may be supported by rhodopsin phototrophy in Verrucomicrobiota. Under culture conditions, we show that rhodopsin phototrophy occurs under carbon starvation, with light-mediated energy generation supporting sugar transport into the cells. Overall, this study suggests that photoheterotrophic Verrucomicrobiota may occupy an ecological niche where energy harvested from light enables bacterial motility toward organic matter and supports nutrient uptake.

Helicobacter pylori Modulates Heptose Metabolite Biosynthesis and Heptose-Dependent Innate Immune Host Cell Activation by Multiple Mechanisms.

Heptose metabolites including ADP-d-glycero-β-d-manno-heptose (ADP-heptose) are involved in bacterial lipopolysaccharide and cell envelope biosynthesis. Recently, heptoses were also identified to have potent proinflammatory activity on human cells as novel microbe-associated molecular patterns. The gastric pathogenic bacterium Helicobacter pylori produces heptose metabolites, which it transports into human cells through its Cag type 4 secretion system. Using H. pylori as a model, we have addressed the question of how proinflammatory ADP-heptose biosynthesis can be regulated by bacteria. We have characterized the interstrain variability and regulation of heptose biosynthesis genes and the modulation of heptose metabolite production by H. pylori, which impact cell-autonomous proinflammatory human cell activation. HldE, a central enzyme of heptose metabolite biosynthesis, showed strong sequence variability between strains and was also variably expressed between strains. Amounts of gene transcripts in the hldE gene cluster displayed intrastrain and interstrain differences, were modulated by host cell contact and the presence of the cag pathogenicity island, and were affected by carbon starvation regulator A (CsrA). We reconstituted four steps of the H. pylori lipopolysaccharide (LPS) heptose biosynthetic pathway in vitro using recombinant purified GmhA, HldE, and GmhB proteins. On the basis of one- and two-dimensional nuclear magnetic resonance (NMR) spectroscopy and mass spectrometry, the structures of major reaction products were identified as β-d-ADP-heptose and β-heptose-1-monophosphate. A proinflammatory heptose-monophosphate variant was also identified for the first time as a novel cell-active product in H. pylori bacteria. Separate purified HldE subdomains and variant HldE allowed us to uncover additional strain variation in generating heptose metabolites. IMPORTANCE Bacterial heptose metabolites, intermediates of lipopolysaccharide (LPS) biosynthesis, are novel microbe-associated molecular patterns (MAMPs) that activate proinflammatory signaling. In the gastric pathogen Helicobacter pylori, heptoses are transferred into host cells by the Cag type IV secretion system, which is also involved in carcinogenesis. Little is known about how H. pylori, which is highly strain variable, regulates heptose biosynthesis and downstream host cell activation. We report here that the regulation of proinflammatory heptose production by H. pylori is strain specific. Heptose gene cluster activity is modulated by the presence of an active cag pathogenicity island (cagPAI), contact with human cells, and the carbon starvation regulator A. Reconstitution with purified biosynthesis enzymes and purified bacterial lysates allowed us to biochemically characterize heptose pathway products, identifying a heptose-monophosphate variant as a novel proinflammatory metabolite. These findings emphasize that the bacteria use heptose biosynthesis to fine-tune inflammation and also highlight opportunities to mine the heptose biosynthesis pathway as a potential therapeutic target against infection, inflammation, and cancer.

Proteome Dynamics of Persulfidation in Leaf Tissue under Light/Dark Conditions and Carbon Deprivation.

Hydrogen sulfide (H2S) acts as a signaling molecule in plants, bacteria, and mammals, regulating various physiological and pathological processes. The molecular mechanism by which hydrogen sulfide exerts its action involves the posttranslational modification of cysteine residues to form a persulfidated thiol motif. This research aimed to study the regulation of protein persulfidation. We used a label-free quantitative approach to measure the protein persulfidation profile in leaves under different growth conditions such as light regimen and carbon deprivation. The proteomic analysis identified a total of 4599 differentially persulfidated proteins, of which 1115 were differentially persulfidated between light and dark conditions. The 544 proteins that were more persulfidated in the dark were analyzed, and showed significant enrichment in functions and pathways related to protein folding and processing in the endoplasmic reticulum. Under light conditions, the persulfidation profile changed, and the number of differentially persulfidated proteins increased up to 913, with the proteasome and ubiquitin-dependent and ubiquitin-independent catabolic processes being the most-affected biological processes. Under carbon starvation conditions, a cluster of 1405 proteins was affected by a reduction in their persulfidation, being involved in metabolic processes that provide primary metabolites to essential energy pathways and including enzymes involved in sulfur assimilation and sulfide production.

Hello darkness, my old friend: 3-KETOACYL-COENZYME A SYNTHASE4 is a branch point in the regulation of triacylglycerol synthesis in Arabidopsis thaliana.

Plant lipids are important as alternative sources of carbon and energy when sugars or starch are limited. Here, we applied combined heat and darkness or extended darkness to a panel of ∼300 Arabidopsis (Arabidopsis thaliana) accessions to study lipid remodeling under carbon starvation. Natural allelic variation at 3-KETOACYL-COENZYME A SYNTHASE4 (KCS4), a gene encoding an enzyme involved in very long chain fatty acid (VLCFA) synthesis, underlies the differential accumulation of polyunsaturated triacylglycerols (puTAGs) under stress. Ectopic expression of KCS4 in yeast and plants proved that KCS4 is a functional enzyme localized in the endoplasmic reticulum with specificity for C22 and C24 saturated acyl-CoA. Allelic mutants and transient overexpression in planta revealed the differential role of KCS4 alleles in VLCFA synthesis and leaf wax coverage, puTAG accumulation, and biomass. Moreover, the region harboring KCS4 is under high selective pressure and allelic variation at KCS4 correlates with environmental parameters from the locales of Arabidopsis accessions. Our results provide evidence that KCS4 plays a decisive role in the subsequent fate of fatty acids released from chloroplast membrane lipids under carbon starvation. This work sheds light on both plant response mechanisms and the evolutionary events shaping the lipidome under carbon starvation.

Silencing GmATG10 results in activation of immune responses in soybean

Autophagy is a highly conserved mechanism for material degradation and recycling in eukaryote cells, and plays important roles in growth, development, stress tolerance and immune responses. ATG10 plays a key role in autophagosome formation. To understand the function of ATG10 in soybean, two homologous GmATG10 genes, namely GmATG10a and GmATG10b, were silenced simultaneously by bean pod mottle virus (BPMV) induced gene silencing. The carbon starvation induced by dark treatment and Western blotting analysis of GmATG8 accumulation level indicated that concurrent silencing GmATG10a/10b resulted in the impairment of autophagy in soybean; disease resistance and kinase assays demonstrated that GmATG10a/10b participated in the immune responses by negatively regulating the activation of GmMPK3/6, indicating that GmATG10a/10b plays a negative regulatory role in immune response in soybean.

Sugar infusion into trees: A novel method to study tree carbon relations and its regulations.

Many carbon-related physiological questions in plants such as carbon (C) limitation or starvation have not yet been resolved thoroughly due to the lack of suitable experimental methodology. As a first step towards resolving these problems, we conducted infusion experiments with bonsai trees (Ficus microcarpa) and young maple trees (Acer pseudoplatanus) in greenhouse, and with adult Scots pine trees (Pinus sylvestris) in the field, that were "fed" with 13C-labelled glucose either through the phloem or the xylem. We then traced the 13C-signal in plant organic matter and respiration to test whether trees can take up and metabolize exogenous sugars infused. Ten weeks after infusion started, xylem but not phloem infusion significantly increased the δ13C values in both aboveground and belowground tissues of the bonsai trees in the greenhouse, whereas xylem infusion significantly increased xylem δ13C values and phloem infusion significantly increased phloem δ13C values of the adult pines in the field experiment, compared to the corresponding controls. The respiration measurement experiment with young maple trees showed significantly increased δ13C-values in shoot respired CO2 at the time of four weeks after xylem infusion started. Our results clearly indicate that trees do translocate and metabolize exogenous sugars infused, and because the phloem layer is too thin, and thus xylem infusion can be better operated than phloem infusion. This tree infusion method developed here opens up new avenues and has great potential to be used for research on the whole plant C balance and its regulation in response to environmental factors and extreme stress conditions.

Identification of genetic and biochemical mechanisms associated with heat shock and heat stress adaptation in grain amaranths.

Heat stress is poised to become a major factor negatively affecting plant performance worldwide. In terms of world food security, increased ambient temperatures are poised to reduce yields in cereals and other economically important crops. Grain amaranths are known to be productive under poor and/or unfavorable growing conditions that significantly affect cereals and other crops. Several physiological and biochemical attributes have been recognized to contribute to this favorable property, including a high water-use efficiency and the activation of a carbon starvation response. This study reports the behavior of the three grain amaranth species to two different stress conditions: short-term exposure to heat shock (HS) conditions using young plants kept in a conditioned growth chamber or long-term cultivation under severe heat stress in greenhouse conditions. The latter involved exposing grain amaranth plants to daylight temperatures that hovered around 50°C, or above, for at least 4h during the day and to higher than normal nocturnal temperatures for a complete growth cycle in the summer of 2022 in central Mexico. All grain amaranth species showed a high tolerance to HS, demonstrated by a high percentage of recovery after their return to optimal growing conditions. The tolerance observed coincided with increased expression levels of unknown function genes previously shown to be induced by other (a)biotic stress conditions. Included among them were genes coding for RNA-binding and RNA-editing proteins, respectively. HS tolerance was also in accordance with favorable changes in several biochemical parameters usually induced in plants in response to abiotic stresses. Conversely, exposure to a prolonged severe heat stress seriously affected the vegetative and reproductive development of all three grain amaranth species, which yielded little or no seed. The latter data suggested that the usually stress-tolerant grain amaranths are unable to overcome severe heat stress-related damage leading to reproductive failure.

Climate warming causes photobiont degradation and carbon starvation in a boreal climate sentinel lichen.

The long-term potential for acclimation by lichens to changing climates is poorly known, despite their prominent roles in forested ecosystems. Although often considered "extremophiles," lichens may not readily acclimate to novel climates well beyond historical norms. In a previous study (Smith et al., 2018), Evernia mesomorpha transplants in a whole-ecosystem climate change experiment showed drastic mass loss after 1 yr of warming and drying; however, the causes of this mass loss were not addressed. We examined the causes of this warming-induced mass loss by measuring physiological, functional, and reproductive attributes of lichen transplants. Severe loss of mass and physiological function occurred above +2°C of experimental warming. Loss of algal symbionts ("bleaching") and turnover in algal community compositions increased with temperature and were the clearest impacts of experimental warming. Enhanced CO2 had no significant physiological or symbiont composition effects. The functional loss of algal photobionts led to significant loss of mass and specific thallus mass (STM), which in turn reduced water-holding capacity (WHC). Although algal genotypes remained detectable in thalli exposed to higher stress, within-thallus photobiont communities shifted in composition toward greater diversity. The strong negative impacts of warming and/or lower humidity on Evernia mesomorpha were driven by a loss of photobiont activity. Analogous to the effects of climate change on corals, the balance of symbiont carbon metabolism in lichens is central to their resilience to changing conditions.

Effects of concentration and frequency of CO2 supply on productivity of marine microalga Isochrysis galbana

Demand for the high fucoxanthin and docosahexaenoic acid (DHA) content found in the marine microalga Isochrysis galbana is rising annually, and it is essential to devise large-scale cultivation systems that are low in energy and cost. Continuous CO2 supply occupies a considerable portion of total electricity consumption for microalgae cultivation. While an intermittent, high-concentration CO2 supply/agitation has the potential to reduce the electricity consumption required for aeration, it may cause carbon starvation and sedimentation of cells when the supply frequency is insufficient. This study investigated the effects of concentration and frequency of CO2 supply on productivity of I. galbana. Semi-continuous cultivation was conducted under different conditions of CO2 supply (0.04 (air) to 10 % concentration and continuous or intermittent (1 min every 9 min) supply). Under continuous air supply (control), biomass productivity reached 0.18 g L−1 day−1. A continuous supply of CO2 at 5 % showed the highest biomass productivity (0.33 g L−1 day−1). Productivity was only 0.12 g L−1 day−1 with a continuous CO2 supply at 10 % concentration. It was confirmed that growth was suppressed under conditions of a continuous supply of CO2 at high concentrations. The study found that intermittent CO2 concentrations of 5 % and 10 % showed productivity of 0.29 and 0.24 g L−1 day−1, respectively. These productivities were higher than that in the control. Under conditions of continuous CO2 supply, 0.61 mg/g dry weight (DW) of a higher fucoxanthin content was achieved with air supply than was found at concentrations of 5 % and 10 % (0.18 and 0.11 mg/g DW, respectively). However, under intermittent supply conditions, the fucoxanthin reached the higher content of 0.56 mg/g DW under 5% supply. Under conditions of high CO2 concentrations, continuous supply inhibited the accumulation of fucoxanthin, while an intermittent supply promoted its accumulation.

A research program-linked, course-based undergraduate research experience that allows undergraduates to participate in current research on mycobacterial gene regulation.

Undergraduate instructional biology laboratories are typically taught within two paradigms. Some labs focus on protocols and techniques delivered in "cookbook" format with defined experimental outcomes. There is increasing momentum to alternatively employ student-driven, open-ended, and discovery-based strategies, often via course-based undergraduate research experiences (CUREs) using crowd-sourcing initiatives. A fraction of students also participate in funded research in faculty research labs, where they have opportunities to work on projects designed to expand the frontiers of human knowledge. These experiences are widely recognized as valuable but are not scalable, as most institutions have many more undergraduates than research lab positions. We sought to address this gap through our department's curriculum by creating an opportunity for students to participate in the real-world research process within a laboratory course. We conceived, developed, and delivered an authentic, guided research experience to students in an upper-level molecular biology laboratory course. We refer to this model as a "research program-linked CURE." The research questions come directly from a faculty member's research lab and evolve along with that research program. Students study post-transcriptional regulation in mycobacteria. We use current molecular biology methodologies to test hypotheses like "UTRs affect RNA and protein expression levels," "there is functional redundancy among RNA helicases," and "carbon starvation alters mRNA 5' end chemistries." We conducted standard assessments and developed a customized "Skills and Concepts Inventory" survey to gauge how well the course met our student learning outcomes. We report the results of our assessments and describe challenges addressed during development and execution of the course, including organizing activities to fit within an instructional lab, balancing breadth with depth, and maintaining authenticity while giving students the experience of obtaining interpretable and novel results. Our data suggest student learning was enhanced through this truly authentic research approach. Further, students were able to perceive they were participants and contributors within an active research paradigm. Students reported increases in their self-identification as scientists, and a positive impact on their career trajectories. An additional benefit was reciprocation back to the funded research laboratory, by funneling course alumni, results, materials, and protocols.

Transcriptome and Deletion Mutant Analyses Revealed that an RpoH Family Sigma Factor Is Essential for Photosystem Production in Roseateles depolymerans under Carbon Starvation.

Roseateles depolymerans is an obligately aerobic bacterium that produces a photosynthetic apparatus only under the scarcity of carbon substrates. We herein examined changes in the transcriptomes of R. depolymerans cells to clarify the expression of photosynthesis genes and their upstream regulatory factors under carbon starvation. Transcriptomes 0, 1, and 6‍ ‍h after the depletion of a carbon substrate indicated that transcripts showing the greatest variations (a 500-fold increase [6 h/0 h]) were light-harvesting proteins (PufA and PufB). Moreover, loci with more than 50-fold increases (6 h/0‍ ‍h) were fully related to the photosynthetic gene cluster. Among 13 sigma factor genes, the transcripts of a sigma 70 family sigma factor related to RpoH (SP70) increased along photosynthesis genes under starvation; therefore, a knockout experiment of SP70 was performed. ΔSP70 mutants were found to lack photosynthetic pigments (carotenoids and bacteriochlo-rophyll a) regardless of carbon starvation. We also examined the effects of heat stress on ΔSP70 mutants, and found that SP70 was also related to heat stress tolerance, similar to other RpoH sigma factors (while heat stress did not trigger photosystem production). The deficient accumulation of photosynthetic pigments and the heat stress tolerance of ΔSP70 mutants were both complemented by the introduction of an intact SP70 gene. Furthermore, the transcription of photosynthetic gene operons (puf, puh, and bch) was markedly reduced in the ΔSP70 mutant. The RpoH homologue SP70 was concluded to be a sigma factor that is essential for the transcription of photosynthetic gene operons in R. depolymerans.

Concurrent time course of xylem hydraulic dysfunction and non-structural carbohydrates under contrasting water deficits and nitrogen supplies in poplar

We compared the temporal dynamics of growth, leaf gas exchange, xylem hydraulic dysfunction and non-structural carbohydrates (NSC) of two poplar hybrid genotypes subjected to a differential nitrogen (N) supply under contrasting water deficits. Moderate water deficit generated intermediate embolism rates (45–70%) but had marginal effects on NSC. Severe water deficit generated progressive hydraulic failure and NSC reduction, leading to tree death within 90 days. At death, NSC in perennial tissues were not entirely depleted because of remaining soluble sugars. Higher N availability primarily affected growth and NSC dynamics (mainly starch), not embolism dynamics. The faster growing genotype benefiting most from N addition ceased growth and photosynthesis almost simultaneously, with no effect of N addition on process cessation, leading to progressive starch depletion during severe drought. In contrast, the slower growing genotype ceased growth before photosynthesis, leading to a transient increase in starch concentration during early drought stages, but this tended to be suppressed by higher N availability. Altogether, our findings indicate that carbon starvation alone is unlikely in hydraulically vulnerable species such as poplar even under prolonged moderate water deficit, but starch relative depletion covaries in time with progressive hydraulic failure under lethal water deficit. Nutritional status has the potential to shape drought responses, primarily through adjustments in carbon source-sink relations rather than in intrinsic xylem hydraulics, in a genotype-dependent manner. Exploring how hydraulic and carbon safety margins can be coordinated within species would be valuable in understanding how variation in drought response strategies can be exploited in breeding/selection.

Gut colonization by Bacteroides requires translation by an EF-G paralog lacking GTPase activity.

Protein synthesis is crucial for cell growth and survival yet one of the most energy-consuming cellular processes. How, then, do cells sustain protein synthesis under starvation conditions when energy is limited? To accelerate the translocation of mRNA-tRNAs through the ribosome, bacterial elongation factor G (EF-G) hydrolyzes energy-rich guanosine triphosphate (GTP) for every amino acid incorporated into a protein. Here, we identify an EF-G paralog-EF-G2-that supports translocation without hydrolyzing GTP in the gut commensal bacterium Bacteroides thetaiotaomicron. EF-G2's singular ability to sustain protein synthesis, albeit at slow rates, is crucial for bacterial gut colonization. EF-G2 is ~10-fold more abundant than canonical EF-G1 in bacteria harvested from murine ceca and, unlike EF-G1, specifically accumulates during carbon starvation. Moreover, we uncover a 26-residue region unique to EF-G2 that is essential for protein synthesis, EF-G2 dissociation from the ribosome, and responsible for the absence of GTPase activity. Our findings reveal how cells curb energy consumption while maintaining protein synthesis to advance fitness in nutrient-fluctuating environments.

Mechanistic investigations of corrosion and localized corrosion of X80 steel in seawater comprising sulfate-reducing bacteria under continuous carbon starvation

Sulfate-reducing bacteria (SRB) corrosion is one of the main reasons leading to the service failure of engineering materials in the marine environment. An understanding of SRB corrosion especially under continuous organic carbon starvation is required. In this work, SRB corrosion of X80 pipeline steel under continuous organic carbon starvation was studied using mass loss, electrochemical measurements, and surface analysis. The effects of initial SRB concentrations on steel corrosion were also studied and discussed. The results indicated that more than 99% population of SRB die after 21 d of testing in various conditions, but there are still amounts of SRB survivors. SRB survivors with continued organic carbon starvation have a better adaptive ability and accelerated steel corrosion. Both the uniform and localized corrosion rates are also proportional to the initial SRB concentration as well as SRB survivors. The localized corrosion rate and density of corrosion pits of specimen corresponding to an initial SRB count of 108 cells/mL reach bigger values of (0.693 ± 0.114) mm/y and (1.94 ± 0.39)×104 pits/cm2, respectively, which are induced by the combination of the sessile and planktonic SRB.

Effects of Water Deficit Stress on Growth Parameters of Robinia pseudoacacia L. Selected Clones under In Vitro Conditions

Rapid screening methods for drought-resistant genotypes are urgently needed in tree improvement programs in the face of current climate change. We used a plant tissue culture technique to assess the phenotypic response of three highly productive genotypes of Robinia pseudoacacia to water deficit induced by mannitol and sucrose in a range of water potentials from 0 MPa to −1.5 MPa in an eight-week experiment. Our study showed genotype-specific responses to induced drought stress, indicating the potential for tree improvement in productivity and stress tolerance. Considering that all plantlets were constantly supplied with carbon, from the medium during the drought-induced experiment, our results suggest that hydraulic failure rather than carbon starvation may be the main cause of drought-induced mortality. Furthermore, our results showed different metabolic pathways of sucrose depending on the concentration of sucrose in the medium and different responses to osmoticum (mannitol vs. sucrose) and its concentration among the clones tested. We believe, that for large-scale breeding programs wanting to select for drought-tolerant genotypes, the use of culture media containing 90 gL−1 mannitol or 90 gL−1 sucrose at an early selection stage should provide satisfactory screening results. However, lab-based screening should be supported by further field trials, preferably at multiple sites, to assess the long-term impact and phenotypic stability of the early selection strategies.

Stress Responses Elicited by Glucose Withdrawal in Aspergillus fumigatus.

Glucose is a widely used carbon source in laboratory practice to culture Aspergillus fumigatus, however, glucose availability is often low in its “natural habitats”, including the human body. We used a physiological−transcriptomical approach to reveal differences between A. fumigatus Af293 cultures incubated on glucose, glucose and peptone, peptone (carbon limitation), or without any carbon source (carbon starvation). Autolytic cell wall degradation was upregulated by both carbon starvation and limitation. The importance of autolytic cell wall degradation in the adaptation to carbon stress was also highlighted by approximately 12.4% of the A. fumigatus genomes harboring duplication of genes involved in N-acetyl glucosamine utilization. Glucose withdrawal increased redox imbalance, altered both the transcription of antioxidative enzyme genes and oxidative stress tolerance, and downregulated iron acquisition, but upregulated heme protein genes. Transcriptional activity of the Gliotoxin cluster was low in all experiments, while the Fumagillin cluster showed substantial activity both on glucose and under carbon starvation, and the Hexadehydro-astechrome cluster only on glucose. We concluded that glucose withdrawal substantially modified the physiology of A. fumigatus, including processes that contribute to virulence. This may explain the challenge of predicting the in vivo behavior of A. fumigatus based on data from glucose rich cultures.

HYPHAEdelity: a quantitative image analysis tool for assessing peripheral whole colony filamentation.

The yeast Saccharomyces cerevisiae, also known as brewer's yeast, can undergo a reversible stress-responsive transition from individual ellipsoidal cells to chains of elongated cells in response to nitrogen- or carbon starvation. Whole colony morphology is frequently used to evaluate phenotypic switching response; however, quantifying two-dimensional top-down images requires each pixel to be characterized as belonging to the colony or background. While feasible for a small number of colonies, this labor-intensive assessment process is impracticable for larger datasets. The software tool HYPHAEdelity has been developed to semi-automate the assessment of two-dimensional whole colony images and quantify the magnitude of peripheral whole colony yeast filamentation using image analysis tools intrinsic to the OpenCV Python library. The software application functions by determining the total area of filamentous growth, referred to as the f-measure, by subtracting the area of the inner colony boundary from the outer-boundary area associated with hyphal projections. The HYPHAEdelity application was validated against automated and manually pixel-counted two-dimensional top-down images of S. cerevisiae colonies exhibiting varying degrees of filamentation. HYPHAEdelity's f-measure results were comparable to areas determined through a manual pixel enumeration method and found to be more accurate than other whole colony filamentation software solutions.