Corn bioethanol production generates large volumes of animal feed coproducts with nutritional value largely determined by their fiber and protein content. Here, we combined microscopic and biochemical lignin analysis with untargeted proteomics to characterize corn flour (CF) feedstock and the downstream dried distiller’s grains with solubles (DDGS) and corn fermented protein (CFP) fractions from an industrial bioethanol plant. DDGS contained up to 8

Monocarboxylate transporter 2 (MCT2; SLC16A7) is a high-affinity pyruvate transporter implicated in cancer metabolism. However, its role in lung cancer progression and the tumor microenvironment remains unclear. This study examined the effects of MCT2 reduction on tumor growth and cell-type-specific transcriptional changes within the tumor microenvironment. MCT2 loxP/loxP mice were crossed with mCre-Tg mice, and MCT2 deletion was induced by tamoxifen. Control (CO) mice received vehicle treatment. TC1 cells (100,000 cells/mouse) were injected subcutaneously, and tumors were harvested after 24 days. Single-nucleus RNA sequencing (snRNA-seq) was performed on isolated tumor nuclei (4000 nuclei/sample; n = 3 per group) using the 10x Genomics Chromium platform. Data were processed with Cell Ranger v3.0.2 and Seurat v5.2.1, followed by differential expression and pathway enrichment analyses integrated with macrophage bulk RNA-seq data. Tumors in mice with systemic MCT2 reduction grew significantly faster than those in control mice, demonstrating an association between host MCT2 reduction and increased tumor growth. Transcriptomic analysis generated high-quality profiles from 6864 CO and 10,055 KO nuclei. Clustering identified 12 cellular populations and cell types. MCT2 reduction altered pathways involved in glycolysis, the tricarboxylic acid cycle, oxidative phosphorylation, and fatty acid metabolism across multiple populations. Macrophages showed prominent transcriptional changes, including enrichment of MAPK, PI3K-Akt, IgSF-CAM, ECM, and cytokine–cytokine signaling pathways. These findings were supported by macrophage bulk RNA-seq data. Systemic MCT2 reduction was associated with increased tumor growth and broad transcriptional alterations within the tumor micro-environment. Differences in metabolic and immune-related transcriptional programs, particularly in macrophages, identify potential mechanisms associated with tumor progression that warrant further functional investigation.

ABSTRACT In nature, plants are subjected to multiple environmental stress factors simultaneously or sequentially. Recent studies revealed that when three or more stress factors impact a plant simultaneously (termed ‘multifactorial stress combination’; MFSC), plant survival declines, even if the intensity of each individual stress involved in the MFSC is low. We previously identified RAP2.3 as a key transcription factor (TF) required for Arabidopsis thaliana survival, specifically under a MFSC of salt+excess light+heat stress ( i.e., S+EL+HS). Here we report that RAP2.3 is required for the expression of SIGMA3, a nuclear-encoded factor that directs plastid RNA polymerase to specific plastid promoters, and MYB51, a key stress response TF involved in glucosinolate metabolism and oxidative stress responses, specifically during a MFSC of S+EL+HS. Like rap2.3 mutants, myb51 and sig3 mutants display significantly low survival rate specifically under the MFSC of S+EL+HS. Based on MYB51 gene regulatory network analysis and characterization of jasmonic acid (JA) mutants, we further reveal that suppression of JA signaling could play an important role in promoting plant survival under conditions of S+EL+HS. Our findings uncover an additional layer of the response of plants to MFSC, as well as identify potential targets for breeding crops with enhanced tolerance to climate change.

Cancer immunoprevention leverages the immune system's surveillance mechanisms to mitigate tumor development. Vaccines that constitute a tumor antigen and an immune adjuvant are perceived as immunoprevention modalities. However, relevant tumor antigens are unknown for non-viral cancers, which constitute most human cancers. Our group has recently shown that SA-4-1BBL, a novel agonist of CD137 receptor, but not antibodies, shows immunoprevention efficacy against various tumors. Advanced bioinformatics analyses of bulk RNA-seq data were conducted to elucidate mechanisms underlying cancer immunoprevention. Mice received subcutaneous injections of SA-4-1BBL or agonistic 3H3 antibody, and the injection-site tissue (IS) and draining lymph nodes (LN) were analyzed for differential gene expression. SA-4-1BBL induced a compartmentalized and temporally dynamic immune program characterized by early effector activation at IS and sustained immune regulation in draining LN. K-means clustering of 4564 DEGs identified eight functionally distinct clusters. IS-enriched clusters contained activation genes for CD4+ T and NK cells, including Cd28, Klra1, Cd4, Cd40, and Cd40l, while LN clusters were enriched for regulatory genes (Tnfaip3, Irf5, Col1a2) that ensure immune priming and homeostatic restraint for a balanced response. SA-4-1BBL generated a more selective and durable activation of adaptive immunity, TCR signaling, Th1/Th2 differentiation, and NK cytotoxicity. 3H3 activated broader innate inflammatory programs, including Toll-like receptor and neurodegeneration-linked pathways. IMPRes analysis showed that SA-4-1BBL activates sequential immune-regulatory circuits centered on Stat1, Cd247, and Ifng and modulates the CD151-TGF-β axis. These findings demonstrate that SA-4-1BBL elicits a balanced immune response, ensuring both safety and efficacy in preventing cancer development.

Aging is a multifaceted process impacting physiological, genomic, metabolic, and immune functions. This study investigates the role of luminal fecal exosomes (LFEs) in age-associated metabolic dysfunction. We analyzed LFEs from young (3-month) and old (24-month) male and female C57BL/6 mice to characterize age-related differences in exosomal proteomic and miRNA cargos. To explore interactions between LFEs and the gut microbiome, naïve young mice were gavage fed with LFEs from old donors, followed by 16S rRNA sequencing. Gut permeability in vitro and in vivo and systemic metabolic effects were assessed using ECIS, 3D microfluidic models, and insulin sensitivity assays. Bioinformatic analyses identified specific proteins and miRNAs linked to insulin resistance and barrier dysfunction. Heatmaps and principal component analysis revealed distinct differences in LFE profiles between young and old mice. Notably, LFEs from old mice impaired gut barrier integrity and metabolic function in young recipients, with reciprocal effects noted in older mice when receiving LFEs from young mice. Multi-omics profiling, including proteomics and miRNA sequencing, identified age-dependent and gender-related changes in LFE cargo, encompassing host- and GM-derived proteins and miRNAs. These age-specific profiles were associated with pathways implicated in cancer, neurobehavioral changes, and metabolic dysfunction. Our findings highlight that LFEs from old mice are enriched with proteins and miRNAs involved in insulin resistance and gut barrier disruption. Together, these findings identify gut luminal exosomes as age-dependent mediators of microbiome-host communication that contribute to intestinal barrier dysfunction and metabolic decline.

Extracellular ATP (eATP) and L-Glutamic acid (L-Glu) are important damage-associated molecular pattern (DAMP) molecules released from cells during injury. Both molecules trigger wound-associated signal transduction pathways, as well as the enhanced production of reactive oxygen species (ROS) by the RESPIRATORY BURST OXIDASE HOMOLOG D (RBOHD) protein. However, whether eATP and L-Glu trigger overlapping or distinct pathways is mostly unknown. Here, we report that Arabidopsis (Arabidopsis thaliana) responses to eATP or L-Glu are distinct from each other in terms of tissue specificity and transcriptomic responses. Thus, although both DAMPs trigger the expression of multiple wounding and hormone response transcripts in systemic tissues, eATP and L-Glu-induced transcripts have little overlap between them. We further show that wounding of different tissues may result in ROS responses that are controlled by different DAMP receptors. Thus, activation of ROS production following injury of non-vascular tissues primarily depended on the eATP receptors PURINORECEPTOR 2 KINASE 1 and 2 (P2K1P2K2), while activation of ROS responses in vascular tissues following injury primarily depended on the L-Glu receptors GLU-LIKE RECEPTORS 3.3 and 3.6 (GLR3.3GLR3.6). Interestingly, we found that in the absence of the GLR3.3GLR3.6 receptors (ie, in the glr3.3glr3.6 double mutant), the ROS response to eATP application is enhanced. This finding suggests that the L-Glu pathway may suppress the eATP pathway during wounding. Taken together, our findings suggest that the DAMP molecules eATP and L-Glu have complex interactions that appear to be both partly complementary and partly antagonistic, as well as tissue dependent.

Introduction Accurate cell type annotation remains a major bottleneck in plant single-cell RNA sequencing (scRNA-seq), where existing tools are often adapted from animal studies and perform sub-optimally on plant data. The lack of plant-specific computational frameworks limits the construction of plant cell atlases and downstream biological discovery. Objectives We develop and evaluate scPlantAnnotate, a Transformer-based reference annotation framework tailored for plant scRNA-seq data, and benchmark it against state-of-the-art deep learning and conventional methods across multiple plant species. Methods Species-specific scPlantAnnotate models were trained using curated datasets from Arabidopsis thaliana, Zea mays, Oryza sativa, and Glycine max. We compared scPlantAnnotate with leading baselines under both standard random-split evaluation and a more stringent leave-one-dataset-out setting, which tests robustness to completely unseen datasets and tissue types. Results scPlantAnnotate consistently outperforms existing approaches across all four species under random-split evaluation. In the leave-one-dataset-out setting for A. thaliana, where performance drops markedly for all methods due to strong batch effects and dataset heterogeneity, scPlantAnnotate nonetheless achieves the highest Accuracy, Macro-F1, Balanced Accuracy, and Macro-AUROC on average and ranks first on most held-out datasets. These results demonstrate improved robustness to dataset shifts, a critical yet underexplored challenge in plant scRNA-seq analysis. A freely accessible web server enables users to annotate their own datasets using pretrained models. Conclusion scPlantAnnotate provides a plant-specific, Transformer-based framework for single-cell annotation that delivers state-of-the-art performance and enhanced robustness to unseen datasets. By addressing limitations of existing tools and enabling scalable reference-based annotation, scPlantAnnotate supports the development of comprehensive plant cell atlases and facilitates broader use of single-cell genomics in plant biology.

Genomic resources for underutilized crops like sorghum [Sorghum bicolor (L.) Moench] often lag behind major staples, hindering efforts to link genetic diversity to agronomic traits. Improvements in genomic sequencing and bioinformatics have advanced crop genomics, yet species-specific challenges persist in the lack of tailored genomic resources, limiting precision breeding and applied genomics applications. This study presents the addition of a SorghumHub to the KBCommons web portal, an applied genomics platform for Sorghum. The core focus of this framework is the Sorghum Allele Catalog Tool, with a web-based interface enabling researchers to query, visualize, and download allelic variants from curated sorghum datasets of 988 resequenced accessions. The newly improved Allele Catalog Tool now features the ability to connect variant positions with phenotype information collected for Plant Introduction (PI) accessions in the Germplasm Resource Information Network (GRIN). On the SorghumHub, users can also find the Protein Sequence Logos, a new tool complementary to the Allele Catalog Tool, which allows users to graphically explore missense amino acid sequence changes for conservation and frequency. These custom genomic tools available on the SorghumHub enable users to query genomic variants, allele frequency, and trait-associated genotype–phenotype relationships leveraging data produced through our group’s computational pipelines for variant calling and processing into curated Allele Catalog datasets. This research work demonstrates practical application of Sorghum Allele Catalog Tool and Protein Sequence Logos by highlighting different candidate alleles for the predicted flowering time (FT-like) gene SbFT12 (Sobic.006G047700) in Sorghum bicolor; additionally, we discovered a relatively rare missense allele of the betaine aldehyde dehydrogenase (BADH2) gene that may provide a new avenue for fragrant/scented sorghum. The tools in SorghumHub allow researchers to optimize the accession selection process with efficient trait targeting and bridge the gaps between genomics and breeding. This approach can assist breeders and researchers in the development of resilient and high yielding crop varieties with broader positive outcomes for the agricultural industry and global food security.

The frequency, intensity, and duration of environmental stressors, such as droughts, heat waves, floods, and pollution, are increasing due to anthropogenic activities, subjecting plants to compound conditions of ‘multifactorial stress combination (MFSC)’. These can have contrasting or additive effects on plant physiological performance and impact different ecosystems and agro-ecosystems worldwide. Using an integrative physiological, genetic, hormonal, and transcriptomic analysis, we studied the response of Arabidopsis thaliana (L.) Heynh to a combination of water deficit (WD), heat stress (HS) and/or elevated levels of CO2 (eCO2). Our findings identified unique, as well as common, responses to MFSC. We further reveal that stomatal aperture regulation under conditions of MFSC is controlled by a combination of different pathways that is different than that activated in plants under less complex stress conditions. Under a combination of WD+HS+eCO2, for example, stomatal closure requires the function of the S-type anion channel SLAH3, but not the S-type anion channel SLAC1, or many other regulators typically found to control stomatal aperture under simpler conditions of abiotic stress/stress combinations. Our findings highlight the complexity of plant physiological and molecular responses to conditions of MFSC and open the way for future studies focused on how climate change and increased pollution impact different ecosystems and agro-ecosystems worldwide.

Soybean [Glycine max (L.) Merr.] cultivars are presumed to be highly homogeneous; however, intra-cultivar variation has been reported in previous studies. The objective of this study was to characterize intra-cultivar variation and utilize it to identify sublines that out-perform source cultivars for agronomic and seed composition traits. Additionally, we aimed to identify quantitative trait loci (QTLs) and candidate genes underlying these traits. Multiple sublines derived from high-yielding soybean cultivars outperformed source cultivars for both agronomic and seed composition-related traits. Across subline populations, protein content QTLs were identified on soybean chromosomes 1, 10, 14, and 19, with the effect sizes ranging from 3 to 7 g kg-1. Additionally, two QTLs on chromosomes 10 and 16 were identified for oil content with effect sizes of 5 and 4 g kg-1, respectively. These protein and oil QTLs were non-pleiotropic for seed composition and yield, offering a valuable resource for soybean breeders to improve seed composition without negatively impacting grain yield. An additional QTL located on chromosome 13 was associated with plant height. Through investigating functional impacts of genetic variation, we were able to identify candidate genes underlying QTL. A candidate gene, Glyma.13G196000, for plant height was identified containing a similar to 40 bp deletion encompassing the stop codon. Overall, this study provides a framework for characterization of genomic regions associated with important quantitative traits in soybean. Furthermore, our results demonstrate the potential of exploiting intra-cultivar variation for agronomic and seed quality trait improvement.

A combination of drought and heat stress is highly disruptive to crop yields worldwide. Previous studies of this stress combination were primarily conducted under controlled growth conditions, revealing that it resulted in the activation of unique molecular and physiological stress responses. However, whether similar responses to a combination of drought and heat stress occur under field conditions remains largely unknown. Here we report on a two-year field study of the transcriptomic and physiological responses of vegetative and reproductive tissues of soybean (Glycine max) to water deficit, heat treatment, and their combination. Our findings reveal that the transcriptomic responses of soybeans grown in the field are different from those grown under controlled growth conditions. These differences were especially noticeable in plants subjected to the heat or water deficit treatments, and less in plants subjected to the combination of these stresses. In addition, we report that differential transpiration between leaves and pods, that was originally discovered in plants grown under controlled growth conditions, occurs in field-grown soybeans in response to heat stress, as well as heat stress combined with water deficit. Our findings identify novel transcriptomic patterns unique to field-grown plants and highlight the need for more omics and physiological studies of crops grown under field conditions.

Zika virus (ZIKV) is the lone member of Flavivirus family known to cause congenital glaucoma following in utero exposure. The molecular mechanisms of ZIKV-induced glaucoma remain elusive, with no known therapeutic modalities. Autophagy plays a dual role in viral infections and glaucoma pathogenesis. However, how ZIKV interferes with autophagic pathways in the trabecular meshwork (TM) and whether autophagy contributes to ZIKV-induced ocular complications remains elusive. Here, we investigated the role of autophagy in ZIKV ocular pathogenesis utilizing a primary human trabecular meshwork cells (HTMC) and IFNAR1-/- mouse model of ocular infection. Our results show that ZIKV permissively infects HTMC, elicits a dysregulated cytokine response, and triggers autophagic activity in TM and the anterior segment of the IFNAR1-/- mouse eyes. Although ZIKV initially activates autophagy in TM, it impairs the overall autophagic flux during infection by modulating the VPS39, a HOPS complex, and STX17, a SNARE complex protein, to inhibit autophagosomal maturation. ZIKV utilizes late endosomes/lysosomes for its replication and egress in TM. We further demonstrated that pharmacologic inhibition of autophagy at the autophagosomal-lysosomal fusion stage by hydroxychloroquine (HCQ), or bafilomycin A1 (Baf-A1), restricts viral replication in TM and ZIKV-induced ocular pathology in mice. Using RNA-seq analysis, we uncovered that autophagy modulation modulates ZIKV-altered host transcriptomes governing critical cellular pathways, including host immunity, antigen processing and presentation, and cell death. Our study is the first to demonstrate the mechanistic insight by which ZIKV manipulates autophagic activity in the ocular milieu, and precise modulation of autophagy could be a potential therapeutic avenue to treat or prevent ZIKV-induced ocular complications. IMPORTANCE:In utero exposure to Zika virus (ZIKV) causes congenital glaucoma; however, the molecular mechanisms of ZIKV-induced glaucoma remain unknown. Here, we demonstrated that ZIKV activates the autophagic activity in the trabecular meshwork and impairs the overall autophagic flux during infection to avoid autolysosomal maturation and uses late-endosomes/lysosomes for their egress. Inhibition of autophagy at late stages, specifically at the autophagosomal-lysosomal fusion stage, restricts viral replication and ZIKV-induced ocular complications.

Plants use sophisticated signaling networks to communicate with each other. This process is thought to support the overall health and resilience of plant communities, but could also reflect eavesdropping between plants, used for competition. Here we reveal that plants that physically touch each other aboveground are more resilient to stress, and that this phenomenon is dependent on the ability of plants to exchange electric, calcium, and reactive oxygen species (ROS) signals with each other. We further separate electric-from calcium/ROS-dependent plant-to-plant signals and transcriptional landscapes and show that the function of several calcium/ROS-dependent transcripts is required for stress acclimation. Our study reveals that plants that live together and physically touch each other establish an aboveground community-wide signaling network that enhances their collective resilience to stress. ### Competing Interest Statement The authors have declared no competing interest. U.S. National Science Foundation, https://ror.org/021nxhr62, IOS-2414183, IOS-2110017, IOS-2343815

The frequency, intensity and duration of global change factors and/or environmental stressors, such as droughts, heat waves, floods, and pollution, are increasing due to anthropogenic activities, subjecting plants to compound conditions of ‘Global Change Factor combination’ (GCFc), or ‘stress combination’. These can have contrasting or additive effects on plant physiological performance and impact different ecosystems and agro-ecosystems worldwide. Using an integrative physiological, genetic, hormonal, and transcriptomic analysis, we studied the response of Arabidopsis thaliana (L.) Heynh to a combination of water deficit (WD), heat stress (HS) and/or elevated levels of CO2 (eCO2). Our findings reveal high specificity in plant responses to GCFc. We further reveal that stomatal aperture regulation under conditions of GCFc is controlled by a blend of unique and shared regulators, that together determine the specific aperture size under each different set of GCFc conditions. Under a combination of WD, HS, and eCO2 (WD+HS+eCO2), for example, stomatal closure required the function of nitric oxide, OPEN STOMATA 1 and the S-type anion channel SLAH3, but not the S-type anion channel SLAC1, or many other regulators typically found to control stomatal aperture under less complex conditions, including HS+WD. Our findings highlight the complexity of plant physiological and molecular responses to conditions of stress combination and open the way for future studies of plant responses to stress combination, crucial to our understanding of how GCFc impacts different ecosystems and agro-ecosystems worldwide. In addition, our study provides an initial definition for a ‘stomatal hierarchical stress code’ that could apply to future studies. ### Competing Interest Statement The authors have declared no competing interest. National Science Foundation, https://ror.org/021nxhr62, IOS-2414183; IOS-2110017, IOS-2343815

Serotonin reuptake inhibitors (SSRIs) are commonly prescribed to pregnant women experiencing depression. Such drugs, however, might adversely affect placenta and fetal brain development. Parietal trophoblast giant cells (pTGCs) in the mouse placenta are postulated to internalize maternal serotonin (5-HT) via transport through SERT, encoded by Slc6a4, and to provide the initial source of 5-HT to the emerging brain via the placental-brain axis. Genetic deletion of Slc6a4 in pTGCs has been hypothesized to impact placental and fetal brain development. A transgenic mouse line with high-affinity SERT, encoded by Slc6a4, was selectively deleted by pairing mice with Cre recombinase linked to Prl2c2, with LoxP sites flanking the Slc6a4 gene. PRL2C2 is solely expressed by pTGCs and other giant cells of the placenta. To compare placental and fetal brain development in selective Slc6a4 KO and WT mice, 5-HT content in the placenta and fetal brains of conceptuses was measured. No significant differences in 5-HT content were evident between knockout (KO) and wild-type (WT) placentas or fetal brains. However, there were significantly fewer pTGCs in KO placentas compared to WT (p ≤ 0.05). Sexually dimorphic differences in gene expression were evident in the placenta and fetal brain between KO and WT counterparts, with female conceptuses showing the most dramatic responses, including decrease in Prl7a2, Prl5a1, Prl3a1, Slc28a3, and Ceacam 15 in female placental samples. These findings suggest that ablation of Slc6a4 in pTGC disrupts the placenta-brain axis in a sex-dependent manner. The results might have important clinical ramifications for pregnant women being treated with SSRIs.

Multifactorial stress combination (MFSC) is emerging as a major constraint to crop productivity under different climate change scenarios. While the physiological impacts of MFSC have been previously characterized in different plant species, the molecular and metabolic effects of MFSC remain poorly defined. Here, we used an integrative multi-omics approach to dissect the response of tomato (Solanum lycopersicum) plants to an MFSC of up to 6 low-intensity abiotic stressors. Our analysis uncovered a complexity-dependent molecular program in tomato. Transcriptomic analysis identified a core set of 194 transcripts commonly altered across all stress conditions, along with 155 transcription factors (TFs) specifically regulated under high-complexity conditions (4-, 5-, and 6-stress combinations). Focusing on heat-associated MFSC responses, we identified 103 transcripts uniquely responsive to these conditions, including 2 TFs (Zinc finger TF 32 and a B3 family protein) that may act as master regulators of all heat-associated MFSCs. Metabolomic profiling revealed a pronounced reprogramming of primary metabolism under MFSC, marked by decreased levels of tricarboxylic acid intermediates and accumulation of sugars, γ-aminobutyric acid, and branched-chain amino acids, suggesting a trade-off that favors osmoprotection and redox homeostasis over energy-intensive processes. Comparative analyses across tomato, Arabidopsis, Chlamydomonas, rice, and soybean highlighted a conserved molecular signature associated with MFSC. Integrated omics correlation analysis uncovered functional links among phytohormone signaling, photosynthetic efficiency, and key MFSC-related transcripts and metabolic hubs. Together, we reveal a coordinated and complexity-dependent molecular program in tomato, offering insights into plant adaptation to MFSC and identifying candidate regulatory and metabolic markers for engineering climate-resilient crops.

How biological systems respond to stress is a fundamental question in biology, primarily addressed using the reductionist approach of applying one stress condition at a time. In nature, however, organisms experience a multitude of stresses, simultaneously or sequentially, questioning the validity of the reductionist approach for predicting plant responses to stress under natural conditions. Here, we reveal that in the flowering plant Arabidopsis thaliana, the transcriptional regulator bHLH35 is required for plant survival under a specific set of stress conditions that includes a combination of salinity, excess light, and heat, occurring simultaneously (but not for each of these stresses applied individually or in any other combination). Under these conditions, bHLH35 interacts with NAC069 and binds the promoter of LBD31, also specifically required for survival under the 3-stress combination. Our findings uncover a high degree of specificity in the response of organisms to stress, a specificity that would not have been revealed using the reductionist approach, and one that should be taken into consideration when developing agronomically important crops with heightened resilience to climate change.

Global change factors associated with climate change and increased pollution are subjecting plants, microbes, and different ecosystems to conditions of multifactorial stress combination (MFSC). While recent studies have centered on the effects of MFSC on different plants and ecosystems, much less is known about how these conditions impact unicellular organisms. Here, we report on the physiological and proteomic responses of wild-type and respiratory burst oxidase homolog 1 (rbo1) mutant cells of Chlamydomonas reinhardtii to an MFSC of 5 different abiotic stresses. While several similarities were found between plant and C. reinhardtii responses to MFSC, our work revealed that MFSC induces aggregation in the unicellular organism C. reinhardtii. We further show that MFSC-induced aggregation is enhanced in the rbo1 mutant and can be triggered by H2O2. As aggregation typically leads to reduced growth, respiration, and photosynthesis, enhanced aggregation of unicellular organisms in different ecosystems subjected to MFSC could explain part of the negative impacts of MFSC on the services they provide. Our findings shed light on the response of unicellular organisms to MFSC and suggest that one of the main drivers leading to multicellularity during evolution was early conditions of MFSC under an oxygenated environment.