Biology subjects

Cerny, M.

Publications and source records attributed to Cerny, M..

14 recordsLinked to original sources

bioRxiv2026

Glycogen removal enhances ethylene production in Synechocystis sp. PCC 6803 but exposes a 2-oxoglutarate-GS-GOGAT-phosphate constraint

Cyanobacteria are promising platforms for converting CO2 directly into value-added chemicals. However, redirecting fixed carbon toward a product requires understanding of how cells balance the energetic and redox supplies and demands - and how efficiently metabolic flux can be channelled into product formation rather than being dissipated or excreted. Glycogen, synthesized via ADP-glucose pyrophosphorylase (GlgC), is the major native carbon and energy sink, and its removal forces cells to redirect excess carbon and reducing power into alternative pathways. As a target product we used ethylene, a major industrial building block currently derived almost entirely from fossil feedstocks, which can be produced photoautotrophically by the ethylene-forming enzyme (EFE) at the expense of 2-oxoglutarate (2-OG), a central node linking carbon and nitrogen metabolism. Here, we compared wild-type Synechocystis sp. PCC 6803 with strains carrying either a complete ({Delta}glgC) or attenuated, light-responsive (LGS; psbA2-driven glgC) glycogen synthesis block, with an EFE-expressing strain (EFE), and with the combined {Delta}glgC+EFE strain. All strains were cultivated across a light gradient (35-300 mol photons m-2 s-1) under nitrogen-replete conditions. Integrating measurements of growth and ethylene production rates, chlorophyll-fluorescence analysis, quantitative proteomics, FTIR profiling, and overflow-metabolite analysis revealed that partial suppression of glycogen synthesis (LGS) enhanced growth under low irradiance but became detrimental above a critical light threshold. EFE expression in the {Delta}glgC background increased ethylene production in a light-dependent manner but further compromised growth. Both strains with EFE increased photosynthetic electron transport consistent with the carbon-sink effect, yet this electron surplus was not channelled proportionally into product or biomass. Instead, continuous 2-OG drain by EFE constrained flux through the GS-GOGAT node and induced a secondary inorganic phosphate limitation in {Delta}glgC+EFE that activated photoprotection and limited productivity. These results point to the 2-OG/GS-GOGAT/Pi axis as a key design constraint determining whether the surplus photosynthetic flux gets converted into product, dissipated through photoprotection, or released as overflow metabolites.

plant biology↗

bioRxiv2026

Cytokinin Mediates Photosynthesis and Senescence Action Through CHLOROPLAST IMPORT APPARATUS 2 (CIA2) and CIA2-like (CIL)

Progression of leaf senescence is regulated by internal factors like the plant hormone cytokinin (CK) that acts to delay senescence and retain photosynthesis by maintaining chloroplast-functionality. While initial CK-signaling components controlling senescence have been described, downstream targets regulating physiological performance have remained less characterized. Here we present the CHLOROPLAST IMPORT APARATUS 2 (CIA2) and CIA2-like (CIL) transcription factors (TFs) as mediators of CK senescence activity, enabling chloroplast-function. Utilizing loss-of-function mutants, we show CIA2 and CIL negatively regulate senescence progression and alter physiological performance in Arabidopsis thaliana. Integrated transcriptomic-metabolomic analysis shows that CIA2 and CIL action is distinct from other known chloroplast-related TFs (like GLKs/GATAs) and can boost antioxidant response over senescence. We find that CIA2 and CIL are essential in relaying CK activity during chloroplast biogenesis, photosynthesis, rubisco function, and metabolite accumulation. In turn, ARR10/12 CK signaling components were needed for CK induction of CIA2 and CIL themselves - a cascade that induced chloroplast- and photosynthesis-associated nuclear genes, to boost chloroplast-function and metabolism. We also show that CK acts to repress senescence through accumulation of the polyamine, spermidine, in a CIA2/CIL dependent manner. This work presents a CIA2/CIL-cascade as a new anterograde signal for CK antisenescence effects, indicating a mechanism through which CK prolongs leaf function. This fundamental understanding of CK action has implications towards engineering plants with elevated photosynthesis performance and delayed senescence.

plant biology↗

bioRxiv2026

Nutrient Availability Modulates Beneficial Effect of Bacterial Community Volatiles and Contact-Dependent Interactions Differently

Plant growth-promoting bacteria enhance plant performance, yet how different modes of plant-microbe interaction shape nutrient-specific host responses remains poorly understood. In particular, it is unclear how direct bacterial contact and volatile-mediated interactions originating from the same bacterial community differentially regulate plant nutrient acquisition pathways. Here, we investigated how a 16-member synthetic bacterial community (16SC) affects plant growth, nutrient status, signaling, and metabolite profiles under full nutrient supply as well as nitrogen (N), sulfur (S), and phosphorus (P) limitation in Arabidopsis thaliana. We show that volatile organic compounds (VOCs) emitted by the 16SC promote shoot growth under nitrate limitation and full nutrient conditions, whereas this growth promotion is lost under sulfur- and phosphorus-limiting conditions. In contrast, direct interaction (DBC) between plants and the 16SC abolishes growth promotion under all three nutrient-limiting conditions. These nutrient-dependent phenotypes correlate with distinct regulation of nutrient transporters and key transcriptional regulators involved in N (NRT1;1 / NLP7), S (SULTR1;2 / SLIM1/EIL3), and P (PHO2 / PHR1) signaling pathways. Genetic analyses using nutrient transporter mutants revealed that VOC-induced growth promotion requires functional NRT1;1 and SULTR1;2 transporters, whereas growth promotion mediated by direct bacterial contact is retained in the corresponding mutants. This uncoupling of VOC- and contact-dependent effects indicates that distinct host regulatory pathways underlie bacterial community growth promotion depending on the interaction mode. Together, our findings demonstrate that bacterial community-mediated plant growth promotion is strongly shaped by nutrient context and interaction mode, and that volatile-mediated and contact-dependent mechanisms engage separable host nutrient regulatory networks.

plant biology↗

bioRxiv2026

Cytokinin N-conjugate Form Activity, Metabolism, and Signaling During Leaf Senescence

Cytokinin (CK) N-glucosides are the most abundant CK metabolites in Arabidopsis and most angiosperms, yet their role in cytokinin activity and response is unclear. Here, we examined metabolomic, transcriptomic, and proteomic profiles of seven CK N-glucoside conjugates in detached Arabidopsis leaves across a 144-hour dark-induced senescence (DIS) timecourse. All tested N-glucosides were found to undergo a slow conversion to their corresponding base forms at position-dependent rates, with N9-glucosides releasing base faster than their corresponding N7-glucosides. Conversion during DIS was strictly isoform-specific and not accompanied by coordinated induction of CK biosynthesis genes, arguing against de novo synthesis as the source of accumulated base. Despite progressive base accumulation, N-glucoside-treated leaves produced substantially fewer Differentially Expressed Genes than direct base application at comparable base concentrations, revealing a disconnect between hormone presence and transcriptional output. Unbiased model comparison identified the base:glucoside ratio as a stronger predictor of CK-Two Component Signaling (TCS) gene expression than absolute base concentration, though modulated by base-type-specific receptor affinities. Early proteomic profiling further revealed a coordinated response shared across N-glucosides but largely absent from base treatments. Together, these findings support that CK N-glucosides as kinetically slow, position-dependent reservoirs whose presence in abundance modulate activation of CK-TCS elicited by bioactive forms. HighlightsPhysiology, metabolomic, transcriptomic, and proteomic findings here support CK N-glucosides as kinetically slow, position-dependent reservoirs whose presence in abundance modulate activation of CK-TCS elicited by bioactive forms.

plant biology↗

bioRxiv2026

Cytokinin Senescence Delay Is Shaped by Receptor Specificity and Metabolic Stability

Each of the four different cytokinin (CK) base forms, trans Zeatin (tZ), isopentenyladenine (iP), dihydrozeatin (DHZ), and cis Zeatin (cZ) have distinct chemical metabolism and affinity to the CK Histidine Kinase (CHK) receptors. However, it remains unclear how specific biochemical features of each form such receptor specificity or metabolic differences drives distinct tissue-specific physiological hormone output in response to application of these CK bases. Here, we show that CK receptor preference and metabolic persistence together shape isoform-specific CK signaling strength, including tissue-dependent hormone responses in Arabidopsis leaf versus root assays. Physiological, genetic, and multi-omics integration was used to show that tZ and iP anti senescence activity is matched by DHZ through a distinct receptor metabolic mechanism. DHZ requires Arabidopsis Histidine Kinase 3 (AHK3) signaling to be fully effective in a leaf Dark Induced Senescence (DIS) assay and where it overcomes its lower receptor affinity through higher metabolic persistence, accumulating at levels [~]2.5-fold above tZ and iP early in a senescence time course. Together, these findings provide a framework for integration of receptor preference and metabolic stability to determine CK isoform activity.

plant biology↗

bioRxiv2025

Citizen science and detailed searches reveal widespread occurrence of hybrid-like Syrian x Great-spotted woodpeckers

The Syrian Woodpecker (hereafter SW) expansion into Europe since the end of the 19th century has enabled its hybridisation with the Great spotted Woodpecker (hereafter GW). This phenomenon, however, has generally been considered sporadic. Here, we examined the occurrence of mixed pairs and hybrids based on our own field observations and citizen science data. Interspecific pairs and individuals of SW and GW were found to be generally scarce but occurred in all parts of the area investigated, not only at the edges of the SWs range. The proportion of hybrid occurrence in woodpecker populations was found to be similar across the whole SWs range in Europe and in both urban and rural populations. However, on a local level, hybrids seemed to be more abundant in certain regions, for example, E Austria, Moravia, SE Poland, W Russia, S Ukraine, Bulgaria, E Greece and W Turkey. Generally, females were 1.4 times more frequent among hybrids. Three major phenotypical hybrid forms were identified: GW-like, SW-like, and intermediate, with first two forms being more frequent in the central and south of Europe respectively. Whereas proportion of the intermediate form was relatively constant across the whole range of hybrid occurrence. The results indicate that hybrids of the two species have become significant part of woodpecker populations and thus should be considered in research and nature conservation projects.

zoology↗

bioRxiv2025

Multilevel analysis of response to plant growth promoting and pathogenic bacteria in Arabidopsis roots and the role of CYP71A27 in this response

Understanding how plants distinguish between commensal and pathogenic microorganisms is one of the major challenges in the plant microbe interaction research. We previously identified a gene encoding CYP71A27 connected to camalexin, which is necessary for a plant growth promoting (PGP) activity of a number of bacterial strains. To dissect its function, we compared multilevel responses in roots of wild type Arabidopsis and the cyp71A27 mutant to two bacterial strains, a PGP Pseudomonas fluorescens CH267 and a pathogen Burkholderia glumeae PG1. We show that incubation with these bacteria leads to significant and distinct transcriptional reprogramming. This is accompanied by a proteome remodelling in both shoots and roots and profound changes in accumulation of many metabolites, primarily sugars, amino acids, and TCA cycle intermediates, but also by alterations in the ionome. We then analysed the mutant cyp71a27 and identified number of genes and proteins differently regulated, particularly after interaction with the PGP strain, but only a very mild impact of the mutation on root metabolites and exudates. We analysed a variety of mutants in genes differentially regulated by Pseudomonas sp. CH267 in cyp71a27 and revealed that their response to this PGP bacterial strain is similar to cyp71a27. Thus, it seems that CYP71A27 is a non-canonical P-450 without a metabolic function but active in signalling pointing to a regulatory role of the CYP71A27 gene, particularly in interaction with a plant growth promoting bacteria.

plant biology↗

bioRxiv2024

Light quantity impacts early response to cold and cold acclimation in young leaves of Arabidopsis

Plant reactions to stress vary with development stage and fitness. This study assessed the relationship between light and chilling stress in Arabidopsis acclimation. By analyzing the transcriptome and proteome responses of expanding leaves subjected to varying light intensity and cold, 2251 and 2064 early response genes and proteins were identified, respectively. Many of these represent as yet unknown part of early response to cold, illustrating development-dependent response to stress and a duality in plant adaptations. While standard light promoted photosynthetic upregulation, plastid maintenance, and increased resilience, low light triggered a unique metabolic shift, prioritizing ribosome biogenesis and lipid metabolism and attenuating expression of genes associated with plant immunity. The comparison of early response in young leaves with that in expanded ones showed striking differences, suggesting a sacrifice of expanded leaves to support young ones. Validations of selected DEGs in mutant background confirmed a role of HSP90-1, transcription factor FLZ13, and Phospholipase A1 (PLIP) in response to cold, and the PLIP family emerged as crucial in promoting acclimation and freezing stress tolerance. The findings highlight the dynamic mechanisms that enable plants to adapt to challenging environments and pave the way for the development of genetically modified crops with enhanced freezing tolerance.

plant biology↗

bioRxiv2024

Expanding the CarD interaction network: CrsL is a novel transcription factor in Mycobacterium smegmatis

Bacterial transcription regulation is critical for adaptation and survival. CarD is an essential transcription factor in mycobacteria involved in regulation of gene expression. We searched for CarD interaction partners in the model organism Mycobacterium smegmatis and identified two proteins: ApeB (MSMEG_5828) and an uncharacterized protein, which we named CrsL (MSMEG_5890). While ApeB interacted with CarD only when CarD was overexpressed, CrsL associated with CarD at its physiological levels. CrsL is a 5.7 kDa protein shown by NMR to be intrinsically disordered. CrsL homologs are present in actinobacteria including pathogenic species such as Mycobacterium tuberculosis. CrsL directly interacts with CarD and binds RNAP. ChIP-seq showed that CrsL associates with promoters of actively transcribed genes and [~]75 % of these regions are also associated with CarD. RNA-seq showed [~]50% and [~]66% overlap in differentially expressed genes between CrsL and CarD knockdowns during exponential and stationary phases, respectively. CrsL represses expression of DesA desaturase (MSMEG_5773) and DEAD/DEAH-box RNA helicase MSMEG_1930, which are important for adaptation to cold stress. Furthermore, CrsL promotes the growth of M. smegmatis at elevated temperature. In summary, this study identifies CrsL as a novel actinobacterial transcription factor and provides a basis for its further investigation.

microbiology↗

bioRxiv2024

Diversity and impact of single-stranded RNA viruses in Czech Heterobasidion populations

Heterobasidion annosum sensu lato comprises some of the most devastating conifer pathogens conifers. Exploring virocontrol as a potential strategy to mitigate economic losses caused by these fungi holds promise for the future. In this study, we conducted a comprehensive screening for viruses in a 98 H. annosum s.l. specimens from different regions of Czechia aiming to identify viruses inducing hypovirulence. Initial examination for dsRNA presence was followed by RNA-Seq analyses using pooled RNA libraries constructed from H. annosum and Heterobasidion parviporum, with diverse bioinformatic pipelines employed for virus discovery. Our study uncovered 25 distinct ssRNA viruses, including two ourmia-like viruses, one mitovirus, one fusarivirus, one tobamo-like virus, one cogu-like virus, one bisegmented narna-like virus and one segment of another narna-like virus, and 17 ambi-like viruses, for which hairpin and hammerhead ribozymes were detected. Coinfections of up to 10 viruses were observed in six Heterobasidion isolates, while another six harbored a single virus. 73% of the isolates analyzed by RNA-Seq were virus-free. These findings show that the virome of Heterobasidion populations in Czechia is highly diverse and differs from that in the boreal region. We further investigated the host effects of certain identified viruses through comparisons of the mycelial growth rate and proteomic analyses and found that certain tested viruses caused growth reductions of up to 22% and significant alterations in the host proteome profile. Their intraspecific transmission rates ranged from 0% to 33%. Further studies are needed to fully understand the biocontrol potential of these viruses in planta. ImportanceO_LIFirst report of a fusarivirus, a tobamo- and a cogu-like virus in Heterobasidion C_LIO_LICertain viruses caused mycelial growth reduction in H. annosum host strains C_LIO_LIViral infections lead to proteome changes in Heterobasidion C_LI

microbiology↗

bioRxiv2023

Lack of Purple Acid Phosphatase SlPAP26b compromises the phosphorous starvation response in tomato independent of SlPHR1 and SlPHL1

The scarcity of soil phosphorus (P), an essential macronutrient, often limits plant growth and development. Enhanced secretion of intracellular and secretory acid phosphatases is essential to maintain cellular inorganic P (Pi) homeostasis in plants. Herein, using transcriptomics and proteomics approach, we observed upregulation of several purple acid phosphatases (PAPs), including SlPAP1, SlPAP10b, SlPAP12, SlPAP15, SlPAP17b, SlPAP26a, and SlPAP26b in Pi-deficient tomato seedlings. Higher transcript levels of SlPAP17b and SlPAP26b in the older senescing leaves than the younger leaves indicated active involvement of these PAPs in Pi remobilization. Subsequent detailed characterization of SlPAP17b, SlPAP26a, and SlPAP26b revealed a prominent role of SlPAP26b in Pi homeostasis. Silencing of SlPAP26b led to an exacerbated P starvation response as these plants exhibited smaller shoots, lower soluble Pi, total P levels, and higher sucrose than their EV controls under Pi deprivation. SlPAP26b-silenced plants also showed misregulation of P starvation inducible genes such as phosphate transporters and glycerolipid remodellers, even under Pi-sufficient conditions. Whereas SlPAP26b levels were induced by external sucrose, its expression was found to be independent of the Myb class master regulators of P starvation response, SlPHR1 and SlPHL1. Altogether, this study identifies a prominent role of SlPAP26b in the Pi compensation network in tomato seedlings.

plant biology↗

bioRxiv2020

COP1 destabilizes DELLA proteins in Arabidopsis

DELLA transcriptional regulators are central components in the control of plant body form in response to the environment. This is considered to be mediated by changes in the metabolism of the hormones gibberellins (GAs), which promote the degradation of DELLAs. However, here we show that warm temperature or shade reduced the stability of a GA-insensitive DELLA allele in Arabidopsis. Furthermore, the degradation of DELLA induced by the warmth anticipated changes in GA levels and depended on the E3 ubiquitin ligase CONSTITUTIVELY PHOTOMORPHOGENIC1 (COP1). COP1 enhanced the degradation of normal and GA-insensitive DELLA alleles when co-expressed in N. benthamiana. DELLA proteins physically interacted with COP1 in yeast, mammalian and plant cells. This interaction was enhanced by the COP1 complex partner SUPRESSOR OF phyA-105 1 (SPA1). The level of ubiquitination of DELLA was enhanced by COP1 and COP1 ubiquitinated DELLA proteins in vitro. We propose that DELLAs are destabilized not only by the canonical GA-dependent pathway but also by COP1 and that this control is relevant for growth responses to shade and warm temperature. SignificanceDELLA proteins are plant-specific transcriptional regulators that act as signaling hubs at the interface between the environment and the transcriptional networks that control growth. DELLAs are destabilized by the growth-promoting hormone gibberellin, whose levels are very sensitive to environmental changes. Here we describe an alternative pathway to destabilize these proteins. We show that DELLAs are substrate of COP1, an E3 ubiquitin ligase that increases its nuclear activity to promote growth in response to shade or warmth. Our results also show that the destabilization of DELLAs by COP1 precedes the action of gibberellins, suggesting the existence of a sequential mechanism to control the stability of these proteins.

plant biology↗

bioRxiv2018

Effect of methylene blue on the formation of oxidized phospholipid vesicles

Soybean phosphatidylcholine, which is rich in linoleic acid, was oxidized with singlet oxygen through photosensitization with methylene blue. This compound facilitates the oxidation of phospholipids relative to the reaction with free unsaturated fatty acids. A response surface methodology was used to control oxidation, with methylene blue concentration and the amount of available air as independent variables. The conjugated diene-to triene ratio was then monitored. Hydroperoxide yield dependent principally on the amount of air, whereas photosensitizer concentration strongly influenced the size and zeta potential of vesicles formed by the sonication of oxidized phospholipids in water. Methylene blue plays an important role in the surface charge expression and ion permeability of these vesicles.

bioengineering↗

bioRxiv2017

An algorithm for template-based prediction of secondary structures of individual RNA sequences

While understanding the structure of RNA molecules is vital for deciphering their functions, determining RNA structures experimentally is exceptionally hard. At the same time, extant approaches to computational RNA structure prediction have limited applicability and reliability. In this paper we provide a method to solve a simpler yet still biologically relevant problem: prediction of secondary RNA structure using structure of different molecules as a template.\n\nOur method identifies conserved and unconserved subsequences within an RNA molecule. For conserved subsequences, the template structure is directly transferred into the generated structure and combined with de-novo predicted structure for the unconserved subsequences with low evolutionary conservation. The method also determines, when the generated structure is unreliable.\n\nThe method is validated using experimentally identified structures. The accuracy of the method exceeds that of classical prediction algorithms and constrained prediction methods. This is demonstrated by comparison using large number of heterogeneous RNAs. The presented method is fast and robust, and useful for various applications requiring knowledge of secondary structures of individual RNA sequences.

bioinformatics↗