Biology subjects

Schmitz, C.

Publications and source records attributed to Schmitz, C..

At least 19 recordsLinked to original sources

bioRxiv2026

Pathogenic MAPK8IP3 variants drive distinct motor and behavioral phenotypes in humans and mice

Pathogenic variants in MAPK8IP3 (JIP3) cause Neurodevelopmental Disorder with or without variable Brain Abnormalities (NEDBA), characterized by cognitive impairment, global developmental delay, motor dysfunction, abnormal muscle tone, behavioral dysregulation including autism and attention deficit hyperactivity disorder (ADHD), seizures, and structural brain abnormalities. While more than 30 largely de novo MAPK8IP3 variants have been reported, the functional impact of variants across JIP3 protein structural domains is poorly defined. To address this knowledge gap, we compare clinical features of individuals with a truncating (p.E27X) or one of two missense (p.R578C and p.R1146C) variants from distinct JIP3 functional protein domains to corresponding knock-in mouse models. Our findings showed that all individuals, regardless of variant type, exhibited delays in language and gross motor function, but individual variants were associated with distinct motor, cognitive, and psychiatric symptoms. Corresponding homozygous p.E27X and p.R1147C variant mice resulted in embryonic lethality, consistent with essential roles for JIP3 in early neurodevelopment. Behavioral characterization of viable heterozygous mice revealed variant-specific locomotor, motor coordination, and hindlimb clasping defects that closely recapitulate clinical observations. Heterozygous p.R579C mice exhibited reduced locomotion, impaired motor performance, and hypertonia-like clasping, mirroring human motor deficits. In contrast, p.R1147C mice displayed hyperactivity, hindlimb clasping, and decreased brain weight, paralleling human clinical features. Together, our findings demonstrate that while distinct MAPK8IP3 variants lead to some shared phenotypes, they are also associated with distinct phenotypes that could reveal domain-specific aspects of JIP3 function. This work establishes the first domain-resolved in vivo rodent models of NEDBA and provides a validated translational platform for mechanistic investigation and preclinical therapeutic testing.

neuroscience↗

bioRxiv2026

Detergent selection as a determinant of lysate melting behavior and drug-target hit-calling in PISA

Thermal proteome profiling (TPP) and proteome integral solubility alteration (PISA) assays measure drug-target interactions by monitoring protein thermal stability across the proteome. While detergents are routinely used in lysate-based thermal profiling, the field lacks consensus on whether detergents should be present during the melting step or only added afterward as an extraction buffer, and whether detergent identity matters for this choice. Here, we evaluate how commonly used detergents and the timing of their use in thermal stability workflows affect proteome-wide thermal stability and PISA hit calling in TF-1 lysates. We find that NP-40 and DDM produce highly correlated melting profiles when used exclusively as post-melt extraction buffers, but diverge substantially when present during the melting step. DDM in particular prevents the thermally-induced loss in solubility of large classes of proteins, such as cell surface proteins, and these effects propagate directly into PISA hit calling. Performing the PISA melt in DDM versus NP-40 results in the gain and loss of distinct drug-target interactions for both the PAK4 inhibitor PF-3758309 and the PLK1 inhibitor volasertib. Notably, DDM enables detection of a volasertib-TMEM97 interaction that was previously not detected in NP-40. However, we also find that the stabilization effects of DDM mask the identification of some known PISA hits for these drugs. We further introduce a four-parameter logistic model of protein melting to aid in modeling of these findings and a linear regression framework for PISA hit calling that outperforms pairwise t-tests in low-replicate settings. Together, these results establish detergent selection as a tunable experimental variable in thermal profiling and suggest that some drug-target engagements previously attributed exclusively to intact-cell context may be recoverable in lysates with appropriate buffer conditions.

biochemistry↗

bioRxiv2026

Human lung γδ T cells maintain functionality during inflammatory lung disease

{gamma}{delta} T cells provide mucosal defense against infection while also contributing to tissue repair. However, data regarding the effect of the human lung environment on {gamma}{delta} T cell functionality remains limited. To address whether lung inflammation impacts {gamma}{delta} T cell functionality, we analyzed lung and matched hilar lymph node (LN) tissue from deceased donors and patients with interstitial lung disease (ILD). We performed high-parameter spectral flow cytometry to examine the expression pattern of phenotypic biomarkers and assess ex vivo function. We identified lung-specific enrichment of {gamma}{delta} T cells with an effector memory phenotype relative to matched regional LN. We then used an ex vivo stimulation approach to interrogate the capacity to protect against infection (granzyme B [GzmB], interferon-{gamma} [IFN{gamma}] and tumor necrosis factor [TNF]) and promote epithelial cell proliferation (amphiregulin [AREG]). We found that {gamma}{delta} T cells in lung and LN from deceased donors had similar functional properties. While {gamma}{delta} T cell populations from ILD lungs largely maintained cytokine production capacity, expression was diminished relative to LN counterparts. Importantly, lung {gamma}{delta} T cells maintained polyfunctional GzmB, IFN{gamma} and TNF expression across cohorts. Overall, we report human lung {gamma}{delta} T cells are regionally distinct with conserved functionality in a fibrotic environment.

immunology↗

bioRxiv2026

Distinct SMA beta bursts support the development of anticipatory postural control in children

To produce smooth and coordinated movements, the brain generates anticipatory postural adjustments (APA) - feedforward motor commands that counteract predicted postural disturbances before they occur. Although APA emerge early in life, their maturation extends into early adulthood, yet little is known about the underlying neural mechanisms. Using magnetoencephalography during a naturalistic bimanual load-lifting task, we investigated anticipatory postural control in children aged 7-12 years. In this task, lifting a load from the contralateral postural forearm induces upward elbow rotation, which is anticipatorily counteracted by inhibition of Biceps brachii postural activity, stabilizing forearm posture. Surprisingly, despite greater variability in inhibition timing and less efficient forearm stabilization in children, the mechanism of anticipatory inhibition was consistent with that recently described in adults: it was mediated by transient oscillatory beta bursts (19-24 Hz), associated with reduced supplementary motor area (SMA) excitability indexed by high-gamma (90-130 Hz) power suppression, and preceded by directed influences from lateral prefrontal and ventral premotor cortex. However, burst frequency was lower in children, and an additional APA mechanism was recruited: while 19-24 Hz bursts were linked to immediate SMA suppression and stronger muscle inhibition, higher-frequency bursts (24-29 Hz) were associated with delayed ([~]100 ms) SMA suppression mediated by alpha activity (8 Hz), which attenuated post-inhibition forearm instability. These results reveal that children in middle childhood already engage adult-like neural mechanisms for anticipatory inhibition, while additionally recruiting a compensatory mechanism to counteract postural imprecision, providing new insights into the neural basis of APA maturation and its alteration in neurodevelopmental conditions.

neuroscience↗

bioRxiv2025

Explosive shock wave exposure leads to age-accelerated motor and sensory decline in C. elegans

BackgroundMilitary and law enforcement personnel are routinely exposed to shock waves from weapon systems and explosive devices during training and operations. Even when such shock wave exposure results in mild traumatic brain injury (mTBI) without abnormalities on routine imaging, persistent neurological symptoms may occur. Yet, the biological processes that link an acute blast wave insult to progressive neuronal dysfunction remain poorly defined. MethodsWe established Caenorhabditis elegans nematodes as a genetically accessible animal model for blast-related mTBI (br-mTBI). Animals were exposed in gelatin to shock waves generated by a custom-built shock wave generator (SWG), which produces explosive shock waves with an abrupt overpressure peak followed by a negative pressure phase. The resulting pressure-time curves are very similar to the profiles of conventional explosives in free-field setups. To enable mechanistic studies under controlled laboratory conditions, we developed a complementary platform using a medical radial extracorporeal shock wave therapy (rESWT) device. Motor behavior, mechanosensory function, neuronal morphology and cytoskeletal integrity were analyzed during aging. ResultsSWG-derived shock waves induced immediate but reversible motor and sensory deficits, followed by an accelerated age-dependent decline in movement and touch sensitivity. The rESWT platform accurately reproduced these phenotypes. Touch receptor neurons showed progressive structural abnormalities, including degeneration, alongside acute PTL-1/Tau mislocalization consistent with cytoskeletal injury. ConclusionsDefined shock wave exposure is sufficient to provoke long-term neuronal functional decline in C. elegans, accompanied by structural deterioration and premature neurodegeneration. This tractable model enables lifelong phenotyping and mechanistic dissection of how an acute shock wave insult progresses to chronic neuronal dysfunction, and provides a scalable platform for identifying molecular and pharmacological modifiers that promote resilience.

neuroscience↗

bioRxiv2025

CRISPR-DA: Adapting CRISPR gRNA design for detection assays

CRISPR-Cas systems offer a viable alternative to traditional detection and diagnosis methods. However, their effectiveness relies heavily on the selection of appropriate guide RNA sequences. Existing gRNA design tools were primarily developed for gene editing and are not always directly applicable to CRISPR-based detection assays. In particular, alignmentbased methods are still used to estimate gRNA specificity, even though they can miss a substantial portion of off-target sites. In this work, we introduce CRISPR-DA, a CRISPR gRNA design tool for detection assays. We show that it provides a better assessment of gRNA specificity than BLAST, which detected only 33.27 % and 0.43 % of cross-species off-targets in two datasets. Additionally, CRISPR-DA ran two and six times faster than BLAST on these datasets, respectively. Our method incorporates advances from gene-editing guide RNA design tools, including uncertainty-informed guide RNA design, to improve the selection of guides with high on-target activity. CRISPR-DA is available at https://github.com/bmds-lab/CRISPR-DA.

bioinformatics↗

bioRxiv2025

Shared latent representations of speech production for cross-patient speech decoding

Speech brain-computer interfaces (BCIs) can restore communication in individuals with neuromotor disorders who are unable to speak. However, current speech BCIs limit patient usability and successful deployment by requiring large volumes of patient-specific data collected over long periods of time. A promising solution to facilitate usability and accelerate their successful deployment is to combine data from multiple patients. This has proven difficult, however, due to differences in user neuroanatomy, varied placement of electrode arrays, and sparse sampling of targeted anatomy. Here, by aligning patient-specific neural data to a shared latent space, we show that speech BCIs can be trained on data combined across patients. Using canonical correlation analysis and high-density micro-electrocorticography (ECoG), we uncovered shared neural latent dynamics with preserved micro-scale speech information. This approach enabled cross-patient decoding models to achieve improved performance relative to patient-specific models facilitated by the high resolution and broad coverage of ECoG. Our findings support future speech BCIs that are more accurate and rapidly deployable, ultimately improving the quality of life for people with impaired communication from neuromotor disorders.

neuroscience↗

bioRxiv2025

Structural basis and mechanism of action of NrdR, a bacterial master regulator of ribonucleotide reduction

Ribonucleotide reductases (RNRs) are the essential enzymes responsible for synthesizing dNTPs, the building blocks of DNA. In bacteria, the entire RNR network is controlled by the master regulator NrdR. As a regulator of an essential pathway with no eukaryotic equivalent, NrdR is a promising antimicrobial target. Recent studies have outlined a mechanism of action for NrdR, in which ATP and dATP induce changes in the protein quaternary structure, regulating RNR repression. However, due to a lack of functional studies linking the known structures to their biological roles, the role of NrdR is not yet fully understood. Here, we conducted a comprehensive study of NrdR in Escherichia coli and Pseudomonas aeruginosa. We delimited the NrdR regulon, combining transcriptomics and motif-based sequence analysis. We crystallized NrdR and obtained structural data on nucleotide binding and its relation to the stability of protein-protein interfaces involved in NrdR oligomerization. We examined the quaternary structures of NrdR using SEC-MALS and atomic force microscopy and correlated structure to function using point mutations, EMSAs, and in vitro transcription assays. Overall, our results demonstrate the mechanism used by NrdR to modulate its quaternary structure and activity and provide structural data that pave the way for targeted antimicrobial therapies.

molecular biology↗

bioRxiv2025

High-beta bursts in SMA mediate anticipatory muscle inhibition in a bimanual motor task

In motor networks, inhibition has been associated with oscillatory activity in the mu (8-12 Hz) or beta (13-30 Hz) bands, yet how these rhythms ultimately influence muscle activity remains unclear. The Bimanual Load-Lifting Task (BLLT) elicits anticipatory inhibition of the elbow flexors in the load-supporting arm during voluntary load-lifting, providing a suitable model to study oscillatory mechanisms of muscle inhibition. We recorded magnetoencephalography (MEG) in adult participants performing the BLLT. Optimal postural stabilization occurred when Biceps brachii inhibition preceded unloading by [~]10-40 ms. Stronger muscle inhibition within this time window was associated with reduced high-gamma (90-130 Hz) power, potentially reflecting reduced excitability, and increased high-beta power in the contralateral Supplementary Motor Area (SMA), with high-gamma power reduction partially mediating the effect of high-beta power on anticipatory inhibition. Furthermore, trials containing beta bursts (22-28 Hz) exhibited both optimally timed anticipatory inhibition and concurrent reduction in high-gamma power. These findings suggest that optimal anticipatory inhibition in the load-supporting elbow flexor is linked to reduced SMA excitability mediated by inhibitory beta bursts. More broadly, they provide new evidence linking beta bursts to the inhibitory control of muscle function, and contribute to a better understanding of motor anticipation.

neuroscience↗

bioRxiv2024

Evaluation of near-infrared light therapy for the treatment of neurodegenerative diseases: Limited penetration depth into the brain likely hinders efficacy

BackgroundNear-infrared (NIR) light therapy is used to treat various musculoskeletal disorders. It has been proposed that transcranial NIR light treatment may also be beneficial for Alzheimers disease (AD). However, the ability of NIR light to penetrate the scalp and skull efficiently and induce cytoprotective responses in the brain parenchyma has not been sufficiently examined so far. This study aimed to evaluate whether the amount of NIR light that can penetrate through the human skull can cause a biological effect. MethodsThree commercially available devices (a medical laser emitting light at a wavelength of 905 nm and two LED helmets operating at wavelengths of 810 nm and 1070 nm, respectively) were used to measure the NIR light transmittance through human post-mortem skulls with a thermal power sensor. Furthermore, the biological effects of the fraction of light power that passed through the skull were investigated in a human neuronal cell line and in C. elegans. ResultsThe 905 nm laser achieved transmittances of up to 0.31% (173 {micro}W/cm2) of its input power, and the LED helmets 0.71% (41 {micro}W/cm2; 810 nm) and 0.45% (19 {micro}W/cm2; 1070 nm) of their respective input powers. NIR light exposure at a power density of 134 mW/cm2 was sufficient to activate mitochondrial metabolism in cultured human neurons and C. elegans, as demonstrated by increased cytochrome c oxidase activity and induction of mitochondrial chaperones. However, this stimulatory effect was no longer observed when the applied power density was reduced to 2.5 mW/cm2. ConclusionsMore than 99% of the NIR light emitted by the investigated devices was either absorbed or scattered by the human skull. The residual NIR light that would reach underlying brain structures was too weak to elicit biological effects. In conclusion, NIR light treatment is unlikely to be effective to treat brain diseases such as AD due to the low penetrability of the skull.

neuroscience↗

bioRxiv2024

Perception/action coupling is modulated by the age-related motor skills of the agent performing the action

The perception/action coupling, underpinned by Mirror Neurons, allows the understanding of others action goal and intention picked up from cues conveyed by the individuals kinematics and the context of the action. This mechanism can be modulated by familiarity with the observed action, by motor experience and motor expertise for that action, but could it be modulated by the age-related motor skills of the agent performing the action? We used an eye-tracking visual preference paradigm to study the modulation of perception/action coupling in 62 adults when viewing videos of daily actions presented in a forward reading direction or, for the same action, in a backward reading direction. Video actions were performed by young actors aged 4, 8, and 13 years and by adults. We found greater pupil dilation for all backward videos compared to forward videos. Interestingly, pupil dilation was greater for the forward videos with child actors than for those with adolescent/adult actors. An overall comparison of looking times during the preference phase did not reveal a preference for either video but testing for a context effect revealed a preference for backward videos when the last video observed in the exposure phase was forward. This study demonstrated the influence of the agents age-related motor abilities on the observers perception/action coupling and revealed that pupil dilation and the context effect could be relevant cues for exploring this coupling in a non-invasive, passive experimental setup that is particularly suitable for exploring perception/action coupling in very young children with a developmental disorder.

neuroscience↗

bioRxiv2024

Perception/action coupling in children with autism: insights from looking time and pupil dilation measurements

The objective of this study was to characterize, through indices extracted from eye-tracking measurements, the spontaneous distinction of videos of daily actions with a variable perception/action coupling, depending on whether, for the same action, the video was presented in the forward reading direction (strong coupling), or in the backward reading direction (weaker coupling). 17 pairs of videos of daily actions performed by adults were viewed by 36 typically developing children and 28 children with ASD aged 7-18 years. During the exposure phase, they watched two videos of the same action (forward and backward) presented successively, before looking at these two videos in competition, in a second visual preference phase. During the exposure phase, all participants paid similar general attention to each of the videos. We found greater pupillary dilation for backward than forward actions in both groups, but significantly less in the ASD group. In the visual preference phase, both groups showed significantly greater looking times for backward actions over forward ones, with no difference between groups. If TD children perceived the kinematics of the backward videos as violating their expectations given the strong perception/action coupling they had already built over that action, on the contrary, the lower increased in pupil dilation found in ASD children could reflect altered perception/action coupling. This study confirms the validity of looking time and pupil dilation as behavioral and physiological markers that could be used in a 10-mn eye-tracking test to explore perception/action coupling in childhood and in ASD. Lay summaryPeople with ASD often have difficulty understanding the actions of others. The fine understanding of actions requires a coupling between the action we observe, and its representation stored in our memory. This process might be challenged in autism. Here we used a 10-mn eye-tracking test to explore perception/action coupling in ASD. Participants were watching videos of daily actions. Looking time and pupil dilation were measured while participants watched videos of daily actions, and were found to be relevant indexes.

neuroscience↗

bioRxiv2024

Dsg2 truncation causes a lethal barrier breakdown in mice

Inflammatory bowel diseases (IBD) such as Crohns disease (CD) have a complex aetiology with alterations of both the intestinal epithelial barrier and the IL23/IL17 immune response. Here, we investigated the role of a novel mutation in the desmosomal cadherin desmoglein 2 gene (DSG2) in the pathogenesis of IBD. DSG2 is known to regulate intestinal epithelial barrier integrity. Genetic analysis of a CD patient revealed a novel likely pathogenic DSG2 mutation leading to a truncated protein lacking part of the intracellular domain. We generated an enterocyte-specific mouse model, recapitulating the human mutation to study how the cytoplasmic truncation of Dsg2 affects intestinal barrier properties systemically. Moreover, we analysed the intestinal genetic profile in these mice and compared it to IBD patients. We describe a first CD patient with a rare mutation in the DSG2 gene causing cytoplasmic truncation with affects Dsg2 mobility. Mice with enterocyte-specific Dsg2 truncation suffered from a lethal intestinal barrier defect and presented a skewed IL17 response similar to CD patients. We identified the desmosomal cadherin Dsg2 as a regulator of the skewed IL17 response. These data indicate that desmosomes regulate inflammation similar to psoriasis which explains why the same novel immune therapies are effective for both diseases.

cell biology↗

bioRxiv2024

Unveiling the Molecular Mechanisms of the Type-IX Secretion System's Response Regulator: Structural and Functional Insights

The Type-IX secretion system (T9SS) is a nanomachinery utilized by bacterial pathogens to facilitate infection. The system is regulated by a signaling cascade serving as its activation switch. A pivotal member in this cascade, the response regulator protein PorX, represents a promising drug target to prevent the secretion of virulence factors. Here, we provide a comprehensive characterization of PorX both in vitro and in vivo. First, our structural studies revealed PorX harbours a unique enzymatic effector domain, which, surprisingly, shares structural similarities with the alkaline phosphatase superfamily, involved in nucleotide and lipid signaling pathways. Importantly, such pathways have not been associated with the T9SS until now. Enzymatic characterization of PorXs effector domain revealed a zinc-dependent phosphodiesterase activity, with active site dimensions suitable to accommodate a large substrate. Unlike typical response regulators that dimerize via their receiver domain upon phosphorylation, we found that zinc can also induce conformational changes and promote PorXs dimerization via an unexpected interface. These findings suggest that PorX can serve as a cellular zinc sensor, broadening our understanding of its regulatory mechanisms. Despite the strict conservation of PorX in T9SS-utilizing bacteria, we demonstrate that PorX is essential for virulence factors secretion in Porphyromonas gingivalis and affects metabolic enzymes secretion in the non-pathogenic Flavobacterium johnsoniae, but not for the secretion of gliding adhesins. Overall, this study advances our structural and functional understanding of PorX, highlighting its potential as a druggable target for intervention strategies aimed at disrupting the T9SS and mitigating virulence in pathogenic species.

biochemistry↗

bioRxiv2024

New, biomechanically sound tendon tissue after injection of uncultured, autologous, adipose derived regenerative cells in partial Achilles tendon defects in rabbits

BackgroundCurrent management options for partial tendon tears may not offer future potential to heal tissue and improve clinical results. This study tested the hypothesis that treatment of a partial rabbit common calcaneus tendon (CCT) defect with uncultured, autologous, adipose derived regenerative cells (UA-ADRCs) enables regenerative healing without scar formation, as recently observed in a biopsy of a human supraspinatus tendon. MethodsA full-thickness hole (diameter, 3 mm) was punched into the midsubstance of the right gastrocnemius tendon (GT; which is a part of the CCT) of adult, female New Zealand white rabbits. Immediately thereafter the rabbits were treated by application of an averaged 28.3x106 UA-ADRCs in 0.5 ml lactated Ringers solution (RLS) into the GT defect and surrounding tendon tissue, or underwent sham treatment. Rabbits were sacrificed either four weeks (W4) or twelve weeks (W12) post-treatment, and the CCTs were investigated using histology, immunohistochemistry and non-destructive biomechanical testing. ResultsNewly formed connective tissue was consistent with the formation of new tendon tissue after treatment with UA-ADRCs, and with the formation of scar tissue after sham treatment, at both W4 and W12 post-treatment. Biomechanical testing demonstrated a significantly higher mean percent relaxation after treatment with UA-ADRCs than after sham treatment (p < 0.05), and significant, negative correlations between the peak stress as well as the equilibrium stress and the cross-sectional area of the CCT (p < 0.05) after treatment with UA-ADRCs but not after sham treatment. ConclusionsManagement of partial tendon tears with UA-ADRCs has the potential to be truly "structure-modifying".

bioengineering↗

bioRxiv2024

Leveraging uncertainty quantification to optimise CRISPR guide RNA selection

CRISPR-based genome editing relies on guide RNA sequences to target specific regions of interest. A large number of methods have been developed to predict how efficient different guides are at inducing indels. As more experimental data becomes available, methods based on machine learning have become more prominent. Here, we explore whether quantifying the uncertainty around these predictions can be used to design better guide selection strategies. We demonstrate how using a deep ensemble approach achieves better performance than utilising a single model. This approach can also provide uncertainty quantification. This allows to design, for the first time, strategies that consider uncertainty in guide RNA selection. These strategies achieve precision over 91% and can identify suitable guides for more than 93% of genes in the mouse genome. Our deep ensemble model is available at https://github.com/bmdslab/CRISPR_DeepEnsemble.

bioinformatics↗

bioRxiv2024

Fast and scalable off-target assessment for CRISPR guide RNAs using partial matches

The design of CRISPR-Cas9 guide RNAs is not trivial. In particular, it is crucial to evaluate the risk of unintended, off-target modifications, but this is computationally expensive. To avoid a brute-force approach where each guide RNA is compared against every possible CRISPR target site in the genome, we previously introduced Crackling, a guide RNA design tool that relies on exact matches over 4bp subsequences to approximate a neighbourhood and accelerate off-target scoring by greatly reducing the search space. While this was faster than other existing tools, it still generates large neighbourhoods. Here, we aim to further reduce the search space by requiring more, now non-contiguous, exact matches. The new implementation, called Crackling++, is benchmarked against our initial approach and other off-target evaluation tools. We show that it provides the fastest way to assess candidate guide RNAs. By using memorymapped files, it also scales to the largest genomes. Crackling++ is available at https://github.com/bmds-lab/CracklingPlusPlus under the Berkeley Software Distribution (BSD) 3-Clause license.

bioinformatics↗

bioRxiv2022

Synergism and phytotoxicity: the effects of tank-mix additives on the biological efficacy of Cu2+ against Venturia inaequalis and Podosphaera leucotricha

The wetting behaviour of the spray and biological efficacy of Cu2+ active ingredients in agrochemical formulations may be enhanced by tank-mix additives. We investigated how three BREAK-THRU(R) additives (BT301: biodegradable, BT133 and BT420: bio-based and biodegradable) as tank-mix with commercial copper preparations influence the spray distribution, leaf uptake and biological efficacy of copper additive mixtures against apple scab and apple powdery mildew under controlled conditions. We quantified the synergetic effects of these additives in foliar applications. In addition, we determined the phytotoxic potential and evaluated impacts on photosynthetic activity, non-photochemical quenching and ROS activity. The additives BT301 and BT420 strongly reduced surface tension and contact angle of copper treatments. The fluorescence observations revealed that BT301 achieved better spreading of copper formulation with more complete coverage of the leaf surface than BT420 and BT133, whereas "coffee-ring" spreading was observed with BT133. The additive BT301 showed an increase in relative fluorescence area, indicating higher ROS production as a signal of intra-cellular tissue activity. The photochemical efficiency of photosystem II (Fv/Fm) was not negatively influenced by copper or additive treatment. Thus, we observed no phytotoxic effects of copper-additive mixtures on apple leaves at treatment doses of 4 g Cu2+ L-1. All copper treatments reduced apple scab infestations significantly, by 53-76%. Interestingly, addition of BT301 to copper preparations showed the strongest biological efficacy (83% reduction) against V. inaequalis, whereas addition of BT420 showed the strongest effect against P. leucotricha (89% infection reduction). The synergetic effects of additives on the biological efficacy without phytotoxic effects on plants may have potential for reducing copper loads in horticultural production systems.

pathology↗