Biology subjects

Tarkowski, P.

Publications and source records attributed to Tarkowski, P..

3 recordsLinked to original sources

bioRxiv2025

Vacuolar invertase knockout enhances drought tolerance in potato plants

Drought stress is one of the most critical abiotic constraints limiting crop productivity worldwide, exacerbated by ongoing climate change and increasingly frequent extreme weather events. Stomatal regulation and osmoprotective sugar accumulation are critical adaptive mechanisms for plant survival under drought stress. Here, we characterize the enhanced drought resilience observed in CRISPR/Cas9-mediated potato, mutants in their vacuolar invertase gene (StVInv). Knockout plants exhibited improved performance under progressive drought stress and during rewatering drought, maintaining higher stomatal conductance, elevated transpiration rates, and superior photosynthetic efficiency compared to wild-type (WT) plants. These improved performance under similar transpiration rate led to higher agronomic water-use efficiency (AWUE) in stvinv plants resulting in greater biomass production despite reduced water availability. Metabolomic profiling revealed distinct adaptive strategies; stvinv plants preferentially accumulated galactinol and raffinose, indicating enhanced raffinose family oligosaccharide (RFO) metabolism. Furthermore, stvinv plants displayed lower levels of abscisic acid (ABA) and its catabolites under drought, suggesting a moderated ABA response facilitating a more risk-taking growth strategy that supports sustained growth and physiological stability. Our findings identify targeted metabolic and hormonal adjustments underlying drought resilience in potato plant, offering promising strategies for enhancing crop performance under water-limited conditions.

plant biology↗

bioRxiv2025

CLPC2 plays specific roles in CLP complex-mediated regulation of growth, photosynthesis, embryogenesis and response to growth-promoting microbial compounds

In Arabidopsis, exposure to growth-promoting microbial volatile compounds (VCs) enhances CLPC2 levels. This chaperone forms part of the CLP protease complex, which ensures the correct functioning of essential processes in plastids. Previous studies indicated considerable functional redundancy of CLPC2 with its dominant paralogue CLPC1. However, the function and action mechanism of CLPC2 still remain unknown. Here we found that CLPC2-lacking clpc2-2 mutants were unresponsive to microbial VCs, whereas clpc1-1 knockout mutants exhibited a WT-like response to VCs when grown on sucrose-containing medium. Unlike clpc1-1, clpc2-2 plants presented a fully functional photosystem II and lower than WT stomatal conductance. Furthermore, clpc2-2 plants, but not clpc1-1 plants, produced wrinkled seeds with delayed embryonic development and reduced postgerminative establishment rates that resembled those of mutants lacking P and R components of the CLP proteolytic core. Proteomic analyses revealed that knocking out of CLPC2 enhanced the levels of chloroplastic proteins that are essential for growth, embryo development and seedling establishment. These changes differed from those promoted by the lack of CLPC1, but partially resembled those promoted by CLPPR core inactivation. Nearly 40% of the proteins differentially accumulated by the lack of CLPC2 were VC-responsive. Notably, 35S promoter-driven CLPC2 expression promoted changes in the proteome similar to those promoted by the lack of CLPC1. Collectively, our findings highlighted contrasting functional and molecular specificities for CLPC1 and CLPC2, and provided strong evidence that CLPC2 plays specific roles in CLP complex-mediated regulation of plant growth, photosynthesis, embryogenesis, postgerminative seedling establishment and microbial VC responsiveness.

plant biology↗

bioRxiv2023

In Vitro Safety Signals for Potential Clinical Development of the Anti-Inflammatory Pregnane X Receptor Agonist FKK6

Background and purposeBased on the mimicry of microbial metabolites, functionalized indoles were demonstrated as the ligands and agonists of the pregnane X receptor (PXR). The lead indole, FKK6, displayed PXR-dependent protective effects in DSS-induced colitis in mice and in vitro cytokine-treated intestinal organoid cultures. Here, we performed the initial in vitro pharmacological profiling of FKK6. Experimental approachA complex series of cell-free and cell-based assays were employed. The organic synthesis, and advanced analytical chemistry methods were used. Key resultsFKK6-PXR interactions were characterized by hydrogen-deuterium exchange mass spectrometry. Screening FKK6 against potential cellular off-targets revealed high PXR selectivity. FKK6 has poor aqueous solubility but was highly soluble in simulated gastric and intestinal fluids. FKK6 was bound to plasma proteins and chemically stable in plasma. The partition coefficient of FKK6 was 2.70, and FKK6 moderately partitioned into red blood cells. In Caco2 cells, FKK6 displayed high permeability (A-B: 22.8 x 10-6 cm.s-1) and no active efflux. These data are indicative of essentially complete in vivo absorption of FKK6. FKK6 was rapidly metabolized by cytochromes P450, notably by CYP3A4 in human liver microsomes. Two oxidized FKK6 derivatives, including N6-oxide and C19-phenol, were detected, and these metabolites had 5-7 x lower potency as PXR agonists than FKK6. This implies that despite high intestinal absorption, FKK6 is rapidly eliminated by the liver, and its PXR effects are predicted to be predominantly in the intestines. Conclusion and implicationsThe PXR ligand and agonist FKK6 has a suitable pharmacological profile supporting its potential preclinical development. BULLET POINT SUMMARYWhat is already known: O_LIMicrobial metabolite mimic FKK6 is a hPXR agonist with anti-inflammatory properties in mice and human. C_LIO_LIThe in vitro PXR binding, absorption, and metabolism have not been completely characterized. C_LI What this study adds: O_LIPXR selectivity with unique binding mode, high intestinal cell permeability, rapid and complex microsomal metabolism. C_LIO_LIInitial testing for predicted metabolites shows reduced potency as PXR agonists. C_LI Clinical significance: O_LIPXR effects of FKK6 are predicted to be predominantly in the intestines. C_LIO_LIFKK6 has a suitable pharmacological profile supporting its potential preclinical development. C_LI

biochemistry↗