TY  - CONF
AU  - Todorović, Nataša
AU  - Živić, Miroslav
AU  - Stevanović, Katarina
AU  - Pajić, Tanja
AU  - Cvetić Antić, Tijana
AU  - Atlagić, Kristina
AU  - Krmpot, Aleksandar
PY  - 2023
UR  - https://microbiology.bg.ac.rs/workshop2023/
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6248
AB  - The use of chemical fungicides is detrimental for soil and aquatic ecosystems.
Although highly effective, they raise concerns about impact on the environment.
Therefore, the need to reduce the use of chemical fungicides and protect
the soil and water, pose the challenge to find novel, eco-friendly, but also
more efficient, means of crop protection from phytopathogenic fungi. We are
aiming to utilize the native Trichoderma species, as biological control agent,
for controlling or eliminating the phytopathogenic fungi. Our research will be
based on cell nano-surgery of fungal cell wall using ultrashort laser pulses and
subsequent patch clamping on the released protoplast membrane. This will
enable the studies of activity and potency of specific class of antimicrobial short
peptides, peptaibols, released by Trichoderma species, which act by forming
ion channels in pathogen membrane as one of the main mechanisms of their
fungicide action. The result of this project will be the developed instrumentation
and unique method based on nano-surgery and patch clamp for studies of ionic
channels in native fungal plasma membrane. Testing a range of autochthonous
Trichoderma sp. isolates will enable formulation of their combinations that work
best. The selected Trichoderma strains with the highest standardized peptaibol
activity in specific combination of conditions, can be used as a starting point for
development of new, more potent biocontrol agents.
PB  - Belgrade: University of Belgrade, Faculty of Biology
C3  - Book of Abstracts: ICGEB Workshop: Trends in microbial solutions for sustainable agriculture; 2023 Sep13-15; Belgrade, Serbia
T1  - Novel research project – BioPhysFun for advancement of characterization of Trichoderma as biological control agent
SP  - 84
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_6248
ER  - 

@conference{
author = "Todorović, Nataša and Živić, Miroslav and Stevanović, Katarina and Pajić, Tanja and Cvetić Antić, Tijana and Atlagić, Kristina and Krmpot, Aleksandar",
year = "2023",
abstract = "The use of chemical fungicides is detrimental for soil and aquatic ecosystems.
Although highly effective, they raise concerns about impact on the environment.
Therefore, the need to reduce the use of chemical fungicides and protect
the soil and water, pose the challenge to find novel, eco-friendly, but also
more efficient, means of crop protection from phytopathogenic fungi. We are
aiming to utilize the native Trichoderma species, as biological control agent,
for controlling or eliminating the phytopathogenic fungi. Our research will be
based on cell nano-surgery of fungal cell wall using ultrashort laser pulses and
subsequent patch clamping on the released protoplast membrane. This will
enable the studies of activity and potency of specific class of antimicrobial short
peptides, peptaibols, released by Trichoderma species, which act by forming
ion channels in pathogen membrane as one of the main mechanisms of their
fungicide action. The result of this project will be the developed instrumentation
and unique method based on nano-surgery and patch clamp for studies of ionic
channels in native fungal plasma membrane. Testing a range of autochthonous
Trichoderma sp. isolates will enable formulation of their combinations that work
best. The selected Trichoderma strains with the highest standardized peptaibol
activity in specific combination of conditions, can be used as a starting point for
development of new, more potent biocontrol agents.",
publisher = "Belgrade: University of Belgrade, Faculty of Biology",
journal = "Book of Abstracts: ICGEB Workshop: Trends in microbial solutions for sustainable agriculture; 2023 Sep13-15; Belgrade, Serbia",
title = "Novel research project – BioPhysFun for advancement of characterization of Trichoderma as biological control agent",
pages = "84",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_6248"
}

Todorović, N., Živić, M., Stevanović, K., Pajić, T., Cvetić Antić, T., Atlagić, K.,& Krmpot, A.. (2023). Novel research project – BioPhysFun for advancement of characterization of Trichoderma as biological control agent. in Book of Abstracts: ICGEB Workshop: Trends in microbial solutions for sustainable agriculture; 2023 Sep13-15; Belgrade, Serbia
Belgrade: University of Belgrade, Faculty of Biology., 84.
https://hdl.handle.net/21.15107/rcub_ibiss_6248

Todorović N, Živić M, Stevanović K, Pajić T, Cvetić Antić T, Atlagić K, Krmpot A. Novel research project – BioPhysFun for advancement of characterization of Trichoderma as biological control agent. in Book of Abstracts: ICGEB Workshop: Trends in microbial solutions for sustainable agriculture; 2023 Sep13-15; Belgrade, Serbia. 2023;:84.
https://hdl.handle.net/21.15107/rcub_ibiss_6248 .

Todorović, Nataša, Živić, Miroslav, Stevanović, Katarina, Pajić, Tanja, Cvetić Antić, Tijana, Atlagić, Kristina, Krmpot, Aleksandar, "Novel research project – BioPhysFun for advancement of characterization of Trichoderma as biological control agent" in Book of Abstracts: ICGEB Workshop: Trends in microbial solutions for sustainable agriculture; 2023 Sep13-15; Belgrade, Serbia (2023):84,
https://hdl.handle.net/21.15107/rcub_ibiss_6248 .

TY  - CONF
AU  - Stevanović, Katarina
AU  - Pajić, Tanja
AU  - Krmpot, Aleksandar
AU  - Rabasović, Mihailo
AU  - Žižić, Milan
AU  - Živić, Miroslav
AU  - Todorović, Nataša
PY  - 2023
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6246
AB  - The success of patch-clamp giga-ohm seal formation on filamentous fungi Phycomyces blakesleeanus protoplasts was investigated to evaluate their usefulness in ion channel studies on filamentous fungi. Protoplasts were obtained by laser-mediated nanosurgery of the cell wall stained with Calcofluor White. To enable a successful seal formation, it is critical to prevent cell wall regeneration. Since wall integrity responses in fungi involve kinase-dependent pathways, we aimed to sufficiently reduce intracellular ATP availability using the respiratory inhibitor azide. The effect of azide on phosphate metabolites of Phycomyces blakesleeanus, as determined by 31P NMR spectroscopy, was a reduction in intracellular ATP accompanied by a decrease in long chain polyphosphates. Subsequently, all seal formation measurements were performed in the presence of azide, and protoplast viability was confirmed by cytoplasmic streaming. The success of seal formation depends on the size of the protoplasts, as larger protoplasts are more prone to successful seal formation. It was also found that the laser power used in nanosurgery could influence the success of seal formation, as higher values were associated with a lower success rate. The protoplasts released by laser nanosurgery produced by our newly developed method are a good model system for patch clamp on filamentous fungal membranes because their plasma membrane can easily form high-quality seals with the patch pipette.
PB  - Kragujevac: Institute for Information Technologies, University of Belgrade
C3  - Book of Proceedings: 2nd International Conference on Chemo and Bioinformatics ICCBIKG 2023; 2023 Sep 28-29; Kragujevac, Serbia
T1  - Patch clamp pipette giga seal forming success on the nanosurgery-obtained filamentous fungi protoplasts
DO  - 10.46793/ICCBI23.221S
SP  - 221
EP  - 224
ER  - 

@conference{
author = "Stevanović, Katarina and Pajić, Tanja and Krmpot, Aleksandar and Rabasović, Mihailo and Žižić, Milan and Živić, Miroslav and Todorović, Nataša",
year = "2023",
abstract = "The success of patch-clamp giga-ohm seal formation on filamentous fungi Phycomyces blakesleeanus protoplasts was investigated to evaluate their usefulness in ion channel studies on filamentous fungi. Protoplasts were obtained by laser-mediated nanosurgery of the cell wall stained with Calcofluor White. To enable a successful seal formation, it is critical to prevent cell wall regeneration. Since wall integrity responses in fungi involve kinase-dependent pathways, we aimed to sufficiently reduce intracellular ATP availability using the respiratory inhibitor azide. The effect of azide on phosphate metabolites of Phycomyces blakesleeanus, as determined by 31P NMR spectroscopy, was a reduction in intracellular ATP accompanied by a decrease in long chain polyphosphates. Subsequently, all seal formation measurements were performed in the presence of azide, and protoplast viability was confirmed by cytoplasmic streaming. The success of seal formation depends on the size of the protoplasts, as larger protoplasts are more prone to successful seal formation. It was also found that the laser power used in nanosurgery could influence the success of seal formation, as higher values were associated with a lower success rate. The protoplasts released by laser nanosurgery produced by our newly developed method are a good model system for patch clamp on filamentous fungal membranes because their plasma membrane can easily form high-quality seals with the patch pipette.",
publisher = "Kragujevac: Institute for Information Technologies, University of Belgrade",
journal = "Book of Proceedings: 2nd International Conference on Chemo and Bioinformatics ICCBIKG 2023; 2023 Sep 28-29; Kragujevac, Serbia",
title = "Patch clamp pipette giga seal forming success on the nanosurgery-obtained filamentous fungi protoplasts",
doi = "10.46793/ICCBI23.221S",
pages = "221-224"
}

Stevanović, K., Pajić, T., Krmpot, A., Rabasović, M., Žižić, M., Živić, M.,& Todorović, N.. (2023). Patch clamp pipette giga seal forming success on the nanosurgery-obtained filamentous fungi protoplasts. in Book of Proceedings: 2nd International Conference on Chemo and Bioinformatics ICCBIKG 2023; 2023 Sep 28-29; Kragujevac, Serbia
Kragujevac: Institute for Information Technologies, University of Belgrade., 221-224.
https://doi.org/10.46793/ICCBI23.221S

Stevanović K, Pajić T, Krmpot A, Rabasović M, Žižić M, Živić M, Todorović N. Patch clamp pipette giga seal forming success on the nanosurgery-obtained filamentous fungi protoplasts. in Book of Proceedings: 2nd International Conference on Chemo and Bioinformatics ICCBIKG 2023; 2023 Sep 28-29; Kragujevac, Serbia. 2023;:221-224.
doi:10.46793/ICCBI23.221S .

Stevanović, Katarina, Pajić, Tanja, Krmpot, Aleksandar, Rabasović, Mihailo, Žižić, Milan, Živić, Miroslav, Todorović, Nataša, "Patch clamp pipette giga seal forming success on the nanosurgery-obtained filamentous fungi protoplasts" in Book of Proceedings: 2nd International Conference on Chemo and Bioinformatics ICCBIKG 2023; 2023 Sep 28-29; Kragujevac, Serbia (2023):221-224,
https://doi.org/10.46793/ICCBI23.221S . .

TY  - CONF
AU  - Vitkovac, Aleksandra
AU  - Pajić, Tanja
AU  - Bukumira, Marta
AU  - Stanić, Marina
AU  - Rabasović, Mihailo
AU  - Todorović, Nataša
PY  - 2023
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6247
AB  - Adaptive changes in mitochondrial morphology are associated with changes in the mitochondrial function and metabolic fitness of eukaryotic cells. We previously described in young hyphae of the filamentous fungus Phycomyces blakesleeanus a dramatic effect of an increase in ambient temperature during growth: a 3°C warmer environment compared with a control temperature of 22°C resulted in the appearance of long elongated (“tubular”) mitochondria accompanied by an increase in lipid droplet density. Here, we examined how cooler ambient temperature (18°C) during growth affects mitochondrial morphology in P. blakesleeanus compared with the control grown at 22°C. We used two-photon fluorescence imaging (TPEF) of live hyphae stained with the vital mitochondrial dye rhodamine 123. Extraction of relevant parameters (number, size, and shape of mitochondria) from TPEF images was performed using the Ilastik machine learning-based software. The suitability of the Ilastik analysis was compared with the Particle Analysis (ImageJ). Cold treatment resulted in the appearance of tubular mitochondrial morphology that was absent in the control group. Tubular mitochondrial morphology appears to be an adaptive feature that occurs in both warmer and colder conditions and is likely part of the stress response.
PB  - Kragujevac: Institute for Information Technologies, University of Belgrade
C3  - Book of Proceedings: 2nd International Conference on Chemo and Bioinformatics ICCBIKG 2023; 2023 Sep 28-29; Kragujevac, Serbia
T1  - Slight cooling during growth induced changes in filamentous fungi hypha mitochondrial morphology
DO  - 10.46793/ICCBI23.334V
SP  - 334
EP  - 337
ER  - 

@conference{
author = "Vitkovac, Aleksandra and Pajić, Tanja and Bukumira, Marta and Stanić, Marina and Rabasović, Mihailo and Todorović, Nataša",
year = "2023",
abstract = "Adaptive changes in mitochondrial morphology are associated with changes in the mitochondrial function and metabolic fitness of eukaryotic cells. We previously described in young hyphae of the filamentous fungus Phycomyces blakesleeanus a dramatic effect of an increase in ambient temperature during growth: a 3°C warmer environment compared with a control temperature of 22°C resulted in the appearance of long elongated (“tubular”) mitochondria accompanied by an increase in lipid droplet density. Here, we examined how cooler ambient temperature (18°C) during growth affects mitochondrial morphology in P. blakesleeanus compared with the control grown at 22°C. We used two-photon fluorescence imaging (TPEF) of live hyphae stained with the vital mitochondrial dye rhodamine 123. Extraction of relevant parameters (number, size, and shape of mitochondria) from TPEF images was performed using the Ilastik machine learning-based software. The suitability of the Ilastik analysis was compared with the Particle Analysis (ImageJ). Cold treatment resulted in the appearance of tubular mitochondrial morphology that was absent in the control group. Tubular mitochondrial morphology appears to be an adaptive feature that occurs in both warmer and colder conditions and is likely part of the stress response.",
publisher = "Kragujevac: Institute for Information Technologies, University of Belgrade",
journal = "Book of Proceedings: 2nd International Conference on Chemo and Bioinformatics ICCBIKG 2023; 2023 Sep 28-29; Kragujevac, Serbia",
title = "Slight cooling during growth induced changes in filamentous fungi hypha mitochondrial morphology",
doi = "10.46793/ICCBI23.334V",
pages = "334-337"
}

Vitkovac, A., Pajić, T., Bukumira, M., Stanić, M., Rabasović, M.,& Todorović, N.. (2023). Slight cooling during growth induced changes in filamentous fungi hypha mitochondrial morphology. in Book of Proceedings: 2nd International Conference on Chemo and Bioinformatics ICCBIKG 2023; 2023 Sep 28-29; Kragujevac, Serbia
Kragujevac: Institute for Information Technologies, University of Belgrade., 334-337.
https://doi.org/10.46793/ICCBI23.334V

Vitkovac A, Pajić T, Bukumira M, Stanić M, Rabasović M, Todorović N. Slight cooling during growth induced changes in filamentous fungi hypha mitochondrial morphology. in Book of Proceedings: 2nd International Conference on Chemo and Bioinformatics ICCBIKG 2023; 2023 Sep 28-29; Kragujevac, Serbia. 2023;:334-337.
doi:10.46793/ICCBI23.334V .

Vitkovac, Aleksandra, Pajić, Tanja, Bukumira, Marta, Stanić, Marina, Rabasović, Mihailo, Todorović, Nataša, "Slight cooling during growth induced changes in filamentous fungi hypha mitochondrial morphology" in Book of Proceedings: 2nd International Conference on Chemo and Bioinformatics ICCBIKG 2023; 2023 Sep 28-29; Kragujevac, Serbia (2023):334-337,
https://doi.org/10.46793/ICCBI23.334V . .