Characterization of outer membrane vesicles of plant growth promoting bacteria Paraburkholderia phytofirmans PsJN
Аутори
Nešić, SofijaPantelić, Ana
Samardžić, Jelena
Nikolić, Dragana
Остала ауторства
Dimkić, IvicaVenturi, Vittorio
Конференцијски прилог (Објављена верзија)
Метаподаци
Приказ свих података о документуАпстракт
Plant growth promoting bacteria (PGPB) are microorganisms present in plant
rhizosphere that stimulate plant growth, as well as prime plants against abiotic
and biotic stress. PGPB are also located in plant intercellular space and can directly
interfere with plant pathogens. Many different mechanisms of interaction
between plants and PGPB has been shown, but the interaction through outer
membrane vesicles (OMVs) has only been proposed. OMVs are membrane-enclosed
nanostructures produced by Gram-negative bacteria. Size of OMVs can
range from 20 to 300 nm, depending on the strain. In this research, we determined
optimal method for isolation of OMVs produced by Paraburkholderia
phytofirmans PsJN, characterized the size range of OMVs, their concentration
and origin. OMVs from PGPB PsJN have been isolated us\ing two methodological
approaches: 1) differential centrifugation, ultrafiltration, and purification
using Optiprep density gradient, and 2) OMV isolation by the commercial
E...xoBacteria OMV Isolation Kit that uses an affinity-based column system. Size
and concentration of isolated OMVs were analyzed using Nanoparticle Tracking
Analysis (NTA). As the OMVs are derived from outer membranes of bacteria, their
origin has been proven using an essay for detection of lipopolysaccharides. In
addition, protein profiles of OMV from PsJN were visualized by SDS-PAGE. Comparing
different methods for isolation and purification of vesicles from PsJN we
concluded that for our PGPB strain the commercial ExoBacteria OMV Isolation
Kit provides the highest yield. Quantification of OMV using NTA of OMVs produced
by P. phytofirmans PsJN, isolated with the commercial kit, showed a mean
particle numeration of 4.05*109 particles/ml. Size of OMVs isolated from PsJN
ranged from 50 to 200 nm. Detection of lipopolysaccharides confirmed that
isolated vesicles originated from outer membranes of PsJN. These findings are
the important first step in our research of the role of OMVs in PGPB interaction
with plants.
Кључне речи:
PGPB / extracellular vesicles / outer membrane vesicles / OMVИзвор:
ICGEB WORKSHOP: Trends in microbial solutions for sustainable agriculture, 2023, 72-72Издавач:
- Belgrade : Faculty of Biology
Напомена:
- Book of abstract: ICGEB WORKSHOP; Trends in microbial solutions for sustainable agriculture, 13 – 15 September 2023. Belgrade, Serbia
Институција/група
Institut za molekularnu genetiku i genetičko inženjerstvoTY - CONF AU - Nešić, Sofija AU - Pantelić, Ana AU - Samardžić, Jelena AU - Nikolić, Dragana PY - 2023 UR - https://imagine.imgge.bg.ac.rs/handle/123456789/2158 AB - Plant growth promoting bacteria (PGPB) are microorganisms present in plant rhizosphere that stimulate plant growth, as well as prime plants against abiotic and biotic stress. PGPB are also located in plant intercellular space and can directly interfere with plant pathogens. Many different mechanisms of interaction between plants and PGPB has been shown, but the interaction through outer membrane vesicles (OMVs) has only been proposed. OMVs are membrane-enclosed nanostructures produced by Gram-negative bacteria. Size of OMVs can range from 20 to 300 nm, depending on the strain. In this research, we determined optimal method for isolation of OMVs produced by Paraburkholderia phytofirmans PsJN, characterized the size range of OMVs, their concentration and origin. OMVs from PGPB PsJN have been isolated us\ing two methodological approaches: 1) differential centrifugation, ultrafiltration, and purification using Optiprep density gradient, and 2) OMV isolation by the commercial ExoBacteria OMV Isolation Kit that uses an affinity-based column system. Size and concentration of isolated OMVs were analyzed using Nanoparticle Tracking Analysis (NTA). As the OMVs are derived from outer membranes of bacteria, their origin has been proven using an essay for detection of lipopolysaccharides. In addition, protein profiles of OMV from PsJN were visualized by SDS-PAGE. Comparing different methods for isolation and purification of vesicles from PsJN we concluded that for our PGPB strain the commercial ExoBacteria OMV Isolation Kit provides the highest yield. Quantification of OMV using NTA of OMVs produced by P. phytofirmans PsJN, isolated with the commercial kit, showed a mean particle numeration of 4.05*109 particles/ml. Size of OMVs isolated from PsJN ranged from 50 to 200 nm. Detection of lipopolysaccharides confirmed that isolated vesicles originated from outer membranes of PsJN. These findings are the important first step in our research of the role of OMVs in PGPB interaction with plants. PB - Belgrade : Faculty of Biology C3 - ICGEB WORKSHOP: Trends in microbial solutions for sustainable agriculture T1 - Characterization of outer membrane vesicles of plant growth promoting bacteria Paraburkholderia phytofirmans PsJN EP - 72 SP - 72 UR - https://hdl.handle.net/21.15107/rcub_imagine_2158 ER -
@conference{ author = "Nešić, Sofija and Pantelić, Ana and Samardžić, Jelena and Nikolić, Dragana", year = "2023", abstract = "Plant growth promoting bacteria (PGPB) are microorganisms present in plant rhizosphere that stimulate plant growth, as well as prime plants against abiotic and biotic stress. PGPB are also located in plant intercellular space and can directly interfere with plant pathogens. Many different mechanisms of interaction between plants and PGPB has been shown, but the interaction through outer membrane vesicles (OMVs) has only been proposed. OMVs are membrane-enclosed nanostructures produced by Gram-negative bacteria. Size of OMVs can range from 20 to 300 nm, depending on the strain. In this research, we determined optimal method for isolation of OMVs produced by Paraburkholderia phytofirmans PsJN, characterized the size range of OMVs, their concentration and origin. OMVs from PGPB PsJN have been isolated us\ing two methodological approaches: 1) differential centrifugation, ultrafiltration, and purification using Optiprep density gradient, and 2) OMV isolation by the commercial ExoBacteria OMV Isolation Kit that uses an affinity-based column system. Size and concentration of isolated OMVs were analyzed using Nanoparticle Tracking Analysis (NTA). As the OMVs are derived from outer membranes of bacteria, their origin has been proven using an essay for detection of lipopolysaccharides. In addition, protein profiles of OMV from PsJN were visualized by SDS-PAGE. Comparing different methods for isolation and purification of vesicles from PsJN we concluded that for our PGPB strain the commercial ExoBacteria OMV Isolation Kit provides the highest yield. Quantification of OMV using NTA of OMVs produced by P. phytofirmans PsJN, isolated with the commercial kit, showed a mean particle numeration of 4.05*109 particles/ml. Size of OMVs isolated from PsJN ranged from 50 to 200 nm. Detection of lipopolysaccharides confirmed that isolated vesicles originated from outer membranes of PsJN. These findings are the important first step in our research of the role of OMVs in PGPB interaction with plants.", publisher = "Belgrade : Faculty of Biology", journal = "ICGEB WORKSHOP: Trends in microbial solutions for sustainable agriculture", title = "Characterization of outer membrane vesicles of plant growth promoting bacteria Paraburkholderia phytofirmans PsJN", pages = "72-72", url = "https://hdl.handle.net/21.15107/rcub_imagine_2158" }
Nešić, S., Pantelić, A., Samardžić, J.,& Nikolić, D.. (2023). Characterization of outer membrane vesicles of plant growth promoting bacteria Paraburkholderia phytofirmans PsJN. in ICGEB WORKSHOP: Trends in microbial solutions for sustainable agriculture Belgrade : Faculty of Biology., 72-72. https://hdl.handle.net/21.15107/rcub_imagine_2158
Nešić S, Pantelić A, Samardžić J, Nikolić D. Characterization of outer membrane vesicles of plant growth promoting bacteria Paraburkholderia phytofirmans PsJN. in ICGEB WORKSHOP: Trends in microbial solutions for sustainable agriculture. 2023;:72-72. https://hdl.handle.net/21.15107/rcub_imagine_2158 .
Nešić, Sofija, Pantelić, Ana, Samardžić, Jelena, Nikolić, Dragana, "Characterization of outer membrane vesicles of plant growth promoting bacteria Paraburkholderia phytofirmans PsJN" in ICGEB WORKSHOP: Trends in microbial solutions for sustainable agriculture (2023):72-72, https://hdl.handle.net/21.15107/rcub_imagine_2158 .