TY  - JOUR
AU  - Popržen, Tatjana
AU  - Nikolić, Ivan
AU  - Krstić-Milošević, Dijana
AU  - Uzelac, Branka
AU  - Trifunović-Momčilov, Milana
AU  - Marković, Marija
AU  - Radulović, Olga
PY  - 2023
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6442
AB  - The rhizosphere represents a center of complex and dynamic interactions between plants and microbes, resulting in various positive effects on plant growth and development. However, less is known about the effects of indole-3-acetic acid (IAA) on aquatic plants. In this study, we report the characterization of four Pseudomonas strains isolated from the rhizosphere of the common duckweed (Lemna minor) with IAA-degradation and -utilization ability. Our results confirm previous reports on the negative effect of IAA on aquatic plants, contrary to the effect on terrestrial plants. P. putida A3-104/5 demonstrated particularly beneficial traits, as it exhibited not only IAA-degrading and -producing activity but also a positive effect on the doubling time of duckweeds in the presence of IAA, positive chemotaxis in the presence of IAA, increased tolerance to oxidative stress in the presence of IAA and increased biofilm formation related to IAA. Similarly, P. gessardii C31-106/3 significantly shortened the doubling time of duckweeds in the presence of IAA, while having a neutral effect in the absence of IAA. These traits are important in the context of plant–bacteria interactions and highlight the role of IAA as a common metabolite in these interactions, especially in aquatic environments where plants are facing unique challenges compared to their terrestrial counterparts. We conclude that IAA-degrading and -producing strains presented in this study might regulate IAA effects on aquatic plants and confer evolutionary benefits under adverse conditions (e.g., under oxidative stress, excess of IAA or nutrient scarcity).
PB  - Basel : MDPI
T2  - International Journal of Molecular Sciences
T1  - Characterization of the IAA-producing and -degrading Pseudomonas strains Regulating Growth of the Common Duckweed (Lemna minor L.)
IS  - 24
VL  - 24
DO  - 10.3390/ijms242417207
SP  - 17207
ER  - 

@article{
author = "Popržen, Tatjana and Nikolić, Ivan and Krstić-Milošević, Dijana and Uzelac, Branka and Trifunović-Momčilov, Milana and Marković, Marija and Radulović, Olga",
year = "2023",
abstract = "The rhizosphere represents a center of complex and dynamic interactions between plants and microbes, resulting in various positive effects on plant growth and development. However, less is known about the effects of indole-3-acetic acid (IAA) on aquatic plants. In this study, we report the characterization of four Pseudomonas strains isolated from the rhizosphere of the common duckweed (Lemna minor) with IAA-degradation and -utilization ability. Our results confirm previous reports on the negative effect of IAA on aquatic plants, contrary to the effect on terrestrial plants. P. putida A3-104/5 demonstrated particularly beneficial traits, as it exhibited not only IAA-degrading and -producing activity but also a positive effect on the doubling time of duckweeds in the presence of IAA, positive chemotaxis in the presence of IAA, increased tolerance to oxidative stress in the presence of IAA and increased biofilm formation related to IAA. Similarly, P. gessardii C31-106/3 significantly shortened the doubling time of duckweeds in the presence of IAA, while having a neutral effect in the absence of IAA. These traits are important in the context of plant–bacteria interactions and highlight the role of IAA as a common metabolite in these interactions, especially in aquatic environments where plants are facing unique challenges compared to their terrestrial counterparts. We conclude that IAA-degrading and -producing strains presented in this study might regulate IAA effects on aquatic plants and confer evolutionary benefits under adverse conditions (e.g., under oxidative stress, excess of IAA or nutrient scarcity).",
publisher = "Basel : MDPI",
journal = "International Journal of Molecular Sciences",
title = "Characterization of the IAA-producing and -degrading Pseudomonas strains Regulating Growth of the Common Duckweed (Lemna minor L.)",
number = "24",
volume = "24",
doi = "10.3390/ijms242417207",
pages = "17207"
}

Popržen, T., Nikolić, I., Krstić-Milošević, D., Uzelac, B., Trifunović-Momčilov, M., Marković, M.,& Radulović, O.. (2023). Characterization of the IAA-producing and -degrading Pseudomonas strains Regulating Growth of the Common Duckweed (Lemna minor L.). in International Journal of Molecular Sciences
Basel : MDPI., 24(24), 17207.
https://doi.org/10.3390/ijms242417207

Popržen T, Nikolić I, Krstić-Milošević D, Uzelac B, Trifunović-Momčilov M, Marković M, Radulović O. Characterization of the IAA-producing and -degrading Pseudomonas strains Regulating Growth of the Common Duckweed (Lemna minor L.). in International Journal of Molecular Sciences. 2023;24(24):17207.
doi:10.3390/ijms242417207 .

Popržen, Tatjana, Nikolić, Ivan, Krstić-Milošević, Dijana, Uzelac, Branka, Trifunović-Momčilov, Milana, Marković, Marija, Radulović, Olga, "Characterization of the IAA-producing and -degrading Pseudomonas strains Regulating Growth of the Common Duckweed (Lemna minor L.)" in International Journal of Molecular Sciences, 24, no. 24 (2023):17207,
https://doi.org/10.3390/ijms242417207 . .

TY  - JOUR
AU  - Popović, Zorica
AU  - Vidaković, Vera
AU  - Mijalković, Tatjana
AU  - Krstić-Milošević, Dijana
PY  - 2023
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5889
AB  - Phytochemical profiling of six natural populations of Gentianella austriaca was performed
by HPLC identification and quantification of a number of secondary metabolites, and evaluation of time series of peak areas by chemometric analysis. Phytochemical analysis of G. austriaca revealed the presence of iridoids, flavone-C-glucosides and xanthones. Twelve secondary metabolites were identified in the aerial parts, roots and seeds, including swertiamarin (SWM), gentiopicrin (GP), sweroside (SWZ), isoorientin (ISOOR), swertisin (SWE), demethylbellidifolin-8-O-glucoside (DMB-8-O-glc), bellidifolin-8-O-glucoside (BDF-8-O-glc), mangiferin (MGF), corymbiferin (CBF), corymbiferin-1-O-glucoside (CBF-1-O-glc), bellidifolin (BDF) and campestroside. Multivariate statistical analyses showed relatively low variability among populations according to secondary metabolite content. However, some pharmacologically important compounds were found in higher amounts in a few populations, which could be useful for conservation and future biotechnological procedures.
PB  - Basel: MDPI
T2  - Plants
T1  - Population-Related Variability in Qualitative and Quantitative Secondary Metabolite Profile of Gentianella austriaca (A. & J. Kern.) Holub
IS  - 13
VL  - 12
DO  - 10.3390/plants12132434
SP  - 2434
ER  - 

@article{
author = "Popović, Zorica and Vidaković, Vera and Mijalković, Tatjana and Krstić-Milošević, Dijana",
year = "2023",
abstract = "Phytochemical profiling of six natural populations of Gentianella austriaca was performed
by HPLC identification and quantification of a number of secondary metabolites, and evaluation of time series of peak areas by chemometric analysis. Phytochemical analysis of G. austriaca revealed the presence of iridoids, flavone-C-glucosides and xanthones. Twelve secondary metabolites were identified in the aerial parts, roots and seeds, including swertiamarin (SWM), gentiopicrin (GP), sweroside (SWZ), isoorientin (ISOOR), swertisin (SWE), demethylbellidifolin-8-O-glucoside (DMB-8-O-glc), bellidifolin-8-O-glucoside (BDF-8-O-glc), mangiferin (MGF), corymbiferin (CBF), corymbiferin-1-O-glucoside (CBF-1-O-glc), bellidifolin (BDF) and campestroside. Multivariate statistical analyses showed relatively low variability among populations according to secondary metabolite content. However, some pharmacologically important compounds were found in higher amounts in a few populations, which could be useful for conservation and future biotechnological procedures.",
publisher = "Basel: MDPI",
journal = "Plants",
title = "Population-Related Variability in Qualitative and Quantitative Secondary Metabolite Profile of Gentianella austriaca (A. & J. Kern.) Holub",
number = "13",
volume = "12",
doi = "10.3390/plants12132434",
pages = "2434"
}

Popović, Z., Vidaković, V., Mijalković, T.,& Krstić-Milošević, D.. (2023). Population-Related Variability in Qualitative and Quantitative Secondary Metabolite Profile of Gentianella austriaca (A. & J. Kern.) Holub. in Plants
Basel: MDPI., 12(13), 2434.
https://doi.org/10.3390/plants12132434

Popović Z, Vidaković V, Mijalković T, Krstić-Milošević D. Population-Related Variability in Qualitative and Quantitative Secondary Metabolite Profile of Gentianella austriaca (A. & J. Kern.) Holub. in Plants. 2023;12(13):2434.
doi:10.3390/plants12132434 .

Popović, Zorica, Vidaković, Vera, Mijalković, Tatjana, Krstić-Milošević, Dijana, "Population-Related Variability in Qualitative and Quantitative Secondary Metabolite Profile of Gentianella austriaca (A. & J. Kern.) Holub" in Plants, 12, no. 13 (2023):2434,
https://doi.org/10.3390/plants12132434 . .

TY  - JOUR
AU  - Banjac, Nevena
AU  - Krstić-Milošević, Dijana
AU  - Mijalković, Tatjana
AU  - Petrović, Mirjana
AU  - Ćosić, Tatjana
AU  - Stanišić, Mariana
AU  - Vinterhalter, Branka
PY  - 2023
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5698
AB  - Eruca sativa is known in traditional medicine for its therapeutic effects, while young plants
are used as a salad or green food. Recently, the consumption of rocket has increased considerably,
so it has become very important for breeders. Plant tissue culture provides a platform to overcome
the problems in improving this species. In the present study, an efficient protocol for in vitro shoot
regeneration and propagation of recalcitrant rocket variety Domaća rukola was studied. Murashige
and Skoog (MS, 1962) medium containing 0.1 mg/L BA and frequent subculture over a period
of three weeks proved to be optimal for shoot multiplication with a multiplication index of over 3
and only 8.72% of hyperhydrated shoots without necrosis. Different concentrations of 2,4-D, BA, or
TDZ in combination with NAA, with or without the presence of AgNO3, were tested for de novo
shoot organogenesis (DNSO) from seedling explants. The hypocotyl explants cultured on MS with a
combination of TDZ1.0+NAA0.1+AgNO3 5.0 mg/L regenerated viable shoots with the highest rate
(25.38%) and an average number of 2.18 shoots per regenerating explant. Somatic embryogenesis
from immature zygotic embryos proved to be the best way to regenerate a recalcitrant rocket cultivar.
The highest embryogenic efficiency was achieved in explants cultured on MS medium containing
1.0 mg/L 2,4-D with a frequency of 76.64% and 5.13 mean number of regenerated somatic embryos
per explant, which were further converted into normal plants. Additionally, in vitro-produced rocket
shoots could serve as a possible promising source for the production of flavonoid kaempferol with
proven antioxidant properties.
PB  - Basel: MDPI
T2  - Horticulturae
T1  - In Vitro Shoot Multiplication and Regeneration of the Recalcitrant Rocket (Eruca sativa Mill.) Variety Domaća Rukola
IS  - 5
VL  - 9
DO  - 10.3390/horticulturae9050533
SP  - 533
ER  - 

@article{
author = "Banjac, Nevena and Krstić-Milošević, Dijana and Mijalković, Tatjana and Petrović, Mirjana and Ćosić, Tatjana and Stanišić, Mariana and Vinterhalter, Branka",
year = "2023",
abstract = "Eruca sativa is known in traditional medicine for its therapeutic effects, while young plants
are used as a salad or green food. Recently, the consumption of rocket has increased considerably,
so it has become very important for breeders. Plant tissue culture provides a platform to overcome
the problems in improving this species. In the present study, an efficient protocol for in vitro shoot
regeneration and propagation of recalcitrant rocket variety Domaća rukola was studied. Murashige
and Skoog (MS, 1962) medium containing 0.1 mg/L BA and frequent subculture over a period
of three weeks proved to be optimal for shoot multiplication with a multiplication index of over 3
and only 8.72% of hyperhydrated shoots without necrosis. Different concentrations of 2,4-D, BA, or
TDZ in combination with NAA, with or without the presence of AgNO3, were tested for de novo
shoot organogenesis (DNSO) from seedling explants. The hypocotyl explants cultured on MS with a
combination of TDZ1.0+NAA0.1+AgNO3 5.0 mg/L regenerated viable shoots with the highest rate
(25.38%) and an average number of 2.18 shoots per regenerating explant. Somatic embryogenesis
from immature zygotic embryos proved to be the best way to regenerate a recalcitrant rocket cultivar.
The highest embryogenic efficiency was achieved in explants cultured on MS medium containing
1.0 mg/L 2,4-D with a frequency of 76.64% and 5.13 mean number of regenerated somatic embryos
per explant, which were further converted into normal plants. Additionally, in vitro-produced rocket
shoots could serve as a possible promising source for the production of flavonoid kaempferol with
proven antioxidant properties.",
publisher = "Basel: MDPI",
journal = "Horticulturae",
title = "In Vitro Shoot Multiplication and Regeneration of the Recalcitrant Rocket (Eruca sativa Mill.) Variety Domaća Rukola",
number = "5",
volume = "9",
doi = "10.3390/horticulturae9050533",
pages = "533"
}

Banjac, N., Krstić-Milošević, D., Mijalković, T., Petrović, M., Ćosić, T., Stanišić, M.,& Vinterhalter, B.. (2023). In Vitro Shoot Multiplication and Regeneration of the Recalcitrant Rocket (Eruca sativa Mill.) Variety Domaća Rukola. in Horticulturae
Basel: MDPI., 9(5), 533.
https://doi.org/10.3390/horticulturae9050533

Banjac N, Krstić-Milošević D, Mijalković T, Petrović M, Ćosić T, Stanišić M, Vinterhalter B. In Vitro Shoot Multiplication and Regeneration of the Recalcitrant Rocket (Eruca sativa Mill.) Variety Domaća Rukola. in Horticulturae. 2023;9(5):533.
doi:10.3390/horticulturae9050533 .

Banjac, Nevena, Krstić-Milošević, Dijana, Mijalković, Tatjana, Petrović, Mirjana, Ćosić, Tatjana, Stanišić, Mariana, Vinterhalter, Branka, "In Vitro Shoot Multiplication and Regeneration of the Recalcitrant Rocket (Eruca sativa Mill.) Variety Domaća Rukola" in Horticulturae, 9, no. 5 (2023):533,
https://doi.org/10.3390/horticulturae9050533 . .

TY  - CONF
AU  - Ćosić, Tatjana
AU  - Devrnja, Nina
AU  - Krstić Milošević, Dijana
AU  - Božić, Dragana
AU  - Jančić, Ivan
AU  - Raspor, Martin
AU  - Maksimović, Zoran
PY  - 2022
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5365
AB  - In Serbia, citral chemotype of Thymus pannonicus All. (Lamiaceae) can be only found on Mt
Vršačke planine, as dominant form, where is traditionally used for medicinal purposes. To overcome
difficulties in harvesting due to distribution in inaccessible areas, and to prevent possible
overexploitation from its natural habitat, finding the means for alternative cultivation of this plant
for its prospective beneficial use in human health care was set as objective of this study. Efficient
system for in vitro shoot propagation of T. pannonicus was established for the first time. Different
types of auxin (indole-3-acetic acid, IAA; indole-3-butyric acid, IBA; 1-naphthaleneacetic acid, NAA)
were used for testing the process of rhizogenesis in vitro. Best response was detected on IBA and
NAA for rhizogenesis frequency and number of formed roots, respectively. Rooted plants were
successfully acclimatized. In vitro shoots cultivated on PGR free medium (E1) and medium with 1
mg L-1 IAA (E2) were extracted with 80% methanol and analysed by HPLC. Presence of rosmarinic
acid (RA) as dominant component was indicated in both extracts, with E2 containing higher RA
amount (45.116 mg g-1DW). Antimicrobial activity of extracts was tested using broth microdilution
method against nine strains of laboratory control pathogenic microorganisms. Both extracts
exerted significant antimicrobial activity against all tested pathogens with minimal inhibitory concen-
trations (MIC) in the range of 6.25-50 mg mL-1, while E2 was more effective against Gram positive
bacteria (6.25-12.5 mg mL-1). Presented results could give inducement for potential greater
commercial use of this significant species in the future.
PB  - Belgrade: Serbian Plant Physiology Society
C3  - 4th International Conference on Plant Biology [and] 23rd SPPS Meeting; 2022 Oct 6-8; Belgrade, Serbia
T1  - Potential of Thymus pannonicus All. in vitro culture: micropropagation, rosmarinic acid production and antimicrobial activity
SP  - 104
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_5365
ER  - 

@conference{
author = "Ćosić, Tatjana and Devrnja, Nina and Krstić Milošević, Dijana and Božić, Dragana and Jančić, Ivan and Raspor, Martin and Maksimović, Zoran",
year = "2022",
abstract = "In Serbia, citral chemotype of Thymus pannonicus All. (Lamiaceae) can be only found on Mt
Vršačke planine, as dominant form, where is traditionally used for medicinal purposes. To overcome
difficulties in harvesting due to distribution in inaccessible areas, and to prevent possible
overexploitation from its natural habitat, finding the means for alternative cultivation of this plant
for its prospective beneficial use in human health care was set as objective of this study. Efficient
system for in vitro shoot propagation of T. pannonicus was established for the first time. Different
types of auxin (indole-3-acetic acid, IAA; indole-3-butyric acid, IBA; 1-naphthaleneacetic acid, NAA)
were used for testing the process of rhizogenesis in vitro. Best response was detected on IBA and
NAA for rhizogenesis frequency and number of formed roots, respectively. Rooted plants were
successfully acclimatized. In vitro shoots cultivated on PGR free medium (E1) and medium with 1
mg L-1 IAA (E2) were extracted with 80% methanol and analysed by HPLC. Presence of rosmarinic
acid (RA) as dominant component was indicated in both extracts, with E2 containing higher RA
amount (45.116 mg g-1DW). Antimicrobial activity of extracts was tested using broth microdilution
method against nine strains of laboratory control pathogenic microorganisms. Both extracts
exerted significant antimicrobial activity against all tested pathogens with minimal inhibitory concen-
trations (MIC) in the range of 6.25-50 mg mL-1, while E2 was more effective against Gram positive
bacteria (6.25-12.5 mg mL-1). Presented results could give inducement for potential greater
commercial use of this significant species in the future.",
publisher = "Belgrade: Serbian Plant Physiology Society",
journal = "4th International Conference on Plant Biology [and] 23rd SPPS Meeting; 2022 Oct 6-8; Belgrade, Serbia",
title = "Potential of Thymus pannonicus All. in vitro culture: micropropagation, rosmarinic acid production and antimicrobial activity",
pages = "104",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_5365"
}

Ćosić, T., Devrnja, N., Krstić Milošević, D., Božić, D., Jančić, I., Raspor, M.,& Maksimović, Z.. (2022). Potential of Thymus pannonicus All. in vitro culture: micropropagation, rosmarinic acid production and antimicrobial activity. in 4th International Conference on Plant Biology [and] 23rd SPPS Meeting; 2022 Oct 6-8; Belgrade, Serbia
Belgrade: Serbian Plant Physiology Society., 104.
https://hdl.handle.net/21.15107/rcub_ibiss_5365

Ćosić T, Devrnja N, Krstić Milošević D, Božić D, Jančić I, Raspor M, Maksimović Z. Potential of Thymus pannonicus All. in vitro culture: micropropagation, rosmarinic acid production and antimicrobial activity. in 4th International Conference on Plant Biology [and] 23rd SPPS Meeting; 2022 Oct 6-8; Belgrade, Serbia. 2022;:104.
https://hdl.handle.net/21.15107/rcub_ibiss_5365 .

Ćosić, Tatjana, Devrnja, Nina, Krstić Milošević, Dijana, Božić, Dragana, Jančić, Ivan, Raspor, Martin, Maksimović, Zoran, "Potential of Thymus pannonicus All. in vitro culture: micropropagation, rosmarinic acid production and antimicrobial activity" in 4th International Conference on Plant Biology [and] 23rd SPPS Meeting; 2022 Oct 6-8; Belgrade, Serbia (2022):104,
https://hdl.handle.net/21.15107/rcub_ibiss_5365 .

TY  - CONF
AU  - Tovilović-Kovačević, Gordana
AU  - Krstić-Milošević, Dijana
AU  - Vinterhalter, Branka
AU  - Toljić, Mina
AU  - Perović, Vladimir
AU  - Trajković, Vladimir
AU  - Harhaji-Trajković, Ljubica
AU  - Zogović, Nevena
PY  - 2022
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5317
AB  - Глиобластом je најчешћи и најагресивнији тип тумора централног нервног система код одраслих. Циљ ове студије је био да се процени антиглиомски потенцијал екстраката коренова Gentiana dinarica у култури U251 ћелија хуманог глиобластома. Метанолни екстракти су добијени из нетрансформисаних коренова G. dinarica (екстракт 1, Е1) и трансгених коренова добијених коришћењем два соја Agrobacterium rhizogenes: A4M70GUS (екстракт 2, Е2) и 15834/PI (екстракт 3, Е3). Трансформацијом коренова са A. rhizogenes стимулисана је продукција ксантона, секундарних метаболита са доказаним антиканцерским ефектом. За разлику од Е1 и Е2, Е3 је снажно инхибирао раст U251 ћелија, изазвао застој ћелијског циклуса у G2/M фази и повећао експресију маркера диференцијације – астроцитног глијалног фибриларног киселог протеина (GFAP) и неуронског β-тубулина. Е3 је стимулисао Akt/mTOR-зависну аутофагију, на шта је указивало повећање нивоа аутофагног маркера LC3-II протеина и појачана деградација селективне аутофагне мете протеина p62. Инхибиција аутофагије је спречила експресију маркера диференцијације, без утицаја на застој у ћелијском циклусу. Е3 је повећао и нивое оксидативног стреса у ћелији, а антиоксиданси N-ацетил цистеин (NAC) и витамин Е су инхибирали и аутофагију и диференцијацију U251 ћелија изазвану Е3. Активна компонента Е3 је највероватније ксантонски агликон норсверцијанин, најзаступљеније једињење у Е3. Норсверцијанин је, као и Е3, зауставио пролиферацију U251 ћелија у G2/M фази ћелијског циклуса и изазвао диференцијацију, аутофагију и оксидативни стрес. Резултати ове студије указују да би Е3 и норсверцијанин могли бити кандидати за диференцијациону терапију глиобластома.
AB  - Glioblastom je najčešći i najagresivniji tip tumora centralnog nervnog sistema kod odraslih. Cilj ove studije je bio da se proceni antigliomski potencijal ekstrakata korenova Gentiana dinarica u kulturi U251 ćelija humanog glioblastoma. Metanolni ekstrakti su dobijeni iz netransformisanih korenova G. dinarica (ekstrakt 1, E1) i transgenih korenova dobijenih korišćenjem dva soja Agrobacterium rhizogenes: A4M70GUS (ekstrakt 2, E2) i 15834/PI (ekstrakt 3, E3). Transformacijom korenova sa A. rhizogenes stimulisana je produkcija ksantona, sekundarnih metabolita sa dokazanim antikancerskim efektom. Za razliku od E1 i E2, E3 je snažno inhibirao rast U251 ćelija, izazvao zastoj ćelijskog ciklusa u G2/M fazi i povećao ekspresiju markera diferencijacije – astrocitnog glijalnog fibrilarnog kiselog proteina (GFAP) i neuronskog β-tubulina. E3 je stimulisao Akt/mTOR-zavisnu autofagiju, na šta je ukazivalo povećanje nivoa autofagnog markera LC3-II proteina i pojačana degradacija selektivne autofagne mete proteina p62. Inhibicija autofagije je sprečila ekspresiju markera diferencijacije, bez uticaja na zastoj u ćelijskom ciklusu. E3 je povećao i nivoe oksidativnog stresa u ćeliji, a antioksidansi N-acetil cistein (NAC) i vitamin E su inhibirali i autofagiju i diferencijaciju U251 ćelija izazvanu E3. Aktivna komponenta E3 je najverovatnije ksantonski aglikon norsvercijanin, najzastupljenije jedinjenje u E3. Norsvercijanin je, kao i E3, zaustavio proliferaciju U251 ćelija u G2/M fazi ćelijskog ciklusa i izazvao diferencijaciju, autofagiju i oksidativni stres. Rezultati ove studije ukazuju da bi E3 i norsvercijanin mogli biti kandidati za diferencijacionu terapiju glioblastoma.
PB  - Belgrade: Serbian Biological Society
C3  - Knjiga sažetaka: Treći Kongres biologa Srbije: Osnovna i primenjena istraživanja: Metodika nastave; 2022 Sep 21-25; Zlatibor, Serbia
T1  - Antigliomski efekat ekstrakta korena Gentiana dinarica Beck. obogaćenog ksantonima
T1  - Антиглиомски ефекат екстракта корена Gentiana dinarica Beck. обогаћеног ксантонима
SP  - 280
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_5317
ER  - 

@conference{
author = "Tovilović-Kovačević, Gordana and Krstić-Milošević, Dijana and Vinterhalter, Branka and Toljić, Mina and Perović, Vladimir and Trajković, Vladimir and Harhaji-Trajković, Ljubica and Zogović, Nevena",
year = "2022",
abstract = "Глиобластом je најчешћи и најагресивнији тип тумора централног нервног система код одраслих. Циљ ове студије је био да се процени антиглиомски потенцијал екстраката коренова Gentiana dinarica у култури U251 ћелија хуманог глиобластома. Метанолни екстракти су добијени из нетрансформисаних коренова G. dinarica (екстракт 1, Е1) и трансгених коренова добијених коришћењем два соја Agrobacterium rhizogenes: A4M70GUS (екстракт 2, Е2) и 15834/PI (екстракт 3, Е3). Трансформацијом коренова са A. rhizogenes стимулисана је продукција ксантона, секундарних метаболита са доказаним антиканцерским ефектом. За разлику од Е1 и Е2, Е3 је снажно инхибирао раст U251 ћелија, изазвао застој ћелијског циклуса у G2/M фази и повећао експресију маркера диференцијације – астроцитног глијалног фибриларног киселог протеина (GFAP) и неуронског β-тубулина. Е3 је стимулисао Akt/mTOR-зависну аутофагију, на шта је указивало повећање нивоа аутофагног маркера LC3-II протеина и појачана деградација селективне аутофагне мете протеина p62. Инхибиција аутофагије је спречила експресију маркера диференцијације, без утицаја на застој у ћелијском циклусу. Е3 је повећао и нивое оксидативног стреса у ћелији, а антиоксиданси N-ацетил цистеин (NAC) и витамин Е су инхибирали и аутофагију и диференцијацију U251 ћелија изазвану Е3. Активна компонента Е3 је највероватније ксантонски агликон норсверцијанин, најзаступљеније једињење у Е3. Норсверцијанин је, као и Е3, зауставио пролиферацију U251 ћелија у G2/M фази ћелијског циклуса и изазвао диференцијацију, аутофагију и оксидативни стрес. Резултати ове студије указују да би Е3 и норсверцијанин могли бити кандидати за диференцијациону терапију глиобластома., Glioblastom je najčešći i najagresivniji tip tumora centralnog nervnog sistema kod odraslih. Cilj ove studije je bio da se proceni antigliomski potencijal ekstrakata korenova Gentiana dinarica u kulturi U251 ćelija humanog glioblastoma. Metanolni ekstrakti su dobijeni iz netransformisanih korenova G. dinarica (ekstrakt 1, E1) i transgenih korenova dobijenih korišćenjem dva soja Agrobacterium rhizogenes: A4M70GUS (ekstrakt 2, E2) i 15834/PI (ekstrakt 3, E3). Transformacijom korenova sa A. rhizogenes stimulisana je produkcija ksantona, sekundarnih metabolita sa dokazanim antikancerskim efektom. Za razliku od E1 i E2, E3 je snažno inhibirao rast U251 ćelija, izazvao zastoj ćelijskog ciklusa u G2/M fazi i povećao ekspresiju markera diferencijacije – astrocitnog glijalnog fibrilarnog kiselog proteina (GFAP) i neuronskog β-tubulina. E3 je stimulisao Akt/mTOR-zavisnu autofagiju, na šta je ukazivalo povećanje nivoa autofagnog markera LC3-II proteina i pojačana degradacija selektivne autofagne mete proteina p62. Inhibicija autofagije je sprečila ekspresiju markera diferencijacije, bez uticaja na zastoj u ćelijskom ciklusu. E3 je povećao i nivoe oksidativnog stresa u ćeliji, a antioksidansi N-acetil cistein (NAC) i vitamin E su inhibirali i autofagiju i diferencijaciju U251 ćelija izazvanu E3. Aktivna komponenta E3 je najverovatnije ksantonski aglikon norsvercijanin, najzastupljenije jedinjenje u E3. Norsvercijanin je, kao i E3, zaustavio proliferaciju U251 ćelija u G2/M fazi ćelijskog ciklusa i izazvao diferencijaciju, autofagiju i oksidativni stres. Rezultati ove studije ukazuju da bi E3 i norsvercijanin mogli biti kandidati za diferencijacionu terapiju glioblastoma.",
publisher = "Belgrade: Serbian Biological Society",
journal = "Knjiga sažetaka: Treći Kongres biologa Srbije: Osnovna i primenjena istraživanja: Metodika nastave; 2022 Sep 21-25; Zlatibor, Serbia",
title = "Antigliomski efekat ekstrakta korena Gentiana dinarica Beck. obogaćenog ksantonima, Антиглиомски ефекат екстракта корена Gentiana dinarica Beck. обогаћеног ксантонима",
pages = "280",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_5317"
}

Tovilović-Kovačević, G., Krstić-Milošević, D., Vinterhalter, B., Toljić, M., Perović, V., Trajković, V., Harhaji-Trajković, L.,& Zogović, N.. (2022). Antigliomski efekat ekstrakta korena Gentiana dinarica Beck. obogaćenog ksantonima. in Knjiga sažetaka: Treći Kongres biologa Srbije: Osnovna i primenjena istraživanja: Metodika nastave; 2022 Sep 21-25; Zlatibor, Serbia
Belgrade: Serbian Biological Society., 280.
https://hdl.handle.net/21.15107/rcub_ibiss_5317

Tovilović-Kovačević G, Krstić-Milošević D, Vinterhalter B, Toljić M, Perović V, Trajković V, Harhaji-Trajković L, Zogović N. Antigliomski efekat ekstrakta korena Gentiana dinarica Beck. obogaćenog ksantonima. in Knjiga sažetaka: Treći Kongres biologa Srbije: Osnovna i primenjena istraživanja: Metodika nastave; 2022 Sep 21-25; Zlatibor, Serbia. 2022;:280.
https://hdl.handle.net/21.15107/rcub_ibiss_5317 .

Tovilović-Kovačević, Gordana, Krstić-Milošević, Dijana, Vinterhalter, Branka, Toljić, Mina, Perović, Vladimir, Trajković, Vladimir, Harhaji-Trajković, Ljubica, Zogović, Nevena, "Antigliomski efekat ekstrakta korena Gentiana dinarica Beck. obogaćenog ksantonima" in Knjiga sažetaka: Treći Kongres biologa Srbije: Osnovna i primenjena istraživanja: Metodika nastave; 2022 Sep 21-25; Zlatibor, Serbia (2022):280,
https://hdl.handle.net/21.15107/rcub_ibiss_5317 .

TY  - CONF
AU  - Filipović, Biljana
AU  - Šiler, Branislav
AU  - Banjanac, Tijana
AU  - Nestorović Živković, Jasmina
AU  - Skorić, Marijana
AU  - Krstić Milošević, Dijana
AU  - Mišić, Danijela
PY  - 2022
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6197
AB  - Centaurium erythraea Rafn (common centaury) is an important medicinal plant species whose various health effects have been ascribed to its key specialized bioactive metabolites (mostly to secoiridoids and xanthones). Assessment of species diversity through the analysis of ploidy level and genetic variation is important for the selection of high-yielding genotypes for sustainable production of pharmacologically important specialized metabolites. In this work, we scored ploidy level of C. erythraea germplasm stored in ex situ seed collection containing seeds of more than 40 populations originating at locations across the central Balkan Peninsula. Furthermore, we selected two neighboring populations, one of the diploid and one of the tetraploid genome background, and compared growth parameters and productivity among and within diploid and tetraploid genotypes. Plants belonging to the diploid population showed different growth patterns as compared to tetraploid plants after one month of growth under in vitro conditions. To evaluate the content of iridoids (loganin and loganic acid), secoiridoids (secologanin, sweroside, gentiopicrin, and swertiamarin), and xanthones (demethyleustomin, methylbellidifolin, eustomin, and decussatin) in diploid and tetraploid genotypes, a targeted UHPLC/DAD/(±)HESI-MS2 analysis was performed for methanol extracts of shoots and roots, separately. One-month-old shoots of all diploid genotypes contained significantly higher amounts of total iridoids and xanthones than tetraploids, primarily due to high amounts of sweroside, decussatin, and methylbellidifolin. The observed variation both among and within diploid and tetraploid genotypes should be taken into consideration when estimating strategies for biotechnological improvement and for unraveling molecular background of specialized metabolites biosynthesis.
PB  - Belgrade: Serbian Plant Physiology Society
C3  - Book of Abstracts: 4th International Conference on Plant Biology [and] 23rd SPPS Meeting; 2022 Oct 6-8; Belgrade, Serbia
T1  - Ploidy profiling of Centaurium erythraea germplasm representing the diversity of the central Balkans: in vitro screening for highly productive diploid and tetraploid genotypes
SP  - 32
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_6197
ER  - 

@conference{
author = "Filipović, Biljana and Šiler, Branislav and Banjanac, Tijana and Nestorović Živković, Jasmina and Skorić, Marijana and Krstić Milošević, Dijana and Mišić, Danijela",
year = "2022",
abstract = "Centaurium erythraea Rafn (common centaury) is an important medicinal plant species whose various health effects have been ascribed to its key specialized bioactive metabolites (mostly to secoiridoids and xanthones). Assessment of species diversity through the analysis of ploidy level and genetic variation is important for the selection of high-yielding genotypes for sustainable production of pharmacologically important specialized metabolites. In this work, we scored ploidy level of C. erythraea germplasm stored in ex situ seed collection containing seeds of more than 40 populations originating at locations across the central Balkan Peninsula. Furthermore, we selected two neighboring populations, one of the diploid and one of the tetraploid genome background, and compared growth parameters and productivity among and within diploid and tetraploid genotypes. Plants belonging to the diploid population showed different growth patterns as compared to tetraploid plants after one month of growth under in vitro conditions. To evaluate the content of iridoids (loganin and loganic acid), secoiridoids (secologanin, sweroside, gentiopicrin, and swertiamarin), and xanthones (demethyleustomin, methylbellidifolin, eustomin, and decussatin) in diploid and tetraploid genotypes, a targeted UHPLC/DAD/(±)HESI-MS2 analysis was performed for methanol extracts of shoots and roots, separately. One-month-old shoots of all diploid genotypes contained significantly higher amounts of total iridoids and xanthones than tetraploids, primarily due to high amounts of sweroside, decussatin, and methylbellidifolin. The observed variation both among and within diploid and tetraploid genotypes should be taken into consideration when estimating strategies for biotechnological improvement and for unraveling molecular background of specialized metabolites biosynthesis.",
publisher = "Belgrade: Serbian Plant Physiology Society",
journal = "Book of Abstracts: 4th International Conference on Plant Biology [and] 23rd SPPS Meeting; 2022 Oct 6-8; Belgrade, Serbia",
title = "Ploidy profiling of Centaurium erythraea germplasm representing the diversity of the central Balkans: in vitro screening for highly productive diploid and tetraploid genotypes",
pages = "32",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_6197"
}

Filipović, B., Šiler, B., Banjanac, T., Nestorović Živković, J., Skorić, M., Krstić Milošević, D.,& Mišić, D.. (2022). Ploidy profiling of Centaurium erythraea germplasm representing the diversity of the central Balkans: in vitro screening for highly productive diploid and tetraploid genotypes. in Book of Abstracts: 4th International Conference on Plant Biology [and] 23rd SPPS Meeting; 2022 Oct 6-8; Belgrade, Serbia
Belgrade: Serbian Plant Physiology Society., 32.
https://hdl.handle.net/21.15107/rcub_ibiss_6197

Filipović B, Šiler B, Banjanac T, Nestorović Živković J, Skorić M, Krstić Milošević D, Mišić D. Ploidy profiling of Centaurium erythraea germplasm representing the diversity of the central Balkans: in vitro screening for highly productive diploid and tetraploid genotypes. in Book of Abstracts: 4th International Conference on Plant Biology [and] 23rd SPPS Meeting; 2022 Oct 6-8; Belgrade, Serbia. 2022;:32.
https://hdl.handle.net/21.15107/rcub_ibiss_6197 .

Filipović, Biljana, Šiler, Branislav, Banjanac, Tijana, Nestorović Živković, Jasmina, Skorić, Marijana, Krstić Milošević, Dijana, Mišić, Danijela, "Ploidy profiling of Centaurium erythraea germplasm representing the diversity of the central Balkans: in vitro screening for highly productive diploid and tetraploid genotypes" in Book of Abstracts: 4th International Conference on Plant Biology [and] 23rd SPPS Meeting; 2022 Oct 6-8; Belgrade, Serbia (2022):32,
https://hdl.handle.net/21.15107/rcub_ibiss_6197 .

TY  - CONF
AU  - Vinterhalter, Branka
AU  - Krstić-Milošević, Dijana
AU  - Petrović, Mirjana
AU  - Devrnja, Nina
AU  - Banjac, Nevena
PY  - 2022
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5754
AB  - Eruca sativa (rocket, Brassicaceae), is known in traditional medicine for its therapeutic, while young plants are used as salad, vegetable, or green foods. Genetic improvement of the rocket has been limited due to the appearance of biennial plants and the requirement for isolation barriers. Plant tissue culture offers a platform to overcome existing barriers to improving this species. The present study aimed to achieve a feasible protocol for shoot regeneration and propagation of rocket cv. Domaća rukola and to evaluate the use of AgNO3 to improve regeneration efficiency. Proliferation of shoots from seedling epicotyl, shoot organogenesis from root, hypocotyl and cotyledon seedling explants, and somatic embryogenesis from immature zygotic embryos were studied. Murashige and Skoog (MS) basal medium in combination with a low KIN concentration of 0.1 mg l-1 with frequent subculture over a period of 3 weeks was found to be optimal for shoot multiplication with a multiplication index of almost 3 and only 9.3% of vitrified shoots without necrosis. Different concentrations of 2,4-D, BA or TDZ in combination with NAA and with or without AgNO3 were tested for shoot regeneration from seedling explants. As a result, the hypocotyl explants cultured on MS with a combination of TDZ 1.0+NAA 0.1+AgNO3 5.0 mg l-1 provided healthy shoots with a satisfactory regeneration rate of 25.4% and 2.2 mean number of shoots per regenerating explant. Immature zygotic embryos cultured on MS medium containing 1.0 mg l-1 2.4-D showed the highest regeneration frequency (78.76%) as well as the number of regenerated somatic embryos per explant (5.13) with low incidence of callusing and necrosis. KIN at 0.5 mg l-1 enabled the best conversion of somatic embryos into healthy plants (22.5%). The results provide an opportunity to use established regeneration protocols in rocket breeding improvement programs.
PB  - Ljubljana: Slovenian Society of Plant Biology
C3  - Book of abstracts: Plants in Changing Environment: International conference of the Slovenian Society of Plant Biology; 2022 Sep 15-16; Ljubljana, Slovenia
T1  - In vitro shoot propagation and regeneration of rocket (Eruca sativa Mill.) using organogenesis and somatic embryogenesis
SP  - 18
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_5754
ER  - 

@conference{
author = "Vinterhalter, Branka and Krstić-Milošević, Dijana and Petrović, Mirjana and Devrnja, Nina and Banjac, Nevena",
year = "2022",
abstract = "Eruca sativa (rocket, Brassicaceae), is known in traditional medicine for its therapeutic, while young plants are used as salad, vegetable, or green foods. Genetic improvement of the rocket has been limited due to the appearance of biennial plants and the requirement for isolation barriers. Plant tissue culture offers a platform to overcome existing barriers to improving this species. The present study aimed to achieve a feasible protocol for shoot regeneration and propagation of rocket cv. Domaća rukola and to evaluate the use of AgNO3 to improve regeneration efficiency. Proliferation of shoots from seedling epicotyl, shoot organogenesis from root, hypocotyl and cotyledon seedling explants, and somatic embryogenesis from immature zygotic embryos were studied. Murashige and Skoog (MS) basal medium in combination with a low KIN concentration of 0.1 mg l-1 with frequent subculture over a period of 3 weeks was found to be optimal for shoot multiplication with a multiplication index of almost 3 and only 9.3% of vitrified shoots without necrosis. Different concentrations of 2,4-D, BA or TDZ in combination with NAA and with or without AgNO3 were tested for shoot regeneration from seedling explants. As a result, the hypocotyl explants cultured on MS with a combination of TDZ 1.0+NAA 0.1+AgNO3 5.0 mg l-1 provided healthy shoots with a satisfactory regeneration rate of 25.4% and 2.2 mean number of shoots per regenerating explant. Immature zygotic embryos cultured on MS medium containing 1.0 mg l-1 2.4-D showed the highest regeneration frequency (78.76%) as well as the number of regenerated somatic embryos per explant (5.13) with low incidence of callusing and necrosis. KIN at 0.5 mg l-1 enabled the best conversion of somatic embryos into healthy plants (22.5%). The results provide an opportunity to use established regeneration protocols in rocket breeding improvement programs.",
publisher = "Ljubljana: Slovenian Society of Plant Biology",
journal = "Book of abstracts: Plants in Changing Environment: International conference of the Slovenian Society of Plant Biology; 2022 Sep 15-16; Ljubljana, Slovenia",
title = "In vitro shoot propagation and regeneration of rocket (Eruca sativa Mill.) using organogenesis and somatic embryogenesis",
pages = "18",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_5754"
}

Vinterhalter, B., Krstić-Milošević, D., Petrović, M., Devrnja, N.,& Banjac, N.. (2022). In vitro shoot propagation and regeneration of rocket (Eruca sativa Mill.) using organogenesis and somatic embryogenesis. in Book of abstracts: Plants in Changing Environment: International conference of the Slovenian Society of Plant Biology; 2022 Sep 15-16; Ljubljana, Slovenia
Ljubljana: Slovenian Society of Plant Biology., 18.
https://hdl.handle.net/21.15107/rcub_ibiss_5754

Vinterhalter B, Krstić-Milošević D, Petrović M, Devrnja N, Banjac N. In vitro shoot propagation and regeneration of rocket (Eruca sativa Mill.) using organogenesis and somatic embryogenesis. in Book of abstracts: Plants in Changing Environment: International conference of the Slovenian Society of Plant Biology; 2022 Sep 15-16; Ljubljana, Slovenia. 2022;:18.
https://hdl.handle.net/21.15107/rcub_ibiss_5754 .

Vinterhalter, Branka, Krstić-Milošević, Dijana, Petrović, Mirjana, Devrnja, Nina, Banjac, Nevena, "In vitro shoot propagation and regeneration of rocket (Eruca sativa Mill.) using organogenesis and somatic embryogenesis" in Book of abstracts: Plants in Changing Environment: International conference of the Slovenian Society of Plant Biology; 2022 Sep 15-16; Ljubljana, Slovenia (2022):18,
https://hdl.handle.net/21.15107/rcub_ibiss_5754 .

TY  - CONF
AU  - Krstić-Milošević, Dijana
AU  - Banjac, Nevena
AU  - Janković, Teodora
AU  - Devrnja, Nina
AU  - Eler, Klemen
AU  - Vinterhalter, Branka
PY  - 2022
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5560
AB  - Gentiana clusii Perr. & Song. (Gentianaceae), is a rare plant species found in the Alps, the Pyrenees, the Black Forest Mountains and the Carpathians. G. clusii is endemic for Europe which implies the need for its conservation. G. clusii plants were collected at fruitful stage in June 2015 at Velika Planina Mountain, Republic of Slovenia. Seeds of G. clusii were germinated on MS basal medium. Shoots cultured on WPM + 0.5 mg/L KIN were used as starting material in propagation and rooting experiments. Medium with IBA at 0.5 mg/L was chosen as optimal for rooting. HPLC screening of secondary metabolites of wild plants collected in nature and in vitro plants revealed the presence of two main classes of compounds typical for gentians – secoiridoids and xanthones. Secoiridoids gentiopicrin and sweroside were identified in both aerial parts of wild plants and shoots of in vitro plants, while root extracts contained amarogentin in addition. Aerial parts of wild plants contained gentiacaulein-1-O-primeveroside as dominant xanthone glycoside. HPLC also revealed the presence of gentiakochianin-1-O-primeveroside, norswertianin-1-O-primeveroside, 1,8- dihydroxy-3-methoxy-7-O-primeverosyl xanthone, and xanthone aglycons- norswertianin, gentiacaulein, and gentiakochianin. HPLC analysis shown that in vitro culture considerably promoted the production of secondary metabolites in G. clusii. The selected protocol for shoot propagation (WPM + 0.5 mg/L KIN) increased the content of sweroside, gentiopicrin and norswertianin-1-O-primeveroside for more than 2-fold compared with the wild plants. IBA promoted norswertianin-1-O-primeveroside and norswertianin production in root cultures; their contents were enhanced 6.4- and 18.6-fold, respectively, compared with the wild plants. The extract of these roots displayed high antioxidant capacity. The established shoot and root propagation protocols facilitate in vitro conservation of G. clusii, and provides a promising tool for the large scale production of valuable secoiridoids and xanthones.
PB  - Ljubljana: Slovenian Society of Plant Biology
C3  - Book of abstracts: Plants in Changing Environment: International conference of the Slovenian Society of Plant Biology; 2022 Sep 15-16; Ljubljana, Slovenia
T1  - Gentiana clusii Perr.&Song. cultured in vitro - secondary metabolite profile and evaluation of antioxidant activity
SP  - 31
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_5560
ER  - 

@conference{
editor = "Baebler, Špela, Dermastia, Marina, Grebenc, Tine, Praprotnik, Eva, Razinger, Jaka, Urbanek Krajnc, Andreja",
author = "Krstić-Milošević, Dijana and Banjac, Nevena and Janković, Teodora and Devrnja, Nina and Eler, Klemen and Vinterhalter, Branka",
year = "2022",
abstract = "Gentiana clusii Perr. & Song. (Gentianaceae), is a rare plant species found in the Alps, the Pyrenees, the Black Forest Mountains and the Carpathians. G. clusii is endemic for Europe which implies the need for its conservation. G. clusii plants were collected at fruitful stage in June 2015 at Velika Planina Mountain, Republic of Slovenia. Seeds of G. clusii were germinated on MS basal medium. Shoots cultured on WPM + 0.5 mg/L KIN were used as starting material in propagation and rooting experiments. Medium with IBA at 0.5 mg/L was chosen as optimal for rooting. HPLC screening of secondary metabolites of wild plants collected in nature and in vitro plants revealed the presence of two main classes of compounds typical for gentians – secoiridoids and xanthones. Secoiridoids gentiopicrin and sweroside were identified in both aerial parts of wild plants and shoots of in vitro plants, while root extracts contained amarogentin in addition. Aerial parts of wild plants contained gentiacaulein-1-O-primeveroside as dominant xanthone glycoside. HPLC also revealed the presence of gentiakochianin-1-O-primeveroside, norswertianin-1-O-primeveroside, 1,8- dihydroxy-3-methoxy-7-O-primeverosyl xanthone, and xanthone aglycons- norswertianin, gentiacaulein, and gentiakochianin. HPLC analysis shown that in vitro culture considerably promoted the production of secondary metabolites in G. clusii. The selected protocol for shoot propagation (WPM + 0.5 mg/L KIN) increased the content of sweroside, gentiopicrin and norswertianin-1-O-primeveroside for more than 2-fold compared with the wild plants. IBA promoted norswertianin-1-O-primeveroside and norswertianin production in root cultures; their contents were enhanced 6.4- and 18.6-fold, respectively, compared with the wild plants. The extract of these roots displayed high antioxidant capacity. The established shoot and root propagation protocols facilitate in vitro conservation of G. clusii, and provides a promising tool for the large scale production of valuable secoiridoids and xanthones.",
publisher = "Ljubljana: Slovenian Society of Plant Biology",
journal = "Book of abstracts: Plants in Changing Environment: International conference of the Slovenian Society of Plant Biology; 2022 Sep 15-16; Ljubljana, Slovenia",
title = "Gentiana clusii Perr.&Song. cultured in vitro - secondary metabolite profile and evaluation of antioxidant activity",
pages = "31",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_5560"
}

Baebler, Š., Dermastia, M., Grebenc, T., Praprotnik, E., Razinger, J., Urbanek Krajnc, A., Krstić-Milošević, D., Banjac, N., Janković, T., Devrnja, N., Eler, K.,& Vinterhalter, B.. (2022). Gentiana clusii Perr.&Song. cultured in vitro - secondary metabolite profile and evaluation of antioxidant activity. in Book of abstracts: Plants in Changing Environment: International conference of the Slovenian Society of Plant Biology; 2022 Sep 15-16; Ljubljana, Slovenia
Ljubljana: Slovenian Society of Plant Biology., 31.
https://hdl.handle.net/21.15107/rcub_ibiss_5560

Baebler Š, Dermastia M, Grebenc T, Praprotnik E, Razinger J, Urbanek Krajnc A, Krstić-Milošević D, Banjac N, Janković T, Devrnja N, Eler K, Vinterhalter B. Gentiana clusii Perr.&Song. cultured in vitro - secondary metabolite profile and evaluation of antioxidant activity. in Book of abstracts: Plants in Changing Environment: International conference of the Slovenian Society of Plant Biology; 2022 Sep 15-16; Ljubljana, Slovenia. 2022;:31.
https://hdl.handle.net/21.15107/rcub_ibiss_5560 .

Baebler, Špela, Dermastia, Marina, Grebenc, Tine, Praprotnik, Eva, Razinger, Jaka, Urbanek Krajnc, Andreja, Krstić-Milošević, Dijana, Banjac, Nevena, Janković, Teodora, Devrnja, Nina, Eler, Klemen, Vinterhalter, Branka, "Gentiana clusii Perr.&Song. cultured in vitro - secondary metabolite profile and evaluation of antioxidant activity" in Book of abstracts: Plants in Changing Environment: International conference of the Slovenian Society of Plant Biology; 2022 Sep 15-16; Ljubljana, Slovenia (2022):31,
https://hdl.handle.net/21.15107/rcub_ibiss_5560 .

TY  - JOUR
AU  - Popović, Zorica
AU  - Krstić Milošević, Dijana
AU  - Marković, Milena
AU  - Vidaković, Vera
AU  - Bojović, Srđan
PY  - 2021
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/4122
AB  - Natural populations of Gentiana asclepiadea L., located at two mountainous sites, were HPLC-analyzed regarding the contents of six representative secondary metabolites. The contents of swertiamarin (SWM), gentiopicrin (GP), sweroside (SWZ), mangiferin (MGF), isoorientin (ISOOR), and isovitexin (ISOV) were determined in six populations (three per study site), and separately for aboveground and belowground plant parts. PCA showed a clear separation of four groups according to the contents of the analyzed secondary metabolites. Out of six analyzed compounds, five were present in all samples and only one (SWZ) was found in Golija populations (belowground parts) but not in Vlasina populations, and its presence can be indicative of the geolocation of populations. Clear separation of groups was mostly affected by the different contents of chemical compounds in plant parts (aboveground versus belowground) and by the differences related to population origin (higher content of SWM and GP in belowground parts of individuals from Vlasina populations and higher content of MGF and ISOOR of individuals from Golija populations). The results of this study contribute to the spatiochemical profiling of G. asclepiadea populations and a better understanding of inter- and intrapopulation variability of pharmacologically important compounds.
PB  - Basel, Switzerland: MDPI
T2  - Plants
T1  - Gentiana asclepiadea L. from Two High Mountainous Habitats: Inter- and Intrapopulation Variability Based on Species’ Phytochemistry
IS  - 1
VL  - 10
DO  - 10.3390/plants10010140
SP  - 140
ER  - 

@article{
author = "Popović, Zorica and Krstić Milošević, Dijana and Marković, Milena and Vidaković, Vera and Bojović, Srđan",
year = "2021",
abstract = "Natural populations of Gentiana asclepiadea L., located at two mountainous sites, were HPLC-analyzed regarding the contents of six representative secondary metabolites. The contents of swertiamarin (SWM), gentiopicrin (GP), sweroside (SWZ), mangiferin (MGF), isoorientin (ISOOR), and isovitexin (ISOV) were determined in six populations (three per study site), and separately for aboveground and belowground plant parts. PCA showed a clear separation of four groups according to the contents of the analyzed secondary metabolites. Out of six analyzed compounds, five were present in all samples and only one (SWZ) was found in Golija populations (belowground parts) but not in Vlasina populations, and its presence can be indicative of the geolocation of populations. Clear separation of groups was mostly affected by the different contents of chemical compounds in plant parts (aboveground versus belowground) and by the differences related to population origin (higher content of SWM and GP in belowground parts of individuals from Vlasina populations and higher content of MGF and ISOOR of individuals from Golija populations). The results of this study contribute to the spatiochemical profiling of G. asclepiadea populations and a better understanding of inter- and intrapopulation variability of pharmacologically important compounds.",
publisher = "Basel, Switzerland: MDPI",
journal = "Plants",
title = "Gentiana asclepiadea L. from Two High Mountainous Habitats: Inter- and Intrapopulation Variability Based on Species’ Phytochemistry",
number = "1",
volume = "10",
doi = "10.3390/plants10010140",
pages = "140"
}

Popović, Z., Krstić Milošević, D., Marković, M., Vidaković, V.,& Bojović, S.. (2021). Gentiana asclepiadea L. from Two High Mountainous Habitats: Inter- and Intrapopulation Variability Based on Species’ Phytochemistry. in Plants
Basel, Switzerland: MDPI., 10(1), 140.
https://doi.org/10.3390/plants10010140

Popović Z, Krstić Milošević D, Marković M, Vidaković V, Bojović S. Gentiana asclepiadea L. from Two High Mountainous Habitats: Inter- and Intrapopulation Variability Based on Species’ Phytochemistry. in Plants. 2021;10(1):140.
doi:10.3390/plants10010140 .

Popović, Zorica, Krstić Milošević, Dijana, Marković, Milena, Vidaković, Vera, Bojović, Srđan, "Gentiana asclepiadea L. from Two High Mountainous Habitats: Inter- and Intrapopulation Variability Based on Species’ Phytochemistry" in Plants, 10, no. 1 (2021):140,
https://doi.org/10.3390/plants10010140 . .

TY  - JOUR
AU  - Krstić Milošević, Dijana
AU  - Banjac, Nevena
AU  - Janković, Teodora
AU  - Vinterhalter, Dragan
AU  - Vinterhalter, Branka
PY  - 2021
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/4310
AB  - In vitro shoot culture of the endangered medicinal plant Gentianella lutescens was established from epicotyl explants cultured on MS basal medium with 0.2 mg L−1 6-benzylaminopurine (BA) and evaluated for xanthones content for the first time. Five shoot lines were obtained and no significant variations in multiplication rate, shoot elongation, and xanthones profile were found among them. The highest rooting rate (33.3%) was achieved by shoots treated for 2 days with 5 mg L−1 indole-3-butyric acid (IBA) followed by cultivation in liquid PGR-free ½ MS medium for 60 days. HPLC analysis revealed the lower content of xanthones—mangiferin, bellidifolin, demethylbellidifolin, demethylbellidifolin-8-O-glucoside and bellidifolin-8-O-glucoside—in in vitro cultured shoots compared to wild growing plants. The increasing concentration of sucrose, sorbitol and abiotic elicitors salicylic acid (SA), jasmonic acid (JA) and methyl jasmonate (MeJA) altered shoot growth and xanthone production. Sucrose and sorbitol applied at the highest concentration of 233.6 mM increased dry matter percentage, while SA at 100 μM promoted shoot growth 2-fold. The increased sucrose concentration enhanced accumulation of xanthones in shoot cultures 2–3-fold compared to the control shoots. Elicitors at 100–300 μM increased the accumulation of mangiferin, demethylbellidifolin-8-O-glucoside, and bellidifolin-8-O-glucoside almost equally, while MeJA at the highest concentration of 500 μM enhanced amount of aglycones demethylbellidifolin and bellidifolin 7-fold compared to the control. The obtained results facilitate conservation of G. lutescens and pave the way for further research on large-scale shoot propagation and production of pharmacologically active xanthones.
PB  - Basel: MDPI
T2  - Plants
T1  - Gentianella lutescens subsp. carpatica J. Holub.: Shoot Propagation In Vitro and Effect of Sucrose and Elicitors on Xanthones Production
IS  - 8
VL  - 10
DO  - 10.3390/plants10081651
SP  - 1651
ER  - 

@article{
author = "Krstić Milošević, Dijana and Banjac, Nevena and Janković, Teodora and Vinterhalter, Dragan and Vinterhalter, Branka",
year = "2021",
abstract = "In vitro shoot culture of the endangered medicinal plant Gentianella lutescens was established from epicotyl explants cultured on MS basal medium with 0.2 mg L−1 6-benzylaminopurine (BA) and evaluated for xanthones content for the first time. Five shoot lines were obtained and no significant variations in multiplication rate, shoot elongation, and xanthones profile were found among them. The highest rooting rate (33.3%) was achieved by shoots treated for 2 days with 5 mg L−1 indole-3-butyric acid (IBA) followed by cultivation in liquid PGR-free ½ MS medium for 60 days. HPLC analysis revealed the lower content of xanthones—mangiferin, bellidifolin, demethylbellidifolin, demethylbellidifolin-8-O-glucoside and bellidifolin-8-O-glucoside—in in vitro cultured shoots compared to wild growing plants. The increasing concentration of sucrose, sorbitol and abiotic elicitors salicylic acid (SA), jasmonic acid (JA) and methyl jasmonate (MeJA) altered shoot growth and xanthone production. Sucrose and sorbitol applied at the highest concentration of 233.6 mM increased dry matter percentage, while SA at 100 μM promoted shoot growth 2-fold. The increased sucrose concentration enhanced accumulation of xanthones in shoot cultures 2–3-fold compared to the control shoots. Elicitors at 100–300 μM increased the accumulation of mangiferin, demethylbellidifolin-8-O-glucoside, and bellidifolin-8-O-glucoside almost equally, while MeJA at the highest concentration of 500 μM enhanced amount of aglycones demethylbellidifolin and bellidifolin 7-fold compared to the control. The obtained results facilitate conservation of G. lutescens and pave the way for further research on large-scale shoot propagation and production of pharmacologically active xanthones.",
publisher = "Basel: MDPI",
journal = "Plants",
title = "Gentianella lutescens subsp. carpatica J. Holub.: Shoot Propagation In Vitro and Effect of Sucrose and Elicitors on Xanthones Production",
number = "8",
volume = "10",
doi = "10.3390/plants10081651",
pages = "1651"
}

Krstić Milošević, D., Banjac, N., Janković, T., Vinterhalter, D.,& Vinterhalter, B.. (2021). Gentianella lutescens subsp. carpatica J. Holub.: Shoot Propagation In Vitro and Effect of Sucrose and Elicitors on Xanthones Production. in Plants
Basel: MDPI., 10(8), 1651.
https://doi.org/10.3390/plants10081651

Krstić Milošević D, Banjac N, Janković T, Vinterhalter D, Vinterhalter B. Gentianella lutescens subsp. carpatica J. Holub.: Shoot Propagation In Vitro and Effect of Sucrose and Elicitors on Xanthones Production. in Plants. 2021;10(8):1651.
doi:10.3390/plants10081651 .

Krstić Milošević, Dijana, Banjac, Nevena, Janković, Teodora, Vinterhalter, Dragan, Vinterhalter, Branka, "Gentianella lutescens subsp. carpatica J. Holub.: Shoot Propagation In Vitro and Effect of Sucrose and Elicitors on Xanthones Production" in Plants, 10, no. 8 (2021):1651,
https://doi.org/10.3390/plants10081651 . .

TY  - JOUR
AU  - Vinterhalter, Branka
AU  - Banjac, Nevena
AU  - Vinterhalter, Dragan
AU  - Krstić Milošević, Dijana
PY  - 2021
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/4309
AB  - The hairy root clones of Gentiana dinarica cl-B, cl-D, cl-3, and cl-14 were cultivated in parallel in diverse simple bioreactors, including temporary immersion systems RITA® (TIS RITA®), bubble column bioreactors (BCB), and Erlenmeyer flasks (EF), and evaluated for biomass production and xanthone content. The obtained results showed that TIS RITA® and BCB containing ½ MS medium with 4% sucrose provided equally good growth conditions in which the majority of the clones displayed the higher percentage of dry matter (DM%), and xanthones norswertianin-1-O-primeveroside (nor-1-O-prim) and norswertianin production than those cultivated in EF. Thin and well branched hairy root clone cl-B grown in BCB for 7 weeks was superior regarding all growth parameters tested, including growth index (19.97), dry weight (2.88 g), and DM% (25.70%) compared to all other clones. Cl-B cultured in TIS RITA® contained the highest amount of nor-1-O-prim (56.82 mg per vessel). In BCB with constant aeration, cl-B accumulated the highest norswertianin content reaching 18.08 mg/vessel. The optimized conditions for cultivation of selected G. dinarica hairy root clones in highly aerated TIS RITA® and BCB systems contribute to the development of bioreactor technology designed for the large scale commercial production of xanthones nor-1-O-prim and norswertianin.
PB  - Basel: MDPI
T2  - Plants
T1  - Xanthones Production in Gentiana dinarica Beck Hairy Root Cultures Grown in Simple Bioreactors
IS  - 8
VL  - 10
DO  - 10.3390/plants10081610
SP  - 1610
ER  - 

@article{
author = "Vinterhalter, Branka and Banjac, Nevena and Vinterhalter, Dragan and Krstić Milošević, Dijana",
year = "2021",
abstract = "The hairy root clones of Gentiana dinarica cl-B, cl-D, cl-3, and cl-14 were cultivated in parallel in diverse simple bioreactors, including temporary immersion systems RITA® (TIS RITA®), bubble column bioreactors (BCB), and Erlenmeyer flasks (EF), and evaluated for biomass production and xanthone content. The obtained results showed that TIS RITA® and BCB containing ½ MS medium with 4% sucrose provided equally good growth conditions in which the majority of the clones displayed the higher percentage of dry matter (DM%), and xanthones norswertianin-1-O-primeveroside (nor-1-O-prim) and norswertianin production than those cultivated in EF. Thin and well branched hairy root clone cl-B grown in BCB for 7 weeks was superior regarding all growth parameters tested, including growth index (19.97), dry weight (2.88 g), and DM% (25.70%) compared to all other clones. Cl-B cultured in TIS RITA® contained the highest amount of nor-1-O-prim (56.82 mg per vessel). In BCB with constant aeration, cl-B accumulated the highest norswertianin content reaching 18.08 mg/vessel. The optimized conditions for cultivation of selected G. dinarica hairy root clones in highly aerated TIS RITA® and BCB systems contribute to the development of bioreactor technology designed for the large scale commercial production of xanthones nor-1-O-prim and norswertianin.",
publisher = "Basel: MDPI",
journal = "Plants",
title = "Xanthones Production in Gentiana dinarica Beck Hairy Root Cultures Grown in Simple Bioreactors",
number = "8",
volume = "10",
doi = "10.3390/plants10081610",
pages = "1610"
}

Vinterhalter, B., Banjac, N., Vinterhalter, D.,& Krstić Milošević, D.. (2021). Xanthones Production in Gentiana dinarica Beck Hairy Root Cultures Grown in Simple Bioreactors. in Plants
Basel: MDPI., 10(8), 1610.
https://doi.org/10.3390/plants10081610

Vinterhalter B, Banjac N, Vinterhalter D, Krstić Milošević D. Xanthones Production in Gentiana dinarica Beck Hairy Root Cultures Grown in Simple Bioreactors. in Plants. 2021;10(8):1610.
doi:10.3390/plants10081610 .

Vinterhalter, Branka, Banjac, Nevena, Vinterhalter, Dragan, Krstić Milošević, Dijana, "Xanthones Production in Gentiana dinarica Beck Hairy Root Cultures Grown in Simple Bioreactors" in Plants, 10, no. 8 (2021):1610,
https://doi.org/10.3390/plants10081610 . .

TY  - JOUR
AU  - Devrnja, Nina
AU  - Krstić Milošević, Dijana
AU  - Janošević, Dušica
AU  - Tešević, Vele
AU  - Vinterhalter, Branka
AU  - Savić, Jelena
AU  - Ćalić, Dušica
PY  - 2021
UR  - http://link.springer.com/10.1007/s00709-020-01588-9
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/4038
AB  - In this study, tansy (Tanacetum vulgare L.) in vitro culture was established from seeds collected from natural populations. The multiplication of plantlets was conducted through shoot tips that exhibited potent apical growth and regeneration capacities on basal medium (BM), without the addition of any plant growth regulators (PGRs). PGRs were also omitted for the establishment and cultivation of tansy root cultures. Both abaxial and adaxial leaf surfaces of in vitro micropropagated plantlets were covered with glandular biseriate trichomes. Histochemical staining showed that glandular secretions were rich in lipid and terpene compounds, confirmed by GC-MS analysis of essential oil (EO). In the total EO, similar portions of oxygenated monoterpenes (38.5% m/m) and oxygenated sesquiterpenes (22.6% m/m) were detected. Chemical profiles of methanol extracts of in vitro cultured tansy shoots and roots varied in quantity and quality from those obtained from wild-growingtansy. HPLC analysis indicated that the methanol extracts of in vitro cultured roots were the richest in 3,5-O-dicaffeoylquinic acid (3,5-O-DCQA), in which the concentration was 6 times higher (10.220 mg/g DW) than that in the extract obtained from roots of wild-growing tansy (1.684 mg/g DW). This result is noticeable in the manner of industrial production of biologically active 3,5-O-DCQA that has been shown to have antioxidant, hepatoprotective, antiviral, antimutagenic, and immunomodulatory activity. Biotechnological interventions on secondary metabolite production taking place in trichomes could further enhance the production of some important tansy metabolites and further investigation will be directed toward the elucidation of the pharmaceutical potential of tansy in vitro obtained metabolites, as mixtures or single moieties.
PB  - Springer
T2  - Protoplasma
T1  - In vitro cultivation of tansy (Tanacetum vulgare L.): a tool for the production of potent pharmaceutical agents
IS  - 3
VL  - 258
DO  - 10.1007/s00709-020-01588-9
SP  - 587
EP  - 599
ER  - 

@article{
author = "Devrnja, Nina and Krstić Milošević, Dijana and Janošević, Dušica and Tešević, Vele and Vinterhalter, Branka and Savić, Jelena and Ćalić, Dušica",
year = "2021",
abstract = "In this study, tansy (Tanacetum vulgare L.) in vitro culture was established from seeds collected from natural populations. The multiplication of plantlets was conducted through shoot tips that exhibited potent apical growth and regeneration capacities on basal medium (BM), without the addition of any plant growth regulators (PGRs). PGRs were also omitted for the establishment and cultivation of tansy root cultures. Both abaxial and adaxial leaf surfaces of in vitro micropropagated plantlets were covered with glandular biseriate trichomes. Histochemical staining showed that glandular secretions were rich in lipid and terpene compounds, confirmed by GC-MS analysis of essential oil (EO). In the total EO, similar portions of oxygenated monoterpenes (38.5% m/m) and oxygenated sesquiterpenes (22.6% m/m) were detected. Chemical profiles of methanol extracts of in vitro cultured tansy shoots and roots varied in quantity and quality from those obtained from wild-growingtansy. HPLC analysis indicated that the methanol extracts of in vitro cultured roots were the richest in 3,5-O-dicaffeoylquinic acid (3,5-O-DCQA), in which the concentration was 6 times higher (10.220 mg/g DW) than that in the extract obtained from roots of wild-growing tansy (1.684 mg/g DW). This result is noticeable in the manner of industrial production of biologically active 3,5-O-DCQA that has been shown to have antioxidant, hepatoprotective, antiviral, antimutagenic, and immunomodulatory activity. Biotechnological interventions on secondary metabolite production taking place in trichomes could further enhance the production of some important tansy metabolites and further investigation will be directed toward the elucidation of the pharmaceutical potential of tansy in vitro obtained metabolites, as mixtures or single moieties.",
publisher = "Springer",
journal = "Protoplasma",
title = "In vitro cultivation of tansy (Tanacetum vulgare L.): a tool for the production of potent pharmaceutical agents",
number = "3",
volume = "258",
doi = "10.1007/s00709-020-01588-9",
pages = "587-599"
}

Devrnja, N., Krstić Milošević, D., Janošević, D., Tešević, V., Vinterhalter, B., Savić, J.,& Ćalić, D.. (2021). In vitro cultivation of tansy (Tanacetum vulgare L.): a tool for the production of potent pharmaceutical agents. in Protoplasma
Springer., 258(3), 587-599.
https://doi.org/10.1007/s00709-020-01588-9

Devrnja N, Krstić Milošević D, Janošević D, Tešević V, Vinterhalter B, Savić J, Ćalić D. In vitro cultivation of tansy (Tanacetum vulgare L.): a tool for the production of potent pharmaceutical agents. in Protoplasma. 2021;258(3):587-599.
doi:10.1007/s00709-020-01588-9 .

Devrnja, Nina, Krstić Milošević, Dijana, Janošević, Dušica, Tešević, Vele, Vinterhalter, Branka, Savić, Jelena, Ćalić, Dušica, "In vitro cultivation of tansy (Tanacetum vulgare L.): a tool for the production of potent pharmaceutical agents" in Protoplasma, 258, no. 3 (2021):587-599,
https://doi.org/10.1007/s00709-020-01588-9 . .

TY  - CHAP
AU  - Tovilović-Kovačević, Gordana
AU  - Zogović, Nevena
AU  - Krstić Milošević, Dijana
PY  - 2020
UR  - https://radar.ibiss.bg.ac.rs/123456789/3896
AB  - Gentianaceae have a long history of use as traditional remedies for treatment of various ailments. The medicinal properties of crude herbal drug are attributed to bitter glycosides, flavonoids, and xanthones, the main plant secondary metabolites. These plant-derived molecules, especially naturally occurring xanthones, possess a broad spectrum of bioactivity like anticarcinogenic, antimicrobial, neuroprotective, antidiabetic, cardio-protective. The most of Gentianaceae species are rare and endangered by uncontrolled overharvesting and influences of various environmental factors (habitat loss, climate change, and invasive species spreading). Decline of Gentianaceae species poses a high risk to the loss of enormous diversity of potentially bioactive compounds. In this chapter we will summarize pharmacological activities of identified secondary metabolites from endangered species belonging to four Gentianaceae genera (Gentiana, Gentianella, Centaurium, Swertia), as well as importance of biodiversity conservation in context of their biotherapeutic potential.
PB  - Academic Press
T2  - Biodiversity and Biomedicine Our Future
T1  - Secondary metabolites from endangered Gentiana, Gentianella, Centaurium, and Swertia species (Gentianaceae): promising natural biotherapeutics
DO  - 10.1016/B978-0-12-819541-3.00019-0
SP  - 335
EP  - 384
ER  - 

@inbook{
author = "Tovilović-Kovačević, Gordana and Zogović, Nevena and Krstić Milošević, Dijana",
year = "2020",
abstract = "Gentianaceae have a long history of use as traditional remedies for treatment of various ailments. The medicinal properties of crude herbal drug are attributed to bitter glycosides, flavonoids, and xanthones, the main plant secondary metabolites. These plant-derived molecules, especially naturally occurring xanthones, possess a broad spectrum of bioactivity like anticarcinogenic, antimicrobial, neuroprotective, antidiabetic, cardio-protective. The most of Gentianaceae species are rare and endangered by uncontrolled overharvesting and influences of various environmental factors (habitat loss, climate change, and invasive species spreading). Decline of Gentianaceae species poses a high risk to the loss of enormous diversity of potentially bioactive compounds. In this chapter we will summarize pharmacological activities of identified secondary metabolites from endangered species belonging to four Gentianaceae genera (Gentiana, Gentianella, Centaurium, Swertia), as well as importance of biodiversity conservation in context of their biotherapeutic potential.",
publisher = "Academic Press",
journal = "Biodiversity and Biomedicine Our Future",
booktitle = "Secondary metabolites from endangered Gentiana, Gentianella, Centaurium, and Swertia species (Gentianaceae): promising natural biotherapeutics",
doi = "10.1016/B978-0-12-819541-3.00019-0",
pages = "335-384"
}

Tovilović-Kovačević, G., Zogović, N.,& Krstić Milošević, D.. (2020). Secondary metabolites from endangered Gentiana, Gentianella, Centaurium, and Swertia species (Gentianaceae): promising natural biotherapeutics. in Biodiversity and Biomedicine Our Future
Academic Press., 335-384.
https://doi.org/10.1016/B978-0-12-819541-3.00019-0

Tovilović-Kovačević G, Zogović N, Krstić Milošević D. Secondary metabolites from endangered Gentiana, Gentianella, Centaurium, and Swertia species (Gentianaceae): promising natural biotherapeutics. in Biodiversity and Biomedicine Our Future. 2020;:335-384.
doi:10.1016/B978-0-12-819541-3.00019-0 .

Tovilović-Kovačević, Gordana, Zogović, Nevena, Krstić Milošević, Dijana, "Secondary metabolites from endangered Gentiana, Gentianella, Centaurium, and Swertia species (Gentianaceae): promising natural biotherapeutics" in Biodiversity and Biomedicine Our Future (2020):335-384,
https://doi.org/10.1016/B978-0-12-819541-3.00019-0 . .

TY  - JOUR
AU  - Krstić Milošević, Dijana
AU  - Banjac, Nevena
AU  - Janković, Teodora
AU  - Eler, Klemen
AU  - Vinterhalter, Branka
PY  - 2020
UR  - http://www.ncbi.nlm.nih.gov/pubmed/32763798
UR  - https://radar.ibiss.bg.ac.rs/123456789/3847
AB  - Shoot and root in vitro culture of endemic European species Gentiana clusii was established for the first time. The effects of different concentrations of benzyl adenine (BA), 6-phurphurylaminopurine (KIN), indole-3-butyric acid (IBA) and naphthalene acetic acid (NAA) on shoot propagation and rooting of G. clusii were investigated. The optimal in vitro conditions for shoot propagation and long-term maintenance were achieved using woody plant medium (WPM) supplemented with 0.5 mg l-1 KIN, and subsequent application of IBA at 0.5 mg l-1 significantly improved rooting of these shoots. Root culture was established from excised root tips cultured in ½ MS liquid media with increasing concentrations of IBA (0.1-1.0 mg l-1). A high root growth rate and considerable biomass yield were obtained by addition of 1.0 mg l-1 IBA. HPLC analysis revealed that in vitro culture considerably promoted the production of secondary metabolites in G. clusii. The selected protocol for shoot propagation (WPM + 0.5 mg l-1 KIN) increased the content of sweroside, gentiopicrin and norswertianin-1-O-primeveroside (N-1-P) for more than 2-fold compared with the wild plants. IBA promoted N-1-P and norswertianin production in root cultures; their contents were enhanced 6.4- and 18.6-fold, respectively, compared with the wild plants. The extract of these roots displayed the highest antioxidant capacity (IC50 = 66.57 μg ml-1). The established shoot and root propagation protocols facilitate in vitro conservation of G. clusii, and provides a promising tool for the large scale production of valuable secoiridoids and xanthones.
PB  - Elsevier Masson SAS
T2  - Plant Physiology and Biochemistry
T1  - Gentiana clusii Perr.&Song.: Enhanced production of secondary metabolites by in vitro propagation.
VL  - 154
DO  - 10.1016/j.plaphy.2020.07.007
SP  - 735
EP  - 744
ER  - 

@article{
author = "Krstić Milošević, Dijana and Banjac, Nevena and Janković, Teodora and Eler, Klemen and Vinterhalter, Branka",
year = "2020",
abstract = "Shoot and root in vitro culture of endemic European species Gentiana clusii was established for the first time. The effects of different concentrations of benzyl adenine (BA), 6-phurphurylaminopurine (KIN), indole-3-butyric acid (IBA) and naphthalene acetic acid (NAA) on shoot propagation and rooting of G. clusii were investigated. The optimal in vitro conditions for shoot propagation and long-term maintenance were achieved using woody plant medium (WPM) supplemented with 0.5 mg l-1 KIN, and subsequent application of IBA at 0.5 mg l-1 significantly improved rooting of these shoots. Root culture was established from excised root tips cultured in ½ MS liquid media with increasing concentrations of IBA (0.1-1.0 mg l-1). A high root growth rate and considerable biomass yield were obtained by addition of 1.0 mg l-1 IBA. HPLC analysis revealed that in vitro culture considerably promoted the production of secondary metabolites in G. clusii. The selected protocol for shoot propagation (WPM + 0.5 mg l-1 KIN) increased the content of sweroside, gentiopicrin and norswertianin-1-O-primeveroside (N-1-P) for more than 2-fold compared with the wild plants. IBA promoted N-1-P and norswertianin production in root cultures; their contents were enhanced 6.4- and 18.6-fold, respectively, compared with the wild plants. The extract of these roots displayed the highest antioxidant capacity (IC50 = 66.57 μg ml-1). The established shoot and root propagation protocols facilitate in vitro conservation of G. clusii, and provides a promising tool for the large scale production of valuable secoiridoids and xanthones.",
publisher = "Elsevier Masson SAS",
journal = "Plant Physiology and Biochemistry",
title = "Gentiana clusii Perr.&Song.: Enhanced production of secondary metabolites by in vitro propagation.",
volume = "154",
doi = "10.1016/j.plaphy.2020.07.007",
pages = "735-744"
}

Krstić Milošević, D., Banjac, N., Janković, T., Eler, K.,& Vinterhalter, B.. (2020). Gentiana clusii Perr.&Song.: Enhanced production of secondary metabolites by in vitro propagation.. in Plant Physiology and Biochemistry
Elsevier Masson SAS., 154, 735-744.
https://doi.org/10.1016/j.plaphy.2020.07.007

Krstić Milošević D, Banjac N, Janković T, Eler K, Vinterhalter B. Gentiana clusii Perr.&Song.: Enhanced production of secondary metabolites by in vitro propagation.. in Plant Physiology and Biochemistry. 2020;154:735-744.
doi:10.1016/j.plaphy.2020.07.007 .

Krstić Milošević, Dijana, Banjac, Nevena, Janković, Teodora, Eler, Klemen, Vinterhalter, Branka, "Gentiana clusii Perr.&Song.: Enhanced production of secondary metabolites by in vitro propagation." in Plant Physiology and Biochemistry, 154 (2020):735-744,
https://doi.org/10.1016/j.plaphy.2020.07.007 . .

TY  - JOUR
AU  - Popović, Zorica
AU  - Krstić Milošević, Dijana
AU  - Marković, Milena
AU  - Vidaković, Vera
AU  - Matić, Rada
AU  - Janković, Jovana
AU  - Bojović, Srđan
PY  - 2020
UR  - https://www.tandfonline.com/doi/abs/10.1080/11263504.2020.1785966?journalCode=tplb20
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/3815
AB  - Variability in the contents of representative secondary metabolites in Gentiana pneumonanthe individuals was investigated in relation to population origin, plant parts and developmental stage. Chemical analyses (HPLC) of six secondary metabolites (swertiamarin (SWM), gentiopicrin (GP), sweroside (SWZ), mangiferin (MGF), isoorientin (ISOOR), and isovitexin (ISOV)) were performed for individuals from six populations located within a 3 × 3 km area at Maljen Mt., and separately for below- and above ground plant parts in three developmental stages. Different plant parts showed pronounced differences in the contents of the six secondary metabolites and a high level of variability during the plant ontogeny. The highest contents of three dominant compounds (SWM, GP, and MGF) were detected in different developmental stages. Both below- and above ground plant parts contained GP, MGF, SWZ, and SWM at some developmental stages, whereas ISOOR and ISOV were registered in above ground parts. The full-flowering stage was characterized by the highest contents of MGF, GP, and ISOOR in aboveground-, and GP in belowground parts. In the post-flowering stage, a high content of MF was detected in below ground parts. The results of this study contribute to a better understanding of interpopulation, ontogenetic and between-organ variability in rare and threatened G. pneumonanthe.
PB  - London: Taylor & Francis
T2  - Plant Biosystems
T1  - Variability of six secondary metabolites in plant parts and developmental stages in natural populations of rare Gentiana pneumonanthe
DO  - 10.1080/11263504.2020.1785966
ER  - 

@article{
author = "Popović, Zorica and Krstić Milošević, Dijana and Marković, Milena and Vidaković, Vera and Matić, Rada and Janković, Jovana and Bojović, Srđan",
year = "2020",
abstract = "Variability in the contents of representative secondary metabolites in Gentiana pneumonanthe individuals was investigated in relation to population origin, plant parts and developmental stage. Chemical analyses (HPLC) of six secondary metabolites (swertiamarin (SWM), gentiopicrin (GP), sweroside (SWZ), mangiferin (MGF), isoorientin (ISOOR), and isovitexin (ISOV)) were performed for individuals from six populations located within a 3 × 3 km area at Maljen Mt., and separately for below- and above ground plant parts in three developmental stages. Different plant parts showed pronounced differences in the contents of the six secondary metabolites and a high level of variability during the plant ontogeny. The highest contents of three dominant compounds (SWM, GP, and MGF) were detected in different developmental stages. Both below- and above ground plant parts contained GP, MGF, SWZ, and SWM at some developmental stages, whereas ISOOR and ISOV were registered in above ground parts. The full-flowering stage was characterized by the highest contents of MGF, GP, and ISOOR in aboveground-, and GP in belowground parts. In the post-flowering stage, a high content of MF was detected in below ground parts. The results of this study contribute to a better understanding of interpopulation, ontogenetic and between-organ variability in rare and threatened G. pneumonanthe.",
publisher = "London: Taylor & Francis",
journal = "Plant Biosystems",
title = "Variability of six secondary metabolites in plant parts and developmental stages in natural populations of rare Gentiana pneumonanthe",
doi = "10.1080/11263504.2020.1785966"
}

Popović, Z., Krstić Milošević, D., Marković, M., Vidaković, V., Matić, R., Janković, J.,& Bojović, S.. (2020). Variability of six secondary metabolites in plant parts and developmental stages in natural populations of rare Gentiana pneumonanthe. in Plant Biosystems
London: Taylor & Francis..
https://doi.org/10.1080/11263504.2020.1785966

Popović Z, Krstić Milošević D, Marković M, Vidaković V, Matić R, Janković J, Bojović S. Variability of six secondary metabolites in plant parts and developmental stages in natural populations of rare Gentiana pneumonanthe. in Plant Biosystems. 2020;.
doi:10.1080/11263504.2020.1785966 .

Popović, Zorica, Krstić Milošević, Dijana, Marković, Milena, Vidaković, Vera, Matić, Rada, Janković, Jovana, Bojović, Srđan, "Variability of six secondary metabolites in plant parts and developmental stages in natural populations of rare Gentiana pneumonanthe" in Plant Biosystems (2020),
https://doi.org/10.1080/11263504.2020.1785966 . .

TY  - JOUR
AU  - Zdunić, Gordana
AU  - Alimpić Aradski, Ana
AU  - Gođevac, Dejan
AU  - Živković, Jelena
AU  - Duletić Laušević, Sonja
AU  - Krstić Milošević, Dijana
AU  - Šavikin, Katarina
PY  - 2020
UR  - internal-pdf://Aronija 2020.pdf
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/3619
AB  - Chokeberries (Aronia melanocarpa (Michx.) Elliot have recently become one of the most popular and widely used berry fruits, and there are numerous studies dealing with the investigation of their chemical composition and biological activities. Unlike berries, chokeberry leaves were far less investigated. Therefore, the aim of this study was to conduct a bioactivity-based analysis of chokeberry leaves collected after berries were harvested. Chemical analysis of hydroethanolic chokeberry extract and its ethyl acetate, n-butanol, and water fractions was performed. Antioxidant, hypoglycemic, and antineurodegenerative activities of the obtained samples were assessed by in vitro tests. The highest total phenolics content was found in n-butanol fraction (221.5 mg gallic acid equivalents/g dw). Fourteen compounds (six phenolic acids and eight flavonoids) were identified in the samples. Caffeoylquinic acid derivatives and quercetin glycosides were dominant phenolic compounds. The highest content of phenolic acids, as well as flavonoids, was recorded in n-butanol fraction (80.75 mg/g dw and 47.79 mg/g dw, respectively). This fraction was the most active in 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2′-azino-bis (3-ethyl-benzothiazoline-6-sulfonic acid) (ABTS), and ferric reducing antioxidant power (FRAP) assays. Also, the n-butanol fraction was the most effective inhibitor of acetylcholinesterase (AChE) and tyrosinase (TYR) enzymes. All samples showed a better inhibitory effect on α-glucosidase enzyme with inhibition concentrations IC50 = 4.21 - 9.22 μg/mL, than α-amylase enzymes (IC50 = 14.36 - 88.61 μg/mL). Pearson’s correlation between the content of individual phenolics and the results of biological tests was performed. Thus, hyperoside and isoquercitrin showed a significant correlation with DPPH and ABTS tests, caffeoylquinic acids with FRAP test, quercetin 3-O-vicianoside with β-carotene assay, hyperoside and isoquercitrin with α-amylase inhibition activity, and isorhamnetin 3-O-rutinoside with α-glucosidase inhibition assay. Considering the obtained results, chokeberry leaves represent a valuable natural source of phenolic compounds with promising potential for the development of new products with beneficial effects on human health.
T2  - Industrial Crops and Products
T1  - In vitro hypoglycemic, antioxidant and antineurodegenerative activity of chokeberry (Aronia melanocarpa) leaves
VL  - 148
DO  - 10.1016/j.indcrop.2020.112328
SP  - 112328
ER  - 

@article{
author = "Zdunić, Gordana and Alimpić Aradski, Ana and Gođevac, Dejan and Živković, Jelena and Duletić Laušević, Sonja and Krstić Milošević, Dijana and Šavikin, Katarina",
year = "2020",
abstract = "Chokeberries (Aronia melanocarpa (Michx.) Elliot have recently become one of the most popular and widely used berry fruits, and there are numerous studies dealing with the investigation of their chemical composition and biological activities. Unlike berries, chokeberry leaves were far less investigated. Therefore, the aim of this study was to conduct a bioactivity-based analysis of chokeberry leaves collected after berries were harvested. Chemical analysis of hydroethanolic chokeberry extract and its ethyl acetate, n-butanol, and water fractions was performed. Antioxidant, hypoglycemic, and antineurodegenerative activities of the obtained samples were assessed by in vitro tests. The highest total phenolics content was found in n-butanol fraction (221.5 mg gallic acid equivalents/g dw). Fourteen compounds (six phenolic acids and eight flavonoids) were identified in the samples. Caffeoylquinic acid derivatives and quercetin glycosides were dominant phenolic compounds. The highest content of phenolic acids, as well as flavonoids, was recorded in n-butanol fraction (80.75 mg/g dw and 47.79 mg/g dw, respectively). This fraction was the most active in 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2′-azino-bis (3-ethyl-benzothiazoline-6-sulfonic acid) (ABTS), and ferric reducing antioxidant power (FRAP) assays. Also, the n-butanol fraction was the most effective inhibitor of acetylcholinesterase (AChE) and tyrosinase (TYR) enzymes. All samples showed a better inhibitory effect on α-glucosidase enzyme with inhibition concentrations IC50 = 4.21 - 9.22 μg/mL, than α-amylase enzymes (IC50 = 14.36 - 88.61 μg/mL). Pearson’s correlation between the content of individual phenolics and the results of biological tests was performed. Thus, hyperoside and isoquercitrin showed a significant correlation with DPPH and ABTS tests, caffeoylquinic acids with FRAP test, quercetin 3-O-vicianoside with β-carotene assay, hyperoside and isoquercitrin with α-amylase inhibition activity, and isorhamnetin 3-O-rutinoside with α-glucosidase inhibition assay. Considering the obtained results, chokeberry leaves represent a valuable natural source of phenolic compounds with promising potential for the development of new products with beneficial effects on human health.",
journal = "Industrial Crops and Products",
title = "In vitro hypoglycemic, antioxidant and antineurodegenerative activity of chokeberry (Aronia melanocarpa) leaves",
volume = "148",
doi = "10.1016/j.indcrop.2020.112328",
pages = "112328"
}

Zdunić, G., Alimpić Aradski, A., Gođevac, D., Živković, J., Duletić Laušević, S., Krstić Milošević, D.,& Šavikin, K.. (2020). In vitro hypoglycemic, antioxidant and antineurodegenerative activity of chokeberry (Aronia melanocarpa) leaves. in Industrial Crops and Products, 148, 112328.
https://doi.org/10.1016/j.indcrop.2020.112328

Zdunić G, Alimpić Aradski A, Gođevac D, Živković J, Duletić Laušević S, Krstić Milošević D, Šavikin K. In vitro hypoglycemic, antioxidant and antineurodegenerative activity of chokeberry (Aronia melanocarpa) leaves. in Industrial Crops and Products. 2020;148:112328.
doi:10.1016/j.indcrop.2020.112328 .

Zdunić, Gordana, Alimpić Aradski, Ana, Gođevac, Dejan, Živković, Jelena, Duletić Laušević, Sonja, Krstić Milošević, Dijana, Šavikin, Katarina, "In vitro hypoglycemic, antioxidant and antineurodegenerative activity of chokeberry (Aronia melanocarpa) leaves" in Industrial Crops and Products, 148 (2020):112328,
https://doi.org/10.1016/j.indcrop.2020.112328 . .

TY  - JOUR
AU  - Popović, Zorica
AU  - Krstić Milošević, Dijana
AU  - Marković, Milena
AU  - Matić, Rada
AU  - Vidaković, Vera
AU  - Bojović, Srđan
PY  - 2019
UR  - https://onlinelibrary.wiley.com/doi/abs/10.1002/cbdv.201800509
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/3260
AB  - Inter- and intrapopulation variability in six natural populations of the rare species Gentiana pneumonanthe was examined based on morphological and chemical data. Population size and linear morphometric parameters differed significantly among populations, but without a clear connection to habitat conditions, i. e. water supply and light availability. Leaf shape varied from ovate to lanceolate in all populations, and one population was distinctive in having the largest number of leaves of transitional shape. HPLC analyses of six secondary metabolites were performed separately for belowground parts, and aboveground vegetative and reproductive parts of individual plants (6 populations ×7 individuals ×3 plant parts, n=126) in order to examine differences at the population and individual levels. Three secoiridoids (swertiamarin (SWM), sweroside (SWZ), and gentiopicrin (GP)), one xanthone (mangiferin (MGF)), and two flavones (isoorientin (IO) and isovitexin (IV)) were detected and quantified in the analyzed samples: sweroside dominated in the aboveground reproductive part, mangiferin in the aboveground vegetative part, and gentiopicrin in the belowground part. At the population level, differences in contents of the analyzed chemicals among populations were significant only for a few metabolites. At the individual level, a pronounced organ-dependent distribution of secondary metabolites was revealed. The results of this study contribute to a better understanding of natural variability within populations of the rare and threatened G. pneumonanthe, and provide data on the contents and within-plant distribution of secondary metabolites, which are important as pharmacologically active compounds and may be useful for further biotechnological procedures regarding this species.
T2  - Chemistry & Biodiversity
T1  - Chemical and Morphological Inter- and Intrapopulation Variability in Natural Populations of Gentiana pneumonanthe L.
IS  - 2
VL  - 16
DO  - 10.1002/cbdv.201800509
SP  - e1800509
ER  - 

@article{
author = "Popović, Zorica and Krstić Milošević, Dijana and Marković, Milena and Matić, Rada and Vidaković, Vera and Bojović, Srđan",
year = "2019",
abstract = "Inter- and intrapopulation variability in six natural populations of the rare species Gentiana pneumonanthe was examined based on morphological and chemical data. Population size and linear morphometric parameters differed significantly among populations, but without a clear connection to habitat conditions, i. e. water supply and light availability. Leaf shape varied from ovate to lanceolate in all populations, and one population was distinctive in having the largest number of leaves of transitional shape. HPLC analyses of six secondary metabolites were performed separately for belowground parts, and aboveground vegetative and reproductive parts of individual plants (6 populations ×7 individuals ×3 plant parts, n=126) in order to examine differences at the population and individual levels. Three secoiridoids (swertiamarin (SWM), sweroside (SWZ), and gentiopicrin (GP)), one xanthone (mangiferin (MGF)), and two flavones (isoorientin (IO) and isovitexin (IV)) were detected and quantified in the analyzed samples: sweroside dominated in the aboveground reproductive part, mangiferin in the aboveground vegetative part, and gentiopicrin in the belowground part. At the population level, differences in contents of the analyzed chemicals among populations were significant only for a few metabolites. At the individual level, a pronounced organ-dependent distribution of secondary metabolites was revealed. The results of this study contribute to a better understanding of natural variability within populations of the rare and threatened G. pneumonanthe, and provide data on the contents and within-plant distribution of secondary metabolites, which are important as pharmacologically active compounds and may be useful for further biotechnological procedures regarding this species.",
journal = "Chemistry & Biodiversity",
title = "Chemical and Morphological Inter- and Intrapopulation Variability in Natural Populations of Gentiana pneumonanthe L.",
number = "2",
volume = "16",
doi = "10.1002/cbdv.201800509",
pages = "e1800509"
}

Popović, Z., Krstić Milošević, D., Marković, M., Matić, R., Vidaković, V.,& Bojović, S.. (2019). Chemical and Morphological Inter- and Intrapopulation Variability in Natural Populations of Gentiana pneumonanthe L.. in Chemistry & Biodiversity, 16(2), e1800509.
https://doi.org/10.1002/cbdv.201800509

Popović Z, Krstić Milošević D, Marković M, Matić R, Vidaković V, Bojović S. Chemical and Morphological Inter- and Intrapopulation Variability in Natural Populations of Gentiana pneumonanthe L.. in Chemistry & Biodiversity. 2019;16(2):e1800509.
doi:10.1002/cbdv.201800509 .

Popović, Zorica, Krstić Milošević, Dijana, Marković, Milena, Matić, Rada, Vidaković, Vera, Bojović, Srđan, "Chemical and Morphological Inter- and Intrapopulation Variability in Natural Populations of Gentiana pneumonanthe L." in Chemistry & Biodiversity, 16, no. 2 (2019):e1800509,
https://doi.org/10.1002/cbdv.201800509 . .

TY  - JOUR
AU  - Stanišić, Mariana
AU  - Ćosić, Tatjana
AU  - Savić, Jelena
AU  - Krstić Milošević, Dijana
AU  - Mišić, Danijela
AU  - Smigocki, Ann
AU  - Ninković, Slavica
AU  - Banjac, Nevena
PY  - 2019
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/3791
AB  - Allelopathic plants exploit their chemical ‘weapons’ to prevail over the competition, suppress neighboring plants and consequently
use the available resources more efficiently. However, the investigation of plant allelopathic interactions in rhizosphere is difficult
to perform because of its high complexity due to interactions of biotic and abiotic factors. Thus, autonomous, aseptic root cultures
of apple (Malus × domestica Borkh.) could facilitate allelopathic studies. We report on the successful genetic transformation of
apple cultivars Melrose, Golden Delicious, Cadel and Gloster using ˇ Agrobacterium rhizogenes (Riker et al. 1930) Conn 1942 strain
15834 and for the first time the establishment of apple autonomous and permanent in vitro hairy root cultures that could be used
as a new tool for apple allelopathic assays. Molecular characterization of transgenic hairy root lines was conducted to elucidate
the possible relationship between expression of T-DNA genes and root growth characteristics that include branching. Similar content of phenolic acids (chlorogenic, caffeic, syringic, p-coumaric and ferulic), glycosilated flavonoids (rutin, quercitrin, isoquercitrin,
kaempferol-3-glucoside) and flavonoid aglycones (quercetin and naringenin), and dihydrochalcone phloridzin, was detected in
untransformed and transgenic apple root tissue by ultra high-performance liquid chromatography with mass spectrometry
(UHPLC/(+/–)HESI–MS/MS) analyses, confirming that genetic transformation did not disturb secondary metabolite production in
apple. Chlorogenic and caffeic acids and dihydrochalcones phloridzin and phloretin were detected as putative allelochemicals
exuded into the growth medium in which transgenic roots were maintained for 4 weeks. Apple hairy root exudates significantly
affected shoot and root development and growth of test plant Arabidopsis thaliana (L.) Heynh. seedlings after 5 or 10 days of
treatment. Additionally, core cell-cycle genes CDKA1;1, CDKB2;1, CYCA3;1 and CYCB2;4 were down regulated in Arabidopsis
shoots suggesting, in part, their role in inhibition of shoot growth. The present work highlighted an autonomous and permanent
in vitro hairy root culture system as a valuable tool for studying allelopathic potential of apple, offering new perspective for allelopathy background elucidation in this important fruit species.
PB  - Oxford:Oxford University Press
T2  - Tree Physiology
T1  - Hairy root culture as a valuable tool for allelopathic studies in apple
IS  - 5
VL  - 39
DO  - doi:10.1093/treephys/tpz006
SP  - 888
EP  - 905
ER  - 

@article{
author = "Stanišić, Mariana and Ćosić, Tatjana and Savić, Jelena and Krstić Milošević, Dijana and Mišić, Danijela and Smigocki, Ann and Ninković, Slavica and Banjac, Nevena",
year = "2019",
abstract = "Allelopathic plants exploit their chemical ‘weapons’ to prevail over the competition, suppress neighboring plants and consequently
use the available resources more efficiently. However, the investigation of plant allelopathic interactions in rhizosphere is difficult
to perform because of its high complexity due to interactions of biotic and abiotic factors. Thus, autonomous, aseptic root cultures
of apple (Malus × domestica Borkh.) could facilitate allelopathic studies. We report on the successful genetic transformation of
apple cultivars Melrose, Golden Delicious, Cadel and Gloster using ˇ Agrobacterium rhizogenes (Riker et al. 1930) Conn 1942 strain
15834 and for the first time the establishment of apple autonomous and permanent in vitro hairy root cultures that could be used
as a new tool for apple allelopathic assays. Molecular characterization of transgenic hairy root lines was conducted to elucidate
the possible relationship between expression of T-DNA genes and root growth characteristics that include branching. Similar content of phenolic acids (chlorogenic, caffeic, syringic, p-coumaric and ferulic), glycosilated flavonoids (rutin, quercitrin, isoquercitrin,
kaempferol-3-glucoside) and flavonoid aglycones (quercetin and naringenin), and dihydrochalcone phloridzin, was detected in
untransformed and transgenic apple root tissue by ultra high-performance liquid chromatography with mass spectrometry
(UHPLC/(+/–)HESI–MS/MS) analyses, confirming that genetic transformation did not disturb secondary metabolite production in
apple. Chlorogenic and caffeic acids and dihydrochalcones phloridzin and phloretin were detected as putative allelochemicals
exuded into the growth medium in which transgenic roots were maintained for 4 weeks. Apple hairy root exudates significantly
affected shoot and root development and growth of test plant Arabidopsis thaliana (L.) Heynh. seedlings after 5 or 10 days of
treatment. Additionally, core cell-cycle genes CDKA1;1, CDKB2;1, CYCA3;1 and CYCB2;4 were down regulated in Arabidopsis
shoots suggesting, in part, their role in inhibition of shoot growth. The present work highlighted an autonomous and permanent
in vitro hairy root culture system as a valuable tool for studying allelopathic potential of apple, offering new perspective for allelopathy background elucidation in this important fruit species.",
publisher = "Oxford:Oxford University Press",
journal = "Tree Physiology",
title = "Hairy root culture as a valuable tool for allelopathic studies in apple",
number = "5",
volume = "39",
doi = "doi:10.1093/treephys/tpz006",
pages = "888-905"
}

Stanišić, M., Ćosić, T., Savić, J., Krstić Milošević, D., Mišić, D., Smigocki, A., Ninković, S.,& Banjac, N.. (2019). Hairy root culture as a valuable tool for allelopathic studies in apple. in Tree Physiology
Oxford:Oxford University Press., 39(5), 888-905.
https://doi.org/doi:10.1093/treephys/tpz006

Stanišić M, Ćosić T, Savić J, Krstić Milošević D, Mišić D, Smigocki A, Ninković S, Banjac N. Hairy root culture as a valuable tool for allelopathic studies in apple. in Tree Physiology. 2019;39(5):888-905.
doi:doi:10.1093/treephys/tpz006 .

Stanišić, Mariana, Ćosić, Tatjana, Savić, Jelena, Krstić Milošević, Dijana, Mišić, Danijela, Smigocki, Ann, Ninković, Slavica, Banjac, Nevena, "Hairy root culture as a valuable tool for allelopathic studies in apple" in Tree Physiology, 39, no. 5 (2019):888-905,
https://doi.org/doi:10.1093/treephys/tpz006 . .

TY  - JOUR
AU  - Banjac, Nevena
AU  - Vinterhalter, Branka
AU  - Krstić Milošević, Dijana
AU  - Milojević, Jelena
AU  - Tubić, Ljiljana
AU  - Ghalawenji, Nabil
AU  - Zdravković-Korać, Snežana
PY  - 2019
UR  - http://link.springer.com/10.1007/s11240-019-01595-6
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/3310
AB  - Brassica oleracea var. acephala is an important leafy vegetable that has been widely consumed as a high-nutrient, low-calorie food. Because of the plant’s biennial and self-incompatibility nature, biotechnological approaches are alternative way for propagation and breeding improvements. Since tissue culture studies have been focused in other B. oleracea representatives, the aim of the present study was to achieve effective regeneration protocol distinctive for collard greens, and evaluate the total phenolic content and antioxidant activity of regenerants. The effect of 3 cytokinins [thidiazuron (TDZ), 6-benzyladenine (BA) and 6-furfuryladenine (kinetin, KIN)] at increasing concentrations (0, 5, 7.5, 10, 20 or 30 µM) in combination with tenfold lower concentration of 1-naphtaleneacetic acid (NAA) (0, 0.5, 0.75, 1, 2 or 3 µM, respectively) on the regeneration from hypocotyl slices was studied. Histological analysis revealed the two regeneration pathways, somatic embryogenesis and shoot organogenesis, simultaneously occurred in the same explant, regardless of the cytokinin/NAA combinations used. The regeneration frequency of 95.9%, with 7.5 morphogenic structures regenerated per explant, and the healthy appearance of regenerated plants indicated the optimal combination 20 µM TDZ + 2 µM NAA. TDZ at 5 µM provided the high somatic embryo proliferation rate by generation of secondary embryos (7.79) along with the lowest rate of their abnormalities. Embryo-developed plants were successfully acclimatised (above 90%). The plants regenerated and proliferated on TDZ-containing media had higher total phenolic content that correlated with the highest free radical scavenging activity (IC50 = 19.09 µg ml− 1).
T2  - Plant Cell, Tissue and Organ Culture (PCTOC)
T1  - Somatic embryogenesis and shoot organogenesis from the hypocotyl slices and free radical scavenging activity of regenerants of collard greens (Brassica oleracea L. var. acephala)
DO  - 10.1007/s11240-019-01595-6
ER  - 

@article{
author = "Banjac, Nevena and Vinterhalter, Branka and Krstić Milošević, Dijana and Milojević, Jelena and Tubić, Ljiljana and Ghalawenji, Nabil and Zdravković-Korać, Snežana",
year = "2019",
abstract = "Brassica oleracea var. acephala is an important leafy vegetable that has been widely consumed as a high-nutrient, low-calorie food. Because of the plant’s biennial and self-incompatibility nature, biotechnological approaches are alternative way for propagation and breeding improvements. Since tissue culture studies have been focused in other B. oleracea representatives, the aim of the present study was to achieve effective regeneration protocol distinctive for collard greens, and evaluate the total phenolic content and antioxidant activity of regenerants. The effect of 3 cytokinins [thidiazuron (TDZ), 6-benzyladenine (BA) and 6-furfuryladenine (kinetin, KIN)] at increasing concentrations (0, 5, 7.5, 10, 20 or 30 µM) in combination with tenfold lower concentration of 1-naphtaleneacetic acid (NAA) (0, 0.5, 0.75, 1, 2 or 3 µM, respectively) on the regeneration from hypocotyl slices was studied. Histological analysis revealed the two regeneration pathways, somatic embryogenesis and shoot organogenesis, simultaneously occurred in the same explant, regardless of the cytokinin/NAA combinations used. The regeneration frequency of 95.9%, with 7.5 morphogenic structures regenerated per explant, and the healthy appearance of regenerated plants indicated the optimal combination 20 µM TDZ + 2 µM NAA. TDZ at 5 µM provided the high somatic embryo proliferation rate by generation of secondary embryos (7.79) along with the lowest rate of their abnormalities. Embryo-developed plants were successfully acclimatised (above 90%). The plants regenerated and proliferated on TDZ-containing media had higher total phenolic content that correlated with the highest free radical scavenging activity (IC50 = 19.09 µg ml− 1).",
journal = "Plant Cell, Tissue and Organ Culture (PCTOC)",
title = "Somatic embryogenesis and shoot organogenesis from the hypocotyl slices and free radical scavenging activity of regenerants of collard greens (Brassica oleracea L. var. acephala)",
doi = "10.1007/s11240-019-01595-6"
}

Banjac, N., Vinterhalter, B., Krstić Milošević, D., Milojević, J., Tubić, L., Ghalawenji, N.,& Zdravković-Korać, S.. (2019). Somatic embryogenesis and shoot organogenesis from the hypocotyl slices and free radical scavenging activity of regenerants of collard greens (Brassica oleracea L. var. acephala). in Plant Cell, Tissue and Organ Culture (PCTOC).
https://doi.org/10.1007/s11240-019-01595-6

Banjac N, Vinterhalter B, Krstić Milošević D, Milojević J, Tubić L, Ghalawenji N, Zdravković-Korać S. Somatic embryogenesis and shoot organogenesis from the hypocotyl slices and free radical scavenging activity of regenerants of collard greens (Brassica oleracea L. var. acephala). in Plant Cell, Tissue and Organ Culture (PCTOC). 2019;.
doi:10.1007/s11240-019-01595-6 .

Banjac, Nevena, Vinterhalter, Branka, Krstić Milošević, Dijana, Milojević, Jelena, Tubić, Ljiljana, Ghalawenji, Nabil, Zdravković-Korać, Snežana, "Somatic embryogenesis and shoot organogenesis from the hypocotyl slices and free radical scavenging activity of regenerants of collard greens (Brassica oleracea L. var. acephala)" in Plant Cell, Tissue and Organ Culture (PCTOC) (2019),
https://doi.org/10.1007/s11240-019-01595-6 . .

TY  - JOUR
AU  - Stanišić, Mariana
AU  - Ćosić, Tatjana
AU  - Savić, Jelena
AU  - Krstić Milošević, Dijana
AU  - Mišić, Danijela
AU  - Smigocki, Ann
AU  - Ninković, Slavica
AU  - Banjac, Nevena
PY  - 2019
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/3791
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/3792
AB  - Allelopathic plants exploit their chemical ‘weapons’ to prevail over the competition, suppress neighboring plants and consequentlyuse the available resources more efficiently. However, the investigation of plant allelopathic interactions in rhizosphere is difficultto perform because of its high complexity due to interactions of biotic and abiotic factors. Thus, autonomous, aseptic root culturesof apple (Malus × domestica Borkh.) could facilitate allelopathic studies. We report on the successful genetic transformation ofapple cultivars Melrose, Golden Delicious, Cadel and Gloster using ˇ Agrobacterium rhizogenes (Riker et al. 1930) Conn 1942 strain15834 and for the first time the establishment of apple autonomous and permanent in vitro hairy root cultures that could be usedas a new tool for apple allelopathic assays. Molecular characterization of transgenic hairy root lines was conducted to elucidatethe possible relationship between expression of T-DNA genes and root growth characteristics that include branching. Similar content of phenolic acids (chlorogenic, caffeic, syringic, p-coumaric and ferulic), glycosilated flavonoids (rutin, quercitrin, isoquercitrin,kaempferol-3-glucoside) and flavonoid aglycones (quercetin and naringenin), and dihydrochalcone phloridzin, was detected inuntransformed and transgenic apple root tissue by ultra high-performance liquid chromatography with mass spectrometry(UHPLC/(+/–)HESI–MS/MS) analyses, confirming that genetic transformation did not disturb secondary metabolite production inapple. Chlorogenic and caffeic acids and dihydrochalcones phloridzin and phloretin were detected as putative allelochemicalsexuded into the growth medium in which transgenic roots were maintained for 4 weeks. Apple hairy root exudates significantlyaffected shoot and root development and growth of test plant Arabidopsis thaliana (L.) Heynh. seedlings after 5 or 10 days oftreatment. Additionally, core cell-cycle genes CDKA1;1, CDKB2;1, CYCA3;1 and CYCB2;4 were down regulated in Arabidopsisshoots suggesting, in part, their role in inhibition of shoot growth. The present work highlighted an autonomous and permanentin vitro hairy root culture system as a valuable tool for studying allelopathic potential of apple, offering new perspective for allelopathy background elucidation in this important fruit species.
PB  - Oxford: Oxford University Press
T2  - Tree Physiology
T1  - Hairy root culture as a valuable tool for allelopathic studies in apple
IS  - 5
VL  - 39
DO  - doi:10.1093/treephys/tpz006
SP  - 888
EP  - 905
ER  - 

@article{
author = "Stanišić, Mariana and Ćosić, Tatjana and Savić, Jelena and Krstić Milošević, Dijana and Mišić, Danijela and Smigocki, Ann and Ninković, Slavica and Banjac, Nevena",
year = "2019",
abstract = "Allelopathic plants exploit their chemical ‘weapons’ to prevail over the competition, suppress neighboring plants and consequentlyuse the available resources more efficiently. However, the investigation of plant allelopathic interactions in rhizosphere is difficultto perform because of its high complexity due to interactions of biotic and abiotic factors. Thus, autonomous, aseptic root culturesof apple (Malus × domestica Borkh.) could facilitate allelopathic studies. We report on the successful genetic transformation ofapple cultivars Melrose, Golden Delicious, Cadel and Gloster using ˇ Agrobacterium rhizogenes (Riker et al. 1930) Conn 1942 strain15834 and for the first time the establishment of apple autonomous and permanent in vitro hairy root cultures that could be usedas a new tool for apple allelopathic assays. Molecular characterization of transgenic hairy root lines was conducted to elucidatethe possible relationship between expression of T-DNA genes and root growth characteristics that include branching. Similar content of phenolic acids (chlorogenic, caffeic, syringic, p-coumaric and ferulic), glycosilated flavonoids (rutin, quercitrin, isoquercitrin,kaempferol-3-glucoside) and flavonoid aglycones (quercetin and naringenin), and dihydrochalcone phloridzin, was detected inuntransformed and transgenic apple root tissue by ultra high-performance liquid chromatography with mass spectrometry(UHPLC/(+/–)HESI–MS/MS) analyses, confirming that genetic transformation did not disturb secondary metabolite production inapple. Chlorogenic and caffeic acids and dihydrochalcones phloridzin and phloretin were detected as putative allelochemicalsexuded into the growth medium in which transgenic roots were maintained for 4 weeks. Apple hairy root exudates significantlyaffected shoot and root development and growth of test plant Arabidopsis thaliana (L.) Heynh. seedlings after 5 or 10 days oftreatment. Additionally, core cell-cycle genes CDKA1;1, CDKB2;1, CYCA3;1 and CYCB2;4 were down regulated in Arabidopsisshoots suggesting, in part, their role in inhibition of shoot growth. The present work highlighted an autonomous and permanentin vitro hairy root culture system as a valuable tool for studying allelopathic potential of apple, offering new perspective for allelopathy background elucidation in this important fruit species.",
publisher = "Oxford: Oxford University Press",
journal = "Tree Physiology",
title = "Hairy root culture as a valuable tool for allelopathic studies in apple",
number = "5",
volume = "39",
doi = "doi:10.1093/treephys/tpz006",
pages = "888-905"
}

Stanišić, M., Ćosić, T., Savić, J., Krstić Milošević, D., Mišić, D., Smigocki, A., Ninković, S.,& Banjac, N.. (2019). Hairy root culture as a valuable tool for allelopathic studies in apple. in Tree Physiology
Oxford: Oxford University Press., 39(5), 888-905.
https://doi.org/doi:10.1093/treephys/tpz006

Stanišić M, Ćosić T, Savić J, Krstić Milošević D, Mišić D, Smigocki A, Ninković S, Banjac N. Hairy root culture as a valuable tool for allelopathic studies in apple. in Tree Physiology. 2019;39(5):888-905.
doi:doi:10.1093/treephys/tpz006 .

Stanišić, Mariana, Ćosić, Tatjana, Savić, Jelena, Krstić Milošević, Dijana, Mišić, Danijela, Smigocki, Ann, Ninković, Slavica, Banjac, Nevena, "Hairy root culture as a valuable tool for allelopathic studies in apple" in Tree Physiology, 39, no. 5 (2019):888-905,
https://doi.org/doi:10.1093/treephys/tpz006 . .

TY  - JOUR
AU  - Trifunović-Momčilov, Milana
AU  - Krstić Milošević, Dijana
AU  - Trifunović, Snežana
AU  - Ćirić, Ana
AU  - Glamočlija, Jasmina
AU  - Jevremović, Slađana
AU  - Subotić, Angelina
PY  - 2019
UR  - http://eemj.eu/index.php/EEMJ/article/view/3962
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/3592
AB  - Common centaury, Centaurium erythraea Rafn., represent the best known and the most investigated medicinal plant species of genus Centaurium. Centaury has been used for centuries in traditional medicine. Secondary metabolites such as bitter secoiridoid glucosides (gentiopicrin, swertiamarin and sweroside), xanthones (eustomin and demethyleustomin), and phenolic acids are the main constituents responsible for the therapeutic properties of centaury. Previous investigation showed that overexpression of the AtCKX genes in transgenic centaury plants did not result in a decrease of total cytokinin (CK) content, but in an altered CK profile leading to a decline of bioactive, the most important physiologically active group of CKs.The aim of this study was to investigate antibacterial and antifungal activity of transgenic centaury methanol extracts as well as pure secoiridoid and xanthone compounds on four Gram positive, four Gram negative bacteria and eight species of microfungi. All tested methanol extracts of control and transgenic AtCKX1 centaury shoots and roots showed better antibacterial activity, while pure compounds (gentiopicrin, swertiamarin, eustomin and demethyleustomin) showed better antifungal activity. The results obtained in this work suggest that centaury methanol extracts and pure compounds represent potential antimicrobials confirming the possibility of using these compounds in agronomy, veterinary, medicine or food industry.
PB  - Gheorghe Asachi Technical University of Iasi, Romania
T2  - Environmental Engineering and Management Journal
T1  - Antimicrobial activity, antioxidant potential and total phenolic content of transgenic ATCKX1 centaury (Centaurium erythraea Rafn.) Plants grown in vitro
IS  - 9
VL  - 18
SP  - 2063
EP  - 2072
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_3592
ER  - 

@article{
author = "Trifunović-Momčilov, Milana and Krstić Milošević, Dijana and Trifunović, Snežana and Ćirić, Ana and Glamočlija, Jasmina and Jevremović, Slađana and Subotić, Angelina",
year = "2019",
abstract = "Common centaury, Centaurium erythraea Rafn., represent the best known and the most investigated medicinal plant species of genus Centaurium. Centaury has been used for centuries in traditional medicine. Secondary metabolites such as bitter secoiridoid glucosides (gentiopicrin, swertiamarin and sweroside), xanthones (eustomin and demethyleustomin), and phenolic acids are the main constituents responsible for the therapeutic properties of centaury. Previous investigation showed that overexpression of the AtCKX genes in transgenic centaury plants did not result in a decrease of total cytokinin (CK) content, but in an altered CK profile leading to a decline of bioactive, the most important physiologically active group of CKs.The aim of this study was to investigate antibacterial and antifungal activity of transgenic centaury methanol extracts as well as pure secoiridoid and xanthone compounds on four Gram positive, four Gram negative bacteria and eight species of microfungi. All tested methanol extracts of control and transgenic AtCKX1 centaury shoots and roots showed better antibacterial activity, while pure compounds (gentiopicrin, swertiamarin, eustomin and demethyleustomin) showed better antifungal activity. The results obtained in this work suggest that centaury methanol extracts and pure compounds represent potential antimicrobials confirming the possibility of using these compounds in agronomy, veterinary, medicine or food industry.",
publisher = "Gheorghe Asachi Technical University of Iasi, Romania",
journal = "Environmental Engineering and Management Journal",
title = "Antimicrobial activity, antioxidant potential and total phenolic content of transgenic ATCKX1 centaury (Centaurium erythraea Rafn.) Plants grown in vitro",
number = "9",
volume = "18",
pages = "2063-2072",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_3592"
}

Trifunović-Momčilov, M., Krstić Milošević, D., Trifunović, S., Ćirić, A., Glamočlija, J., Jevremović, S.,& Subotić, A.. (2019). Antimicrobial activity, antioxidant potential and total phenolic content of transgenic ATCKX1 centaury (Centaurium erythraea Rafn.) Plants grown in vitro. in Environmental Engineering and Management Journal
Gheorghe Asachi Technical University of Iasi, Romania., 18(9), 2063-2072.
https://hdl.handle.net/21.15107/rcub_ibiss_3592

Trifunović-Momčilov M, Krstić Milošević D, Trifunović S, Ćirić A, Glamočlija J, Jevremović S, Subotić A. Antimicrobial activity, antioxidant potential and total phenolic content of transgenic ATCKX1 centaury (Centaurium erythraea Rafn.) Plants grown in vitro. in Environmental Engineering and Management Journal. 2019;18(9):2063-2072.
https://hdl.handle.net/21.15107/rcub_ibiss_3592 .

Trifunović-Momčilov, Milana, Krstić Milošević, Dijana, Trifunović, Snežana, Ćirić, Ana, Glamočlija, Jasmina, Jevremović, Slađana, Subotić, Angelina, "Antimicrobial activity, antioxidant potential and total phenolic content of transgenic ATCKX1 centaury (Centaurium erythraea Rafn.) Plants grown in vitro" in Environmental Engineering and Management Journal, 18, no. 9 (2019):2063-2072,
https://hdl.handle.net/21.15107/rcub_ibiss_3592 .

TY  - JOUR
AU  - Vinterhalter, Branka
AU  - Savić, Jelena
AU  - Zdravković-Korać, Snežana
AU  - Banjac, Nevena
AU  - Vinterhalter, Dragan
AU  - Krstić Milošević, Dijana
PY  - 2019
UR  - https://www.sciencedirect.com/science/article/pii/S0926669018311282?via%3Dihub
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/3230
AB  - Production of innovative drugs from natural products in controlled conditions plays an important role in modern pharmacology in order to tackle global health challenges. The potential of Gentiana utriculosa hairy roots, obtained by transformation with Agrobacterium rhizogenes A4M70GUS, as well as that of the shoots regenerated from them via somatic embryogenesis, for xanthone production was investigated. Gentiana utriculosa was shown to be a new source of xanthones, medicinal raw materials for different pharmaceutical applications, among which decussatin has been recently recognized as a prospective hepatoprotective and antiulcer compound. Decussatin and decussatin-1-O-primeveroside were detected in both hairy roots and transgenic shoots, while mangiferin, present in nontransformed plants, was not detected. Quantitative HPLC analysis revealed up to 4.5-fold higher decussatin production in hairy roots line 9 compared with the plants from the nature, and this clone was selected. It showed stable growth after more than two years of continuous subcultivation. Cultivation of hairy roots on solid medium was favorable for biomass production, while liquid culture was beneficial for decussatin and decussatin-1-O-primeveroside accumulation. Cytokinins kinetin or N6-benzyladenine promoted somatic embryo maturation and germination as well as multiplication of obtained plantlets, while active charcoal reduced hyperhydricity. Plants regenerated from selected hairy root line 9 comprised two TL-DNA inserts, as confirmed by Southern blot analysis. They accumulated at least 2.5-fold more decussatin then nontransformed plants. Therefore, these plants could be valuable material to create xanthone high-yielding cultivars of G. utriculosa.
T2  - Industrial Crops and Products
T1  - Agrobacterium rhizogenes-mediated transformation of Gentiana utriculosa L. and xanthones decussatin-1-O-primeveroside and decussatin accumulation in hairy roots and somatic embryo-derived transgenic plants
VL  - 130
DO  - 10.1016/J.INDCROP.2018.12.066
SP  - 216
EP  - 229
ER  - 

@article{
author = "Vinterhalter, Branka and Savić, Jelena and Zdravković-Korać, Snežana and Banjac, Nevena and Vinterhalter, Dragan and Krstić Milošević, Dijana",
year = "2019",
abstract = "Production of innovative drugs from natural products in controlled conditions plays an important role in modern pharmacology in order to tackle global health challenges. The potential of Gentiana utriculosa hairy roots, obtained by transformation with Agrobacterium rhizogenes A4M70GUS, as well as that of the shoots regenerated from them via somatic embryogenesis, for xanthone production was investigated. Gentiana utriculosa was shown to be a new source of xanthones, medicinal raw materials for different pharmaceutical applications, among which decussatin has been recently recognized as a prospective hepatoprotective and antiulcer compound. Decussatin and decussatin-1-O-primeveroside were detected in both hairy roots and transgenic shoots, while mangiferin, present in nontransformed plants, was not detected. Quantitative HPLC analysis revealed up to 4.5-fold higher decussatin production in hairy roots line 9 compared with the plants from the nature, and this clone was selected. It showed stable growth after more than two years of continuous subcultivation. Cultivation of hairy roots on solid medium was favorable for biomass production, while liquid culture was beneficial for decussatin and decussatin-1-O-primeveroside accumulation. Cytokinins kinetin or N6-benzyladenine promoted somatic embryo maturation and germination as well as multiplication of obtained plantlets, while active charcoal reduced hyperhydricity. Plants regenerated from selected hairy root line 9 comprised two TL-DNA inserts, as confirmed by Southern blot analysis. They accumulated at least 2.5-fold more decussatin then nontransformed plants. Therefore, these plants could be valuable material to create xanthone high-yielding cultivars of G. utriculosa.",
journal = "Industrial Crops and Products",
title = "Agrobacterium rhizogenes-mediated transformation of Gentiana utriculosa L. and xanthones decussatin-1-O-primeveroside and decussatin accumulation in hairy roots and somatic embryo-derived transgenic plants",
volume = "130",
doi = "10.1016/J.INDCROP.2018.12.066",
pages = "216-229"
}

Vinterhalter, B., Savić, J., Zdravković-Korać, S., Banjac, N., Vinterhalter, D.,& Krstić Milošević, D.. (2019). Agrobacterium rhizogenes-mediated transformation of Gentiana utriculosa L. and xanthones decussatin-1-O-primeveroside and decussatin accumulation in hairy roots and somatic embryo-derived transgenic plants. in Industrial Crops and Products, 130, 216-229.
https://doi.org/10.1016/J.INDCROP.2018.12.066

Vinterhalter B, Savić J, Zdravković-Korać S, Banjac N, Vinterhalter D, Krstić Milošević D. Agrobacterium rhizogenes-mediated transformation of Gentiana utriculosa L. and xanthones decussatin-1-O-primeveroside and decussatin accumulation in hairy roots and somatic embryo-derived transgenic plants. in Industrial Crops and Products. 2019;130:216-229.
doi:10.1016/J.INDCROP.2018.12.066 .

Vinterhalter, Branka, Savić, Jelena, Zdravković-Korać, Snežana, Banjac, Nevena, Vinterhalter, Dragan, Krstić Milošević, Dijana, "Agrobacterium rhizogenes-mediated transformation of Gentiana utriculosa L. and xanthones decussatin-1-O-primeveroside and decussatin accumulation in hairy roots and somatic embryo-derived transgenic plants" in Industrial Crops and Products, 130 (2019):216-229,
https://doi.org/10.1016/J.INDCROP.2018.12.066 . .

TY  - JOUR
AU  - Bojović, Katarina
AU  - Stanković, Biljana
AU  - Kotur, Nikola
AU  - Krstić Milošević, Dijana
AU  - Gašić, Vladimir
AU  - Pavlović, Sonja
AU  - Zukić, Branka
AU  - Ignjatović, Đurđica
PY  - 2019
UR  - https://www.tandfonline.com/doi/full/10.1080/1028415X.2017.1352121
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/2823
AB  - Gastrointestinal disturbances, nutritional deficiencies, and food intolerances are frequently observed in children with neurodevelopmental disorders (NDD). To reveal possible association of celiac disease risk variants (HLA-DQ), lactose intolerance associated variant (LCT-13910C > T) as well as variant associated with vitamin D function (VDR FokI) with NDD, polymerase chain reaction-based methodology was used. Additionally, intestinal peptide permeability was estimated in NDD patients and healthy children by measuring the level of peptides in urine using high-performance liquid chromatography. Levels of opioid peptides, casomorphin 8, and gluten exorphin C were significantly elevated in urine samples of NDD patients (P = 0.004 and P = 0.005, respectively), but no association of genetic risk variants for celiac disease and lactose intolerance with NDD was found. Our results indicate that increased intestinal peptide permeability observed in analyzed NDD patients is not associated with genetic predictors of celiac disease or lactose intolerance. We have also found that FF genotype of VDR FokI and lower serum levels of vitamin D (25-OH) showed association with childhood autism (CHA), a subgroup of NDD. We hypothesize that vitamin D might be important for the development of CHA.
T2  - Nutritional Neuroscience
T1  - Genetic predictors of celiac disease, lactose intolerance, and vitamin D function and presence of peptide morphins in urine of children with neurodevelopmental disorders
IS  - 1
VL  - 22
DO  - 10.1080/1028415X.2017.1352121
SP  - 40
EP  - 50
ER  - 

@article{
author = "Bojović, Katarina and Stanković, Biljana and Kotur, Nikola and Krstić Milošević, Dijana and Gašić, Vladimir and Pavlović, Sonja and Zukić, Branka and Ignjatović, Đurđica",
year = "2019",
abstract = "Gastrointestinal disturbances, nutritional deficiencies, and food intolerances are frequently observed in children with neurodevelopmental disorders (NDD). To reveal possible association of celiac disease risk variants (HLA-DQ), lactose intolerance associated variant (LCT-13910C > T) as well as variant associated with vitamin D function (VDR FokI) with NDD, polymerase chain reaction-based methodology was used. Additionally, intestinal peptide permeability was estimated in NDD patients and healthy children by measuring the level of peptides in urine using high-performance liquid chromatography. Levels of opioid peptides, casomorphin 8, and gluten exorphin C were significantly elevated in urine samples of NDD patients (P = 0.004 and P = 0.005, respectively), but no association of genetic risk variants for celiac disease and lactose intolerance with NDD was found. Our results indicate that increased intestinal peptide permeability observed in analyzed NDD patients is not associated with genetic predictors of celiac disease or lactose intolerance. We have also found that FF genotype of VDR FokI and lower serum levels of vitamin D (25-OH) showed association with childhood autism (CHA), a subgroup of NDD. We hypothesize that vitamin D might be important for the development of CHA.",
journal = "Nutritional Neuroscience",
title = "Genetic predictors of celiac disease, lactose intolerance, and vitamin D function and presence of peptide morphins in urine of children with neurodevelopmental disorders",
number = "1",
volume = "22",
doi = "10.1080/1028415X.2017.1352121",
pages = "40-50"
}

Bojović, K., Stanković, B., Kotur, N., Krstić Milošević, D., Gašić, V., Pavlović, S., Zukić, B.,& Ignjatović, Đ.. (2019). Genetic predictors of celiac disease, lactose intolerance, and vitamin D function and presence of peptide morphins in urine of children with neurodevelopmental disorders. in Nutritional Neuroscience, 22(1), 40-50.
https://doi.org/10.1080/1028415X.2017.1352121

Bojović K, Stanković B, Kotur N, Krstić Milošević D, Gašić V, Pavlović S, Zukić B, Ignjatović Đ. Genetic predictors of celiac disease, lactose intolerance, and vitamin D function and presence of peptide morphins in urine of children with neurodevelopmental disorders. in Nutritional Neuroscience. 2019;22(1):40-50.
doi:10.1080/1028415X.2017.1352121 .

Bojović, Katarina, Stanković, Biljana, Kotur, Nikola, Krstić Milošević, Dijana, Gašić, Vladimir, Pavlović, Sonja, Zukić, Branka, Ignjatović, Đurđica, "Genetic predictors of celiac disease, lactose intolerance, and vitamin D function and presence of peptide morphins in urine of children with neurodevelopmental disorders" in Nutritional Neuroscience, 22, no. 1 (2019):40-50,
https://doi.org/10.1080/1028415X.2017.1352121 . .

TY  - JOUR
AU  - Tovilović-Kovačević, Gordana
AU  - Krstić Milošević, Dijana
AU  - Vinterhalter, Branka
AU  - Toljić, Mina
AU  - Perović, Vladimir
AU  - Trajković, Vladimir
AU  - Harhaji-Trajković, Ljubica
AU  - Zogović, Nevena
PY  - 2018
UR  - https://www.sciencedirect.com/science/article/pii/S0944711318300874?via%3Dihub
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/3125
AB  - BACKGROUND Glioblastoma multiforme (GMB) is the most malignant of all brain tumors with poor prognosis. Anticancer potential of xanthones, bioactive compounds found in Gentiana dinarica, is well-documented. Transformation of G. dinarica roots with Agrobacterium rhizogenes provides higher xanthones accumulation, which enables better exploitation of these anticancer compounds. HYPOTHESIS/PURPOSE The aim of this study was to investigate antiglioma effect of three different G. dinarica extracts: E1—derived from untransformed roots, E2—derived from roots transformed using A. rhizogenes strain A4M70GUS, and E3—derived from roots transformed using A. rhizogenes strain 15834/PI. Further, mechanisms involved in anticancer potential of the most potent extract were examined in detail, and its active component was determined. METHODS The cell viability was assessed using MTT and crystal violet test. Cell cycle analysis, the expression of differentiation markers, the levels of autophagy, and oxidative stress were analyzed by flow cytometry. Autophagy and related signaling pathways were assessed by immunoblotting. RESULTS E3, in contrast to E1 and E2, strongly reduced growth of U251 human glioblastoma cells, triggered cell cycle arrest in G2/M phase, changed cellular morphology, and increased expression of markers of differentiated astrocytes (glial fibrillary acidic protein) and neurons (β-tubulin). E3 stimulated autophagy, as demonstrated by enhanced intracellular acidification, increased microtubule-associated light chain 3B (LC3-I) conversion to autophagosome associated LC3-II, and decreased level of selective autophagy target p62. Induction of autophagy was associated with Akt-dependent inhibition of main autophagy suppressor mammalian target of rapamycin (mTOR). Both genetic and pharmacological inhibition of autophagy suppressed the expression of differentiation markers, but had no effect on cell cycle arrest in E3-treated cells. E3 stimulated oxidative stress, and antioxidants vitamin E and N-acetyl cysteine inhibited autophagy and differentiation of E3-treated U251 cells. The most prevalent compound of E3, xanthone aglycone norswertianin, also arrested glioblastoma cell proliferation in G2/M phase and induced glioblastoma cell differentiation through induction of autophagy and oxidative stress. CONCLUSION These results indicate that E3 and its main active component norswertianin may serve as a potential candidate for differentiation therapy of glioblastoma.
T2  - Phytomedicine
T1  - Xanthone-rich extract from Gentiana dinarica transformed roots and its active component norswertianin induce autophagy and ROS-dependent differentiation of human glioblastoma cell line
VL  - 47
DO  - 10.1016/J.PHYMED.2018.03.052
SP  - 151
EP  - 160
ER  - 

@article{
author = "Tovilović-Kovačević, Gordana and Krstić Milošević, Dijana and Vinterhalter, Branka and Toljić, Mina and Perović, Vladimir and Trajković, Vladimir and Harhaji-Trajković, Ljubica and Zogović, Nevena",
year = "2018",
abstract = "BACKGROUND Glioblastoma multiforme (GMB) is the most malignant of all brain tumors with poor prognosis. Anticancer potential of xanthones, bioactive compounds found in Gentiana dinarica, is well-documented. Transformation of G. dinarica roots with Agrobacterium rhizogenes provides higher xanthones accumulation, which enables better exploitation of these anticancer compounds. HYPOTHESIS/PURPOSE The aim of this study was to investigate antiglioma effect of three different G. dinarica extracts: E1—derived from untransformed roots, E2—derived from roots transformed using A. rhizogenes strain A4M70GUS, and E3—derived from roots transformed using A. rhizogenes strain 15834/PI. Further, mechanisms involved in anticancer potential of the most potent extract were examined in detail, and its active component was determined. METHODS The cell viability was assessed using MTT and crystal violet test. Cell cycle analysis, the expression of differentiation markers, the levels of autophagy, and oxidative stress were analyzed by flow cytometry. Autophagy and related signaling pathways were assessed by immunoblotting. RESULTS E3, in contrast to E1 and E2, strongly reduced growth of U251 human glioblastoma cells, triggered cell cycle arrest in G2/M phase, changed cellular morphology, and increased expression of markers of differentiated astrocytes (glial fibrillary acidic protein) and neurons (β-tubulin). E3 stimulated autophagy, as demonstrated by enhanced intracellular acidification, increased microtubule-associated light chain 3B (LC3-I) conversion to autophagosome associated LC3-II, and decreased level of selective autophagy target p62. Induction of autophagy was associated with Akt-dependent inhibition of main autophagy suppressor mammalian target of rapamycin (mTOR). Both genetic and pharmacological inhibition of autophagy suppressed the expression of differentiation markers, but had no effect on cell cycle arrest in E3-treated cells. E3 stimulated oxidative stress, and antioxidants vitamin E and N-acetyl cysteine inhibited autophagy and differentiation of E3-treated U251 cells. The most prevalent compound of E3, xanthone aglycone norswertianin, also arrested glioblastoma cell proliferation in G2/M phase and induced glioblastoma cell differentiation through induction of autophagy and oxidative stress. CONCLUSION These results indicate that E3 and its main active component norswertianin may serve as a potential candidate for differentiation therapy of glioblastoma.",
journal = "Phytomedicine",
title = "Xanthone-rich extract from Gentiana dinarica transformed roots and its active component norswertianin induce autophagy and ROS-dependent differentiation of human glioblastoma cell line",
volume = "47",
doi = "10.1016/J.PHYMED.2018.03.052",
pages = "151-160"
}

Tovilović-Kovačević, G., Krstić Milošević, D., Vinterhalter, B., Toljić, M., Perović, V., Trajković, V., Harhaji-Trajković, L.,& Zogović, N.. (2018). Xanthone-rich extract from Gentiana dinarica transformed roots and its active component norswertianin induce autophagy and ROS-dependent differentiation of human glioblastoma cell line. in Phytomedicine, 47, 151-160.
https://doi.org/10.1016/J.PHYMED.2018.03.052

Tovilović-Kovačević G, Krstić Milošević D, Vinterhalter B, Toljić M, Perović V, Trajković V, Harhaji-Trajković L, Zogović N. Xanthone-rich extract from Gentiana dinarica transformed roots and its active component norswertianin induce autophagy and ROS-dependent differentiation of human glioblastoma cell line. in Phytomedicine. 2018;47:151-160.
doi:10.1016/J.PHYMED.2018.03.052 .

Tovilović-Kovačević, Gordana, Krstić Milošević, Dijana, Vinterhalter, Branka, Toljić, Mina, Perović, Vladimir, Trajković, Vladimir, Harhaji-Trajković, Ljubica, Zogović, Nevena, "Xanthone-rich extract from Gentiana dinarica transformed roots and its active component norswertianin induce autophagy and ROS-dependent differentiation of human glioblastoma cell line" in Phytomedicine, 47 (2018):151-160,
https://doi.org/10.1016/J.PHYMED.2018.03.052 . .

TY  - CONF
AU  - Stanišić, Mariana
AU  - Ćosić, Tatjana
AU  - Savić, Jelena
AU  - Krstić Milošević, Dijana
AU  - Mišić, Danijela
AU  - Smigocki, Ann
AU  - Ninković, Slavica
AU  - Banjac, Nevena
PY  - 2018
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5009
AB  - Since the investigation of plant allelopathic interactions in the rhizosphere is difficult to perform
because of its high complexity, microbial and environmental factors interactions, the generation
of autonomous root culture could be appropriate system for the studies of allelopathic potential of apple (Malus x domestica Borkh.). Thus, we have focused our efforts on the generation
of transformed hairy-roots in apple cv. Golden Delicious using the 15834 strain of Agrobacterium
rhizogenes. The agrobacteria inoculation by dipping the basal cut off end of the stem of in vitro
grown apple shoots into bacterial suspension proved to be the best option for transformation,
providing efficiency of 39.3%. Molecular characterization of hairy roots by RT-PCR analyses confirmed
their transgenic nature. In vitro conditions for apple hairy root growth and maintenance
were optimized using different mineral solutions and auxins. The liquid half-strength MS medium
with the addition of IBA at 0.025 mg L-1 was chosen for the initial stimulation of growth. For their
prolonged cultivation and maintenance, the alternation of IBA-containing and IBA-free media is
recommendable. Fast-growing lines continuously maintained in IBA-free medium were used for
further research. Potential allelochemicals including phenolic acids (chlorogenic, caffeic, syringic,
p-coumaric and ferulic), glycosilated flavonoids (phloridzin, rutin, quercitrin, isoquercitrin, kaempferol-
3-glucoside) and flavonoid aglycons (floretin, quercetin and naringenin) have been detected
by UHPLC/DAD/(+/-)HESI-MS/MS analyses in the growth medium, in which these transformed
roots were maintained for 4 weeks, as well as in the root tissue itself. The obtained hairy-root
cultures offer a complementary system for studying and validating the allelopathic potential of
apple roots.
PB  - Belgrade: Serbian Plant Physiology Society
C3  - 3rd International Conference on Plant Biology (22nd SPPS Meeting); 2018 Jun 9-12; Belgrade, Serbia
T1  - Transgenic hairy root in vitro culture system of apple as a tool for allelopathic studies
SP  - 55
EP  - 56
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_5009
ER  - 

@conference{
author = "Stanišić, Mariana and Ćosić, Tatjana and Savić, Jelena and Krstić Milošević, Dijana and Mišić, Danijela and Smigocki, Ann and Ninković, Slavica and Banjac, Nevena",
year = "2018",
abstract = "Since the investigation of plant allelopathic interactions in the rhizosphere is difficult to perform
because of its high complexity, microbial and environmental factors interactions, the generation
of autonomous root culture could be appropriate system for the studies of allelopathic potential of apple (Malus x domestica Borkh.). Thus, we have focused our efforts on the generation
of transformed hairy-roots in apple cv. Golden Delicious using the 15834 strain of Agrobacterium
rhizogenes. The agrobacteria inoculation by dipping the basal cut off end of the stem of in vitro
grown apple shoots into bacterial suspension proved to be the best option for transformation,
providing efficiency of 39.3%. Molecular characterization of hairy roots by RT-PCR analyses confirmed
their transgenic nature. In vitro conditions for apple hairy root growth and maintenance
were optimized using different mineral solutions and auxins. The liquid half-strength MS medium
with the addition of IBA at 0.025 mg L-1 was chosen for the initial stimulation of growth. For their
prolonged cultivation and maintenance, the alternation of IBA-containing and IBA-free media is
recommendable. Fast-growing lines continuously maintained in IBA-free medium were used for
further research. Potential allelochemicals including phenolic acids (chlorogenic, caffeic, syringic,
p-coumaric and ferulic), glycosilated flavonoids (phloridzin, rutin, quercitrin, isoquercitrin, kaempferol-
3-glucoside) and flavonoid aglycons (floretin, quercetin and naringenin) have been detected
by UHPLC/DAD/(+/-)HESI-MS/MS analyses in the growth medium, in which these transformed
roots were maintained for 4 weeks, as well as in the root tissue itself. The obtained hairy-root
cultures offer a complementary system for studying and validating the allelopathic potential of
apple roots.",
publisher = "Belgrade: Serbian Plant Physiology Society",
journal = "3rd International Conference on Plant Biology (22nd SPPS Meeting); 2018 Jun 9-12; Belgrade, Serbia",
title = "Transgenic hairy root in vitro culture system of apple as a tool for allelopathic studies",
pages = "55-56",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_5009"
}

Stanišić, M., Ćosić, T., Savić, J., Krstić Milošević, D., Mišić, D., Smigocki, A., Ninković, S.,& Banjac, N.. (2018). Transgenic hairy root in vitro culture system of apple as a tool for allelopathic studies. in 3rd International Conference on Plant Biology (22nd SPPS Meeting); 2018 Jun 9-12; Belgrade, Serbia
Belgrade: Serbian Plant Physiology Society., 55-56.
https://hdl.handle.net/21.15107/rcub_ibiss_5009

Stanišić M, Ćosić T, Savić J, Krstić Milošević D, Mišić D, Smigocki A, Ninković S, Banjac N. Transgenic hairy root in vitro culture system of apple as a tool for allelopathic studies. in 3rd International Conference on Plant Biology (22nd SPPS Meeting); 2018 Jun 9-12; Belgrade, Serbia. 2018;:55-56.
https://hdl.handle.net/21.15107/rcub_ibiss_5009 .

Stanišić, Mariana, Ćosić, Tatjana, Savić, Jelena, Krstić Milošević, Dijana, Mišić, Danijela, Smigocki, Ann, Ninković, Slavica, Banjac, Nevena, "Transgenic hairy root in vitro culture system of apple as a tool for allelopathic studies" in 3rd International Conference on Plant Biology (22nd SPPS Meeting); 2018 Jun 9-12; Belgrade, Serbia (2018):55-56,
https://hdl.handle.net/21.15107/rcub_ibiss_5009 .