TY  - GEN
AU  - Filipović, Biljana
AU  - Skorić, Marijana
AU  - Gašić, Uroš
AU  - Dragićević, Milan
AU  - Božunović, Jelena
AU  - Matekalo, Dragana
AU  - Nestorović Živković, Jasmina
AU  - Banjanac, Tijana
AU  - Šiler, Branislav
AU  - Bohanec, Borut
AU  - Milutinović, Milica
AU  - Mišić, Danijela
PY  - 2022
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6326
AB  - Kičica (Centaurium erythraea Rafn) je bogat izvor brojnih bioaktivnih jedinjenja, među kojima su najzastupljeniji sekoiridoidi i ksantoni. Poznato je da na akumulaciju specijalizovanih metabolita u lekovitim biljkama utiču razni faktori (ploidnost i starost biljke, tip biljnog organa, itd.). U cilju ispitivanja optimalne produkcije sekoiridoida i ksantona radi boljeg iskorišćavanja biotehnološkog potencijala kičice praćene su promene biosinteze ovih specijalizovanih metabolita u izdancima i korenovima diploidne i tetraploidne kičice tokom tri meseca in vitro razvića. Rezultati uporedne analize ekspresije gena uključenih u biosintetske puteve sekoiridoida i ksantona i fitohemijskog profilisanja ovih jedinjenja su pokazali da su izdanci glavno mesto njihove akumulacije, kao i da njihov odnos varira tokom razvića. Koordinisana ekspresija biosintetskih gena uključenih u metabolizam iridoida i ksantona je regulisana na nivou transkripcije kako tokom razvića tako i na nivou organa. Povišeni nivoi ekspresije biosintetskih gena, ključnih za regulisanje metaboličkog fluksa, su u korelaciji sa sadržajem iridoida i ksantona. Tako je pojačana ekspresija GES, 8HGO i 7DLH2 praćena povećanom produkcijom iridoida, dok je viši nivo transkripata BS i 3HBL pozitivno korelisan sa akumulacijom heksa-supstituisanih ksantona. Može se zaključiti da je diploidni genotip perspektivniji od tetraploidnog u kontrolisanim in vitro uslovima kako u pogledu produkcije iridoida i ksantona, tako i u pogledu prirasta biomase.
AB  - Кичица (Centaurium erythraea Rafn) је богат извор бројних биоактивних једињења,
међу којима су најзаступљенији секоиридоиди и ксантони.1 Познато је да на
акумулацију специјализованих метаболита у лековитим биљкама утичу разни
фактори (плоидност1 и старост биљке, тип биљног органа, итд.). У циљу
испитивања оптималне продукције секоиридоида и ксантона ради бољег
искоришћавања биотехнолошког потенцијала кичице праћене су промене
биосинтезе ових специјализованих метаболита у изданцима и кореновима
диплоидне и тетраплоидне кичице током три месеца in vitro развића.2 Резултати
упоредне анализе експресије гена укључених у биосинтетске путеве секоиридоида
и ксантона и фитохемијског профилисања ових једињења су показали да су
изданци главно место њихове акумулације, као и да њихов однос варира током
развића. Координисана експресија биосинтетских гена укључених у метаболизам
иридоида и ксантона је регулисана на нивоу транскрипције како током развића
тако и на нивоу органа. Повишени нивои експресије биосинтетских гена, кључних
за регулисање метаболичког флукса, су у корелацији са садржајем иридоида и
ксантона. Тако је појачана експресија GES, 8HGO и 7DLH2 праћена повећаном
продукцијом иридоида, док је виши ниво транскрипата BS и 3HBL позитивно
корелисан са акумулацијом хекса-супституисаних ксантона. Може се закључити да
је диплоидни генотип перспективнији од тетраплоидног у контролисаним in vitro
условима како у погледу продукције иридоида и ксантона, тако и у погледу
прираста биомасе.
PB  - Belgrade: Serbian Biological Society
T2  - Knjiga sažetaka: Treći Kongres biologa Srbije: Osnovna i primenjena istraživanja: Metodika nastave; 2022 Sep 21-25; Zlatibor, Serbia
T1  - Prostorni i vremenski obrazac biosinteze sekoiridoida i ksantona tokom ranog razvića diploidne i tetraploidne kičice
T1  - Просторни и временски образац биосинтезе секоиридоида и ксантона током раног развића диплоидне и тетраплоидне кичице
SP  - 33
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_6326
ER  - 

@misc{
author = "Filipović, Biljana and Skorić, Marijana and Gašić, Uroš and Dragićević, Milan and Božunović, Jelena and Matekalo, Dragana and Nestorović Živković, Jasmina and Banjanac, Tijana and Šiler, Branislav and Bohanec, Borut and Milutinović, Milica and Mišić, Danijela",
year = "2022",
abstract = "Kičica (Centaurium erythraea Rafn) je bogat izvor brojnih bioaktivnih jedinjenja, među kojima su najzastupljeniji sekoiridoidi i ksantoni. Poznato je da na akumulaciju specijalizovanih metabolita u lekovitim biljkama utiču razni faktori (ploidnost i starost biljke, tip biljnog organa, itd.). U cilju ispitivanja optimalne produkcije sekoiridoida i ksantona radi boljeg iskorišćavanja biotehnološkog potencijala kičice praćene su promene biosinteze ovih specijalizovanih metabolita u izdancima i korenovima diploidne i tetraploidne kičice tokom tri meseca in vitro razvića. Rezultati uporedne analize ekspresije gena uključenih u biosintetske puteve sekoiridoida i ksantona i fitohemijskog profilisanja ovih jedinjenja su pokazali da su izdanci glavno mesto njihove akumulacije, kao i da njihov odnos varira tokom razvića. Koordinisana ekspresija biosintetskih gena uključenih u metabolizam iridoida i ksantona je regulisana na nivou transkripcije kako tokom razvića tako i na nivou organa. Povišeni nivoi ekspresije biosintetskih gena, ključnih za regulisanje metaboličkog fluksa, su u korelaciji sa sadržajem iridoida i ksantona. Tako je pojačana ekspresija GES, 8HGO i 7DLH2 praćena povećanom produkcijom iridoida, dok je viši nivo transkripata BS i 3HBL pozitivno korelisan sa akumulacijom heksa-supstituisanih ksantona. Može se zaključiti da je diploidni genotip perspektivniji od tetraploidnog u kontrolisanim in vitro uslovima kako u pogledu produkcije iridoida i ksantona, tako i u pogledu prirasta biomase., Кичица (Centaurium erythraea Rafn) је богат извор бројних биоактивних једињења,
међу којима су најзаступљенији секоиридоиди и ксантони.1 Познато је да на
акумулацију специјализованих метаболита у лековитим биљкама утичу разни
фактори (плоидност1 и старост биљке, тип биљног органа, итд.). У циљу
испитивања оптималне продукције секоиридоида и ксантона ради бољег
искоришћавања биотехнолошког потенцијала кичице праћене су промене
биосинтезе ових специјализованих метаболита у изданцима и кореновима
диплоидне и тетраплоидне кичице током три месеца in vitro развића.2 Резултати
упоредне анализе експресије гена укључених у биосинтетске путеве секоиридоида
и ксантона и фитохемијског профилисања ових једињења су показали да су
изданци главно место њихове акумулације, као и да њихов однос варира током
развића. Координисана експресија биосинтетских гена укључених у метаболизам
иридоида и ксантона је регулисана на нивоу транскрипције како током развића
тако и на нивоу органа. Повишени нивои експресије биосинтетских гена, кључних
за регулисање метаболичког флукса, су у корелацији са садржајем иридоида и
ксантона. Тако је појачана експресија GES, 8HGO и 7DLH2 праћена повећаном
продукцијом иридоида, док је виши ниво транскрипата BS и 3HBL позитивно
корелисан са акумулацијом хекса-супституисаних ксантона. Може се закључити да
је диплоидни генотип перспективнији од тетраплоидног у контролисаним in vitro
условима како у погледу продукције иридоида и ксантона, тако и у погледу
прираста биомасе.",
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 = "Prostorni i vremenski obrazac biosinteze sekoiridoida i ksantona tokom ranog razvića diploidne i tetraploidne kičice, Просторни и временски образац биосинтезе секоиридоида и ксантона током раног развића диплоидне и тетраплоидне кичице",
pages = "33",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_6326"
}

Filipović, B., Skorić, M., Gašić, U., Dragićević, M., Božunović, J., Matekalo, D., Nestorović Živković, J., Banjanac, T., Šiler, B., Bohanec, B., Milutinović, M.,& Mišić, D.. (2022). Prostorni i vremenski obrazac biosinteze sekoiridoida i ksantona tokom ranog razvića diploidne i tetraploidne kičice. in Knjiga sažetaka: Treći Kongres biologa Srbije: Osnovna i primenjena istraživanja: Metodika nastave; 2022 Sep 21-25; Zlatibor, Serbia
Belgrade: Serbian Biological Society., 33.
https://hdl.handle.net/21.15107/rcub_ibiss_6326

Filipović B, Skorić M, Gašić U, Dragićević M, Božunović J, Matekalo D, Nestorović Živković J, Banjanac T, Šiler B, Bohanec B, Milutinović M, Mišić D. Prostorni i vremenski obrazac biosinteze sekoiridoida i ksantona tokom ranog razvića diploidne i tetraploidne kičice. in Knjiga sažetaka: Treći Kongres biologa Srbije: Osnovna i primenjena istraživanja: Metodika nastave; 2022 Sep 21-25; Zlatibor, Serbia. 2022;:33.
https://hdl.handle.net/21.15107/rcub_ibiss_6326 .

Filipović, Biljana, Skorić, Marijana, Gašić, Uroš, Dragićević, Milan, Božunović, Jelena, Matekalo, Dragana, Nestorović Živković, Jasmina, Banjanac, Tijana, Šiler, Branislav, Bohanec, Borut, Milutinović, Milica, Mišić, Danijela, "Prostorni i vremenski obrazac biosinteze sekoiridoida i ksantona tokom ranog razvića diploidne i tetraploidne kičice" in Knjiga sažetaka: Treći Kongres biologa Srbije: Osnovna i primenjena istraživanja: Metodika nastave; 2022 Sep 21-25; Zlatibor, Serbia (2022):33,
https://hdl.handle.net/21.15107/rcub_ibiss_6326 .

TY  - JOUR
AU  - Filipović, Biljana
AU  - Skorić, Marijana
AU  - Gašić, Uroš
AU  - Dragićević, Milan
AU  - Božunović, Jelena
AU  - Matekalo, Dragana
AU  - Nestorović Živković, Jasmina
AU  - Banjanac, Tijana
AU  - Šiler, Branislav
AU  - Bohanec, Borut
AU  - Milutinović, Milica
AU  - Mišić, Danijela
PY  - 2022
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5254
AB  - Centaurium erythraea, used as medical plant from the earliest times, is an immense depot of quite rare bioactive compounds. Secoiridoids (sweroside, swertiamarin, and gentiopicrin) and xanthones (methylbellidifolin and decussatin) are predominant bioactive compounds in C. erythraea. The present study aims at providing new insights into how the content of these bioactive principles can be related with plant ploidy level by characterizing possible differences in their biosynthesis and accumulation between diploid and tetraploid genotypes from both spatial and temporal aspects. In general, shoots are determined as the major site of secoiridoids’ and xanthones’ accumulation, whose ratio vary during the development. Genes involved in iridoid and xanthone metabolic pathways were found to be coordinately regulated at the transcriptional level both during the development and among organs. Biosynthetic gene expression levels were found highly correlated with the content of major compounds from these two classes. Diversification in chemical profiles between tetraploid and diploid genotypes may result from the expression difference between homologous loci correspondent to several key biosynthetic genes, which trigger changes in the two metabolic routes. Thus, enhanced expression of genes coding for geraniol synthase (GES), 8-hydroxygeraniol oxidoreductase (8HGO), and 7-deoxyloganic acid hydrolase (7DLH2) is strongly associated with intensive production of iridoids. Interestingly, transcript levels of beta-glucosidase (CebGLU), a candidate to catalyze the first step in the secoiridoid catabolism, is significantly positively correlated with the content of major secoiridoids. Elevated expression of genes coding for benzophenone synthase (BS) and 3-hydroxybenzoate:CoA ligase (3HBL) appear to account for enhanced production of hexa-substituted xanthones. Regarding content of iridoids and xanthones, a diploid genotype appeared to be more productive than a tetraploid genotype under controlled in vitro conditions, therewithal displaying significantly higher biomass.
PB  - Amsterdam : Elsevier
T2  - Industrial Crops and Products
T1  - Spatial and temporal patterns of secoiridoid and xanthone biosynthetic pathways during early development of Centaurium erythraea Rafn, as altered by ploidy level
VL  - 186
DO  - 10.1016/j.indcrop.2022.115146
SP  - 115146
ER  - 

@article{
author = "Filipović, Biljana and Skorić, Marijana and Gašić, Uroš and Dragićević, Milan and Božunović, Jelena and Matekalo, Dragana and Nestorović Živković, Jasmina and Banjanac, Tijana and Šiler, Branislav and Bohanec, Borut and Milutinović, Milica and Mišić, Danijela",
year = "2022",
abstract = "Centaurium erythraea, used as medical plant from the earliest times, is an immense depot of quite rare bioactive compounds. Secoiridoids (sweroside, swertiamarin, and gentiopicrin) and xanthones (methylbellidifolin and decussatin) are predominant bioactive compounds in C. erythraea. The present study aims at providing new insights into how the content of these bioactive principles can be related with plant ploidy level by characterizing possible differences in their biosynthesis and accumulation between diploid and tetraploid genotypes from both spatial and temporal aspects. In general, shoots are determined as the major site of secoiridoids’ and xanthones’ accumulation, whose ratio vary during the development. Genes involved in iridoid and xanthone metabolic pathways were found to be coordinately regulated at the transcriptional level both during the development and among organs. Biosynthetic gene expression levels were found highly correlated with the content of major compounds from these two classes. Diversification in chemical profiles between tetraploid and diploid genotypes may result from the expression difference between homologous loci correspondent to several key biosynthetic genes, which trigger changes in the two metabolic routes. Thus, enhanced expression of genes coding for geraniol synthase (GES), 8-hydroxygeraniol oxidoreductase (8HGO), and 7-deoxyloganic acid hydrolase (7DLH2) is strongly associated with intensive production of iridoids. Interestingly, transcript levels of beta-glucosidase (CebGLU), a candidate to catalyze the first step in the secoiridoid catabolism, is significantly positively correlated with the content of major secoiridoids. Elevated expression of genes coding for benzophenone synthase (BS) and 3-hydroxybenzoate:CoA ligase (3HBL) appear to account for enhanced production of hexa-substituted xanthones. Regarding content of iridoids and xanthones, a diploid genotype appeared to be more productive than a tetraploid genotype under controlled in vitro conditions, therewithal displaying significantly higher biomass.",
publisher = "Amsterdam : Elsevier",
journal = "Industrial Crops and Products",
title = "Spatial and temporal patterns of secoiridoid and xanthone biosynthetic pathways during early development of Centaurium erythraea Rafn, as altered by ploidy level",
volume = "186",
doi = "10.1016/j.indcrop.2022.115146",
pages = "115146"
}

Filipović, B., Skorić, M., Gašić, U., Dragićević, M., Božunović, J., Matekalo, D., Nestorović Živković, J., Banjanac, T., Šiler, B., Bohanec, B., Milutinović, M.,& Mišić, D.. (2022). Spatial and temporal patterns of secoiridoid and xanthone biosynthetic pathways during early development of Centaurium erythraea Rafn, as altered by ploidy level. in Industrial Crops and Products
Amsterdam : Elsevier., 186, 115146.
https://doi.org/10.1016/j.indcrop.2022.115146

Filipović B, Skorić M, Gašić U, Dragićević M, Božunović J, Matekalo D, Nestorović Živković J, Banjanac T, Šiler B, Bohanec B, Milutinović M, Mišić D. Spatial and temporal patterns of secoiridoid and xanthone biosynthetic pathways during early development of Centaurium erythraea Rafn, as altered by ploidy level. in Industrial Crops and Products. 2022;186:115146.
doi:10.1016/j.indcrop.2022.115146 .

Filipović, Biljana, Skorić, Marijana, Gašić, Uroš, Dragićević, Milan, Božunović, Jelena, Matekalo, Dragana, Nestorović Živković, Jasmina, Banjanac, Tijana, Šiler, Branislav, Bohanec, Borut, Milutinović, Milica, Mišić, Danijela, "Spatial and temporal patterns of secoiridoid and xanthone biosynthetic pathways during early development of Centaurium erythraea Rafn, as altered by ploidy level" in Industrial Crops and Products, 186 (2022):115146,
https://doi.org/10.1016/j.indcrop.2022.115146 . .

TY  - CONF
AU  - Filipović, Biljana
AU  - Šiler, Branislav
AU  - Nestorović Živković, Jasmina
AU  - Banjanac, Tijana
AU  - Skorić, Marijana
AU  - Bohanec, Borut
AU  - Mišić, Danijela
PY  - 2018
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/4386
AB  - Polyploidy, or duplication of the entire genomes, is a widespread mechanism in plants which can lead to changes in cytological, biochemical, physiological and developmental characteristics, including conspicuous changes in morphology and metabolite production. In this study, we examined the impact of ploidy level on regenerative in vitro potential and biosynthetic capacity of C.  erythraea plants. Ten genotypes of diploid (2x) and autotetraploid (4x) centaury, with ploidy levels confirmed by flow cytometry, were randomly selected from in vitro pool of seedlings maintained on hormone-free half-strength MS medium (½ MS). Root explants were excised either from basal,  middle or apical root zone of 3 month-old plants. In vitro morphogenesis was induced on ½ MS medium. The frequency of shoot regeneration and the average number of shoots per root explant were recorded after 5 weeks in culture. Regeneration frequency was genotype dependent,  but not affected by explant ploidy level. Regeneration efficiency declined from the apical to the basal root segment. Regenerated shoots were transferred onto ½ MS medium for rooting. Iridoid glycosides (loganic acid, loganin), secoiridoid glycosides (secologanin, sweroside, swertiamarin,  gentiopicrin) and xantones (decussatin, eustomin, methylbellidifolin, desmethyleustomin) were quantified in shoots and roots of two month-old plants using targeted UHPLC-qqqMS analysis. Interestingly, diploid plants showed higher biosynthetic capacity for the production of secoiridoids and xanthones in both shoots and roots. Furthermore, two month-old shoots of 2x genotypes produced higher shoot and root biomass than 4x plants. Results highlight a great potential of diploid C. erythraea genotypes for biotechnology-based sustainable production of secoiridoids.
PB  - Belgrade: Serbian Plant Physiology Society
C3  - 3rd International Conference on Plant Biology and 22nd SPPS Meeting , 9-12 June 2018, Belgrade
T1  - Diploid and tetraploid Centaurium erythraea Rafn: a comparative study of regenerative in vitro potential and biosynthetic capacity
SP  - 109
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_4386
ER  - 

@conference{
author = "Filipović, Biljana and Šiler, Branislav and Nestorović Živković, Jasmina and Banjanac, Tijana and Skorić, Marijana and Bohanec, Borut and Mišić, Danijela",
year = "2018",
abstract = "Polyploidy, or duplication of the entire genomes, is a widespread mechanism in plants which can lead to changes in cytological, biochemical, physiological and developmental characteristics, including conspicuous changes in morphology and metabolite production. In this study, we examined the impact of ploidy level on regenerative in vitro potential and biosynthetic capacity of C.  erythraea plants. Ten genotypes of diploid (2x) and autotetraploid (4x) centaury, with ploidy levels confirmed by flow cytometry, were randomly selected from in vitro pool of seedlings maintained on hormone-free half-strength MS medium (½ MS). Root explants were excised either from basal,  middle or apical root zone of 3 month-old plants. In vitro morphogenesis was induced on ½ MS medium. The frequency of shoot regeneration and the average number of shoots per root explant were recorded after 5 weeks in culture. Regeneration frequency was genotype dependent,  but not affected by explant ploidy level. Regeneration efficiency declined from the apical to the basal root segment. Regenerated shoots were transferred onto ½ MS medium for rooting. Iridoid glycosides (loganic acid, loganin), secoiridoid glycosides (secologanin, sweroside, swertiamarin,  gentiopicrin) and xantones (decussatin, eustomin, methylbellidifolin, desmethyleustomin) were quantified in shoots and roots of two month-old plants using targeted UHPLC-qqqMS analysis. Interestingly, diploid plants showed higher biosynthetic capacity for the production of secoiridoids and xanthones in both shoots and roots. Furthermore, two month-old shoots of 2x genotypes produced higher shoot and root biomass than 4x plants. Results highlight a great potential of diploid C. erythraea genotypes for biotechnology-based sustainable production of secoiridoids.",
publisher = "Belgrade: Serbian Plant Physiology Society",
journal = "3rd International Conference on Plant Biology and 22nd SPPS Meeting , 9-12 June 2018, Belgrade",
title = "Diploid and tetraploid Centaurium erythraea Rafn: a comparative study of regenerative in vitro potential and biosynthetic capacity",
pages = "109",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_4386"
}

Filipović, B., Šiler, B., Nestorović Živković, J., Banjanac, T., Skorić, M., Bohanec, B.,& Mišić, D.. (2018). Diploid and tetraploid Centaurium erythraea Rafn: a comparative study of regenerative in vitro potential and biosynthetic capacity. in 3rd International Conference on Plant Biology and 22nd SPPS Meeting , 9-12 June 2018, Belgrade
Belgrade: Serbian Plant Physiology Society., 109.
https://hdl.handle.net/21.15107/rcub_ibiss_4386

Filipović B, Šiler B, Nestorović Živković J, Banjanac T, Skorić M, Bohanec B, Mišić D. Diploid and tetraploid Centaurium erythraea Rafn: a comparative study of regenerative in vitro potential and biosynthetic capacity. in 3rd International Conference on Plant Biology and 22nd SPPS Meeting , 9-12 June 2018, Belgrade. 2018;:109.
https://hdl.handle.net/21.15107/rcub_ibiss_4386 .

Filipović, Biljana, Šiler, Branislav, Nestorović Živković, Jasmina, Banjanac, Tijana, Skorić, Marijana, Bohanec, Borut, Mišić, Danijela, "Diploid and tetraploid Centaurium erythraea Rafn: a comparative study of regenerative in vitro potential and biosynthetic capacity" in 3rd International Conference on Plant Biology and 22nd SPPS Meeting , 9-12 June 2018, Belgrade (2018):109,
https://hdl.handle.net/21.15107/rcub_ibiss_4386 .

TY  - JOUR
AU  - Stanišić, Mariana
AU  - Raspor, Martin
AU  - Ninković, Slavica
AU  - Milošević, Snežana
AU  - Ćalić, Dušica
AU  - Bohanec, Borut
AU  - Trifunović-Momčilov, Milana
AU  - Petrić, Marija
AU  - Subotić, Angelina
AU  - Jevremović, Slađana
PY  - 2015
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/2094
UR  - https://s100.copyright.com/AppDispatchServlet?publisherName=ELS&contentID=S025462991400194X&orderBeanReset=true
AB  - Efficient protocols, safe from somaclonal variation, were developed for
   regeneration of Iris sibirica plants via organogenesis and somatic
   embryogenesis from leaf-base explants cultivated on Murashige and Skoog
   media supplemented with thidiazuron (TDZ, 1.0 mg/l) or
   2,4-dichlorophenoxyacetic acid (2,4-D, 1.0 mg/l). The morphogenic
   response and callus formation efficiency differed significantly between
   2,4-D (80.9\%) and TDZ (67\%) morphogenesis induction treatments. TDZ
   induced only organogenic calli, while calli obtained with 2,4-D were
   composed of three types differing in color and consistency: white,
   friable - embryogenic calli (4.5\%, 3.8 mg/explant), green, compact -
   organogenic calli (12.4\%, 48.4 mg/explants) and yellow -
   non-regenerative calli (773\%, 254.4 mg/explant). The cultivation of
   embryogenic calli on medium with 2,4-0 and Kinetin resulted in further
   development of somatic embryos (54 embryos/g of calli) which germinated
   with a frequency of 62\% after being transferred to a medium without
   plant growth regulators. Stable shoot cultures were established by
   transferring organogenic calli with shoot primordia to media with 0.1
   mg/l alpha-naphthaleneacetic acid (NAA) and 1.0 mg/l 6-benzylaminopurine
   (BA), while further cultivation on media of the same composition (TDZ or
   2,4-D) resulted in the reduced growth and rhizogenesis, respectively.
   The TDZ induction treatment resulted in higher number of shoots per
   explant (7.9) than the 2,4-0 treatment (43). After successful rooting
   and ex vitro acclimatization, plants were grown in the field and
   flowered to seed production. Flow cytometry, chromosome counting and
   random amplification of polymorphic DNA (RAPD) analysis indicated no
   evidence of genetic variation in plants regenerated via somatic
   embryogenesis or organogenesis. The results suggest that established
   protocols are safe for use in genetic transformation procedures or
   large-scale production of true-to-type I. sibirica plants. (C) 2014
   SAAB. Published by Elsevier B.V. All rights reserved.
T2  - South African Journal of Botany
T1  - Clonal fidelity of Iris sibirica plants regenerated by somatic
 embryogenesis and organogenesis in leaf-base culture - RAPD and flow
 cytometer analyses
VL  - 96
DO  - 10.1016/j.sajb.2014.10.014
SP  - 42
EP  - 52
ER  - 

@article{
author = "Stanišić, Mariana and Raspor, Martin and Ninković, Slavica and Milošević, Snežana and Ćalić, Dušica and Bohanec, Borut and Trifunović-Momčilov, Milana and Petrić, Marija and Subotić, Angelina and Jevremović, Slađana",
year = "2015",
abstract = "Efficient protocols, safe from somaclonal variation, were developed for
   regeneration of Iris sibirica plants via organogenesis and somatic
   embryogenesis from leaf-base explants cultivated on Murashige and Skoog
   media supplemented with thidiazuron (TDZ, 1.0 mg/l) or
   2,4-dichlorophenoxyacetic acid (2,4-D, 1.0 mg/l). The morphogenic
   response and callus formation efficiency differed significantly between
   2,4-D (80.9\%) and TDZ (67\%) morphogenesis induction treatments. TDZ
   induced only organogenic calli, while calli obtained with 2,4-D were
   composed of three types differing in color and consistency: white,
   friable - embryogenic calli (4.5\%, 3.8 mg/explant), green, compact -
   organogenic calli (12.4\%, 48.4 mg/explants) and yellow -
   non-regenerative calli (773\%, 254.4 mg/explant). The cultivation of
   embryogenic calli on medium with 2,4-0 and Kinetin resulted in further
   development of somatic embryos (54 embryos/g of calli) which germinated
   with a frequency of 62\% after being transferred to a medium without
   plant growth regulators. Stable shoot cultures were established by
   transferring organogenic calli with shoot primordia to media with 0.1
   mg/l alpha-naphthaleneacetic acid (NAA) and 1.0 mg/l 6-benzylaminopurine
   (BA), while further cultivation on media of the same composition (TDZ or
   2,4-D) resulted in the reduced growth and rhizogenesis, respectively.
   The TDZ induction treatment resulted in higher number of shoots per
   explant (7.9) than the 2,4-0 treatment (43). After successful rooting
   and ex vitro acclimatization, plants were grown in the field and
   flowered to seed production. Flow cytometry, chromosome counting and
   random amplification of polymorphic DNA (RAPD) analysis indicated no
   evidence of genetic variation in plants regenerated via somatic
   embryogenesis or organogenesis. The results suggest that established
   protocols are safe for use in genetic transformation procedures or
   large-scale production of true-to-type I. sibirica plants. (C) 2014
   SAAB. Published by Elsevier B.V. All rights reserved.",
journal = "South African Journal of Botany",
title = "Clonal fidelity of Iris sibirica plants regenerated by somatic
 embryogenesis and organogenesis in leaf-base culture - RAPD and flow
 cytometer analyses",
volume = "96",
doi = "10.1016/j.sajb.2014.10.014",
pages = "42-52"
}

Stanišić, M., Raspor, M., Ninković, S., Milošević, S., Ćalić, D., Bohanec, B., Trifunović-Momčilov, M., Petrić, M., Subotić, A.,& Jevremović, S.. (2015). Clonal fidelity of Iris sibirica plants regenerated by somatic
 embryogenesis and organogenesis in leaf-base culture - RAPD and flow
 cytometer analyses. in South African Journal of Botany, 96, 42-52.
https://doi.org/10.1016/j.sajb.2014.10.014

Stanišić M, Raspor M, Ninković S, Milošević S, Ćalić D, Bohanec B, Trifunović-Momčilov M, Petrić M, Subotić A, Jevremović S. Clonal fidelity of Iris sibirica plants regenerated by somatic
 embryogenesis and organogenesis in leaf-base culture - RAPD and flow
 cytometer analyses. in South African Journal of Botany. 2015;96:42-52.
doi:10.1016/j.sajb.2014.10.014 .

Stanišić, Mariana, Raspor, Martin, Ninković, Slavica, Milošević, Snežana, Ćalić, Dušica, Bohanec, Borut, Trifunović-Momčilov, Milana, Petrić, Marija, Subotić, Angelina, Jevremović, Slađana, "Clonal fidelity of Iris sibirica plants regenerated by somatic
 embryogenesis and organogenesis in leaf-base culture - RAPD and flow
 cytometer analyses" in South African Journal of Botany, 96 (2015):42-52,
https://doi.org/10.1016/j.sajb.2014.10.014 . .

TY  - JOUR
AU  - Ćalić, Dušica
AU  - Zdravković-Korać, Snežana
AU  - Bohanec, Borut
AU  - Radojević, Ljiljana
AU  - Vinterhalter, Branka
AU  - Stevović, Svetlana
AU  - Cingel, Aleksandar
AU  - Savić, Jelena
PY  - 2010
UR  - https://www.ajol.info/index.php/ajb/article/view/82495
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6169
AB  - The influence of activated charcoal (AC), abscisic acid (ABA) and polyethylene glycol (PEG) on the maturation and conversion of horse chestnut (Aesculus hippocastanum L.) androgenic embryos were tested. Androgenic embryos originating from microspores and anther culture were maturated over 90 days. Androgenic embryos on media containing PEG (50 g/l), in combination with AC (1 g/l) showed a rapid development of embryos in the cotyledonary stage and lowered percentage of abnormal structures. The best results of androgenic microspore embryo germination were observed on media supplemented with AC alone (99%) and in combination with PEG (100%). Also, the greatest number of androgenic microspore plants (18%) and androgenic anther plants (12%) were formed on media enriched with 1 % AC. Lowest germination percentages of 37 and 39% in microspore culture and 33 and 38% in anther culture were obtained on maturation media with ABA 20 mg/l alone and in combination with AC 1g/l. Flow cytometric analysis showed that most of the androgenic embryos were haploid, corresponding to their microspore origin, while half of these became diploid after maturation for 90 days. All regenerants originating from microspore culture were haploid immediately after germination, but only 10% embryos retained haploidity after 3 years subculturing, while 10.5% were diploid, 73.5% tetraploid and 6% octaploid on hormone-free medium. Unlike those from anther culture, after 3 years of subculturing on hormone-free medium, there were no haploid regenerant from anther culture, while 8.5% were diploid, 81% tetraploid and 10.5% octaploid.
PB  - Nairobi: Academic Journals
T2  - African Journal of Biotechnology
T1  - Effect of activated charcoal, abscisic acid and polyethylene glycol on maturation, germination and conversion of Aesculus hippocastanum androgenic embryos
IS  - 25
VL  - 9
SP  - 3786
EP  - 3793
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_6169
ER  - 

@article{
author = "Ćalić, Dušica and Zdravković-Korać, Snežana and Bohanec, Borut and Radojević, Ljiljana and Vinterhalter, Branka and Stevović, Svetlana and Cingel, Aleksandar and Savić, Jelena",
year = "2010",
abstract = "The influence of activated charcoal (AC), abscisic acid (ABA) and polyethylene glycol (PEG) on the maturation and conversion of horse chestnut (Aesculus hippocastanum L.) androgenic embryos were tested. Androgenic embryos originating from microspores and anther culture were maturated over 90 days. Androgenic embryos on media containing PEG (50 g/l), in combination with AC (1 g/l) showed a rapid development of embryos in the cotyledonary stage and lowered percentage of abnormal structures. The best results of androgenic microspore embryo germination were observed on media supplemented with AC alone (99%) and in combination with PEG (100%). Also, the greatest number of androgenic microspore plants (18%) and androgenic anther plants (12%) were formed on media enriched with 1 % AC. Lowest germination percentages of 37 and 39% in microspore culture and 33 and 38% in anther culture were obtained on maturation media with ABA 20 mg/l alone and in combination with AC 1g/l. Flow cytometric analysis showed that most of the androgenic embryos were haploid, corresponding to their microspore origin, while half of these became diploid after maturation for 90 days. All regenerants originating from microspore culture were haploid immediately after germination, but only 10% embryos retained haploidity after 3 years subculturing, while 10.5% were diploid, 73.5% tetraploid and 6% octaploid on hormone-free medium. Unlike those from anther culture, after 3 years of subculturing on hormone-free medium, there were no haploid regenerant from anther culture, while 8.5% were diploid, 81% tetraploid and 10.5% octaploid.",
publisher = "Nairobi: Academic Journals",
journal = "African Journal of Biotechnology",
title = "Effect of activated charcoal, abscisic acid and polyethylene glycol on maturation, germination and conversion of Aesculus hippocastanum androgenic embryos",
number = "25",
volume = "9",
pages = "3786-3793",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_6169"
}

Ćalić, D., Zdravković-Korać, S., Bohanec, B., Radojević, L., Vinterhalter, B., Stevović, S., Cingel, A.,& Savić, J.. (2010). Effect of activated charcoal, abscisic acid and polyethylene glycol on maturation, germination and conversion of Aesculus hippocastanum androgenic embryos. in African Journal of Biotechnology
Nairobi: Academic Journals., 9(25), 3786-3793.
https://hdl.handle.net/21.15107/rcub_ibiss_6169

Ćalić D, Zdravković-Korać S, Bohanec B, Radojević L, Vinterhalter B, Stevović S, Cingel A, Savić J. Effect of activated charcoal, abscisic acid and polyethylene glycol on maturation, germination and conversion of Aesculus hippocastanum androgenic embryos. in African Journal of Biotechnology. 2010;9(25):3786-3793.
https://hdl.handle.net/21.15107/rcub_ibiss_6169 .

Ćalić, Dušica, Zdravković-Korać, Snežana, Bohanec, Borut, Radojević, Ljiljana, Vinterhalter, Branka, Stevović, Svetlana, Cingel, Aleksandar, Savić, Jelena, "Effect of activated charcoal, abscisic acid and polyethylene glycol on maturation, germination and conversion of Aesculus hippocastanum androgenic embryos" in African Journal of Biotechnology, 9, no. 25 (2010):3786-3793,
https://hdl.handle.net/21.15107/rcub_ibiss_6169 .