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In ornamental geophytes, conventional vegetative propagation is not economically feasible due to very slow development and ineffective methods. It can take several years until a new plant is formed and commercial profitability is achieved. Therefore, micropropagation techniques have been developed to increase the multiplication rate and thus shorten the multiplication and regeneration period. The majority of these techniques rely on the formation of new bulbs and their sprouting. Dormancy is one of the main limiting factors to speed up multiplication in vitro. Bulbous species have a period of bulb dormancy which enables them to survive unfavorable natural conditions. Bulbs grown in vitro also exhibit dormancy, which has to be overcome in order to allow sprouting of bulbs in the next vegetation period. During the period of dormancy, numerous physiological processes occur, many of which have not been elucidated yet. Understanding the process of dormancy will allow us to speed up and improve breeding of geophytes and thereby achieve economic profitability, which is very important for horticulture. This review focuses on recent findings in the area of bulb dormancy initiation and release in fritillaries, with particular emphasis on the effect of plant growth regulators and low-temperature pretreatment on dormancy release in relation to induction of antioxidative enzymes’ activity in vitro.
Marija Marković; Milana Trifunović Momčilov; Branka Uzelac; Slađana Jevremović; Angelina Subotić. Bulb Dormancy In Vitro—Fritillaria meleagris: Initiation, Release and Physiological Parameters. Plants 2021, 10, 902 .
AMA StyleMarija Marković, Milana Trifunović Momčilov, Branka Uzelac, Slađana Jevremović, Angelina Subotić. Bulb Dormancy In Vitro—Fritillaria meleagris: Initiation, Release and Physiological Parameters. Plants. 2021; 10 (5):902.
Chicago/Turabian StyleMarija Marković; Milana Trifunović Momčilov; Branka Uzelac; Slađana Jevremović; Angelina Subotić. 2021. "Bulb Dormancy In Vitro—Fritillaria meleagris: Initiation, Release and Physiological Parameters." Plants 10, no. 5: 902.
The bulb is the main propagation organ of snake’s head fritillary (Fritillaria meleagris L.), a horticulturally attractive and rare geophyte plant species. In this study, we investigated the effect of soaking bulbs in GA3 solution (1, 2, and 3 mg L−1) combined with low-temperature treatment (7 °C) on breaking the dormancy of in vitro bulbs. Sugar status (total soluble sugars, glucose, and fructose content) was analyzed in different parts of the sprouted bulbs. The results showed that the soluble sugar concentration was highest in bulbs soaked in GA3. The main sugar in fritillary bulbs was glucose, while fructose content was much lower. Glucose concentration dramatically increased after bulb chilling (7 °C), and its accumulation was predominantly detected in the lower sprout portion during the first weeks of sprouting. Sugar concentration was significantly lower in nonchilled bulbs, which indicates the importance of low temperature in bulb development and sprouting.
Marija Marković; Milana Trifunović Momčilov; Branka Uzelac; Olga Radulović; Snežana Milošević; Slađana Jevremović; Angelina Subotić. Breaking the Dormancy of Snake’s Head Fritillary (Fritillaria meleagris L.) In Vitro Bulbs—Part 2: Effect of GA3 Soaking and Chilling on Sugar Status in Sprouted Bulbs. Plants 2020, 9, 1573 .
AMA StyleMarija Marković, Milana Trifunović Momčilov, Branka Uzelac, Olga Radulović, Snežana Milošević, Slađana Jevremović, Angelina Subotić. Breaking the Dormancy of Snake’s Head Fritillary (Fritillaria meleagris L.) In Vitro Bulbs—Part 2: Effect of GA3 Soaking and Chilling on Sugar Status in Sprouted Bulbs. Plants. 2020; 9 (11):1573.
Chicago/Turabian StyleMarija Marković; Milana Trifunović Momčilov; Branka Uzelac; Olga Radulović; Snežana Milošević; Slađana Jevremović; Angelina Subotić. 2020. "Breaking the Dormancy of Snake’s Head Fritillary (Fritillaria meleagris L.) In Vitro Bulbs—Part 2: Effect of GA3 Soaking and Chilling on Sugar Status in Sprouted Bulbs." Plants 9, no. 11: 1573.
Bulbs are the main vegetative reproductive organs of Fritillaria meleagris L. In nature, as well as in vitro, they become dormant and require low temperatures for further growth during the next vegetative period. In the present study, using 10 μM of gibberellic acid (GA3), or gibberellin biosynthesis (GA) inhibitors—ancymidol (A) and paclobutrazol (P)—the dynamic changes in soluble sugars, fructose and glucose content, fresh weight and sprouting capacity were investigated. F. meleagris bulbs were cultured on medium with GA3 and GA inhibitors for 1, 2 and 5 weeks at two different temperatures (24 and 7 °C). GA3 improved bulb fresh weight, as well as sprouting percentage at both tested temperatures, compared to the control. The highest fresh weight increase (57.7%) and sprouting rate (29.02%) were achieved when bulbs were grown at 24 °C for 5 weeks. In addition, soluble sugar content was the highest in bulbs grown for 5 weeks on medium supplemented with GA3. The main sugar in fritillary bulbs was glucose, while fructose content was lower. The sensitivity of bulbs to GA inhibitors differed and significantly affected sugar content in bulbs. To our knowledge, this is the first study of the sugar composition in F. meleagris bulbs during breaking of the bulb’s dormancy and its sprouting.
Marija Marković; Milana Trifunović Momčilov; Branka Uzelac; Aleksandar Cingel; Snežana Milošević; Slađana Jevremović; Angelina Subotić. Breaking the Dormancy of Snake’s Head Fritillary (Fritillaria meleagris L.) In Vitro Bulbs—Part 1: Effect of GA3, GA Inhibitors and Temperature on Fresh Weight, Sprouting and Sugar Content. Plants 2020, 9, 1449 .
AMA StyleMarija Marković, Milana Trifunović Momčilov, Branka Uzelac, Aleksandar Cingel, Snežana Milošević, Slađana Jevremović, Angelina Subotić. Breaking the Dormancy of Snake’s Head Fritillary (Fritillaria meleagris L.) In Vitro Bulbs—Part 1: Effect of GA3, GA Inhibitors and Temperature on Fresh Weight, Sprouting and Sugar Content. Plants. 2020; 9 (11):1449.
Chicago/Turabian StyleMarija Marković; Milana Trifunović Momčilov; Branka Uzelac; Aleksandar Cingel; Snežana Milošević; Slađana Jevremović; Angelina Subotić. 2020. "Breaking the Dormancy of Snake’s Head Fritillary (Fritillaria meleagris L.) In Vitro Bulbs—Part 1: Effect of GA3, GA Inhibitors and Temperature on Fresh Weight, Sprouting and Sugar Content." Plants 9, no. 11: 1449.
The aim of this study was to develop a fast, reliable and true-to-type protocol for in vitro plant regeneration and long-term storage of horned pansy (Viola cornuta L). Seed germination over 60% was recorded after 12 weeks of growth at 10 °C or 4 °C. Calli formation and shoot induction were obtained in petiole and hypocotyl culture on half-strength MS mineral salts with full concentration of Na–FeEDTA and vitamins (½MS medium) with 2,4-dichlorophenoxyacetic acid (2,4-D, 0.1 mg/L) and 6-benzylaminopurine (BAP, 2.0 mg/L) and leaf culture on ½MS medium with thidiazuron (TDZ,1.0 mg/L). The highest frequency of adventitious shoot induction (50%) with six shoots/explant was achieved in hypocotyl culture from top hypocotyl segments, close to epicotyl which was grown 8 weeks at 16 h light/8 h dark photoperiod. Subsequent shoot multiplication was achieved on ½MS medium with α-naphthaleneacetic acid (NAA, 0.1 or 0.5 mg/L) and BAP (1.0 mg/L). Rooting of shoots was obtained on ½MS medium with low concentration (0.1 mg/L) of auxins: indole-3-acetic acid (IAA), indole-3-butyric acid (IBA) or NAA, or without growth regulators. In vitro-derived plantlets were acclimatized under greenhouse conditions. All plants developed normally, bloomed and set seeds. Shoot tips were cryopreserved succssefully using modified plant vitrification 3 (PVS3-based vitrification procedure). Cold acclimation for 2 weeks significantly improved shoot regrowth (64%) after thawing in comparison to non-acclimated shoots (39%). Clonal fidelity of regenerated plantlets at ploidy level was confirmed by chromosome counting. The presented protocol can be useful for mass propagation, genetic transformation studies and long-term storage of valuable Viola spp.
Milena Trajković; Dragana Antonic; Aleksandar Cingel; Nabil Ghalawenji; Angelina Subotić; Slađana Jevremović. Advancement in protocol for in vitro seed germination, plant regeneration and cryopreservation of Viola cornuta. 3 Biotech 2019, 9, 17 .
AMA StyleMilena Trajković, Dragana Antonic, Aleksandar Cingel, Nabil Ghalawenji, Angelina Subotić, Slađana Jevremović. Advancement in protocol for in vitro seed germination, plant regeneration and cryopreservation of Viola cornuta. 3 Biotech. 2019; 9 (1):17.
Chicago/Turabian StyleMilena Trajković; Dragana Antonic; Aleksandar Cingel; Nabil Ghalawenji; Angelina Subotić; Slađana Jevremović. 2019. "Advancement in protocol for in vitro seed germination, plant regeneration and cryopreservation of Viola cornuta." 3 Biotech 9, no. 1: 17.
Morphogenesis in vitro is a complex and still poorly defined process. We investigated esterase and peroxidase isoforms detected in bulb scale, during Fritillaria meleagris morphogenesis. Bulbs were grown either at 4 °C or on a medium with an increased concentration of sucrose (4.5%) for 30 days. After these pre-treatments, the bulb scales were further grown on nutrient media that contained different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) and kinetin (KIN) or thidiazuron (TDZ). Regeneration of somatic embryos and bulblets occurred at the same explant. The highest numbers of somatic embryos and bulblets were regenerated on the medium containing 2,4-D and KIN (1mg/L each), while morphogenesis was most successful at a TDZ concentration between 0.5 and 1mg/L. Monitoring of esterases and peroxidases was performed by growing bulb scales on a medium enriched with 2,4-D and KIN or TDZ (1mg/L), and the number and activity of isoforms were followed every 7 days for 4 weeks. In control explants, six isoforms of esterase were observed. Three isoforms of peroxidase were not detected in the control bulb scale, which has not begun its morphogenesis process.
Marija Petrić; Angelina Subotić; Sladjana Jevremović; Milana Trifunović-Momčilov; Vojin Tadić; Marica Grujić; Zoran Vujčić. Esterase and peroxidase isoforms in different stages of morphogenesis in Fritillaria meleagris L. in bulb-scale culture. Comptes Rendus. Biologies 2015, 338, 793 -802.
AMA StyleMarija Petrić, Angelina Subotić, Sladjana Jevremović, Milana Trifunović-Momčilov, Vojin Tadić, Marica Grujić, Zoran Vujčić. Esterase and peroxidase isoforms in different stages of morphogenesis in Fritillaria meleagris L. in bulb-scale culture. Comptes Rendus. Biologies. 2015; 338 (12):793-802.
Chicago/Turabian StyleMarija Petrić; Angelina Subotić; Sladjana Jevremović; Milana Trifunović-Momčilov; Vojin Tadić; Marica Grujić; Zoran Vujčić. 2015. "Esterase and peroxidase isoforms in different stages of morphogenesis in Fritillaria meleagris L. in bulb-scale culture." Comptes Rendus. Biologies 338, no. 12: 793-802.
Phytohormones are important regulators of numerous developmental and physiological processes in plants. Spontaneous morphogenesis of the common centaury (Centaurium erythraea Rafn.) is possible on nutrition medium without addition of any plant growth regulator depending solely on endogenous phytohormone levels. Thus, this plant species represents a very good model system for the investigation of numerous physiological processes under phytohormonal control in vitro. We analysed the total amount of endogenous cytokinins (CKs) including the contents of their individual groups in shoots and roots of C. erythraea plants grown in vitro. The total amount of endogenous CKs was 1.4 times higher in shoots than in roots. Inactive or weakly active N-glucosides found to predominate in both organs of centaury plants, whereas free bases and O-glucosides represented only a small portion of the total CK pool. Consequently, centaury roots showed higher IAA content as well as IAA/free CK base ratios compared to shoots. Centaury tissues also showed increased levels of “stress hormones”. In contrast to SA, considerably higher levels of ABA were found in centaury shoots than in roots. Our results could serve as a basis for understanding and elucidating spontaneous de novo shoot organogenesis and further plant regeneration of C. erythraea in vitro.
Milana Trifunović-Momčilov; Václav Motyka; Ivana Dragićević; Marija Petrić; Sladjana Jevremović; Jiří Malbeck; Josef Holík; Petre I. Dobrev; Angelina Subotić. Endogenous Phytohormones in Spontaneously Regenerated Centaurium erythraea Rafn. Plants Grown In Vitro. Journal of Plant Growth Regulation 2015, 35, 543 -552.
AMA StyleMilana Trifunović-Momčilov, Václav Motyka, Ivana Dragićević, Marija Petrić, Sladjana Jevremović, Jiří Malbeck, Josef Holík, Petre I. Dobrev, Angelina Subotić. Endogenous Phytohormones in Spontaneously Regenerated Centaurium erythraea Rafn. Plants Grown In Vitro. Journal of Plant Growth Regulation. 2015; 35 (2):543-552.
Chicago/Turabian StyleMilana Trifunović-Momčilov; Václav Motyka; Ivana Dragićević; Marija Petrić; Sladjana Jevremović; Jiří Malbeck; Josef Holík; Petre I. Dobrev; Angelina Subotić. 2015. "Endogenous Phytohormones in Spontaneously Regenerated Centaurium erythraea Rafn. Plants Grown In Vitro." Journal of Plant Growth Regulation 35, no. 2: 543-552.
Histochemical localization using the β-Glc Yariv reagent and immunolocalization with arabinogalactan protein (AGP) reactive antibodies (LM2, JIM13, JIM15, JIM16, MAC207) were performed during morphogenic induction in root cultures of Centaurium erythraea Rafn cultured on half-strength MS medium without plant growth regulators. The observations revealed that β-Glc Yariv reagent specifically bound to AGPs in cells of the root epidermis and central cylinder. Monoclonal antibodies recognizing AGPs were localized in epidermal cells and cells of the central cylinder (LM2 , JIM16), vascular tissue (JIM15), globular somatic embryos (LM2, MAC207), and de novo-formed meristematic centers in the root cortex (JIM16). The effect of β-Glc Yariv reagent was investigated after supplementation (0–75 μM) in the culture medium. The morphogenetic potential was increased at lower concentrations (15–25 μM) of treatment with β-Glc Yariv reagent but inhibited (40 %) at the highest concentration. These results implicate that AGPs play a significant role during the development of somatic embryos and adventitious shoots in root cultures of C. erythraea.
Milana Trifunović; Angelina Subotić; Marija Petrić; Sladjana Jevremović. The Role of Arabinogalactan Proteins in Morphogenesis of Centaurium erythraea Rafn In Vitro. The Gentianaceae - Volume 2: Biotechnology and Applications 2015, 113 -138.
AMA StyleMilana Trifunović, Angelina Subotić, Marija Petrić, Sladjana Jevremović. The Role of Arabinogalactan Proteins in Morphogenesis of Centaurium erythraea Rafn In Vitro. The Gentianaceae - Volume 2: Biotechnology and Applications. 2015; ():113-138.
Chicago/Turabian StyleMilana Trifunović; Angelina Subotić; Marija Petrić; Sladjana Jevremović. 2015. "The Role of Arabinogalactan Proteins in Morphogenesis of Centaurium erythraea Rafn In Vitro." The Gentianaceae - Volume 2: Biotechnology and Applications , no. : 113-138.
Arabinogalactan proteins (AGPs) are cell wall hydroxyproline-rich glycoproteins with diverse functions in plant growth and development. In this work the role of AGPs was comparatively studied in four developmental pathways that can be induced from centaury leaf explants—indirect somatic embryogenesis (ISE), indirect and direct shoot development (ISD and DSD) and direct root development (DRD). The addition of β-d-glucosyl Yariv (βGlcY) reagent, a synthetic phenylglycoside that specifically crosslinks AGPs, to the growth medium reduced the number of somatic embryos and adventitious buds formed per explant. The morphogenetic paths can be arranged in order of increasing sensitivity to βGlcY as: DRD (insensitive) CeAGP1–CeAGP4, of which CeAGP1, 2 and 4 code for fasciclin-like AGPs. CeAGP3 is an AG peptide with conserved DUF1070 domain. Expression pattern of these genes indicated specific involvement of CeAGP1 in ISE and unspecific involvement of CeAGP3 in morphogenesis.
Ana D. Simonović; Biljana K. Filipović; Milana M. Trifunović; Saša N. Malkov; Vedrana P. Milinkovic; Slađana B. Jevremović; Angelina R. Subotić. Plant regeneration in leaf culture of Centaurium erythraea Rafn. Part 2: the role of arabinogalactan proteins. Plant Cell, Tissue and Organ Culture (PCTOC) 2015, 121, 721 -739.
AMA StyleAna D. Simonović, Biljana K. Filipović, Milana M. Trifunović, Saša N. Malkov, Vedrana P. Milinkovic, Slađana B. Jevremović, Angelina R. Subotić. Plant regeneration in leaf culture of Centaurium erythraea Rafn. Part 2: the role of arabinogalactan proteins. Plant Cell, Tissue and Organ Culture (PCTOC). 2015; 121 (3):721-739.
Chicago/Turabian StyleAna D. Simonović; Biljana K. Filipović; Milana M. Trifunović; Saša N. Malkov; Vedrana P. Milinkovic; Slađana B. Jevremović; Angelina R. Subotić. 2015. "Plant regeneration in leaf culture of Centaurium erythraea Rafn. Part 2: the role of arabinogalactan proteins." Plant Cell, Tissue and Organ Culture (PCTOC) 121, no. 3: 721-739.
Centaurium erythraea Rafn. is a medicinal plant rich in secoiridoids and xanthones and used for gastrointestinal disorders, fever, anemia and many other conditions. C. erythraea is characterized with extraordinary developmental plasticity and manageability in vitro; thus we propose it as an excellent experimental model system for studies in developmental biology. Hereby we describe regeneration of centaury from leaf explants that can proceed via somatic embryogenesis or organogenesis on inductive media containing 2,4-D and CPPU. In the absence of growth regulators, shoots and roots appeared without callusing, on light or in darkness, respectively. Indirect somatic embryogenesis was induced in the presence of growth regulators occurring both on light and in darkness. Light was obligatory for indirect shoot development, where adventitious buds formed simultaneously with somatic embryos. Dynamic changes of antioxidative activities of superoxide dismutase (SOD), catalase (CAT) and peroxidase (POX) in response to morphogenetic changes were followed during developmental pathways in vitro. Wounding of centaury leaves immediately induced all SOD and CAT isoforms but caused a decrease in POX activity. In control leaves and leaf explants, three Cu/Zn-SOD activities were detected, which gradually decreased on inductive treatments on light, but remained unchanged during growth in darkness. Morphogenetic paths on all hormonal and light treatments where characterized with dynamic changes of CAT activity (comprised of three major CAT isoforms), but generally CAT was reduced during morphogenesis induction. POX activity was strongly induced during morphogenesis in all treatments.
Biljana K. Filipović; Ana D. Simonović; Milana M. Trifunović; Slavica S. Dmitrović; Jelena M. Savic; Slađana B. Jevremović; Angelina R. Subotić. Plant regeneration in leaf culture of Centaurium erythraea Rafn. Part 1: The role of antioxidant enzymes. Plant Cell, Tissue and Organ Culture (PCTOC) 2015, 121, 703 -719.
AMA StyleBiljana K. Filipović, Ana D. Simonović, Milana M. Trifunović, Slavica S. Dmitrović, Jelena M. Savic, Slađana B. Jevremović, Angelina R. Subotić. Plant regeneration in leaf culture of Centaurium erythraea Rafn. Part 1: The role of antioxidant enzymes. Plant Cell, Tissue and Organ Culture (PCTOC). 2015; 121 (3):703-719.
Chicago/Turabian StyleBiljana K. Filipović; Ana D. Simonović; Milana M. Trifunović; Slavica S. Dmitrović; Jelena M. Savic; Slađana B. Jevremović; Angelina R. Subotić. 2015. "Plant regeneration in leaf culture of Centaurium erythraea Rafn. Part 1: The role of antioxidant enzymes." Plant Cell, Tissue and Organ Culture (PCTOC) 121, no. 3: 703-719.
Mariana Stanišić; M. Raspor; Slavica Ninkovic; Snežana Milošević; Dusica Calic; B. Bohanec; M. Trifunović; M. Petric; Angelina Subotić; Sladjana Jevremović. Clonal fidelity of Iris sibirica plants regenerated by somatic embryogenesis and organogenesis in leaf-base culture — RAPD and flow cytometer analyses. South African Journal of Botany 2015, 96, 42 -52.
AMA StyleMariana Stanišić, M. Raspor, Slavica Ninkovic, Snežana Milošević, Dusica Calic, B. Bohanec, M. Trifunović, M. Petric, Angelina Subotić, Sladjana Jevremović. Clonal fidelity of Iris sibirica plants regenerated by somatic embryogenesis and organogenesis in leaf-base culture — RAPD and flow cytometer analyses. South African Journal of Botany. 2015; 96 ():42-52.
Chicago/Turabian StyleMariana Stanišić; M. Raspor; Slavica Ninkovic; Snežana Milošević; Dusica Calic; B. Bohanec; M. Trifunović; M. Petric; Angelina Subotić; Sladjana Jevremović. 2015. "Clonal fidelity of Iris sibirica plants regenerated by somatic embryogenesis and organogenesis in leaf-base culture — RAPD and flow cytometer analyses." South African Journal of Botany 96, no. : 42-52.
The plant hormones cytokinins (CKs) regulate a number of physiological processes. Their homeostasis is controlled by the rate of de novo synthesis and the rate of catabolism. The aim of this work was to analyze the content of total as well as individual groups of endogenous CKs in AtCKX1 and AtCKX2-overexpressing centaury (Centaurium erythraea Rafn.) plants grown in vitro. Transgenic CKX plants represent a suitable model system for studying physiological and morphological processes controlled by CKs. In this work we clearly demonstrate a significant effect of AtCKX transgenes on CK metabolism in transgenic centaury plants. However, shoots and roots of only one AtCKX1 line and three AtCKX2 lines with a significant reduction of bioactive CKs were obtained. We also show that changes in the CKs metabolism considerably affected endogenous indole-3-acetic acid (IAA) levels in plant tissues. All analyzed transgenic AtCKX centaury lines exhibited decreased amount of endogenous IAA in shoots as well as in roots. Consequently, the IAA/bioactive CK forms ratios showed a significant variation in the shoots and roots of all analyzed AtCKX centaury transformants.
Milana Trifunović; Václav Motyka; Aleksandar Cingel; Angelina Subotic; Slađana Jevremović; Marija Petric; Josef Holík; Jiří Malbeck; Petre I. Dobrev; Ivana Č. Dragićević; Sladjana Jevremović. Changes in cytokinin content and altered cytokinin homeostasis in AtCKX1 and AtCKX2-overexpressing centaury (Centaurium erythraea Rafn.) plants grown in vitro. Plant Cell, Tissue and Organ Culture (PCTOC) 2014, 120, 767 -777.
AMA StyleMilana Trifunović, Václav Motyka, Aleksandar Cingel, Angelina Subotic, Slađana Jevremović, Marija Petric, Josef Holík, Jiří Malbeck, Petre I. Dobrev, Ivana Č. Dragićević, Sladjana Jevremović. Changes in cytokinin content and altered cytokinin homeostasis in AtCKX1 and AtCKX2-overexpressing centaury (Centaurium erythraea Rafn.) plants grown in vitro. Plant Cell, Tissue and Organ Culture (PCTOC). 2014; 120 (2):767-777.
Chicago/Turabian StyleMilana Trifunović; Václav Motyka; Aleksandar Cingel; Angelina Subotic; Slađana Jevremović; Marija Petric; Josef Holík; Jiří Malbeck; Petre I. Dobrev; Ivana Č. Dragićević; Sladjana Jevremović. 2014. "Changes in cytokinin content and altered cytokinin homeostasis in AtCKX1 and AtCKX2-overexpressing centaury (Centaurium erythraea Rafn.) plants grown in vitro." Plant Cell, Tissue and Organ Culture (PCTOC) 120, no. 2: 767-777.
Genetic modulation of the carotenogenesis in I. germanica 'Fire Bride' by ectopic expression of a crtB gene causes several flower parts to develop novel orange and pink colors. Flower color in tall bearded irises (Iris germanica L.) is determined by two distinct biochemical pathways; the carotenoid pathway, which imparts yellow, orange and pink hues and the anthocyanin pathway, which produces blue, violet and maroon flowers. Red-flowered I. germanica do not exist in nature and conventional breeding methods have thus far failed to produce them. With a goal of developing iris cultivars with red flowers, we transformed a pink iris I. germanica, 'Fire Bride', with a bacterial phytoene synthase gene (crtB) from Pantoea agglomerans under the control of the promoter region of a gene for capsanthin-capsorubin synthase from Lilium lancifolium (Llccs). This approach aimed to increase the flux of metabolites into the carotenoid biosynthetic pathway and lead to elevated levels of lycopene and darker pink or red flowers. Iris callus tissue ectopically expressing the crtB gene exhibited a color change from yellow to pink-orange and red, due to accumulation of lycopene. Transgenic iris plants, regenerated from the crtB-transgenic calli, showed prominent color changes in the ovaries (green to orange), flower stalk (green to orange), and anthers (white to pink), while the standards and falls showed no significant differences in color when compared to control plants. HPLC and UHPLC analysis confirmed that the color changes were primarily due to the accumulation of lycopene. In this study, we showed that ectopic expression of a crtB can be used to successfully alter the color of certain flower parts in I. germanica 'Fire Bride' and produce new flower traits.
Zoran Jeknić; Stevan Jeknić; Slađana Jevremović; Angelina Subotic; Tony H. H. Chen; Sladjana Jevremović. Alteration of flower color in Iris germanica L. ‘Fire Bride’ through ectopic expression of phytoene synthase gene (crtB) from Pantoea agglomerans. Plant Cell Reports 2014, 33, 1307 -1321.
AMA StyleZoran Jeknić, Stevan Jeknić, Slađana Jevremović, Angelina Subotic, Tony H. H. Chen, Sladjana Jevremović. Alteration of flower color in Iris germanica L. ‘Fire Bride’ through ectopic expression of phytoene synthase gene (crtB) from Pantoea agglomerans. Plant Cell Reports. 2014; 33 (8):1307-1321.
Chicago/Turabian StyleZoran Jeknić; Stevan Jeknić; Slađana Jevremović; Angelina Subotic; Tony H. H. Chen; Sladjana Jevremović. 2014. "Alteration of flower color in Iris germanica L. ‘Fire Bride’ through ectopic expression of phytoene synthase gene (crtB) from Pantoea agglomerans." Plant Cell Reports 33, no. 8: 1307-1321.
Arabinogalactan proteins (AGPs) are a family of highly glycosylated cell surface proteins located at the plasma membrane and plant cell wall. AGPs play important roles in plant growth and development. Yariv phenylglycoside (βGlcY), synthetic red-brown dye that specifically binds and precipitates AGPs, has been used for detection and quantification of AGPs in plant tissue. Graded concentrations of βGlcY (0–75 μM) were used to investigate the effect of this synthetic dye on induction of in vitro morphogenesis in Centaurium erythraea root culture on two nutrient media: ½MS and ½MS + IBA 1.0 μM. Regeneration of C. erythraea shoots on root explants was stimulated on both media supplemented with 25 μM βGlcY after 8 weeks in culture. Quantification of AGPs in different tissues of C. erythraea was determinate with single radial diffusion method. This work emphasizes clear effect of βGlcY on induction of morphogenesis in vitro in C. erythraea root culture.
Milana Trifunović; Vojin Tadić; Marija Petrić; Danijela Jontulović; Sladjana Jevremović; Angelina Subotić. Quantification of arabinogalactan proteins during in vitro morphogenesis induced by β-d-glucosyl Yariv reagent in Centaurium erythraea root culture. Acta Physiologiae Plantarum 2014, 36, 1187 -1195.
AMA StyleMilana Trifunović, Vojin Tadić, Marija Petrić, Danijela Jontulović, Sladjana Jevremović, Angelina Subotić. Quantification of arabinogalactan proteins during in vitro morphogenesis induced by β-d-glucosyl Yariv reagent in Centaurium erythraea root culture. Acta Physiologiae Plantarum. 2014; 36 (5):1187-1195.
Chicago/Turabian StyleMilana Trifunović; Vojin Tadić; Marija Petrić; Danijela Jontulović; Sladjana Jevremović; Angelina Subotić. 2014. "Quantification of arabinogalactan proteins during in vitro morphogenesis induced by β-d-glucosyl Yariv reagent in Centaurium erythraea root culture." Acta Physiologiae Plantarum 36, no. 5: 1187-1195.
Marija Petrić; Sladjana Jevremović; Milana Trifunovic; Vojin Tadic; Snežana Milošević; Angelina Subotić. Activity of antioxidant enzymes during induction of morphogenesis of Fritillaria meleagris in bulb scale culture. TURKISH JOURNAL OF BIOLOGY 2014, 38, 328 -338.
AMA StyleMarija Petrić, Sladjana Jevremović, Milana Trifunovic, Vojin Tadic, Snežana Milošević, Angelina Subotić. Activity of antioxidant enzymes during induction of morphogenesis of Fritillaria meleagris in bulb scale culture. TURKISH JOURNAL OF BIOLOGY. 2014; 38 ():328-338.
Chicago/Turabian StyleMarija Petrić; Sladjana Jevremović; Milana Trifunovic; Vojin Tadic; Snežana Milošević; Angelina Subotić. 2014. "Activity of antioxidant enzymes during induction of morphogenesis of Fritillaria meleagris in bulb scale culture." TURKISH JOURNAL OF BIOLOGY 38, no. : 328-338.
We investigated the effect of low temperature and gibberellic acid (GA3) treatment on dormancy in Fritillaria meleagris L. bulbs. Also, we studied the effect of dormancy breaking on the antioxidant enzymes activity. To overcome dormancy, bulbs require a period (4–8 weeks) of exposure to low temperature. Bulbs regenerated in vitro were grown in the dark on medium without growth regulators at the standard (24 °C) or at low temperatures (4 and 15 °C) for 4, 6, 8 and 10 weeks. Bulbs were collected after 3, 4 and 5 weeks of cooling at 4 °C. To investigate the influence of GA3 on dormancy, bulbs were treated for 24 h with GA3 solutions with 1, 2 and 3 mg l−1 concentrations. During the period of growth of bulbs at 4 °C, regeneration of bulbs was very weak, while at 15 °C the number of regenerated bulbs increased significantly. Improved bulb sprouting was achieved by a short treatment with gibberellin. Low temperature also represents a kind of oxidative stress for the plant. The activity of superoxide dismutase, catalase (CAT) and peroxidase (POX) in bulbs of F. meleagris L. grown in vitro and ex vitro increased with decreasing temperature in contrast to glutathione reductase. POX showed generally lower activity than CAT which indicates that major role in the breaking dormancy and preparing bulbs for sprouting have catalases.
Marija Petrić; Sladjana Jevremović; Milana Trifunović-Momčilov; Vojin Tadić; Snežana Milošević; Milan Dragićević; Angelina Subotić. The effect of low temperature and GA3 treatments on dormancy breaking and activity of antioxidant enzymes in Fritillaria meleagris bulblets cultured in vitro. Acta Physiologiae Plantarum 2013, 35, 3223 -3236.
AMA StyleMarija Petrić, Sladjana Jevremović, Milana Trifunović-Momčilov, Vojin Tadić, Snežana Milošević, Milan Dragićević, Angelina Subotić. The effect of low temperature and GA3 treatments on dormancy breaking and activity of antioxidant enzymes in Fritillaria meleagris bulblets cultured in vitro. Acta Physiologiae Plantarum. 2013; 35 (11):3223-3236.
Chicago/Turabian StyleMarija Petrić; Sladjana Jevremović; Milana Trifunović-Momčilov; Vojin Tadić; Snežana Milošević; Milan Dragićević; Angelina Subotić. 2013. "The effect of low temperature and GA3 treatments on dormancy breaking and activity of antioxidant enzymes in Fritillaria meleagris bulblets cultured in vitro." Acta Physiologiae Plantarum 35, no. 11: 3223-3236.
Cytokinin oxidase/dehydrogenase (CKX) is the only known enzyme involved in cytokinin catabolism. Genes coding for two Arabidopsis CKX isoforms, AtCKX1 and AtCKX2, were introduced separately into a binary cloning vector, immobilized into Agrobacterium tumefaciens strain GV3101, and introduced into root explants of centaury (Centaurium erythraea Rafn.). The integration of each transgene was confirmed by genomic PCR. Of the total transformed explants, 30 and 28.2 % of the transformants carried AtCKX1 and AtCKX2 transgenes, respectively. Of these transformants, 50 % exhibited expression of the AtCKX1 transgene, while 64 % of transformants exhibited expression of the AtCKX2 transgene. For all analysed AtCKX transgenic centaury lines, as well as for untransformed control plants, CKX activity was higher in roots than in shoots. Expression of AtCKX in most transgenic lines contributed to enhanced levels of CKX activity in root tissues; whereas, only a few lines demonstrated increased CKX activity in shoot tissues compared to those of control plants. Moreover, overexpression of AtCKX resulted in reduced morphogenetic potential in transgenic plants, but did not significantly affect biomass production in comparison to untransformed control plants.
Milana Trifunović; Aleksandar Cingel; Ana Simonović; Sladjana Jevremović; Marija Petrić; Ivana Dragićević; Václav Motyka; Petre I. Dobrev; Lenka Zahajská; Angelina Subotić. Overexpression of Arabidopsis cytokinin oxidase/dehydrogenase genes AtCKX1 and AtCKX2 in transgenic Centaurium erythraea Rafn. Plant Cell, Tissue and Organ Culture (PCTOC) 2013, 115, 139 -150.
AMA StyleMilana Trifunović, Aleksandar Cingel, Ana Simonović, Sladjana Jevremović, Marija Petrić, Ivana Dragićević, Václav Motyka, Petre I. Dobrev, Lenka Zahajská, Angelina Subotić. Overexpression of Arabidopsis cytokinin oxidase/dehydrogenase genes AtCKX1 and AtCKX2 in transgenic Centaurium erythraea Rafn. Plant Cell, Tissue and Organ Culture (PCTOC). 2013; 115 (2):139-150.
Chicago/Turabian StyleMilana Trifunović; Aleksandar Cingel; Ana Simonović; Sladjana Jevremović; Marija Petrić; Ivana Dragićević; Václav Motyka; Petre I. Dobrev; Lenka Zahajská; Angelina Subotić. 2013. "Overexpression of Arabidopsis cytokinin oxidase/dehydrogenase genes AtCKX1 and AtCKX2 in transgenic Centaurium erythraea Rafn." Plant Cell, Tissue and Organ Culture (PCTOC) 115, no. 2: 139-150.
S. Jevremovic; A. Subotic; D. Miljkovic; M. Trifunović; M. Petric; A. Cingel. CLONAL FIDELITY OF CHRYSANTHEMUM CULTIVARS AFTER LONG TERM MICROPROPAGATION BY STEM SEGMENT CULTURE. Acta Horticulturae 2012, 211 -216.
AMA StyleS. Jevremovic, A. Subotic, D. Miljkovic, M. Trifunović, M. Petric, A. Cingel. CLONAL FIDELITY OF CHRYSANTHEMUM CULTIVARS AFTER LONG TERM MICROPROPAGATION BY STEM SEGMENT CULTURE. Acta Horticulturae. 2012; (961):211-216.
Chicago/Turabian StyleS. Jevremovic; A. Subotic; D. Miljkovic; M. Trifunović; M. Petric; A. Cingel. 2012. "CLONAL FIDELITY OF CHRYSANTHEMUM CULTIVARS AFTER LONG TERM MICROPROPAGATION BY STEM SEGMENT CULTURE." Acta Horticulturae , no. 961: 211-216.
The orange color of tiger lily (Lolium lancifolium ‘Splendens’) flowers is due, primarily, to the accumulation of two κ-xanthophylls, capsanthin and capsorubin. An enzyme, known as capsanthin-capsorubin synthase (CCS), catalyzes the conversion of antheraxanthin and violaxanthin into capsanthin and capsorubin, respectively. We cloned the gene for capsanthin-capsorubin synthase (Llccs) from flower tepals of L. lancifolium by the rapid amplification of cDNA ends (RACE) with a heterologous non-degenerate primer that was based on the sequence of a gene for lycopene β-cyclase (lcyB). The full-length cDNA of Llccs was 1,785 bp long and contained an open reading frame of 1,425 bp that encoded a polypeptide of 474 amino acids with a predicted N-terminal plastid-targeting sequence. Analysis by reverse transcription–PCR (RT–PCR) revealed that expression of Llccs was spatially and temporally regulated, with expression in flower buds and flowers of L. lancifolium but not in vegetative tissues. Stable overexpression of the Llccs gene in callus tissue of Iris germanica, which accumulates several xanthophylls including violaxanthin, the precursor of capsorubin, resulted in transgenic callus whose color had changed from its normal yellow to red-orange. This novel red-orange coloration was due to the accumulation of two non-native κ-xanthophylls, capsanthin and capsorubin, as confirmed by HPLC and ultraperformance liquid chromatography–tandem mass spectrometry (UPLC-MS/MS) analysis with authentic standards. Cloning of the Llccs gene should advance our understanding of the molecular and genetic mechanisms of the biosynthesis of κ-carotenoids in general and in the genus Lilium in particular, and will facilitate transgenic alterations of the colors of flowers and fruits of many plant species.
Zoran Jeknić; Jeffrey T. Morré; Stevan Jeknić; Sladjana Jevremović; Angelina Subotić; Tony H.H. Chen. Cloning and Functional Characterization of a Gene for Capsanthin-Capsorubin Synthase from Tiger Lily (Lilium lancifolium Thunb. ‘Splendens’). Plant and Cell Physiology 2012, 53, 1899 -1912.
AMA StyleZoran Jeknić, Jeffrey T. Morré, Stevan Jeknić, Sladjana Jevremović, Angelina Subotić, Tony H.H. Chen. Cloning and Functional Characterization of a Gene for Capsanthin-Capsorubin Synthase from Tiger Lily (Lilium lancifolium Thunb. ‘Splendens’). Plant and Cell Physiology. 2012; 53 (11):1899-1912.
Chicago/Turabian StyleZoran Jeknić; Jeffrey T. Morré; Stevan Jeknić; Sladjana Jevremović; Angelina Subotić; Tony H.H. Chen. 2012. "Cloning and Functional Characterization of a Gene for Capsanthin-Capsorubin Synthase from Tiger Lily (Lilium lancifolium Thunb. ‘Splendens’)." Plant and Cell Physiology 53, no. 11: 1899-1912.
Irises are perennial plants widely used as ornamental garden plants or cut flowers. Some species accumulate secondary metabolites, making them highly valuable to the pharmaceutical and perfume industries. Micropropagation of irises has successfully been accomplished by culturing zygotic embryos, different flower parts, and leaf base tissues as starting explants. Plantlets are regenerated via somatic embryogenesis, organogenesis, or both processes at the same time depending on media composition and plant species. A large number of uniform plants are produced by somatic embryogenesis, however, some species have decreased morphogenetic potential overtime. Shoot cultures obtained by organogenesis can be multiplied for many years. Somatic embryogenic tissue can be reestablished from leaf bases of in vitro-grown shoots. The highest number of plants can be obtained by cell suspension cultures. This chapter describes effective in vitro plant regeneration protocols for Iris species from different types of explants by somatic embryogenesis and/or organogenesis suitable for the mass propagation of ornamental and pharmaceutical irises.
Slađana Jevremović; Zoran Jeknić; Angelina Subotic. Micropropagation of Iris sp. Advanced Structural Safety Studies 2012, 11013, 291 -303.
AMA StyleSlađana Jevremović, Zoran Jeknić, Angelina Subotic. Micropropagation of Iris sp. Advanced Structural Safety Studies. 2012; 11013 ():291-303.
Chicago/Turabian StyleSlađana Jevremović; Zoran Jeknić; Angelina Subotic. 2012. "Micropropagation of Iris sp." Advanced Structural Safety Studies 11013, no. : 291-303.
Ornamental plants Impatiens hawkerii and Impatiens walleriana are susceptible to Tomato spotted wilt virus infection that can be eradicated by meristem-tip propagation. Comparison of superoxide dismutase (SOD), catalase (CAT), guaiacol peroxidase (GPX) and ascorbate peroxidase (APX) profiles of healthy, infected and virus-eliminated plants showed that long-term infection did not cause major changes in I. hawkerii, but caused induction of several APX and GPX isoforms, increase in Mn-SOD and APX and a decrease in CAT activity in I. walleriana. This is the first report on the effects of virus elimination by meristem-tip propagation on the antioxidative metabolism in plants.
S. Milošević; A. Simonović; A. Cingel; S. Jevremović; S. Todorović; B. Filipović; A. Subotić. Response of antioxidative enzymes to long-term Tomato spotted wilt virus infection and virus elimination by meristem-tip culture in two Impatiens species. Physiological and Molecular Plant Pathology 2012, 79, 79 -88.
AMA StyleS. Milošević, A. Simonović, A. Cingel, S. Jevremović, S. Todorović, B. Filipović, A. Subotić. Response of antioxidative enzymes to long-term Tomato spotted wilt virus infection and virus elimination by meristem-tip culture in two Impatiens species. Physiological and Molecular Plant Pathology. 2012; 79 ():79-88.
Chicago/Turabian StyleS. Milošević; A. Simonović; A. Cingel; S. Jevremović; S. Todorović; B. Filipović; A. Subotić. 2012. "Response of antioxidative enzymes to long-term Tomato spotted wilt virus infection and virus elimination by meristem-tip culture in two Impatiens species." Physiological and Molecular Plant Pathology 79, no. : 79-88.