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University of Belgrade, Institute for Biological Research "Siniša Stanković", National Institute of Republic of Serbia
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.
Somatic embryogenesis (SE) is a developmental process during which plant somatic cells, under suitable conditions, produce embryogenic cells that develop into somatic embryos (se). SE is the most important method for plant propagation in vitro, having both fundamental and applicative significance. SE can be induced from different tissues and organs, but when se are used as explants, the process is recognized as secondary or cyclic SE. We induced secondary SE in Centaurium erythraea by application of 2,4-dichlorophenoxyacetic acid (2,4-D) and N-(2-chloro-4-pyridyl)-N′-phenylurea (CPPU). A medium containing 0.1 mgL−1 2,4-D and 0.25 mgL−1 CPPU was optimal in terms of the number of primary SE explants forming se, the number of well-developed se per explant, and morphological appearance of the obtained se. These concentrations allowed SE to progress through three cycles, whereas at higher concentrations of 0.2 mgL−1 2,4-D and 0.5 mgL−1 CPPU, only two cycles were achieved. Histological analysis revealed that secondary se are formed both directly and indirectly. Secondary SE readily germinated and converted into plantlets. Induction of cyclic SE contributes to the conservation efforts of this endangered medicinal plant and expands the spectrum of in vitro developmental pathways described in centaury—an emerging model in developmental biology.
Milica Bogdanović; Katarina Ćuković; Angelina Subotić; Milan Dragićević; Ana Simonović; Biljana Filipović; Slađana Todorović. Secondary Somatic Embryogenesis in Centaurium erythraea Rafn. Plants 2021, 10, 199 .
AMA StyleMilica Bogdanović, Katarina Ćuković, Angelina Subotić, Milan Dragićević, Ana Simonović, Biljana Filipović, Slađana Todorović. Secondary Somatic Embryogenesis in Centaurium erythraea Rafn. Plants. 2021; 10 (2):199.
Chicago/Turabian StyleMilica Bogdanović; Katarina Ćuković; Angelina Subotić; Milan Dragićević; Ana Simonović; Biljana Filipović; Slađana Todorović. 2021. "Secondary Somatic Embryogenesis in Centaurium erythraea Rafn." Plants 10, no. 2: 199.
Aquaporins comprise a large group of transmembrane proteins responsible for water transport, which is crucial for plant survival under stress conditions. Despite the vital role of aquaporins, nothing is known about this protein family in Impatiens walleriana, a commercially important horticultural plant, which is sensitive to drought stress. In the present study, attention is given to the molecular characterization of aquaporins in I. walleriana and their expression during drought stress and recovery. We identified four I. walleriana aquaporins: IwPIP1;4, IwPIP2;2, IwPIP2;7 and IwTIP4;1. All of them had conserved NPA motifs (Asparagine-Proline-Alanine), transmembrane helices (TMh), pore characteristics, stereochemical properties and tetrameric structure of holoprotein. Drought stress and recovery treatment affected the aquaporins expression in I. walleriana leaves, which was up- or downregulated depending on stress intensity. Expression of IwPIP2;7 was the most affected of all analyzed I. walleriana aquaporins. At 15% and 5% soil moisture and recovery from 15% and 5% soil moisture, IwPIP2;7 expression significantly decreased and increased, respectively. Aquaporins IwPIP1;4 and IwTIP4;1 had lower expression in comparison to IwPIP2;7, with moderate expression changes in response to drought and recovery, while IwPIP2;2 expression was of significance only in recovered plants. Insight into the molecular structure of I. walleriana aquaporins expanded knowledge about plant aquaporins, while its expression during drought and recovery contributed to I. walleriana drought tolerance mechanisms and re-acclimation.
Marija Đurić; Angelina Subotić; Ljiljana Prokić; Milana Trifunović-Momčilov; Aleksandar Cingel; Milan Dragićević; Ana Simonović; Snežana Milošević. Molecular Characterization and Expression of Four Aquaporin Genes in Impatiens walleriana During Drought Stress and Recovery. Plants 2021, 10, 154 .
AMA StyleMarija Đurić, Angelina Subotić, Ljiljana Prokić, Milana Trifunović-Momčilov, Aleksandar Cingel, Milan Dragićević, Ana Simonović, Snežana Milošević. Molecular Characterization and Expression of Four Aquaporin Genes in Impatiens walleriana During Drought Stress and Recovery. Plants. 2021; 10 (1):154.
Chicago/Turabian StyleMarija Đurić; Angelina Subotić; Ljiljana Prokić; Milana Trifunović-Momčilov; Aleksandar Cingel; Milan Dragićević; Ana Simonović; Snežana Milošević. 2021. "Molecular Characterization and Expression of Four Aquaporin Genes in Impatiens walleriana During Drought Stress and Recovery." Plants 10, no. 1: 154.
Centaurium erythraea (centaury) is a traditionally used medicinal plant, with a spectrum of secondary metabolites with confirmed healing properties. Centaury is an emerging model in plant developmental biology due to its vigorous regenerative potential and great developmental plasticity when cultured in vitro. Hereby, we review nearly two decades of research on somatic embryogenesis (SE) in centaury. During SE, somatic cells are induced by suitable culture conditions to express their totipotency, acquire embryogenic characteristics, and eventually give rise to somatic embryos. When SE is initiated from centaury root explants, the process occurs spontaneously (on hormone-free medium), directly (without the callusing phase), and the somatic embryos are of unicellular origin. SE from leaf explants has to be induced by plant growth regulators and is indirect (preceded by callusing). Histological observations and culture conditions are compared in these two systems. The changes in antioxidative enzymes were followed during SE from the leaf explants. Special focus is given to the role of arabinogalactan proteins during SE, which were analyzed using a variety of approaches. The newest and preliminary results, including centaury transcriptome, novel potential SE markers, and novel types of arabinogalactan proteins, are discussed as perspectives of centaury research.
Ana D. Simonović; Milana M. Trifunović-Momčilov; Biljana K. Filipović; Marija P. Marković; Milica D. Bogdanović; Angelina R. Subotić. Somatic Embryogenesis in Centaurium erythraea Rafn—Current Status and Perspectives: A Review. Plants 2020, 10, 70 .
AMA StyleAna D. Simonović, Milana M. Trifunović-Momčilov, Biljana K. Filipović, Marija P. Marković, Milica D. Bogdanović, Angelina R. Subotić. Somatic Embryogenesis in Centaurium erythraea Rafn—Current Status and Perspectives: A Review. Plants. 2020; 10 (1):70.
Chicago/Turabian StyleAna D. Simonović; Milana M. Trifunović-Momčilov; Biljana K. Filipović; Marija P. Marković; Milica D. Bogdanović; Angelina R. Subotić. 2020. "Somatic Embryogenesis in Centaurium erythraea Rafn—Current Status and Perspectives: A Review." Plants 10, no. 1: 70.
Impatiens walleriana is a valued ornamental plant sensitive to drought stress. We investigated whether the foliar application of 2mM salicylic acid (SA) can protect potted I. walleriana plants from drought stress. The plants were divided into: watered plants, drought-stressed plants, watered plants treated with SA and drought-stressed plants treated with SA. The number of flowers and flower buds, relative water content (RWC), contents of malondialdehyde (MDA) and proline (Pro) and the activities of superoxide dismutases, catalases and peroxidases were recorded at different time points. Three dehydrin sequences were identified in de novo assembled leaf transcriptome: IwDhn1, IwDhn2.1 and IwDhn2.2. Drought stress caused wilting, floral abortion, reduction of RWC and increased MDA—an indicator of lipid peroxidation. In response to drought, Impatiens accumulated Pro and induced chloroplastic Cu/ZnSOD and two peroxidase isoforms. The most remarkable drought response was strong induction of IwDhn2.1 and IwDhn2.2. Rehydration restored RWC, Pro level, Cu/ZnSOD activity and dehydrins expression in drought-stressed plants approximately to the values of watered plants.SA had ameliorating effects on plants exposed to drought, including prevention of wilting, preservation of RWC, increased Pro accumulation, modulation of antioxidative activities and remarkable decrease of lipid peroxidation, but without effects on flowers’ preservation.
Dragana D. Antonić; Angelina R. Subotić; Milan B. Dragićević; Danijel Pantelić; Snežana M. Milošević; Ana D. Simonović; Ivana Momčilović. Effects of Exogenous Salicylic Acid on Drought Response and Characterization of Dehydrins in Impatiens walleriana. Plants 2020, 9, 1589 .
AMA StyleDragana D. Antonić, Angelina R. Subotić, Milan B. Dragićević, Danijel Pantelić, Snežana M. Milošević, Ana D. Simonović, Ivana Momčilović. Effects of Exogenous Salicylic Acid on Drought Response and Characterization of Dehydrins in Impatiens walleriana. Plants. 2020; 9 (11):1589.
Chicago/Turabian StyleDragana D. Antonić; Angelina R. Subotić; Milan B. Dragićević; Danijel Pantelić; Snežana M. Milošević; Ana D. Simonović; Ivana Momčilović. 2020. "Effects of Exogenous Salicylic Acid on Drought Response and Characterization of Dehydrins in Impatiens walleriana." Plants 9, no. 11: 1589.
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.
This study was carried out to examine the drought effect on development, physiological, biochemical and molecular parameters in Impatiens walleriana grown ex vitro. Experiment design included three treatments: Control plants—grown under optimal watering (35%–37% of soil moisture content), drought-stressed plants—non-irrigated to reach 15% and 5% of soil moisture content and recovery plants—rehydrated for four days to reach optimal soil moisture content. Drought reduced fresh weight, total leaf area, as well as dry weight of I. walleriana shoots. Drought up-regulated expression of abscisic acid (ABA) biosynthesis genes 9-cis-epoxycarotenoid dioxygenase 4 (NCED4) and abscisic aldehyde oxidase 2 (AAO2) and catabolic gene ABA 8′-hydroxylase 3 (ABA8ox3) which was followed by increased ABA content in the leaves. Decrement in water potential of shoots during the drought was not accompanied with increased amino acid proline content. We detected an increase in chlorophyll, carotenoid, total polyphenols and flavonols content under drought conditions, as well as malondialdehyde, hydrogen peroxide and DPPH (1,1′-diphenyl-2-picrylhydrazyl) activity. Increased antioxidant enzyme activities (superoxide dismutase, peroxidase and catalase) throughout drought were also determined. Recovery treatment was significant for neutralizing drought effect on growth parameters, shoot water potential, proline content and genes expression.
Marija Đurić; Angelina Subotić; Ljiljana Prokić; Milana Trifunović-Momčilov; Aleksandar Cingel; Milorad Vujičić; Snežana Milošević. Morpho-Physiological and Molecular Evaluation of Drought and Recovery in Impatiens walleriana Grown Ex Vitro. Plants 2020, 9, 1559 .
AMA StyleMarija Đurić, Angelina Subotić, Ljiljana Prokić, Milana Trifunović-Momčilov, Aleksandar Cingel, Milorad Vujičić, Snežana Milošević. Morpho-Physiological and Molecular Evaluation of Drought and Recovery in Impatiens walleriana Grown Ex Vitro. Plants. 2020; 9 (11):1559.
Chicago/Turabian StyleMarija Đurić; Angelina Subotić; Ljiljana Prokić; Milana Trifunović-Momčilov; Aleksandar Cingel; Milorad Vujičić; Snežana Milošević. 2020. "Morpho-Physiological and Molecular Evaluation of Drought and Recovery in Impatiens walleriana Grown Ex Vitro." Plants 9, no. 11: 1559.
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.
Centaurium erythraea Rafn. (common centaury, Gentianaceae) is a medicinal plant with great regeneration potential and developmental plasticity in vitro. Centaury can be regenerated from leaf explants by both somatic embryogenesis (SE) and shoot development (SD). We believe that its regeneration potential and developmental plasticity rest on high activity of certain genes, which may not be active or present in species recalcitrant to in vitro regeneration. However, there are no sequenced Gentianaceae genomes to support investigation of the molecular events during SE or SD. To this end, we have sequenced six centaury transcriptomes (embryogenic calli, globular somatic embryos, cotyledonary somatic embryos, adventitious buds, leaves and roots of in vitro grown plants) and de novo assembled centaury referent transcriptome comprising 105.726 genes. The high quality and completeness transcriptome was functionally annotated against NCBI nt, Swissprot and PFAM databases with KOG and GO enrichment. In addition, 11 housekeeping and functional genes were validated for expression stability in 27 tissue samples representing the processes of SE and SD, plants from nature and wounded tissues using GeNorm and NormFinder. The most stable genes that can be used for expression studies during SE, SD and in vitro manipulations are Ribosomal protein L2 (RPL2) and TATA binding protein 1 (TBP1) in combination with RAS (Rat Sarcoma)-related Nuclear protein (RAN) or Adenosine kinase (AK). These results comprise a complete framework for the search for genes involved in SE and SD, but may also be useful in identifying genes involved in biosynthesis of C. erythraea secondary metabolites.
Katarina Ćuković; Milan Dragićević; Milica Bogdanović; Danijela Paunović; Giorgio Giurato; Biljana Filipović; Angelina Subotić; Slađana Todorović; Ana Simonović. Plant regeneration in leaf culture of Centaurium erythraea Rafn. Part 3: de novo transcriptome assembly and validation of housekeeping genes for studies of in vitro morphogenesis. Plant Cell, Tissue and Organ Culture (PCTOC) 2020, 141, 417 -433.
AMA StyleKatarina Ćuković, Milan Dragićević, Milica Bogdanović, Danijela Paunović, Giorgio Giurato, Biljana Filipović, Angelina Subotić, Slađana Todorović, Ana Simonović. Plant regeneration in leaf culture of Centaurium erythraea Rafn. Part 3: de novo transcriptome assembly and validation of housekeeping genes for studies of in vitro morphogenesis. Plant Cell, Tissue and Organ Culture (PCTOC). 2020; 141 (2):417-433.
Chicago/Turabian StyleKatarina Ćuković; Milan Dragićević; Milica Bogdanović; Danijela Paunović; Giorgio Giurato; Biljana Filipović; Angelina Subotić; Slađana Todorović; Ana Simonović. 2020. "Plant regeneration in leaf culture of Centaurium erythraea Rafn. Part 3: de novo transcriptome assembly and validation of housekeeping genes for studies of in vitro morphogenesis." Plant Cell, Tissue and Organ Culture (PCTOC) 141, no. 2: 417-433.
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.
The genus Centaurium includes about 50 plant species found throughout the northern hemisphere. Plant species Centaurium erythraea Rafn., commonly known as common centaury, has been used for centuries for medical purposes. Centaury is used to treat anemia, jaundice, and gout and to cure febrile conditions and regulate blood sugar. So far, centaury species were genetically transformed mostly using Agrobacterium rhizogenes. Only one report has described A. tumefaciens-mediated gene delivery for the production of transgenic centaury plants. Genetic transformation of centaury using AtCKX genes did not influence the quality but influenced the quantity of xanthones in shoots and roots. The majority of AtCKX transformed centaury lines grown in vitro produced increased eustomin and/or demethyleustomin content than untransformed control plants. This work clearly demonstrates, for the first time, the effect of centaury secoiridoids and xanthones on colorectal cancer cell line (DLD1) and its resistant counterpart (DLD1-TxR). The xanthone eustomin showed the most significant cell growth inhibition effects. Since xanthones are increasingly being used for their pharmacological properties, AtCKX transgenic centaury plants could be used as a useful source of plant material for the production of novel drugs.
Milana Trifunović-Momčilov; Dijana Krstić-Milošević; Snežana Trifunović; Ana Podolski-Renić; Milica Pešić; Angelina Subotić. Secondary Metabolite Profile of Transgenic Centaury (Centaurium erythraea Rafn.) Plants, Potential Producers of Anticancer Compounds. Reference Series in Phytochemistry 2017, 205 -230.
AMA StyleMilana Trifunović-Momčilov, Dijana Krstić-Milošević, Snežana Trifunović, Ana Podolski-Renić, Milica Pešić, Angelina Subotić. Secondary Metabolite Profile of Transgenic Centaury (Centaurium erythraea Rafn.) Plants, Potential Producers of Anticancer Compounds. Reference Series in Phytochemistry. 2017; ():205-230.
Chicago/Turabian StyleMilana Trifunović-Momčilov; Dijana Krstić-Milošević; Snežana Trifunović; Ana Podolski-Renić; Milica Pešić; Angelina Subotić. 2017. "Secondary Metabolite Profile of Transgenic Centaury (Centaurium erythraea Rafn.) Plants, Potential Producers of Anticancer Compounds." Reference Series in Phytochemistry , no. : 205-230.
The aim of this study was to determine the enzymatic profile of esterases and peroxidases during early stages of somatic embryogenesis of Fritillaria meleagris L. Somatic embryogenesis was induced using the leaf base as explant on a medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D). Zymography showed the presence of different moieties, six isoforms of esterases and peroxidases, during morphogenesis as compared to control explants. One isoform of esterases was detected only during the process of somatic embryogenesis, and one isoform was detected in control explants. Analysis of esterases with 1-naphthyl butyrate proved that esterases, which participate in somatic embryogenesis of F. meleagris, belong to the family of aryl esterases. For the first time it was proved that five isoforms of esterases, which are involved in morphogenesis of F. meleagris, belong to the family of aryl esterases, while two isoforms are carboxyl esterases. One isoform of carboxyl esterases was visible in control explants. This is also the first description of peroxidases during the morphogenetic process, and of the difference between aryl and carboxyl esterases. More isoforms of esterases during morphogenesis as compared to control explants are probably responsible for some early physiological process during somatic embryogenesis of F. meleagris.
Marija Petric; Angelina Subotic; Sladjana Jevremovic; Milana Trifunovic-Momcilov; Vojin Tadic; Marica Grujic; Zoran Vujcic. Esterase and peroxidase isoforms during initial stages of somatic embryogenesis in Fritillaria meleagris L. leaf base. Archives of Biological Sciences 2017, 69, 619 -625.
AMA StyleMarija Petric, Angelina Subotic, Sladjana Jevremovic, Milana Trifunovic-Momcilov, Vojin Tadic, Marica Grujic, Zoran Vujcic. Esterase and peroxidase isoforms during initial stages of somatic embryogenesis in Fritillaria meleagris L. leaf base. Archives of Biological Sciences. 2017; 69 (4):619-625.
Chicago/Turabian StyleMarija Petric; Angelina Subotic; Sladjana Jevremovic; Milana Trifunovic-Momcilov; Vojin Tadic; Marica Grujic; Zoran Vujcic. 2017. "Esterase and peroxidase isoforms during initial stages of somatic embryogenesis in Fritillaria meleagris L. leaf base." Archives of Biological Sciences 69, no. 4: 619-625.
The genus Centaurium includes about 50 plant species found throughout the northern hemisphere. Plant species Centaurium erythraea Rafn., commonly known as common centaury, has been used for centuries for medical purposes. Centaury is used to treat anemia, jaundice, and gout and to cure febrile conditions and regulate blood sugar. So far, centaury species were genetically transformed mostly using Agrobacterium rhizogenes. Only one report has described A. tumefaciens-mediated gene delivery for the production of transgenic centaury plants. Genetic transformation of centaury using AtCKX genes did not influence the quality but influenced the quantity of xanthones in shoots and roots. The majority of AtCKX transformed centaury lines grown in vitro produced increased eustomin and/or demethyleustomin content than untransformed control plants. This work clearly demonstrates, for the first time, the effect of centaury secoiridoids and xanthones on colorectal cancer cell line (DLD1) and its resistant counterpart (DLD1-TxR). The xanthone eustomin showed the most significant cell growth inhibition effects. Since xanthones are increasingly being used for their pharmacological properties, AtCKX transgenic centaury plants could be used as a useful source of plant material for the production of novel drugs.
Milana Trifunović-Momčilov; Dijana Krstić-Milošević; Snežana Trifunović; Ana Podolski-Renić; Milica Pešić; Angelina Subotić. Secondary Metabolite Profile of Transgenic Centaury (Centaurium erythraea Rafn.) Plants, Potential Producers of Anticancer Compounds. Reference Series in Phytochemistry 2016, 1 -26.
AMA StyleMilana Trifunović-Momčilov, Dijana Krstić-Milošević, Snežana Trifunović, Ana Podolski-Renić, Milica Pešić, Angelina Subotić. Secondary Metabolite Profile of Transgenic Centaury (Centaurium erythraea Rafn.) Plants, Potential Producers of Anticancer Compounds. Reference Series in Phytochemistry. 2016; ():1-26.
Chicago/Turabian StyleMilana Trifunović-Momčilov; Dijana Krstić-Milošević; Snežana Trifunović; Ana Podolski-Renić; Milica Pešić; Angelina Subotić. 2016. "Secondary Metabolite Profile of Transgenic Centaury (Centaurium erythraea Rafn.) Plants, Potential Producers of Anticancer Compounds." Reference Series in Phytochemistry , no. : 1-26.
The genus Centaurium includes about 50 plant species found throughout the northern hemisphere. Plant species Centaurium erythraea Rafn., commonly known as common centaury, has been used for centuries for medical purposes. Centaury is used to treat anemia, jaundice, and gout and to cure febrile conditions and regulate blood sugar. So far, centaury species were genetically transformed mostly using Agrobacterium rhizogenes. Only one report has described A. tumefaciens-mediated gene delivery for the production of transgenic centaury plants. Genetic transformation of centaury using AtCKX genes did not influence the quality but influenced the quantity of xanthones in shoots and roots. The majority of AtCKX transformed centaury lines grown in vitro produced increased eustomin and/or demethyleustomin content than untransformed control plants. This work clearly demonstrates, for the first time, the effect of centaury secoiridoids and xanthones on colorectal cancer cell line (DLD1) and its resistant counterpart (DLD1-TxR). The xanthone eustomin showed the most significant cell growth inhibition effects. Since xanthones are increasingly being used for their pharmacological properties, AtCKX transgenic centaury plants could be used as a useful source of plant material for the production of novel drugs.
Milana Trifunović-Momčilov; Dijana Krstić-Milošević; Snežana Trifunović; Ana Podolski-Renić; Milica Pešić; Angelina Subotić. Secondary Metabolite Profile of Transgenic Centaury (Centaurium erythraea Rafn.) Plants, Potential Producers of Anticancer Compounds. Reference Series in Phytochemistry 2016, 1 -26.
AMA StyleMilana Trifunović-Momčilov, Dijana Krstić-Milošević, Snežana Trifunović, Ana Podolski-Renić, Milica Pešić, Angelina Subotić. Secondary Metabolite Profile of Transgenic Centaury (Centaurium erythraea Rafn.) Plants, Potential Producers of Anticancer Compounds. Reference Series in Phytochemistry. 2016; ():1-26.
Chicago/Turabian StyleMilana Trifunović-Momčilov; Dijana Krstić-Milošević; Snežana Trifunović; Ana Podolski-Renić; Milica Pešić; Angelina Subotić. 2016. "Secondary Metabolite Profile of Transgenic Centaury (Centaurium erythraea Rafn.) Plants, Potential Producers of Anticancer Compounds." Reference Series in Phytochemistry , no. : 1-26.
Dragana Antonic; Snežana Milošević; Aleksandar Cingel; M. Lojić; M. Trifunović-Momčilov; M. Petric; Angelina Subotić; Ana Simonović. Effects of exogenous salicylic acid on Impatiens walleriana L. grown in vitro under polyethylene glycol-imposed drought. South African Journal of Botany 2016, 105, 226 -233.
AMA StyleDragana Antonic, Snežana Milošević, Aleksandar Cingel, M. Lojić, M. Trifunović-Momčilov, M. Petric, Angelina Subotić, Ana Simonović. Effects of exogenous salicylic acid on Impatiens walleriana L. grown in vitro under polyethylene glycol-imposed drought. South African Journal of Botany. 2016; 105 ():226-233.
Chicago/Turabian StyleDragana Antonic; Snežana Milošević; Aleksandar Cingel; M. Lojić; M. Trifunović-Momčilov; M. Petric; Angelina Subotić; Ana Simonović. 2016. "Effects of exogenous salicylic acid on Impatiens walleriana L. grown in vitro under polyethylene glycol-imposed drought." South African Journal of Botany 105, no. : 226-233.
Over 20% of all protein domains are currently annotated as ?domains of unknown function? or DUFs. In a recently identified Centaurium erythraea arabinogalactan peptide, CeAGP3 (AGN92423), a conserved DUF1070 domain was found. Since identifying functions for DUFs is important in systems biology, we have analyzed the distribution and structure of DUF1070 domain (pfam06376) using a set of bioinformatics tools. There are 271 publically available DUF1070 members from 25 diverse families of vascular plants, and most are short sequences (50-100 aa). The N-terminal signal peptide (Nsp) was found in almost all complete sequences. In 233 sequences, at least two noncontiguous prolines were found as clustered dipeptides predicted to be hydroxylated and glycosylated with type II arabino-3,6-galactans, thus representing AG-II glycomodules. In addition, 35 sequences contained a region rich in basic residues (basic linker, BL). The N-terminal part of the DUF1070 domain is comprised of (part of) AG-II and/or BL, while the highly conserved C-terminus is a region of 26 aa, termed SH26. In 212 sequences, SH26 was a typical glycosylphosphatidylinositol lipid anchor signal peptide (GPIsp), but in 83 cases GPIsp was not predicted due to software constraints. In sequences where both Nsp and GPIsp were predicted, the length of mature peptides could be calculated, and it was 10-16 aa. Our analysis suggests that DUF1070 members are arabinogalactan (AG) peptides, of which the majority are GPI-anchored. DUF1070 is the only conserved domain found in classical arabinogalactan proteins and AG peptides. The SH26 region can be used for mining and annotation of AG peptides.
Ana D. Simonović; Milan B. Dragićević; Milica D. Bogdanović; Milana M. Trifunović-Momčilov; Angelina R. Subotić; Slađana Todorović. DUF1070 as a signature domain of a subclass of arabinogalactan peptides. Archives of Biological Sciences 2016, 68, 737 -746.
AMA StyleAna D. Simonović, Milan B. Dragićević, Milica D. Bogdanović, Milana M. Trifunović-Momčilov, Angelina R. Subotić, Slađana Todorović. DUF1070 as a signature domain of a subclass of arabinogalactan peptides. Archives of Biological Sciences. 2016; 68 (4):737-746.
Chicago/Turabian StyleAna D. Simonović; Milan B. Dragićević; Milica D. Bogdanović; Milana M. Trifunović-Momčilov; Angelina R. Subotić; Slađana Todorović. 2016. "DUF1070 as a signature domain of a subclass of arabinogalactan peptides." Archives of Biological Sciences 68, no. 4: 737-746.
Primary and secondary ammonium assimilation is catalyzed by the glutamine synthetase-glutamate synthase (GS-GOGAT) pathway in plants. The Arabidopsis genome contains five cytosolic GS1 genes (GLN1;1 - GLN1;5), one nuclear gene for chloroplastic GS2 isoform (GLN2), two Fd-GOGAT genes (GLU1 and GLU2) and a GLT1 gene coding for NADH-GOGAT. Even though the regulation of GS and GOGAT isoforms has been extensively studied in response to various environmental and metabolic cues in many plant species, little is known about the effects of phytohormones on their regulation. The objective of this study was to investigate the impact of representative plant growth regulators, kinetin (KIN), abscisic acid (ABA), gibberellic acid (GA3) and 2,4-dichlorophenoxyacetic acid (2,4-D), on the expression of A. thaliana GS and GOGAT genes. The obtained results indicate that GS and GOGAT genes are differentially regulated by growth regulators in shoots and roots. KIN and 2,4-D repressed GS and GOGAT expression in roots, with little effect on transcript levels in shoots. KIN affected all tested genes; 2,4-D was apparently more selective and less potent. ABA induced the expression of GLN1;1 and GLU2 in whole seedlings, while GA3 enhanced the expression of all tested genes in shoots, except GLU2. The observed expression patterns are discussed in relation to physiological roles of investigated plant growth regulators and N-assimilating enzymes.
Milan Dragicevic; Ana Simonovic; Milica Bogdanovic; Angelina Subotic; Nabil Ghalawenji; Ivana Dragicevic; Slađana Todorović. Differential regulation of GS-GOGAT gene expression by plant growth regulators in Arabidopsis seedlings. Archives of Biological Sciences 2016, 68, 399 -404.
AMA StyleMilan Dragicevic, Ana Simonovic, Milica Bogdanovic, Angelina Subotic, Nabil Ghalawenji, Ivana Dragicevic, Slađana Todorović. Differential regulation of GS-GOGAT gene expression by plant growth regulators in Arabidopsis seedlings. Archives of Biological Sciences. 2016; 68 (2):399-404.
Chicago/Turabian StyleMilan Dragicevic; Ana Simonovic; Milica Bogdanovic; Angelina Subotic; Nabil Ghalawenji; Ivana Dragicevic; Slađana Todorović. 2016. "Differential regulation of GS-GOGAT gene expression by plant growth regulators in Arabidopsis seedlings." Archives of Biological Sciences 68, no. 2: 399-404.
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.