TY  - CONF
AU  - Trajković, Milena
AU  - Jeknić, Zoran
AU  - Antonić Reljin, Dragana
AU  - Subotić, Angelina
AU  - Jevremović, Slađana
AU  - Cingel, Aleksandar
PY  - 2018
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6540
AB  - Viola cornuta L. ‘Lutea Splendens’ is a perennial ornamental plant with small yellow flowers that
naturally grows in the Pyrenees in Spain and France. To develop novel cultivars with orange and
red flower colors, we performed Agrobacterium tumefaciens (LBA4404)-mediated transformation
with the binary vector pWBVec10a/P35S::Llccs::TNos that harbored capsanthin-capsorubin synthase
(Llccs) gene from Lilium lancifolium under the control of CaMV35S constitutive promoter and
the nopaline synthase (Nos) terminator. Capsanthin-capsorubin synthase catalyzes the conversion
of anteraxanthin and violaxanthin, two yellow ubiquitous 5-6-epoxy-xanthophylls, into capsanthin
and capsorubin, two red xanthophylls, respectively. Starting with hypocotyl explants, we developed
a transformation protocol with 0.3% shoot regeneration efficiency. Histochemical assay for
β-glucuronidase (GUS) activity showed uidA reporter gene expression in all putative Llccs-transgenic
shoots. The presence of Llccs transgene, hygromycin phosphotransferase (hpt) selectable
marker gene and uidA (GUS) reporter gene in all putative Llccs-transgenic lines were confirmed
by PCR analysis. This is the first report on Agrobacterium-mediated genetic transformation of V.
cornuta L. with the aim to introduce desirable traits into this species.
PB  - Belgrade: Serbian Plant Physiology Society, Institute for Biological Research “Siniša Stanković”, University of Belgrade Faculty of Biology, University of Belgrade
C3  - Book of abstracts. 3rd International Conference on Plant Biology (22nd SPPS Meeting); 2018 Jun 9-12; Belgrade, Serbia
T1  - Agrobacterium-mediated genetic transformation of Viola cornuta L. "Lutea Splendens" with capsanthin-capsorubin synthase gene
SP  - 152
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_6540
ER  - 

@conference{
author = "Trajković, Milena and Jeknić, Zoran and Antonić Reljin, Dragana and Subotić, Angelina and Jevremović, Slađana and Cingel, Aleksandar",
year = "2018",
abstract = "Viola cornuta L. ‘Lutea Splendens’ is a perennial ornamental plant with small yellow flowers that
naturally grows in the Pyrenees in Spain and France. To develop novel cultivars with orange and
red flower colors, we performed Agrobacterium tumefaciens (LBA4404)-mediated transformation
with the binary vector pWBVec10a/P35S::Llccs::TNos that harbored capsanthin-capsorubin synthase
(Llccs) gene from Lilium lancifolium under the control of CaMV35S constitutive promoter and
the nopaline synthase (Nos) terminator. Capsanthin-capsorubin synthase catalyzes the conversion
of anteraxanthin and violaxanthin, two yellow ubiquitous 5-6-epoxy-xanthophylls, into capsanthin
and capsorubin, two red xanthophylls, respectively. Starting with hypocotyl explants, we developed
a transformation protocol with 0.3% shoot regeneration efficiency. Histochemical assay for
β-glucuronidase (GUS) activity showed uidA reporter gene expression in all putative Llccs-transgenic
shoots. The presence of Llccs transgene, hygromycin phosphotransferase (hpt) selectable
marker gene and uidA (GUS) reporter gene in all putative Llccs-transgenic lines were confirmed
by PCR analysis. This is the first report on Agrobacterium-mediated genetic transformation of V.
cornuta L. with the aim to introduce desirable traits into this species.",
publisher = "Belgrade: Serbian Plant Physiology Society, Institute for Biological Research “Siniša Stanković”, University of Belgrade Faculty of Biology, University of Belgrade",
journal = "Book of abstracts. 3rd International Conference on Plant Biology (22nd SPPS Meeting); 2018 Jun 9-12; Belgrade, Serbia",
title = "Agrobacterium-mediated genetic transformation of Viola cornuta L. "Lutea Splendens" with capsanthin-capsorubin synthase gene",
pages = "152",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_6540"
}

Trajković, M., Jeknić, Z., Antonić Reljin, D., Subotić, A., Jevremović, S.,& Cingel, A.. (2018). Agrobacterium-mediated genetic transformation of Viola cornuta L. "Lutea Splendens" with capsanthin-capsorubin synthase gene. in Book of abstracts. 3rd International Conference on Plant Biology (22nd SPPS Meeting); 2018 Jun 9-12; Belgrade, Serbia
Belgrade: Serbian Plant Physiology Society, Institute for Biological Research “Siniša Stanković”, University of Belgrade Faculty of Biology, University of Belgrade., 152.
https://hdl.handle.net/21.15107/rcub_ibiss_6540

Trajković M, Jeknić Z, Antonić Reljin D, Subotić A, Jevremović S, Cingel A. Agrobacterium-mediated genetic transformation of Viola cornuta L. "Lutea Splendens" with capsanthin-capsorubin synthase gene. in Book of abstracts. 3rd International Conference on Plant Biology (22nd SPPS Meeting); 2018 Jun 9-12; Belgrade, Serbia. 2018;:152.
https://hdl.handle.net/21.15107/rcub_ibiss_6540 .

Trajković, Milena, Jeknić, Zoran, Antonić Reljin, Dragana, Subotić, Angelina, Jevremović, Slađana, Cingel, Aleksandar, "Agrobacterium-mediated genetic transformation of Viola cornuta L. "Lutea Splendens" with capsanthin-capsorubin synthase gene" in Book of abstracts. 3rd International Conference on Plant Biology (22nd SPPS Meeting); 2018 Jun 9-12; Belgrade, Serbia (2018):152,
https://hdl.handle.net/21.15107/rcub_ibiss_6540 .

TY  - CONF
AU  - Trajković, Milena
AU  - Jeknić, Zoran
AU  - Antonić Reljin, Dragana
AU  - Subotić, Angelina
AU  - Jevremović, Slađana
AU  - Cingel, Aleksandar
PY  - 2017
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6731
AB  - Introduction: Viola cornuta is a valuable perennial ornamental plant. Development of new traits, such as new flower color with classical breeding suffers from many difficulties, which can be overcome using genetic engineering. With aim to develop protocol for Agrobacterium-mediated transformation of V. cornuta, we used Agrobacterium tumefaciens strain LBA4404 harbouring the superbinary vector pTOK233 carried a GUS reporter gene and hygromycin phosphotransferase selectable marker gene.
Methods: Hypocotyl explants obtained from seedlings were grown on ½MS medium supplemented with 0.1 mg/l 2,4-D and 2.0 mg/l BA for shoot induction. After two days of pre-cultivation, hypocotyl explants were inoculated in bacterial suspension for 15 min and placed on the same culture medium with addition of acetosyringone 100 μM at pH 5.2. After two days of co-cultivation, explants were transferred on shoot induction medium supplemented with cefotaxime and hygromycin B for selection. Regenerated putative transformants were analyzed by PCR for hygromycin phsphotransferase gene presence and by histochemical assay for β-glucuronidase (GUS) activity.
Results: Shoots were obtained within 8 weeks after explants were inoculated with A. tumefaciens, with 2.0% regeneration efficiency. PCR analysis confirmed selectable marker gene presence in twelve out of sixteen (75.0%) independently derived putatively transformed lines that were tested. Additionally, all analyzed lines exhibited a notable β-glucuronidase activity that was not present in untransformed plants.
Conclusion: This is the first report about V. cornuta susceptibility to A. tumefaciens. Presented protocol for genetic transformation can be used for further introduction of desirable traits in V. cornuta cultivars.
PB  - Belgrade: University of Belgrade, Faculty of Biology
C3  - Book of Abstracts: 1st Congress of Molecular Biologists of Serbia: CoMBoS; 2017 Sep 20-21; Belgrade, Serbia
T1  - Agrobacterium-mediated genetic transformation of Viola cornuta
SP  - 83
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_6731
ER  - 

@conference{
author = "Trajković, Milena and Jeknić, Zoran and Antonić Reljin, Dragana and Subotić, Angelina and Jevremović, Slađana and Cingel, Aleksandar",
year = "2017",
abstract = "Introduction: Viola cornuta is a valuable perennial ornamental plant. Development of new traits, such as new flower color with classical breeding suffers from many difficulties, which can be overcome using genetic engineering. With aim to develop protocol for Agrobacterium-mediated transformation of V. cornuta, we used Agrobacterium tumefaciens strain LBA4404 harbouring the superbinary vector pTOK233 carried a GUS reporter gene and hygromycin phosphotransferase selectable marker gene.
Methods: Hypocotyl explants obtained from seedlings were grown on ½MS medium supplemented with 0.1 mg/l 2,4-D and 2.0 mg/l BA for shoot induction. After two days of pre-cultivation, hypocotyl explants were inoculated in bacterial suspension for 15 min and placed on the same culture medium with addition of acetosyringone 100 μM at pH 5.2. After two days of co-cultivation, explants were transferred on shoot induction medium supplemented with cefotaxime and hygromycin B for selection. Regenerated putative transformants were analyzed by PCR for hygromycin phsphotransferase gene presence and by histochemical assay for β-glucuronidase (GUS) activity.
Results: Shoots were obtained within 8 weeks after explants were inoculated with A. tumefaciens, with 2.0% regeneration efficiency. PCR analysis confirmed selectable marker gene presence in twelve out of sixteen (75.0%) independently derived putatively transformed lines that were tested. Additionally, all analyzed lines exhibited a notable β-glucuronidase activity that was not present in untransformed plants.
Conclusion: This is the first report about V. cornuta susceptibility to A. tumefaciens. Presented protocol for genetic transformation can be used for further introduction of desirable traits in V. cornuta cultivars.",
publisher = "Belgrade: University of Belgrade, Faculty of Biology",
journal = "Book of Abstracts: 1st Congress of Molecular Biologists of Serbia: CoMBoS; 2017 Sep 20-21; Belgrade, Serbia",
title = "Agrobacterium-mediated genetic transformation of Viola cornuta",
pages = "83",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_6731"
}

Trajković, M., Jeknić, Z., Antonić Reljin, D., Subotić, A., Jevremović, S.,& Cingel, A.. (2017). Agrobacterium-mediated genetic transformation of Viola cornuta. in Book of Abstracts: 1st Congress of Molecular Biologists of Serbia: CoMBoS; 2017 Sep 20-21; Belgrade, Serbia
Belgrade: University of Belgrade, Faculty of Biology., 83.
https://hdl.handle.net/21.15107/rcub_ibiss_6731

Trajković M, Jeknić Z, Antonić Reljin D, Subotić A, Jevremović S, Cingel A. Agrobacterium-mediated genetic transformation of Viola cornuta. in Book of Abstracts: 1st Congress of Molecular Biologists of Serbia: CoMBoS; 2017 Sep 20-21; Belgrade, Serbia. 2017;:83.
https://hdl.handle.net/21.15107/rcub_ibiss_6731 .

Trajković, Milena, Jeknić, Zoran, Antonić Reljin, Dragana, Subotić, Angelina, Jevremović, Slađana, Cingel, Aleksandar, "Agrobacterium-mediated genetic transformation of Viola cornuta" in Book of Abstracts: 1st Congress of Molecular Biologists of Serbia: CoMBoS; 2017 Sep 20-21; Belgrade, Serbia (2017):83,
https://hdl.handle.net/21.15107/rcub_ibiss_6731 .

TY  - CONF
AU  - Trajković, Milena
AU  - Antonić Reljin, Dragana
AU  - Jeknić, Zoran
AU  - Jeknić, Stevan
AU  - Cingel, Aleksandar
AU  - Subotić, Angelina
AU  - Jevremović, Slađana
PY  - 2015
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6779
AB  - We developed an efficient plant regeneration protocol for a yellow cultivar of Viola cornuta ‘Lutea Splendens’.
Calli and shoots were induced from seedling explants (petiole, leaf, hypocotyls) by one month of culture
on ½-strength MS medium supplemented with 2,4-D (0.1 mg L-1) and BA (2.0 mg L-1). The highest frequency
of shoot induction (12%) was obtained from petiole explants, while the highest frequency of callus formation
(97%) was achieved on hypocotyl explants. The effects of explant origin (top, center, and bottom of hypocotyl)
and light/dark conditions on shoot and callus induction in hypocotyl cultures were also investigated. The
highest shoot induction in hypocotyl cultures (30%) was achieved when explants from the upper section of
the hypocotyls (near the epicotyls) were cultured in light conditions. Shoot multiplication from all explants
was achieved by transferring initially formed shoots to ½strength MS medium supplemented with NAA (0.2
mg L-1), GA3 (2.0 mg L-1), and TDZ (1.0 mg L-1) for four weeks of culture. Developed shoots were then further
multiplied on ½strength MS medium supplemented with NAA (0.5 mg L-1) and BA (1.0 mg L-1). Regenerated
shoots successfully rooted on ½-strength MS medium without growth regulators, acclimatized in a greenhouse
in autumn, and bloomed the following spring. All the regenerated plants exhibited normal morphology
and flower color. This regeneration protocol could be useful for mass propagation using in vitro cultures
and genetic transformation studies using seedling explants.
PB  - Belgrade: Serbian Plant Physiology Society, Institute for Biological Research “Siniša Stanković”, University of Belgrade
C3  - Book of Abstracts: 2nd International Conference on Plant Biology, 21th Symposium of the Serbian Plant Physiology Society, and CОST Action FA1106 Quality Fruit Workshop; 2015 Jun 17-20; Petnica, Serbia
T1  - Efficient plant regeneration of Viola cornuta ‘Lutea Splendens’ L. using seedling explants
SP  - 25
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_6779
ER  - 

@conference{
author = "Trajković, Milena and Antonić Reljin, Dragana and Jeknić, Zoran and Jeknić, Stevan and Cingel, Aleksandar and Subotić, Angelina and Jevremović, Slađana",
year = "2015",
abstract = "We developed an efficient plant regeneration protocol for a yellow cultivar of Viola cornuta ‘Lutea Splendens’.
Calli and shoots were induced from seedling explants (petiole, leaf, hypocotyls) by one month of culture
on ½-strength MS medium supplemented with 2,4-D (0.1 mg L-1) and BA (2.0 mg L-1). The highest frequency
of shoot induction (12%) was obtained from petiole explants, while the highest frequency of callus formation
(97%) was achieved on hypocotyl explants. The effects of explant origin (top, center, and bottom of hypocotyl)
and light/dark conditions on shoot and callus induction in hypocotyl cultures were also investigated. The
highest shoot induction in hypocotyl cultures (30%) was achieved when explants from the upper section of
the hypocotyls (near the epicotyls) were cultured in light conditions. Shoot multiplication from all explants
was achieved by transferring initially formed shoots to ½strength MS medium supplemented with NAA (0.2
mg L-1), GA3 (2.0 mg L-1), and TDZ (1.0 mg L-1) for four weeks of culture. Developed shoots were then further
multiplied on ½strength MS medium supplemented with NAA (0.5 mg L-1) and BA (1.0 mg L-1). Regenerated
shoots successfully rooted on ½-strength MS medium without growth regulators, acclimatized in a greenhouse
in autumn, and bloomed the following spring. All the regenerated plants exhibited normal morphology
and flower color. This regeneration protocol could be useful for mass propagation using in vitro cultures
and genetic transformation studies using seedling explants.",
publisher = "Belgrade: Serbian Plant Physiology Society, Institute for Biological Research “Siniša Stanković”, University of Belgrade",
journal = "Book of Abstracts: 2nd International Conference on Plant Biology, 21th Symposium of the Serbian Plant Physiology Society, and CОST Action FA1106 Quality Fruit Workshop; 2015 Jun 17-20; Petnica, Serbia",
title = "Efficient plant regeneration of Viola cornuta ‘Lutea Splendens’ L. using seedling explants",
pages = "25",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_6779"
}

Trajković, M., Antonić Reljin, D., Jeknić, Z., Jeknić, S., Cingel, A., Subotić, A.,& Jevremović, S.. (2015). Efficient plant regeneration of Viola cornuta ‘Lutea Splendens’ L. using seedling explants. in Book of Abstracts: 2nd International Conference on Plant Biology, 21th Symposium of the Serbian Plant Physiology Society, and CОST Action FA1106 Quality Fruit Workshop; 2015 Jun 17-20; Petnica, Serbia
Belgrade: Serbian Plant Physiology Society, Institute for Biological Research “Siniša Stanković”, University of Belgrade., 25.
https://hdl.handle.net/21.15107/rcub_ibiss_6779

Trajković M, Antonić Reljin D, Jeknić Z, Jeknić S, Cingel A, Subotić A, Jevremović S. Efficient plant regeneration of Viola cornuta ‘Lutea Splendens’ L. using seedling explants. in Book of Abstracts: 2nd International Conference on Plant Biology, 21th Symposium of the Serbian Plant Physiology Society, and CОST Action FA1106 Quality Fruit Workshop; 2015 Jun 17-20; Petnica, Serbia. 2015;:25.
https://hdl.handle.net/21.15107/rcub_ibiss_6779 .

Trajković, Milena, Antonić Reljin, Dragana, Jeknić, Zoran, Jeknić, Stevan, Cingel, Aleksandar, Subotić, Angelina, Jevremović, Slađana, "Efficient plant regeneration of Viola cornuta ‘Lutea Splendens’ L. using seedling explants" in Book of Abstracts: 2nd International Conference on Plant Biology, 21th Symposium of the Serbian Plant Physiology Society, and CОST Action FA1106 Quality Fruit Workshop; 2015 Jun 17-20; Petnica, Serbia (2015):25,
https://hdl.handle.net/21.15107/rcub_ibiss_6779 .

TY  - CONF
AU  - Jeknić, Stevan
AU  - Jevremović, Slađana
AU  - Trajković, Milena
AU  - Subotić, Angelina
AU  - Jeknić, Zoran
PY  - 2015
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6778
AB  - The elusive red iris (I. germanica, I. hollandica) has been a subject of interest for over a century as it does
not exist naturally and efforts with classical breeding methods have failed to produce it. Genetic engineering,
on the other hand, offers alternative ways of introducing this desirable trait by expanding the color-determining
gene pool that is available. We first studied the effects of ectopic expression of a bacterial phytoene synthase
gene (crtB) from Pantoea agglomerans in the pink I. germanica cultivar ‘Fire Bride’. This approach aimed
to increase the flux of metabolites into the carotenoid biosynthetic pathway, and ultimately lead to elevated
accumulation of lycopene, thus generating darker pink or red flowers. CrtB-transgenic plants showed prominent
color changes in the ovaries (green to orange), flower stalk (green to orange), and anthers (white to
pink) when compared to control plants, while the standards and falls showed no significant color change.
Next, we studied the feasibility of using a gene for capsanthin-capsorubin synthase (Llccs) from tiger lily (Lilium
lancifolium ‘Splendens’) in yellow iris flowers to alter their color to red. This approach aimed to produce
the flower-specific accumulation of two red κ-xanthophylls, capsanthin and capsorubin. Llccs-transgenic iriscalli successfully accumulated the novel carotenoids and changed color from yellow to red-orange, but no
plants were regenerated due to the lack of morphogenic response from the cultivar ‘Hot Property’. Efforts are
currently under way to transform different morphogenic yellow cultivars with Llccs to produce red I. germanica,
as well as I. hollandica, flowers.
PB  - Belgrade: Serbian Plant Physiology Society, Institute for Biological Research “Siniša Stanković”, University of Belgrade
C3  - Book of Abstracts: 2nd International Conference on Plant Biology, 21th Symposium of the Serbian Plant Physiology Society, and CОST Action FA1106 Quality Fruit Workshop; 2015 Jun 17-20; Petnica, Serbia
T1  - Recent advancements in the production of a red Iris flower through genetic engineering
SP  - 23
EP  - 24
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_6778
ER  - 

@conference{
author = "Jeknić, Stevan and Jevremović, Slađana and Trajković, Milena and Subotić, Angelina and Jeknić, Zoran",
year = "2015",
abstract = "The elusive red iris (I. germanica, I. hollandica) has been a subject of interest for over a century as it does
not exist naturally and efforts with classical breeding methods have failed to produce it. Genetic engineering,
on the other hand, offers alternative ways of introducing this desirable trait by expanding the color-determining
gene pool that is available. We first studied the effects of ectopic expression of a bacterial phytoene synthase
gene (crtB) from Pantoea agglomerans in the pink I. germanica cultivar ‘Fire Bride’. This approach aimed
to increase the flux of metabolites into the carotenoid biosynthetic pathway, and ultimately lead to elevated
accumulation of lycopene, thus generating darker pink or red flowers. CrtB-transgenic plants showed prominent
color changes in the ovaries (green to orange), flower stalk (green to orange), and anthers (white to
pink) when compared to control plants, while the standards and falls showed no significant color change.
Next, we studied the feasibility of using a gene for capsanthin-capsorubin synthase (Llccs) from tiger lily (Lilium
lancifolium ‘Splendens’) in yellow iris flowers to alter their color to red. This approach aimed to produce
the flower-specific accumulation of two red κ-xanthophylls, capsanthin and capsorubin. Llccs-transgenic iriscalli successfully accumulated the novel carotenoids and changed color from yellow to red-orange, but no
plants were regenerated due to the lack of morphogenic response from the cultivar ‘Hot Property’. Efforts are
currently under way to transform different morphogenic yellow cultivars with Llccs to produce red I. germanica,
as well as I. hollandica, flowers.",
publisher = "Belgrade: Serbian Plant Physiology Society, Institute for Biological Research “Siniša Stanković”, University of Belgrade",
journal = "Book of Abstracts: 2nd International Conference on Plant Biology, 21th Symposium of the Serbian Plant Physiology Society, and CОST Action FA1106 Quality Fruit Workshop; 2015 Jun 17-20; Petnica, Serbia",
title = "Recent advancements in the production of a red Iris flower through genetic engineering",
pages = "23-24",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_6778"
}

Jeknić, S., Jevremović, S., Trajković, M., Subotić, A.,& Jeknić, Z.. (2015). Recent advancements in the production of a red Iris flower through genetic engineering. in Book of Abstracts: 2nd International Conference on Plant Biology, 21th Symposium of the Serbian Plant Physiology Society, and CОST Action FA1106 Quality Fruit Workshop; 2015 Jun 17-20; Petnica, Serbia
Belgrade: Serbian Plant Physiology Society, Institute for Biological Research “Siniša Stanković”, University of Belgrade., 23-24.
https://hdl.handle.net/21.15107/rcub_ibiss_6778

Jeknić S, Jevremović S, Trajković M, Subotić A, Jeknić Z. Recent advancements in the production of a red Iris flower through genetic engineering. in Book of Abstracts: 2nd International Conference on Plant Biology, 21th Symposium of the Serbian Plant Physiology Society, and CОST Action FA1106 Quality Fruit Workshop; 2015 Jun 17-20; Petnica, Serbia. 2015;:23-24.
https://hdl.handle.net/21.15107/rcub_ibiss_6778 .

Jeknić, Stevan, Jevremović, Slađana, Trajković, Milena, Subotić, Angelina, Jeknić, Zoran, "Recent advancements in the production of a red Iris flower through genetic engineering" in Book of Abstracts: 2nd International Conference on Plant Biology, 21th Symposium of the Serbian Plant Physiology Society, and CОST Action FA1106 Quality Fruit Workshop; 2015 Jun 17-20; Petnica, Serbia (2015):23-24,
https://hdl.handle.net/21.15107/rcub_ibiss_6778 .

TY  - JOUR
AU  - Jevremović, Slađana
AU  - Trajković, Milena
AU  - Jeknić, Zoran
AU  - Trifunović-Momčilov, Milana
AU  - Antonić Reljin, Dragana
AU  - Marković, Marija
AU  - Subotić, Angelina
AU  - Radojević, Ljiljana
PY  - 2015
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6411
AB  - An efficient propagation protocol by somatic embryogenesis and organogenesis of Balkan endemic iris, Iris reichenbachii, was achieved and clonal fidelity of regenerated plants evaluated. Both regeneration pathways were induced at the same time in zygotic embryo culture on Murashige & Skoog (MS) medium supplemented with 0.5-5.0 mM 2,4-dichlorophenoxy acetic acid (2,4-D) as the sole hormone. Embryogenic calli were further mantained on medium supplemented with 2,4-D and kinetin (Kn; 0.5 and 5.0 mM, respectively). Organogenic calli were selected and further cultured on MS media supplemented with 1-naphthaleneacetic acid (NAA) and benzyladenine (BA; 0.5 and 4.5 mM, respectively) for shoot initiation. Somatic embryos germinated and shoots rooted on MS plant growth regulator-free medium. Plants regenerated by both procceses were succesfully acclimatized in greenhouse conditions and flowered in the following flowering season. Some alterations in flower morphology were detected among plants regenerated by organogenesis. Flow cytometric analysis revealed that plants with altered morphology of flowers had the same ploidy level and genome size as plants collected from the natural habitat. A tetraploid plant was observed in the population of plants regenerated by somatic embryogenesis induced at a high concentration of 2,4-D (10.0 mM).
AB  - Prikazan je efikasan protokol za propagaciju somatskom embriogenezom i organogenezom Balkanske endemične perunike, Iris reichenbachii i procenjena je klonalna indentičnost dobijenih biljaka. Oba načina za regeneraciju u uslovima in vitro su postignuta u kulturi zrelih zigotskih embriona na Murashige & Skoog (MS) hranljivoj podlozi obogaćenoj sa 2,4-dihlorofenoksi sirćetnom kiselinom 0.5-5.0 mM (2,4-D) kao jedinim regulatorom rastenja. Dobijeni embriogeni kalusi su dalje gajeni na hranljivoj podlozi sa 2,4-D i kinetinom (Kn; 0.5 odnosno 5.0 mM). Formirani organogeni kalusi su dalje gajeni na MS hranljivoj podlozi obogaćenoj sa α-naftilsirćetnom kiselinom (NAA) i benziladeninom (BA; 0.5 odnosno 4.5 mM) kada je došlo do formiranja izdanaka. Klijanje somatskih embriona kao i ožiljavanje formiranih izdanaka postignuto je na MS hranljivoj podlozi bez biljnih regulatora rastenja. Biljke dobijene na oba načina su dalje uspešno aklimatizovane na uslove gajenja u stakleniku i cvetale su sledeće godine u proleće. Uočene su neke promene u morfologiji cvetova kod biljaka dobijenih procesom organogeneze. Na osnovu flow-citometrijske analize pokazano je da su biljke sa izmenjenom morfologijom cvetova imale isti nivo ploidnosti i veličinu genoma kao biljke iz prirode. U populaciji biljaka regenerisanih procesom somatske embriogeneze koja je indukovana na podlozi sa visokom koncentracijom 2,4-D (10.0 mM) jedna biljka je bila tetraploidna.
PB  - Belgrade: Institute of Botany and Botanical Garden Jevremovac
T2  - Botanica Serbica
T1  - In vitro propagation of Iris reichenbachii Heuff. and clonal fidelity of regenerated plants
T1  - In vitro propagacija Iris reichenbachii Heuff. i klonalna identičnost dobijenih biljaka
IS  - 2
VL  - 39
SP  - 129
EP  - 136
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_6411
ER  - 

@article{
author = "Jevremović, Slađana and Trajković, Milena and Jeknić, Zoran and Trifunović-Momčilov, Milana and Antonić Reljin, Dragana and Marković, Marija and Subotić, Angelina and Radojević, Ljiljana",
year = "2015",
abstract = "An efficient propagation protocol by somatic embryogenesis and organogenesis of Balkan endemic iris, Iris reichenbachii, was achieved and clonal fidelity of regenerated plants evaluated. Both regeneration pathways were induced at the same time in zygotic embryo culture on Murashige & Skoog (MS) medium supplemented with 0.5-5.0 mM 2,4-dichlorophenoxy acetic acid (2,4-D) as the sole hormone. Embryogenic calli were further mantained on medium supplemented with 2,4-D and kinetin (Kn; 0.5 and 5.0 mM, respectively). Organogenic calli were selected and further cultured on MS media supplemented with 1-naphthaleneacetic acid (NAA) and benzyladenine (BA; 0.5 and 4.5 mM, respectively) for shoot initiation. Somatic embryos germinated and shoots rooted on MS plant growth regulator-free medium. Plants regenerated by both procceses were succesfully acclimatized in greenhouse conditions and flowered in the following flowering season. Some alterations in flower morphology were detected among plants regenerated by organogenesis. Flow cytometric analysis revealed that plants with altered morphology of flowers had the same ploidy level and genome size as plants collected from the natural habitat. A tetraploid plant was observed in the population of plants regenerated by somatic embryogenesis induced at a high concentration of 2,4-D (10.0 mM)., Prikazan je efikasan protokol za propagaciju somatskom embriogenezom i organogenezom Balkanske endemične perunike, Iris reichenbachii i procenjena je klonalna indentičnost dobijenih biljaka. Oba načina za regeneraciju u uslovima in vitro su postignuta u kulturi zrelih zigotskih embriona na Murashige & Skoog (MS) hranljivoj podlozi obogaćenoj sa 2,4-dihlorofenoksi sirćetnom kiselinom 0.5-5.0 mM (2,4-D) kao jedinim regulatorom rastenja. Dobijeni embriogeni kalusi su dalje gajeni na hranljivoj podlozi sa 2,4-D i kinetinom (Kn; 0.5 odnosno 5.0 mM). Formirani organogeni kalusi su dalje gajeni na MS hranljivoj podlozi obogaćenoj sa α-naftilsirćetnom kiselinom (NAA) i benziladeninom (BA; 0.5 odnosno 4.5 mM) kada je došlo do formiranja izdanaka. Klijanje somatskih embriona kao i ožiljavanje formiranih izdanaka postignuto je na MS hranljivoj podlozi bez biljnih regulatora rastenja. Biljke dobijene na oba načina su dalje uspešno aklimatizovane na uslove gajenja u stakleniku i cvetale su sledeće godine u proleće. Uočene su neke promene u morfologiji cvetova kod biljaka dobijenih procesom organogeneze. Na osnovu flow-citometrijske analize pokazano je da su biljke sa izmenjenom morfologijom cvetova imale isti nivo ploidnosti i veličinu genoma kao biljke iz prirode. U populaciji biljaka regenerisanih procesom somatske embriogeneze koja je indukovana na podlozi sa visokom koncentracijom 2,4-D (10.0 mM) jedna biljka je bila tetraploidna.",
publisher = "Belgrade: Institute of Botany and Botanical Garden Jevremovac",
journal = "Botanica Serbica",
title = "In vitro propagation of Iris reichenbachii Heuff. and clonal fidelity of regenerated plants, In vitro propagacija Iris reichenbachii Heuff. i klonalna identičnost dobijenih biljaka",
number = "2",
volume = "39",
pages = "129-136",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_6411"
}

Jevremović, S., Trajković, M., Jeknić, Z., Trifunović-Momčilov, M., Antonić Reljin, D., Marković, M., Subotić, A.,& Radojević, L.. (2015). In vitro propagation of Iris reichenbachii Heuff. and clonal fidelity of regenerated plants. in Botanica Serbica
Belgrade: Institute of Botany and Botanical Garden Jevremovac., 39(2), 129-136.
https://hdl.handle.net/21.15107/rcub_ibiss_6411

Jevremović S, Trajković M, Jeknić Z, Trifunović-Momčilov M, Antonić Reljin D, Marković M, Subotić A, Radojević L. In vitro propagation of Iris reichenbachii Heuff. and clonal fidelity of regenerated plants. in Botanica Serbica. 2015;39(2):129-136.
https://hdl.handle.net/21.15107/rcub_ibiss_6411 .

Jevremović, Slađana, Trajković, Milena, Jeknić, Zoran, Trifunović-Momčilov, Milana, Antonić Reljin, Dragana, Marković, Marija, Subotić, Angelina, Radojević, Ljiljana, "In vitro propagation of Iris reichenbachii Heuff. and clonal fidelity of regenerated plants" in Botanica Serbica, 39, no. 2 (2015):129-136,
https://hdl.handle.net/21.15107/rcub_ibiss_6411 .

TY  - JOUR
AU  - Jeknić, Zoran
AU  - Morre, Jeffrey T
AU  - Jeknić, Stevan
AU  - Jevremović, Slađana
AU  - Subotić, Angelina
AU  - Chen, Tony HH
PY  - 2012
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/1089
AB  - The orange color of tiger lily (Lolium lancifolium 'Splendens') flowers is due, primarily, to the accumulation of two kappa-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 beta-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 kappa-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 kappa-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.
T2  - Plant and Cell Physiology
T1  - Cloning and Functional Characterization of a Gene for Capsanthin-Capsorubin Synthase from Tiger Lily (Lilium lancifolium Thunb. 'Splendens')
IS  - 11
VL  - 53
SP  - 299
EP  - 1912
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_1089
ER  - 

@article{
author = "Jeknić, Zoran and Morre, Jeffrey T and Jeknić, Stevan and Jevremović, Slađana and Subotić, Angelina and Chen, Tony HH",
year = "2012",
abstract = "The orange color of tiger lily (Lolium lancifolium 'Splendens') flowers is due, primarily, to the accumulation of two kappa-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 beta-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 kappa-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 kappa-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.",
journal = "Plant and Cell Physiology",
title = "Cloning and Functional Characterization of a Gene for Capsanthin-Capsorubin Synthase from Tiger Lily (Lilium lancifolium Thunb. 'Splendens')",
number = "11",
volume = "53",
pages = "299-1912",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_1089"
}

Jeknić, Z., Morre, J. T., Jeknić, S., Jevremović, S., Subotić, A.,& Chen, T. H.. (2012). Cloning and Functional Characterization of a Gene for Capsanthin-Capsorubin Synthase from Tiger Lily (Lilium lancifolium Thunb. 'Splendens'). in Plant and Cell Physiology, 53(11), 299-1912.
https://hdl.handle.net/21.15107/rcub_ibiss_1089

Jeknić Z, Morre JT, Jeknić S, Jevremović S, Subotić A, Chen TH. Cloning and Functional Characterization of a Gene for Capsanthin-Capsorubin Synthase from Tiger Lily (Lilium lancifolium Thunb. 'Splendens'). in Plant and Cell Physiology. 2012;53(11):299-1912.
https://hdl.handle.net/21.15107/rcub_ibiss_1089 .

Jeknić, Zoran, Morre, Jeffrey T, Jeknić, Stevan, Jevremović, Slađana, Subotić, Angelina, Chen, Tony HH, "Cloning and Functional Characterization of a Gene for Capsanthin-Capsorubin Synthase from Tiger Lily (Lilium lancifolium Thunb. 'Splendens')" in Plant and Cell Physiology, 53, no. 11 (2012):299-1912,
https://hdl.handle.net/21.15107/rcub_ibiss_1089 .