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

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

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

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

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

TY  - JOUR
AU  - Savić, Jelena
AU  - Nikolić, Radomirka
AU  - Banjac, Nevena
AU  - Zdravković-Korać, Snežana
AU  - Stupar, Sofija
AU  - Cingel, Aleksandar
AU  - Ćosić, Tatjana
AU  - Raspor, Martin
AU  - Smigocki, Ann
AU  - Ninković, Slavica
PY  - 2019
UR  - https://www.sciencedirect.com/science/article/pii/S0176161719301786?via%3Dihub
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/3490
AB  - Food demands of increasing human population dictate intensification of livestock production, however, environmental stresses could jeopardize producers' efforts. Forage legumes suffer from yield losses and poor nutritional status due to salinity increase of agricultural soils. As tools aimed to reduce negative impacts of biotic or abiotic stresses, proteinase inhibitors (PIs) have been promoted for biotechnological improvements. In order to increase tolerance of Lotus corniculatus L. to salt stress, serine PI, BvSTI, was introduced into this legume using Agrobacterium rhizogenes, with final transformation efficiency of 4.57%. PCR, DNA gel-blot, RT-PCR and in-gel protein activity assays confirmed the presence and activity of BvSTI products in transformed lines. Plants from three selected transgenic lines (21, 73 and 109) showed significant alterations in overall phenotypic appearance, corresponding to differences in BvSTI accumulation. Lines 73 and 109 showed up to 7.3-fold higher number of tillers and massive, up to 5.8-fold heavier roots than in nontransformed controls (NTC). Line 21 was phenotypically similar to NTC, accumulated less BvSTI transcripts and did not exhibit an additional band of recombinant trypsin inhibitor as seen in lines 73 and 109. Exposure of the transgenic lines to NaCl revealed different levels of salt stress susceptibility. The NaCl sensitivity index, based on morphological appearance and chlorophyll concentrations showed that lines 73 and 109 were significantly less affected by salinity than NTC or line 21. High level of BvSTI altered morphology and delayed salt stress related senescence, implicating BvSTI gene as a promising tool for salinity tolerance improvement trials in L. corniculatus.
T2  - Journal of Plant Physiology
T1  - Beneficial implications of sugar beet proteinase inhibitor BvSTI on plant architecture and salt stress tolerance in Lotus corniculatus L.
VL  - 243
DO  - 10.1016/j.jplph.2019.153055
SP  - 153055
ER  - 

@article{
author = "Savić, Jelena and Nikolić, Radomirka and Banjac, Nevena and Zdravković-Korać, Snežana and Stupar, Sofija and Cingel, Aleksandar and Ćosić, Tatjana and Raspor, Martin and Smigocki, Ann and Ninković, Slavica",
year = "2019",
abstract = "Food demands of increasing human population dictate intensification of livestock production, however, environmental stresses could jeopardize producers' efforts. Forage legumes suffer from yield losses and poor nutritional status due to salinity increase of agricultural soils. As tools aimed to reduce negative impacts of biotic or abiotic stresses, proteinase inhibitors (PIs) have been promoted for biotechnological improvements. In order to increase tolerance of Lotus corniculatus L. to salt stress, serine PI, BvSTI, was introduced into this legume using Agrobacterium rhizogenes, with final transformation efficiency of 4.57%. PCR, DNA gel-blot, RT-PCR and in-gel protein activity assays confirmed the presence and activity of BvSTI products in transformed lines. Plants from three selected transgenic lines (21, 73 and 109) showed significant alterations in overall phenotypic appearance, corresponding to differences in BvSTI accumulation. Lines 73 and 109 showed up to 7.3-fold higher number of tillers and massive, up to 5.8-fold heavier roots than in nontransformed controls (NTC). Line 21 was phenotypically similar to NTC, accumulated less BvSTI transcripts and did not exhibit an additional band of recombinant trypsin inhibitor as seen in lines 73 and 109. Exposure of the transgenic lines to NaCl revealed different levels of salt stress susceptibility. The NaCl sensitivity index, based on morphological appearance and chlorophyll concentrations showed that lines 73 and 109 were significantly less affected by salinity than NTC or line 21. High level of BvSTI altered morphology and delayed salt stress related senescence, implicating BvSTI gene as a promising tool for salinity tolerance improvement trials in L. corniculatus.",
journal = "Journal of Plant Physiology",
title = "Beneficial implications of sugar beet proteinase inhibitor BvSTI on plant architecture and salt stress tolerance in Lotus corniculatus L.",
volume = "243",
doi = "10.1016/j.jplph.2019.153055",
pages = "153055"
}

Savić, J., Nikolić, R., Banjac, N., Zdravković-Korać, S., Stupar, S., Cingel, A., Ćosić, T., Raspor, M., Smigocki, A.,& Ninković, S.. (2019). Beneficial implications of sugar beet proteinase inhibitor BvSTI on plant architecture and salt stress tolerance in Lotus corniculatus L.. in Journal of Plant Physiology, 243, 153055.
https://doi.org/10.1016/j.jplph.2019.153055

Savić J, Nikolić R, Banjac N, Zdravković-Korać S, Stupar S, Cingel A, Ćosić T, Raspor M, Smigocki A, Ninković S. Beneficial implications of sugar beet proteinase inhibitor BvSTI on plant architecture and salt stress tolerance in Lotus corniculatus L.. in Journal of Plant Physiology. 2019;243:153055.
doi:10.1016/j.jplph.2019.153055 .

Savić, Jelena, Nikolić, Radomirka, Banjac, Nevena, Zdravković-Korać, Snežana, Stupar, Sofija, Cingel, Aleksandar, Ćosić, Tatjana, Raspor, Martin, Smigocki, Ann, Ninković, Slavica, "Beneficial implications of sugar beet proteinase inhibitor BvSTI on plant architecture and salt stress tolerance in Lotus corniculatus L." in Journal of Plant Physiology, 243 (2019):153055,
https://doi.org/10.1016/j.jplph.2019.153055 . .

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

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

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

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

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

TY  - GEN
AU  - Cingel, Aleksandar
AU  - Savić, Jelena
AU  - Lazarević, Jelica
AU  - Ćosić, Tatjana
AU  - Raspor, Martin
AU  - Smigocki, Ann
AU  - Ninković, Slavica
PY  - 2017
UR  - http://doi.wiley.com/10.1111/1744-7917.12364
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/2529
AB  - Colorado potato beetle (CPB; Leptinotarsa decemlineata Say, Coleoptera: Chrysomelidae) has shown a remarkable adaptability to a variety of control measures. Although oryzacystatin I and II (OCI and OCII) have potential in controlling pests that use cysteine proteinases for food digestion, expression of a single OC gene in potato exhibited a minimal or no effect on CPB fitness traits. The aim of this study was to examine the effect of coexpressed OCI and OCII in potato (Solanum tuberosum L.) cultivars Desiree, Dragačevka and Jelica on CPB larvae. Growth parameters, consumption rates and food utilization, as well as activity of proteases of CPB larvae were assayed. Second and third instar larvae fed on transformed leaves molted earlier and had higher relative growth and consumption rates than larvae fed on nontransformed leaves, while efficiency of food utilization was unaffected. In contrast, fourth instar maximum weight gain and amount of leaves consumed were about 20% lower for the larvae fed on transgenic potato. Analysis of total protease activity of third instar larvae revealed reduction in overall proteolytic activity measured by azocasein hydrolysis, accompanied with inhibition of cysteine proteinase activity 24 h after ingestion of potato leaves expressing OCI and OCII. However, after long-term feeding on transformed leaves proteolytic activities of larvae became similar to the controls. Although feeding on OCI/OCII leaves did not affect larval survival, coexpression of OC genes reduced the development time and thus significantly decreased plant damage caused by CPB larvae.
PB  - Blackwell Publishing Ltd
T2  - Insect Science
T1  - Co-expression of the proteinase inhibitors oryzacystatin I and oryzacystatin II in transgenic potato alters Colorado potato beetle larval development
IS  - 5
VL  - 24
DO  - 10.1111/1744-7917.12364
SP  - 768
EP  - 780
ER  - 

@misc{
author = "Cingel, Aleksandar and Savić, Jelena and Lazarević, Jelica and Ćosić, Tatjana and Raspor, Martin and Smigocki, Ann and Ninković, Slavica",
year = "2017",
abstract = "Colorado potato beetle (CPB; Leptinotarsa decemlineata Say, Coleoptera: Chrysomelidae) has shown a remarkable adaptability to a variety of control measures. Although oryzacystatin I and II (OCI and OCII) have potential in controlling pests that use cysteine proteinases for food digestion, expression of a single OC gene in potato exhibited a minimal or no effect on CPB fitness traits. The aim of this study was to examine the effect of coexpressed OCI and OCII in potato (Solanum tuberosum L.) cultivars Desiree, Dragačevka and Jelica on CPB larvae. Growth parameters, consumption rates and food utilization, as well as activity of proteases of CPB larvae were assayed. Second and third instar larvae fed on transformed leaves molted earlier and had higher relative growth and consumption rates than larvae fed on nontransformed leaves, while efficiency of food utilization was unaffected. In contrast, fourth instar maximum weight gain and amount of leaves consumed were about 20% lower for the larvae fed on transgenic potato. Analysis of total protease activity of third instar larvae revealed reduction in overall proteolytic activity measured by azocasein hydrolysis, accompanied with inhibition of cysteine proteinase activity 24 h after ingestion of potato leaves expressing OCI and OCII. However, after long-term feeding on transformed leaves proteolytic activities of larvae became similar to the controls. Although feeding on OCI/OCII leaves did not affect larval survival, coexpression of OC genes reduced the development time and thus significantly decreased plant damage caused by CPB larvae.",
publisher = "Blackwell Publishing Ltd",
journal = "Insect Science",
title = "Co-expression of the proteinase inhibitors oryzacystatin I and oryzacystatin II in transgenic potato alters Colorado potato beetle larval development",
number = "5",
volume = "24",
doi = "10.1111/1744-7917.12364",
pages = "768-780"
}

Cingel, A., Savić, J., Lazarević, J., Ćosić, T., Raspor, M., Smigocki, A.,& Ninković, S.. (2017). Co-expression of the proteinase inhibitors oryzacystatin I and oryzacystatin II in transgenic potato alters Colorado potato beetle larval development. in Insect Science
Blackwell Publishing Ltd., 24(5), 768-780.
https://doi.org/10.1111/1744-7917.12364

Cingel A, Savić J, Lazarević J, Ćosić T, Raspor M, Smigocki A, Ninković S. Co-expression of the proteinase inhibitors oryzacystatin I and oryzacystatin II in transgenic potato alters Colorado potato beetle larval development. in Insect Science. 2017;24(5):768-780.
doi:10.1111/1744-7917.12364 .

Cingel, Aleksandar, Savić, Jelena, Lazarević, Jelica, Ćosić, Tatjana, Raspor, Martin, Smigocki, Ann, Ninković, Slavica, "Co-expression of the proteinase inhibitors oryzacystatin I and oryzacystatin II in transgenic potato alters Colorado potato beetle larval development" in Insect Science, 24, no. 5 (2017):768-780,
https://doi.org/10.1111/1744-7917.12364 . .

TY  - JOUR
AU  - Cingel, Aleksandar
AU  - Savić, Jelena
AU  - Lazarević, Jelica
AU  - Ćosić, Tatjana
AU  - Raspor, Martin
AU  - Smigocki, Ann
AU  - Ninković, Slavica
PY  - 2016
UR  - http://www.mdpi.com/1422-0067/17/9/1538
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/2528
AB  - Expanding from remote areas of Mexico to a worldwide scale, the ten-striped insect, the Colorado potato beetle (CPB, Leptinotarsa decemlineata Say), has risen from being an innocuous beetle to a prominent global pest. A diverse life cycle, phenotypic plasticity, adaptation to adverse conditions, and capability to detoxify or tolerate toxins make this insect appear to be virtually “indestructible”. With increasing advances in molecular biology, tools of biotechnological warfare were deployed to combat CPB. In the last three decades, genetically modified potato has created a new challenge for the beetle. After reviewing hundreds of scientific papers dealing with CPB control, it became clear that even biotechnological means of control, if used alone, would not defeat the Colorado potato beetle. This control measure once again appears to be provoking the potato beetle to exhibit its remarkable adaptability. Nonetheless, the potential for adaptation to these techniques has increased our knowledge of this pest and thus opened possibilities for devising more sustainable CPB management programs.
T2  - International Journal of Molecular Sciences
T1  - Extraordinary Adaptive Plasticity of Colorado Potato Beetle: “Ten-Striped Spearman” in the Era of Biotechnological Warfare
IS  - 9
VL  - 17
DO  - 10.3390/ijms17091538
SP  - 1538
EP  - 1538
ER  - 

@article{
author = "Cingel, Aleksandar and Savić, Jelena and Lazarević, Jelica and Ćosić, Tatjana and Raspor, Martin and Smigocki, Ann and Ninković, Slavica",
year = "2016",
abstract = "Expanding from remote areas of Mexico to a worldwide scale, the ten-striped insect, the Colorado potato beetle (CPB, Leptinotarsa decemlineata Say), has risen from being an innocuous beetle to a prominent global pest. A diverse life cycle, phenotypic plasticity, adaptation to adverse conditions, and capability to detoxify or tolerate toxins make this insect appear to be virtually “indestructible”. With increasing advances in molecular biology, tools of biotechnological warfare were deployed to combat CPB. In the last three decades, genetically modified potato has created a new challenge for the beetle. After reviewing hundreds of scientific papers dealing with CPB control, it became clear that even biotechnological means of control, if used alone, would not defeat the Colorado potato beetle. This control measure once again appears to be provoking the potato beetle to exhibit its remarkable adaptability. Nonetheless, the potential for adaptation to these techniques has increased our knowledge of this pest and thus opened possibilities for devising more sustainable CPB management programs.",
journal = "International Journal of Molecular Sciences",
title = "Extraordinary Adaptive Plasticity of Colorado Potato Beetle: “Ten-Striped Spearman” in the Era of Biotechnological Warfare",
number = "9",
volume = "17",
doi = "10.3390/ijms17091538",
pages = "1538-1538"
}

Cingel, A., Savić, J., Lazarević, J., Ćosić, T., Raspor, M., Smigocki, A.,& Ninković, S.. (2016). Extraordinary Adaptive Plasticity of Colorado Potato Beetle: “Ten-Striped Spearman” in the Era of Biotechnological Warfare. in International Journal of Molecular Sciences, 17(9), 1538-1538.
https://doi.org/10.3390/ijms17091538

Cingel A, Savić J, Lazarević J, Ćosić T, Raspor M, Smigocki A, Ninković S. Extraordinary Adaptive Plasticity of Colorado Potato Beetle: “Ten-Striped Spearman” in the Era of Biotechnological Warfare. in International Journal of Molecular Sciences. 2016;17(9):1538-1538.
doi:10.3390/ijms17091538 .

Cingel, Aleksandar, Savić, Jelena, Lazarević, Jelica, Ćosić, Tatjana, Raspor, Martin, Smigocki, Ann, Ninković, Slavica, "Extraordinary Adaptive Plasticity of Colorado Potato Beetle: “Ten-Striped Spearman” in the Era of Biotechnological Warfare" in International Journal of Molecular Sciences, 17, no. 9 (2016):1538-1538,
https://doi.org/10.3390/ijms17091538 . .

TY  - JOUR
AU  - Vinterhalter, Branka
AU  - Krstić Milošević, Dijana
AU  - Jankovic, Teodora
AU  - Pljevljakusic, Dejan
AU  - Ninković, Slavica
AU  - Smigocki, Ann
AU  - Vinterhalter, Dragan
PY  - 2015
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/1952
AB  - The induction and establishment of hairy root cultures of Gentiana
   dinarica using two strains of Agrobacterium rhizogenes (A4M70GUS and
   15834/PI) is reported for the first time. Strain 15834/PI had higher
   induction rate of hairy roots (32.15 \%) than strain A4M70GUS (6.12 \%).
   Transgenic nature of the roots was confirmed by GUS assay and PCR
   analysis. Two clones per strain (A4M70GUS-D and -I, and 15834/PI-2 and
   -3) with marked differences in general morphology and growth rate were
   further studied. The methanol extracts of hairy root clones were
   analyzed for xanthones content using HPLC method. The effects of the
   type of carbohydrate source (sucrose, fructose or glucose) at different
   concentrations on the growth parameters (growth index, dry weight,
   fresh/dry weight ratio), phenolic and xanthone production, and free
   radical scavenging activity of the transgenic clones were evaluated.
   Statistical two level factorial design was used to define optimal
   conditions for growth and successful secondary metabolite production in
   G. dinarica hairy root clones. The results showed that clones A4M70GUS-D
   and 15834/PI-3 were the superior ones. These two clones had the highest
   dry weight on 116.8 mM sucrose, producing up to threefold higher amounts
   of total phenolics and norswertianin-1-O-primeveroside than other
   clones, untransformed roots and roots of wild-grown plants.
T2  - Plant Cell Tissue and Organ Culture
T1  - Gentiana dinarica Beck. hairy root cultures and evaluation of factors
 affecting growth and xanthone production
IS  - 3
VL  - 121
DO  - 10.1007/s11240-015-0737-z
SP  - 667
EP  - 679
ER  - 

@article{
author = "Vinterhalter, Branka and Krstić Milošević, Dijana and Jankovic, Teodora and Pljevljakusic, Dejan and Ninković, Slavica and Smigocki, Ann and Vinterhalter, Dragan",
year = "2015",
abstract = "The induction and establishment of hairy root cultures of Gentiana
   dinarica using two strains of Agrobacterium rhizogenes (A4M70GUS and
   15834/PI) is reported for the first time. Strain 15834/PI had higher
   induction rate of hairy roots (32.15 \%) than strain A4M70GUS (6.12 \%).
   Transgenic nature of the roots was confirmed by GUS assay and PCR
   analysis. Two clones per strain (A4M70GUS-D and -I, and 15834/PI-2 and
   -3) with marked differences in general morphology and growth rate were
   further studied. The methanol extracts of hairy root clones were
   analyzed for xanthones content using HPLC method. The effects of the
   type of carbohydrate source (sucrose, fructose or glucose) at different
   concentrations on the growth parameters (growth index, dry weight,
   fresh/dry weight ratio), phenolic and xanthone production, and free
   radical scavenging activity of the transgenic clones were evaluated.
   Statistical two level factorial design was used to define optimal
   conditions for growth and successful secondary metabolite production in
   G. dinarica hairy root clones. The results showed that clones A4M70GUS-D
   and 15834/PI-3 were the superior ones. These two clones had the highest
   dry weight on 116.8 mM sucrose, producing up to threefold higher amounts
   of total phenolics and norswertianin-1-O-primeveroside than other
   clones, untransformed roots and roots of wild-grown plants.",
journal = "Plant Cell Tissue and Organ Culture",
title = "Gentiana dinarica Beck. hairy root cultures and evaluation of factors
 affecting growth and xanthone production",
number = "3",
volume = "121",
doi = "10.1007/s11240-015-0737-z",
pages = "667-679"
}

Vinterhalter, B., Krstić Milošević, D., Jankovic, T., Pljevljakusic, D., Ninković, S., Smigocki, A.,& Vinterhalter, D.. (2015). Gentiana dinarica Beck. hairy root cultures and evaluation of factors
 affecting growth and xanthone production. in Plant Cell Tissue and Organ Culture, 121(3), 667-679.
https://doi.org/10.1007/s11240-015-0737-z

Vinterhalter B, Krstić Milošević D, Jankovic T, Pljevljakusic D, Ninković S, Smigocki A, Vinterhalter D. Gentiana dinarica Beck. hairy root cultures and evaluation of factors
 affecting growth and xanthone production. in Plant Cell Tissue and Organ Culture. 2015;121(3):667-679.
doi:10.1007/s11240-015-0737-z .

Vinterhalter, Branka, Krstić Milošević, Dijana, Jankovic, Teodora, Pljevljakusic, Dejan, Ninković, Slavica, Smigocki, Ann, Vinterhalter, Dragan, "Gentiana dinarica Beck. hairy root cultures and evaluation of factors
 affecting growth and xanthone production" in Plant Cell Tissue and Organ Culture, 121, no. 3 (2015):667-679,
https://doi.org/10.1007/s11240-015-0737-z . .

TY  - JOUR
AU  - Cingel, Aleksandar
AU  - Savic, Jelena
AU  - Ćosić, Tatjana
AU  - Raspor, Martin
AU  - Ghalawenji, Nabil
AU  - Smigocki, Ann
AU  - Ninković, Slavica
PY  - 2015
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/2032
AB  - The evaluation of transgenic plants which is usually carried out under
   controlled conditions in culture rooms and greenhouses can yield
   valuable information about the influence of introduced genes on a
   transgenic plant phenotype. However, an overall assessment of plant
   performance can only be made by testing transgenic plants in the field
   environment. Thus, the effects of pyramided rice cystatin genes OCI and
   OCII on morphological parameters of transgenic potato cv. Desiree,
   Dragacevka and Jelica lines were compared under in vitro, greenhouse,
   and field conditions. All analyzed OC co-expressing transgenic lines
   exhibited normal phenotype, both in vitro and in greenhouse conditions.
   In the field environment, eight of nine OCI/OCII lines were similar to
   the wild-type control plants in their general phenotypic appearance.
   Yield parameters, such as tuber number and tuber weight for these
   phenotypically normal OCI/OCII lines, were also comparable to the
   controls. Only transgenic cv. Jelica line 4 plants exhibited slightly
   reduced growth, atypical leaf morphology and, contrary to the plants of
   other transgenic lines and untransformed controls, failed to flower.
   However, despite the phenotypic and developmental changes under field
   conditions, the OCI/OCII Jelica line 4 did not exhibit a significant
   decrease in tuber yield. Stacking of OCI and OCII genes preserves
   important attributes of the parental lines, confirming that this
   approach could be suitable for improving agronomical traits in potato.
T2  - Archives of Biological Sciences
T1  - Phenotypic performance of transgenic potato (Solanum tuberosum L.) plants with pyramided rice cystatin genes (OCI and OCII)
IS  - 3
VL  - 67
DO  - 10.2298/ABS141201058C
SP  - 957
EP  - 964
ER  - 

@article{
author = "Cingel, Aleksandar and Savic, Jelena and Ćosić, Tatjana and Raspor, Martin and Ghalawenji, Nabil and Smigocki, Ann and Ninković, Slavica",
year = "2015",
abstract = "The evaluation of transgenic plants which is usually carried out under
   controlled conditions in culture rooms and greenhouses can yield
   valuable information about the influence of introduced genes on a
   transgenic plant phenotype. However, an overall assessment of plant
   performance can only be made by testing transgenic plants in the field
   environment. Thus, the effects of pyramided rice cystatin genes OCI and
   OCII on morphological parameters of transgenic potato cv. Desiree,
   Dragacevka and Jelica lines were compared under in vitro, greenhouse,
   and field conditions. All analyzed OC co-expressing transgenic lines
   exhibited normal phenotype, both in vitro and in greenhouse conditions.
   In the field environment, eight of nine OCI/OCII lines were similar to
   the wild-type control plants in their general phenotypic appearance.
   Yield parameters, such as tuber number and tuber weight for these
   phenotypically normal OCI/OCII lines, were also comparable to the
   controls. Only transgenic cv. Jelica line 4 plants exhibited slightly
   reduced growth, atypical leaf morphology and, contrary to the plants of
   other transgenic lines and untransformed controls, failed to flower.
   However, despite the phenotypic and developmental changes under field
   conditions, the OCI/OCII Jelica line 4 did not exhibit a significant
   decrease in tuber yield. Stacking of OCI and OCII genes preserves
   important attributes of the parental lines, confirming that this
   approach could be suitable for improving agronomical traits in potato.",
journal = "Archives of Biological Sciences",
title = "Phenotypic performance of transgenic potato (Solanum tuberosum L.) plants with pyramided rice cystatin genes (OCI and OCII)",
number = "3",
volume = "67",
doi = "10.2298/ABS141201058C",
pages = "957-964"
}

Cingel, A., Savic, J., Ćosić, T., Raspor, M., Ghalawenji, N., Smigocki, A.,& Ninković, S.. (2015). Phenotypic performance of transgenic potato (Solanum tuberosum L.) plants with pyramided rice cystatin genes (OCI and OCII). in Archives of Biological Sciences, 67(3), 957-964.
https://doi.org/10.2298/ABS141201058C

Cingel A, Savic J, Ćosić T, Raspor M, Ghalawenji N, Smigocki A, Ninković S. Phenotypic performance of transgenic potato (Solanum tuberosum L.) plants with pyramided rice cystatin genes (OCI and OCII). in Archives of Biological Sciences. 2015;67(3):957-964.
doi:10.2298/ABS141201058C .

Cingel, Aleksandar, Savic, Jelena, Ćosić, Tatjana, Raspor, Martin, Ghalawenji, Nabil, Smigocki, Ann, Ninković, Slavica, "Phenotypic performance of transgenic potato (Solanum tuberosum L.) plants with pyramided rice cystatin genes (OCI and OCII)" in Archives of Biological Sciences, 67, no. 3 (2015):957-964,
https://doi.org/10.2298/ABS141201058C . .

TY  - JOUR
AU  - Cingel, Aleksandar
AU  - Savic, Jelena
AU  - Vinterhalter, Branka
AU  - Vinterhalter, Dragan
AU  - Kostic, Miroslav
AU  - Šešlija Jovanović, Darka
AU  - Smigocki, Ann
AU  - Ninković, Slavica
PY  - 2015
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/1923
AB  - Plant proteinase inhibitors (PIs) are attractive tools for crop
   improvement and their heterologous expression can enhance insect
   resistance in transgenic plants. PI oryzacystatin II (OCII), isolated
   from rice, showed potential in controlling pests that utilize cysteine
   proteinases for protein digestion. To evaluate the applicability of the
   OCII gene in enhancing plant defence, OCII-transformed potatoes were
   bioassayed for resistance to Colorado potato beetle (Leptinotarsa
   decemlineata Say). Feeding on transformed leaves of potato cultivars
   Desiree and Jelica significantly affected larval growth and development,
   but did not change mortality rates. During the L2 and L3 developmental
   stages larvae consumed the OCII-transformed foliage faster as compared
   to the nontransformed control. Also these larvae reached the prepupal
   stage (end of L4 stage) 2 days earlier than those fed on control leaves.
   However, the total amounts of consumed OCII-transformed leaves were up
   to 23 \% lower than of control, and the maximal weights of prepupal
   larvae were reduced by up to 18 \% as compared to larvae fed on
   nontransformed leaves. The reduction in insect fitness reported in this
   study in combination with other control measures, could lead to improved
   CPB resistance management in potato.
T2  - Transgenic Research
T1  - Growth and development of Colorado potato beetle larvae, Leptinotarsa
 decemlineata, on potato plants expressing the oryzacystatin II
 proteinase inhibitor
IS  - 4
VL  - 24
DO  - 10.1007/s11248-015-9873-9
SP  - 729
EP  - 740
ER  - 

@article{
author = "Cingel, Aleksandar and Savic, Jelena and Vinterhalter, Branka and Vinterhalter, Dragan and Kostic, Miroslav and Šešlija Jovanović, Darka and Smigocki, Ann and Ninković, Slavica",
year = "2015",
abstract = "Plant proteinase inhibitors (PIs) are attractive tools for crop
   improvement and their heterologous expression can enhance insect
   resistance in transgenic plants. PI oryzacystatin II (OCII), isolated
   from rice, showed potential in controlling pests that utilize cysteine
   proteinases for protein digestion. To evaluate the applicability of the
   OCII gene in enhancing plant defence, OCII-transformed potatoes were
   bioassayed for resistance to Colorado potato beetle (Leptinotarsa
   decemlineata Say). Feeding on transformed leaves of potato cultivars
   Desiree and Jelica significantly affected larval growth and development,
   but did not change mortality rates. During the L2 and L3 developmental
   stages larvae consumed the OCII-transformed foliage faster as compared
   to the nontransformed control. Also these larvae reached the prepupal
   stage (end of L4 stage) 2 days earlier than those fed on control leaves.
   However, the total amounts of consumed OCII-transformed leaves were up
   to 23 \% lower than of control, and the maximal weights of prepupal
   larvae were reduced by up to 18 \% as compared to larvae fed on
   nontransformed leaves. The reduction in insect fitness reported in this
   study in combination with other control measures, could lead to improved
   CPB resistance management in potato.",
journal = "Transgenic Research",
title = "Growth and development of Colorado potato beetle larvae, Leptinotarsa
 decemlineata, on potato plants expressing the oryzacystatin II
 proteinase inhibitor",
number = "4",
volume = "24",
doi = "10.1007/s11248-015-9873-9",
pages = "729-740"
}

Cingel, A., Savic, J., Vinterhalter, B., Vinterhalter, D., Kostic, M., Šešlija Jovanović, D., Smigocki, A.,& Ninković, S.. (2015). Growth and development of Colorado potato beetle larvae, Leptinotarsa
 decemlineata, on potato plants expressing the oryzacystatin II
 proteinase inhibitor. in Transgenic Research, 24(4), 729-740.
https://doi.org/10.1007/s11248-015-9873-9

Cingel A, Savic J, Vinterhalter B, Vinterhalter D, Kostic M, Šešlija Jovanović D, Smigocki A, Ninković S. Growth and development of Colorado potato beetle larvae, Leptinotarsa
 decemlineata, on potato plants expressing the oryzacystatin II
 proteinase inhibitor. in Transgenic Research. 2015;24(4):729-740.
doi:10.1007/s11248-015-9873-9 .

Cingel, Aleksandar, Savic, Jelena, Vinterhalter, Branka, Vinterhalter, Dragan, Kostic, Miroslav, Šešlija Jovanović, Darka, Smigocki, Ann, Ninković, Slavica, "Growth and development of Colorado potato beetle larvae, Leptinotarsa
 decemlineata, on potato plants expressing the oryzacystatin II
 proteinase inhibitor" in Transgenic Research, 24, no. 4 (2015):729-740,
https://doi.org/10.1007/s11248-015-9873-9 . .

TY  - JOUR
AU  - Cingel, Aleksandar
AU  - Savić, Jelena
AU  - Ćosić, Tatjana
AU  - Zdravković-Korać, Snežana
AU  - Momčilović, Ivana
AU  - Smigocki, Ann
AU  - Ninković, Slavica
PY  - 2014
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/2181
AB  - Proteinase inhibitors oryzacystatins I and II (OCI and OCII) have shown
   potential in controlling pests that utilize cysteine proteinases for
   protein digestion. In order to achieve an additive effect against
   Colorado potato beetle (CPB, Leptinotarsa decemlineata Say) digestive
   proteinases, we combined two individual OC genes in potato (Solanum
   tuberosum L.) cultivars Desiree, Dragaevka and Jelica. The OC genes were
   stacked into the potato genome by simultaneous co-transformation with
   both OCI and OCII genes or by sequential re-transformation of an
   OCI-expressing transgenic line with an OCII gene. Low constitutive and
   high wound-induced transcript levels of both OCI and OCII genes,
   directed by the inducible pin2 promoter, were detected in all doubly
   transformed lines from all three cultivars. Both recombinant OCs,
   detected by immunoblot, were in an active conformation as shown by their
   strong papain inhibitory activity. Although no significant differences
   in CPB larval mortality were observed when larvae fed on OCI/OCII
   transformed or control potato foliage, expression of both OCI and OCII
   in potato plants caused a reduction in CPB adult body weight and in
   weight of foliage consumed, indicating an additive effect. Overall,
   these results show that pyramiding of the OCI and OCII genes could be an
   effective strategy for breeding cultivars exhibiting elevated levels of
   resistance to CPB.
T2  - Euphytica
T1  - Pyramiding rice cystatin OCI and OCII genes in transgenic potato
 (Solanum tuberosum L.) for resistance to Colorado potato beetle
 (Leptinotarsa decemlineata Say)
IS  - 3
VL  - 198
DO  - 10.1007/s10681-014-1119-z
SP  - 425
EP  - 438
ER  - 

@article{
author = "Cingel, Aleksandar and Savić, Jelena and Ćosić, Tatjana and Zdravković-Korać, Snežana and Momčilović, Ivana and Smigocki, Ann and Ninković, Slavica",
year = "2014",
abstract = "Proteinase inhibitors oryzacystatins I and II (OCI and OCII) have shown
   potential in controlling pests that utilize cysteine proteinases for
   protein digestion. In order to achieve an additive effect against
   Colorado potato beetle (CPB, Leptinotarsa decemlineata Say) digestive
   proteinases, we combined two individual OC genes in potato (Solanum
   tuberosum L.) cultivars Desiree, Dragaevka and Jelica. The OC genes were
   stacked into the potato genome by simultaneous co-transformation with
   both OCI and OCII genes or by sequential re-transformation of an
   OCI-expressing transgenic line with an OCII gene. Low constitutive and
   high wound-induced transcript levels of both OCI and OCII genes,
   directed by the inducible pin2 promoter, were detected in all doubly
   transformed lines from all three cultivars. Both recombinant OCs,
   detected by immunoblot, were in an active conformation as shown by their
   strong papain inhibitory activity. Although no significant differences
   in CPB larval mortality were observed when larvae fed on OCI/OCII
   transformed or control potato foliage, expression of both OCI and OCII
   in potato plants caused a reduction in CPB adult body weight and in
   weight of foliage consumed, indicating an additive effect. Overall,
   these results show that pyramiding of the OCI and OCII genes could be an
   effective strategy for breeding cultivars exhibiting elevated levels of
   resistance to CPB.",
journal = "Euphytica",
title = "Pyramiding rice cystatin OCI and OCII genes in transgenic potato
 (Solanum tuberosum L.) for resistance to Colorado potato beetle
 (Leptinotarsa decemlineata Say)",
number = "3",
volume = "198",
doi = "10.1007/s10681-014-1119-z",
pages = "425-438"
}

Cingel, A., Savić, J., Ćosić, T., Zdravković-Korać, S., Momčilović, I., Smigocki, A.,& Ninković, S.. (2014). Pyramiding rice cystatin OCI and OCII genes in transgenic potato
 (Solanum tuberosum L.) for resistance to Colorado potato beetle
 (Leptinotarsa decemlineata Say). in Euphytica, 198(3), 425-438.
https://doi.org/10.1007/s10681-014-1119-z

Cingel A, Savić J, Ćosić T, Zdravković-Korać S, Momčilović I, Smigocki A, Ninković S. Pyramiding rice cystatin OCI and OCII genes in transgenic potato
 (Solanum tuberosum L.) for resistance to Colorado potato beetle
 (Leptinotarsa decemlineata Say). in Euphytica. 2014;198(3):425-438.
doi:10.1007/s10681-014-1119-z .

Cingel, Aleksandar, Savić, Jelena, Ćosić, Tatjana, Zdravković-Korać, Snežana, Momčilović, Ivana, Smigocki, Ann, Ninković, Slavica, "Pyramiding rice cystatin OCI and OCII genes in transgenic potato
 (Solanum tuberosum L.) for resistance to Colorado potato beetle
 (Leptinotarsa decemlineata Say)" in Euphytica, 198, no. 3 (2014):425-438,
https://doi.org/10.1007/s10681-014-1119-z . .

TY  - JOUR
AU  - Savić, Jelena M.
AU  - Smigocki, Ann
PY  - 2012
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/1087
AB  - Expression pattern of a sugar beet serine proteinase inhibitor gene, BvSTI, was characterized in response to mechanical and fall armyworm (FAW; Spodoptera frugiperda J.E. Smith) induced wounding. BvSTI expression was analyzed in three breeding lines moderately resistant to sugar beet root maggot (Tetanops myopaeformis Roder) and in a susceptible line, F1010. Increased mechanical wound induced levels of BvSTI expression were observed in all resistant lines as compared to F1010 during the first 24 h. The most intensive response to wounding was observed in one of the resistant lines, F1016, with a maximum 5- and 2.5-fold increase of BvSTI transcript levels over non-wounded roots and leaves, respectively. In contrast, slight increase of BvSTI transcript levels in leaves and even an initial decrease in roots were observed in F1010. BvSTI transcript accumulation in F1016 and F1010 tissues wounded by FAW showed a similar gene expression pattern, but it was delayed and less intense than the response incited by abiotic wounding. On the protein level, BvSTI specific antibody confirmed increased accumulation of the 30 kDa BvSTI protein in wounded leaves but not in roots of F1016 and F1010. Using trypsin inhibition assays, the activity of BvSTI was confirmed in F1016 roots and leaves and F1010 leaves. In F1010 roots BvSTI activity was completely lacking. We conclude that BvSTI gene expression was wound induced in the insect resistant germplasm suggesting that BvSTI can be used in biotechnological approaches or in breeding programs for improving insect resistance.
T2  - Euphytica
T1  - Beta vulgaris L. serine proteinase inhibitor gene expression in insect resistant sugar beet
IS  - 2
VL  - 188
SP  - 34
EP  - 198
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_1087
ER  - 

@article{
author = "Savić, Jelena M. and Smigocki, Ann",
year = "2012",
abstract = "Expression pattern of a sugar beet serine proteinase inhibitor gene, BvSTI, was characterized in response to mechanical and fall armyworm (FAW; Spodoptera frugiperda J.E. Smith) induced wounding. BvSTI expression was analyzed in three breeding lines moderately resistant to sugar beet root maggot (Tetanops myopaeformis Roder) and in a susceptible line, F1010. Increased mechanical wound induced levels of BvSTI expression were observed in all resistant lines as compared to F1010 during the first 24 h. The most intensive response to wounding was observed in one of the resistant lines, F1016, with a maximum 5- and 2.5-fold increase of BvSTI transcript levels over non-wounded roots and leaves, respectively. In contrast, slight increase of BvSTI transcript levels in leaves and even an initial decrease in roots were observed in F1010. BvSTI transcript accumulation in F1016 and F1010 tissues wounded by FAW showed a similar gene expression pattern, but it was delayed and less intense than the response incited by abiotic wounding. On the protein level, BvSTI specific antibody confirmed increased accumulation of the 30 kDa BvSTI protein in wounded leaves but not in roots of F1016 and F1010. Using trypsin inhibition assays, the activity of BvSTI was confirmed in F1016 roots and leaves and F1010 leaves. In F1010 roots BvSTI activity was completely lacking. We conclude that BvSTI gene expression was wound induced in the insect resistant germplasm suggesting that BvSTI can be used in biotechnological approaches or in breeding programs for improving insect resistance.",
journal = "Euphytica",
title = "Beta vulgaris L. serine proteinase inhibitor gene expression in insect resistant sugar beet",
number = "2",
volume = "188",
pages = "34-198",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_1087"
}

Savić, J. M.,& Smigocki, A.. (2012). Beta vulgaris L. serine proteinase inhibitor gene expression in insect resistant sugar beet. in Euphytica, 188(2), 34-198.
https://hdl.handle.net/21.15107/rcub_ibiss_1087

Savić JM, Smigocki A. Beta vulgaris L. serine proteinase inhibitor gene expression in insect resistant sugar beet. in Euphytica. 2012;188(2):34-198.
https://hdl.handle.net/21.15107/rcub_ibiss_1087 .

Savić, Jelena M., Smigocki, Ann, "Beta vulgaris L. serine proteinase inhibitor gene expression in insect resistant sugar beet" in Euphytica, 188, no. 2 (2012):34-198,
https://hdl.handle.net/21.15107/rcub_ibiss_1087 .

TY  - CONF
AU  - Ninković, Slavica
AU  - Smigocki, Ann
AU  - Nikolić, Radomirka R
AU  - Banjac, Nevena
AU  - Cingel, Aleksandar
AU  - Savić, Jelena M.
AU  - Đorđević, Tatjana
PY  - 2010
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/1399
C3  - In Vitro Cellular & Developmental Biology-Animal
T1  - Genetic Engineering of Lotus corniculatus L Forage Legume with Proteinase Inhibitor Genes
IS  - null
VL  - 46
EP  - S191
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_1399
ER  - 

@conference{
author = "Ninković, Slavica and Smigocki, Ann and Nikolić, Radomirka R and Banjac, Nevena and Cingel, Aleksandar and Savić, Jelena M. and Đorđević, Tatjana",
year = "2010",
journal = "In Vitro Cellular & Developmental Biology-Animal",
title = "Genetic Engineering of Lotus corniculatus L Forage Legume with Proteinase Inhibitor Genes",
number = "null",
volume = "46",
pages = "S191",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_1399"
}

Ninković, S., Smigocki, A., Nikolić, R. R., Banjac, N., Cingel, A., Savić, J. M.,& Đorđević, T.. (2010). Genetic Engineering of Lotus corniculatus L Forage Legume with Proteinase Inhibitor Genes. in In Vitro Cellular & Developmental Biology-Animal, 46(null).
https://hdl.handle.net/21.15107/rcub_ibiss_1399

Ninković S, Smigocki A, Nikolić RR, Banjac N, Cingel A, Savić JM, Đorđević T. Genetic Engineering of Lotus corniculatus L Forage Legume with Proteinase Inhibitor Genes. in In Vitro Cellular & Developmental Biology-Animal. 2010;46(null):null-S191.
https://hdl.handle.net/21.15107/rcub_ibiss_1399 .

Ninković, Slavica, Smigocki, Ann, Nikolić, Radomirka R, Banjac, Nevena, Cingel, Aleksandar, Savić, Jelena M., Đorđević, Tatjana, "Genetic Engineering of Lotus corniculatus L Forage Legume with Proteinase Inhibitor Genes" in In Vitro Cellular & Developmental Biology-Animal, 46, no. null (2010),
https://hdl.handle.net/21.15107/rcub_ibiss_1399 .

TY  - JOUR
AU  - Cingel, Aleksandar
AU  - Vinterhalter, Branka
AU  - Vinterhalter, Dragan V
AU  - Ćalić-Dragosavac, Dušica D.
AU  - Smigocki, Ann
AU  - Ninković, Slavica
PY  - 2010
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/1360
AB  - An efficient protocol for Agrobacterium-mediated transformation of Serbian potato cultivars Dragacevka and Jelica, enabling the introduction of oryzacystatin genes OCI and OCII, was established. Starting with leaf explants, a two-stage transformation protocol combining procedures of Webb and Wenzler provided high shoot regeneration efficiency: 84 - 89% for Dragacevka cultivar and 60 - 68% for Jelica cultivar as compared to 76 - 86% for Desiree, the most frequently used cultivar in transformation experiments. PCR analysis of a small sample of putative transformants showed a nptII integration frequency of 90.9, 76.9 and 86.4% for Dragacevka, Jelica and Desiree, respectively. Regeneration and transformation efficiency was strongly genotype-dependent.
T2  - African Journal of Biotechnology
T1  - Agrobacterium-mediated transformation of two Serbian potato cultivars (Solanum tuberosum L. cv. Dragacevka and cv. Jelica)
IS  - 30
VL  - 9
EP  - 4650
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_1360
ER  - 

@article{
author = "Cingel, Aleksandar and Vinterhalter, Branka and Vinterhalter, Dragan V and Ćalić-Dragosavac, Dušica D. and Smigocki, Ann and Ninković, Slavica",
year = "2010",
abstract = "An efficient protocol for Agrobacterium-mediated transformation of Serbian potato cultivars Dragacevka and Jelica, enabling the introduction of oryzacystatin genes OCI and OCII, was established. Starting with leaf explants, a two-stage transformation protocol combining procedures of Webb and Wenzler provided high shoot regeneration efficiency: 84 - 89% for Dragacevka cultivar and 60 - 68% for Jelica cultivar as compared to 76 - 86% for Desiree, the most frequently used cultivar in transformation experiments. PCR analysis of a small sample of putative transformants showed a nptII integration frequency of 90.9, 76.9 and 86.4% for Dragacevka, Jelica and Desiree, respectively. Regeneration and transformation efficiency was strongly genotype-dependent.",
journal = "African Journal of Biotechnology",
title = "Agrobacterium-mediated transformation of two Serbian potato cultivars (Solanum tuberosum L. cv. Dragacevka and cv. Jelica)",
number = "30",
volume = "9",
pages = "4650",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_1360"
}

Cingel, A., Vinterhalter, B., Vinterhalter, D. V., Ćalić-Dragosavac, D. D., Smigocki, A.,& Ninković, S.. (2010). Agrobacterium-mediated transformation of two Serbian potato cultivars (Solanum tuberosum L. cv. Dragacevka and cv. Jelica). in African Journal of Biotechnology, 9(30).
https://hdl.handle.net/21.15107/rcub_ibiss_1360

Cingel A, Vinterhalter B, Vinterhalter DV, Ćalić-Dragosavac DD, Smigocki A, Ninković S. Agrobacterium-mediated transformation of two Serbian potato cultivars (Solanum tuberosum L. cv. Dragacevka and cv. Jelica). in African Journal of Biotechnology. 2010;9(30):null-4650.
https://hdl.handle.net/21.15107/rcub_ibiss_1360 .

Cingel, Aleksandar, Vinterhalter, Branka, Vinterhalter, Dragan V, Ćalić-Dragosavac, Dušica D., Smigocki, Ann, Ninković, Slavica, "Agrobacterium-mediated transformation of two Serbian potato cultivars (Solanum tuberosum L. cv. Dragacevka and cv. Jelica)" in African Journal of Biotechnology, 9, no. 30 (2010),
https://hdl.handle.net/21.15107/rcub_ibiss_1360 .

TY  - JOUR
AU  - Uzelac, Branka
AU  - Popović, Zorica
AU  - Mijović, Aleksandar
AU  - Budimir, Snežana
AU  - Janošević, Dušica A.
AU  - Smigocki, Ann
AU  - Ninković, Slavica
PY  - 2008
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/1512
AB  - Background and Purpose: Introduction of a foreign gene into plant genome may induce morphological and physiological alterations in transgenic plants. The purpose of this study was to assess the impact of gene introduction on morphology and photosynthetic activity of two transgenic alfalfa shoot cultures. Material and Methods: In vitro maintained shoot cultures of alfalfa (Medicago sativa L. cv. Zajecarska 83) that were transformed with the oryzacystatin II (OC-II) gene and propagated on growth regulator-free medium were subjected to analysis of morphological characteristics and photosynthetic activity. For analysis of morphological characteristics, the length of the main axis, the number of axillary and adventitious shoots, total number of leaves and the number of senesced leaves as well as plant dry mass were determined. Net photosynthetic rate was determined as CO(2) influx using a LI-6200 closed photosynthesis system. Photosynthetic function was assessed as the rate of basic chlorophyll fluorescence and determined with a Plant Stress Meter by method of induced fluorometry. Chlorophyll content in leaf samples was determined spectrophotometrically. Results: The most striking feature of transformed cultures was reduced apical dominance and the absence of adventitious roots. In comparison with Control, main axis length was also reduced. In addition, a general den-ease in photosynthesis in transgenic shoots was also observed. Conclusion: Both morphology and the key processes in photosynthesis were modified in transgenic shoots. However, the fact that transgenic plants carry a gene that confers pest resistance gives potential agronomic value to at least some of the clones whose morphological characteristics and photosynthetic activity were minimally, disturbed.
T2  - Periodicum Biologorum
T1  - Growth habit and photosynthetic activity of shoot cultures of Medicago sativa L. transformed with the oryzacystatin II gene
IS  - 3
VL  - 110
EP  - 235
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_1512
ER  - 

@article{
author = "Uzelac, Branka and Popović, Zorica and Mijović, Aleksandar and Budimir, Snežana and Janošević, Dušica A. and Smigocki, Ann and Ninković, Slavica",
year = "2008",
abstract = "Background and Purpose: Introduction of a foreign gene into plant genome may induce morphological and physiological alterations in transgenic plants. The purpose of this study was to assess the impact of gene introduction on morphology and photosynthetic activity of two transgenic alfalfa shoot cultures. Material and Methods: In vitro maintained shoot cultures of alfalfa (Medicago sativa L. cv. Zajecarska 83) that were transformed with the oryzacystatin II (OC-II) gene and propagated on growth regulator-free medium were subjected to analysis of morphological characteristics and photosynthetic activity. For analysis of morphological characteristics, the length of the main axis, the number of axillary and adventitious shoots, total number of leaves and the number of senesced leaves as well as plant dry mass were determined. Net photosynthetic rate was determined as CO(2) influx using a LI-6200 closed photosynthesis system. Photosynthetic function was assessed as the rate of basic chlorophyll fluorescence and determined with a Plant Stress Meter by method of induced fluorometry. Chlorophyll content in leaf samples was determined spectrophotometrically. Results: The most striking feature of transformed cultures was reduced apical dominance and the absence of adventitious roots. In comparison with Control, main axis length was also reduced. In addition, a general den-ease in photosynthesis in transgenic shoots was also observed. Conclusion: Both morphology and the key processes in photosynthesis were modified in transgenic shoots. However, the fact that transgenic plants carry a gene that confers pest resistance gives potential agronomic value to at least some of the clones whose morphological characteristics and photosynthetic activity were minimally, disturbed.",
journal = "Periodicum Biologorum",
title = "Growth habit and photosynthetic activity of shoot cultures of Medicago sativa L. transformed with the oryzacystatin II gene",
number = "3",
volume = "110",
pages = "235",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_1512"
}

Uzelac, B., Popović, Z., Mijović, A., Budimir, S., Janošević, D. A., Smigocki, A.,& Ninković, S.. (2008). Growth habit and photosynthetic activity of shoot cultures of Medicago sativa L. transformed with the oryzacystatin II gene. in Periodicum Biologorum, 110(3).
https://hdl.handle.net/21.15107/rcub_ibiss_1512

Uzelac B, Popović Z, Mijović A, Budimir S, Janošević DA, Smigocki A, Ninković S. Growth habit and photosynthetic activity of shoot cultures of Medicago sativa L. transformed with the oryzacystatin II gene. in Periodicum Biologorum. 2008;110(3):null-235.
https://hdl.handle.net/21.15107/rcub_ibiss_1512 .

Uzelac, Branka, Popović, Zorica, Mijović, Aleksandar, Budimir, Snežana, Janošević, Dušica A., Smigocki, Ann, Ninković, Slavica, "Growth habit and photosynthetic activity of shoot cultures of Medicago sativa L. transformed with the oryzacystatin II gene" in Periodicum Biologorum, 110, no. 3 (2008),
https://hdl.handle.net/21.15107/rcub_ibiss_1512 .

TY  - JOUR
AU  - Uzelac, Branka
AU  - Ninković, Slavica
AU  - Smigocki, Ann
AU  - Budimir, Snežana
PY  - 2007
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/1612
AB  - Non-transformed and transformed embryogenic cultures of alfalfa (Medicago saliva L. cv. Zajecarska 83), long-term maintained on growth regulator-free medium, were histologically analyzed. In all examined cultures, somatic embryos at various stages of development were observed and secondary embryos were formed in the cotyledonary, hypocotylary and radicular region of the primary embryos. Detailed histological analysis of the torpedo shape somatic embryo revealed that secondary somatic embryos arose directly from single epidermal cells of hypocotylary axis after an unequal periclinal division. Bipolar proembryos were composed of one smaller cytoplasm rich cell and one larger more vacuolated cell. Further cell division pattern was similar for both non-transformed and transformed embryos. However, multicellular origin of secondary embryos in a direct process and even from callus can not be excluded.
T2  - Biologia Plantarum
T1  - Origin and development of secondary somatic embryos in transformed embryogenic cultures of Medicago sativa
IS  - 1
VL  - 51
EP  - 6
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_1612
ER  - 

@article{
author = "Uzelac, Branka and Ninković, Slavica and Smigocki, Ann and Budimir, Snežana",
year = "2007",
abstract = "Non-transformed and transformed embryogenic cultures of alfalfa (Medicago saliva L. cv. Zajecarska 83), long-term maintained on growth regulator-free medium, were histologically analyzed. In all examined cultures, somatic embryos at various stages of development were observed and secondary embryos were formed in the cotyledonary, hypocotylary and radicular region of the primary embryos. Detailed histological analysis of the torpedo shape somatic embryo revealed that secondary somatic embryos arose directly from single epidermal cells of hypocotylary axis after an unequal periclinal division. Bipolar proembryos were composed of one smaller cytoplasm rich cell and one larger more vacuolated cell. Further cell division pattern was similar for both non-transformed and transformed embryos. However, multicellular origin of secondary embryos in a direct process and even from callus can not be excluded.",
journal = "Biologia Plantarum",
title = "Origin and development of secondary somatic embryos in transformed embryogenic cultures of Medicago sativa",
number = "1",
volume = "51",
pages = "6",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_1612"
}

Uzelac, B., Ninković, S., Smigocki, A.,& Budimir, S.. (2007). Origin and development of secondary somatic embryos in transformed embryogenic cultures of Medicago sativa. in Biologia Plantarum, 51(1).
https://hdl.handle.net/21.15107/rcub_ibiss_1612

Uzelac B, Ninković S, Smigocki A, Budimir S. Origin and development of secondary somatic embryos in transformed embryogenic cultures of Medicago sativa. in Biologia Plantarum. 2007;51(1):null-6.
https://hdl.handle.net/21.15107/rcub_ibiss_1612 .

Uzelac, Branka, Ninković, Slavica, Smigocki, Ann, Budimir, Snežana, "Origin and development of secondary somatic embryos in transformed embryogenic cultures of Medicago sativa" in Biologia Plantarum, 51, no. 1 (2007),
https://hdl.handle.net/21.15107/rcub_ibiss_1612 .