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
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/3792
AB  - Allelopathic plants exploit their chemical ‘weapons’ to prevail over the competition, suppress neighboring plants and consequentlyuse the available resources more efficiently. However, the investigation of plant allelopathic interactions in rhizosphere is difficultto perform because of its high complexity due to interactions of biotic and abiotic factors. Thus, autonomous, aseptic root culturesof apple (Malus × domestica Borkh.) could facilitate allelopathic studies. We report on the successful genetic transformation ofapple cultivars Melrose, Golden Delicious, Cadel and Gloster using ˇ Agrobacterium rhizogenes (Riker et al. 1930) Conn 1942 strain15834 and for the first time the establishment of apple autonomous and permanent in vitro hairy root cultures that could be usedas a new tool for apple allelopathic assays. Molecular characterization of transgenic hairy root lines was conducted to elucidatethe 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 inuntransformed 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 inapple. Chlorogenic and caffeic acids and dihydrochalcones phloridzin and phloretin were detected as putative allelochemicalsexuded into the growth medium in which transgenic roots were maintained for 4 weeks. Apple hairy root exudates significantlyaffected shoot and root development and growth of test plant Arabidopsis thaliana (L.) Heynh. seedlings after 5 or 10 days oftreatment. Additionally, core cell-cycle genes CDKA1;1, CDKB2;1, CYCA3;1 and CYCB2;4 were down regulated in Arabidopsisshoots suggesting, in part, their role in inhibition of shoot growth. The present work highlighted an autonomous and permanentin 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 consequentlyuse the available resources more efficiently. However, the investigation of plant allelopathic interactions in rhizosphere is difficultto perform because of its high complexity due to interactions of biotic and abiotic factors. Thus, autonomous, aseptic root culturesof apple (Malus × domestica Borkh.) could facilitate allelopathic studies. We report on the successful genetic transformation ofapple cultivars Melrose, Golden Delicious, Cadel and Gloster using ˇ Agrobacterium rhizogenes (Riker et al. 1930) Conn 1942 strain15834 and for the first time the establishment of apple autonomous and permanent in vitro hairy root cultures that could be usedas a new tool for apple allelopathic assays. Molecular characterization of transgenic hairy root lines was conducted to elucidatethe 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 inuntransformed 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 inapple. Chlorogenic and caffeic acids and dihydrochalcones phloridzin and phloretin were detected as putative allelochemicalsexuded into the growth medium in which transgenic roots were maintained for 4 weeks. Apple hairy root exudates significantlyaffected shoot and root development and growth of test plant Arabidopsis thaliana (L.) Heynh. seedlings after 5 or 10 days oftreatment. Additionally, core cell-cycle genes CDKA1;1, CDKB2;1, CYCA3;1 and CYCB2;4 were down regulated in Arabidopsisshoots suggesting, in part, their role in inhibition of shoot growth. The present work highlighted an autonomous and permanentin 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  - CONF
AU  - Stanišić, Mariana
AU  - Ćosić, Tatjana
AU  - Savić, Jelena
AU  - Smigocki, Ann
AU  - Ninković, Slavica
AU  - Banjac, Nevena
PY  - 2018
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/4999
AB  - In the previous study, phenolic acids and polyphenolic flavonoids were identified as putative
allelochemicals in apple cv. Golden Delicious hairy roots growth medium. In order to examine
their allelopathic potential, the effect of hairy root exudates on the germination and seedling
growth of the test species Arabidopsis thaliana was elucidated. Arabidopsis seeds were cultivated
in Erlenmeyer flasks containing 25 mL of liquid medium in which apple hairy roots were cultivated
for 4 weeks. The effect of apple hairy root growth medium on Arabidopsis was evaluated after 5
and 10 days of seed cultivation. Apple hairy root exudates had no significant effect on Arabidopsis
germination rate, while growth characteristics of seedlings were significantly affected both after
5 and 10 days of treatment. The roots of treated seedlings were significantly shorter (35.1%), with
lower number (74% decrement) of lateral roots compared to control after10 days of treatment.
The growth and morphology of Arabidopsis shoots and leaves have been affected too. Thus, the
shoots of treated seedlings were almost twice shorter compared to control after 5 days of treatment. At the 10th day, this distinction became more pronounced, when two fully expanded leaves
were visible in treated seedlings, while control had completely developed rosettes with six leaves.
In addition, leaves of treated seedlings were smaller and pale in colour compared to control. qPCR
expression analysis of genes involved in regulation of the cell cycle showed reduction of transcript
levels in roots of treated seedlings, with the highest expression decrease detected for CDKA1 and
CYCB2;4 cyclin.
PB  - Belgrade: Serbian Plant Physiology Society
C3  - 3rd International Conference on Plant Biology and 22nd SPPS Meeting , 9-12 June 2018, Belgrade
T1  - Allelopathic effect of apple hairy root exudates on the germination and seedlings growth of Arabidopsis thaliana in vitro
SP  - 55
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_4999
ER  - 

@conference{
author = "Stanišić, Mariana and Ćosić, Tatjana and Savić, Jelena and Smigocki, Ann and Ninković, Slavica and Banjac, Nevena",
year = "2018",
abstract = "In the previous study, phenolic acids and polyphenolic flavonoids were identified as putative
allelochemicals in apple cv. Golden Delicious hairy roots growth medium. In order to examine
their allelopathic potential, the effect of hairy root exudates on the germination and seedling
growth of the test species Arabidopsis thaliana was elucidated. Arabidopsis seeds were cultivated
in Erlenmeyer flasks containing 25 mL of liquid medium in which apple hairy roots were cultivated
for 4 weeks. The effect of apple hairy root growth medium on Arabidopsis was evaluated after 5
and 10 days of seed cultivation. Apple hairy root exudates had no significant effect on Arabidopsis
germination rate, while growth characteristics of seedlings were significantly affected both after
5 and 10 days of treatment. The roots of treated seedlings were significantly shorter (35.1%), with
lower number (74% decrement) of lateral roots compared to control after10 days of treatment.
The growth and morphology of Arabidopsis shoots and leaves have been affected too. Thus, the
shoots of treated seedlings were almost twice shorter compared to control after 5 days of treatment. At the 10th day, this distinction became more pronounced, when two fully expanded leaves
were visible in treated seedlings, while control had completely developed rosettes with six leaves.
In addition, leaves of treated seedlings were smaller and pale in colour compared to control. qPCR
expression analysis of genes involved in regulation of the cell cycle showed reduction of transcript
levels in roots of treated seedlings, with the highest expression decrease detected for CDKA1 and
CYCB2;4 cyclin.",
publisher = "Belgrade: Serbian Plant Physiology Society",
journal = "3rd International Conference on Plant Biology and 22nd SPPS Meeting , 9-12 June 2018, Belgrade",
title = "Allelopathic effect of apple hairy root exudates on the germination and seedlings growth of Arabidopsis thaliana in vitro",
pages = "55",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_4999"
}

Stanišić, M., Ćosić, T., Savić, J., Smigocki, A., Ninković, S.,& Banjac, N.. (2018). Allelopathic effect of apple hairy root exudates on the germination and seedlings growth of Arabidopsis thaliana in vitro. in 3rd International Conference on Plant Biology and 22nd SPPS Meeting , 9-12 June 2018, Belgrade
Belgrade: Serbian Plant Physiology Society., 55.
https://hdl.handle.net/21.15107/rcub_ibiss_4999

Stanišić M, Ćosić T, Savić J, Smigocki A, Ninković S, Banjac N. Allelopathic effect of apple hairy root exudates on the germination and seedlings growth of Arabidopsis thaliana in vitro. in 3rd International Conference on Plant Biology and 22nd SPPS Meeting , 9-12 June 2018, Belgrade. 2018;:55.
https://hdl.handle.net/21.15107/rcub_ibiss_4999 .

Stanišić, Mariana, Ćosić, Tatjana, Savić, Jelena, Smigocki, Ann, Ninković, Slavica, Banjac, Nevena, "Allelopathic effect of apple hairy root exudates on the germination and seedlings growth of Arabidopsis thaliana in vitro" in 3rd International Conference on Plant Biology and 22nd SPPS Meeting , 9-12 June 2018, Belgrade (2018):55,
https://hdl.handle.net/21.15107/rcub_ibiss_4999 .