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Efekat ekspresije gena za protein-kinazu 1 duvana (NPK1) kod transformisanih biljaka karfiola (Brassica oleracea L. var. botrytis) na razvoj tolerancije prema povišenim koncentracijama NaCI in vitro

Effect of tobacco protein kinase 1 (NPK1) gene expression in transformed cauliflower (Brassica oleracea L. var. botrytis) plants on the development of tolerance to increased concentrations of NaCl in vitro

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2015
4.pdf (4.442Mb)
Authors
Pavlović, Suzana
Contributors
Mitić, Nevena
Janošević, Dušica
Vinterhalter, Branka
Savić, Jelena
Cvetić-Antić, Tijana
Doctoral thesis (Published version)
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Abstract
Biljke su neprekidno izloţene razliĉitim vrstama stresa iz spoljašnje sredine, koji nepovoljno utiĉu na njihov rast i razviće. Povećan salinitet zemljišta je jedan od faktora spoljašnje sredine, koji u velikoj meri ograniĉava produktivnost gajenih biljaka. Karfiol (Brassica oleracea L. var. botrytis), je znaĉajna povrtarska kultura koja je, kao i ostale povrtarske kupusnjaĉe, podloţna uticaju razliĉitih stresnih faktora, pa i povećanom salinitetu usled intenzivnog zalivanja. Uz klasiĉne metode oplemenjivanja, biotehnološke metode mogu biti efikasan pristup za povećanje tolerancije biljaka prema stresu. Mitogenom aktivirane protein-kinaze (MAPK) igraju kljuĉnu ulogu u rastu i razviću biljaka i njihovom odgovoru na biotiĉke i abiotiĉke faktore spoljašnje sredine. Introdukcija gena za komponente MAPK kaskada moţe dovesti do razvoja transgenih biljaka sa povećanom tolerancijom prema stresu, pri ĉemu su MAPKKK najpogodnije za genetiĉki inţenjering jer funkcionišu na poĉetku kaskade koja vod...i aktivaciji više transkripcionih faktora i stres inducibilnih gena. Preduslov za primenu ovih metoda u oplemenjivanju biljaka jeste uspostavljanje efikasnog sistema za regeneraciju biljaka in vitro. Kod ĉetiri varijeteta B. oleracea (karfiola, kupusa, brokolija i kelja) uspostavljen je sistem za regeneraciju pupoljaka in vitro putem organogeneze. Eksplantati hipokotila su kod skoro svih testiranih varijeteta imali najbolji regenerativni potencijal (od 75,0-91,2% sa produkcijom 3,5-7,4 pupoljaka po eksplantatu). Podloge koje su sadrţale N6-benziladenin (BA) su bile optimalne, kako za regeneraciju pupoljaka, tako i za njihovu kasniju multiplikaciju. Regenerisani izdanci su uspešno oţiljeni i aklimatizovani. Kod kupusa i karfiola uspostavljen je i jednostavan protokol za direktnu somatsku embriogenezu iz nezrelih zigotskih embriona gajenih na B5 podlozi bez regulatora rastenja (B5-0), koji je omogućio efikasnu transformaciju karfiola kao, u tom pogledu, problematiĉnog varijeteta. Kod oba varijeteta, zigotski embrioni u kotiledonarnom stadijumu razvoja imali su najveći kapacitet za formiranje somatskih embriona (11,84 kod kupusa i 11,95 kod karfiola)...

Plants are continuously exposed to various types of stress from the environment that adversely affect their growth and development. The increased salinity of the soil is one of the environmetal factors that greatly limits the productivity of the cultivated plants. Cauliflower (Brassica oleracea L. var. botrytis), is an important vegetable crop, which, like other Brassica vegetable, is grown under the influence of various biotic and abiotic stresses, including the increased salinity due to intensive watering. In addition to classical breeding methods, biotechnology methods may be an effective approach to increase plants stress tolerance. The mitogen-activated protein kinases (MAPK) play a key role in the growth and development of plants and their response to biotic and abiotic stresses. Introduction of genes for components of MAPK cascade may lead to the development of transgenic plants with the increased tolerance to stress, wherein MAPKKK are most suitable for genetic engineering, bec...ause they act at the beginning of the cascade that leads to activation of several transcription factors and stress inducible genes. A prerequisite for the application of these methods in plant breeding is establishing an efficient plant regeneration system in vitro. A regenerative system via organogenesis was established in four B. oleracea varieties (cauliflower, cabbage, broccoli, kale). The hypocotyl explants in almost all the tested varieties have the best regenerative potential (from 75.0-91.2% with the production of 3.5-7.4 shoots per explant). The media containing N6-benzyladenine (BA) were optimal both for shoot regeneration and their subsequent multiplication. Regenerated shoots were succesfully rooted and acclimatized. In cabbage and cauliflower a simple protocol was established for the direct somatic embryogenesis from immature zigotic embryos grown on B5 medium without growth regulators (B5-0), which enabled the efficient transformation of cauliflower, in this regard, as a problematic variety. In both varieties, zygotic embryos at the cotyledonary stage of development had the highest embryo-forming capacity (11.84 in cabbage and 11.95 in cauliflower)...

Keywords:
Brassica oleracea L. / Brassica oleracea L. / karfiol / organogeneza / somatska embriogeneza / genetiĉka transformacija / Agrobacterium tumefaciens / NPK1 gen / bar gen / abiotiĉki stres / salinitet / cauliflower / organogenesis / somatic embryogenesis / genetic transformation / Agrobacterium tumefaciens / NPK1 gene / bar gene / abiotic stress / salinity
Source:
2015
Publisher:
  • Univerzitet u Beogradu, Biološki fakultet
Funding / projects:
  • Integrating biotechnology approach in breeding vegetable crops for sustainable agricultural systems (RS-31059)
[ Google Scholar ]
Handle
https://hdl.handle.net/21.15107/rcub_nardus_4622
URI
http://nardus.mpn.gov.rs/handle/123456789/4622
http://eteze.bg.ac.rs/application/showtheses?thesesId=2449
https://fedorabg.bg.ac.rs/fedora/get/o:10460/bdef:Content/download
http://vbs.rs/scripts/cobiss?command=DISPLAY&base=70036&RID=47614991
http://RIVeC.institut-palanka.rs/handle/123456789/7
Collections
  • Doktorske disertacije / Doctoral Thesis
Institution/Community
Institut za povrtarstvo
TY  - THES
AU  - Pavlović, Suzana
PY  - 2015
UR  - http://nardus.mpn.gov.rs/handle/123456789/4622
UR  - http://eteze.bg.ac.rs/application/showtheses?thesesId=2449
UR  - https://fedorabg.bg.ac.rs/fedora/get/o:10460/bdef:Content/download
UR  - http://vbs.rs/scripts/cobiss?command=DISPLAY&base=70036&RID=47614991
UR  - http://RIVeC.institut-palanka.rs/handle/123456789/7
AB  - Biljke su neprekidno izloţene razliĉitim vrstama stresa iz spoljašnje sredine, koji nepovoljno utiĉu na njihov rast i razviće. Povećan salinitet zemljišta je jedan od faktora spoljašnje sredine, koji u velikoj meri ograniĉava produktivnost gajenih biljaka. Karfiol (Brassica oleracea L. var. botrytis), je znaĉajna povrtarska kultura koja je, kao i ostale povrtarske kupusnjaĉe, podloţna uticaju razliĉitih stresnih faktora, pa i povećanom salinitetu usled intenzivnog zalivanja. Uz klasiĉne metode oplemenjivanja, biotehnološke metode mogu biti efikasan pristup za povećanje tolerancije biljaka prema stresu. Mitogenom aktivirane protein-kinaze (MAPK) igraju kljuĉnu ulogu u rastu i razviću biljaka i njihovom odgovoru na biotiĉke i abiotiĉke faktore spoljašnje sredine. Introdukcija gena za komponente MAPK kaskada moţe dovesti do razvoja transgenih biljaka sa povećanom tolerancijom prema stresu, pri ĉemu su MAPKKK najpogodnije za genetiĉki inţenjering jer funkcionišu na poĉetku kaskade koja vodi aktivaciji više transkripcionih faktora i stres inducibilnih gena. Preduslov za primenu ovih metoda u oplemenjivanju biljaka jeste uspostavljanje efikasnog sistema za regeneraciju biljaka in vitro. Kod ĉetiri varijeteta B. oleracea (karfiola, kupusa, brokolija i kelja) uspostavljen je sistem za regeneraciju pupoljaka in vitro putem organogeneze. Eksplantati hipokotila su kod skoro svih testiranih varijeteta imali najbolji regenerativni potencijal (od 75,0-91,2% sa produkcijom 3,5-7,4 pupoljaka po eksplantatu). Podloge koje su sadrţale N6-benziladenin (BA) su bile optimalne, kako za regeneraciju pupoljaka, tako i za njihovu kasniju multiplikaciju. Regenerisani izdanci su uspešno oţiljeni i aklimatizovani. Kod kupusa i karfiola uspostavljen je i jednostavan protokol za direktnu somatsku embriogenezu iz nezrelih zigotskih embriona gajenih na B5 podlozi bez regulatora rastenja (B5-0), koji je omogućio efikasnu transformaciju karfiola kao, u tom pogledu, problematiĉnog varijeteta. Kod oba varijeteta, zigotski embrioni u kotiledonarnom stadijumu razvoja imali su najveći kapacitet za formiranje somatskih embriona (11,84 kod kupusa i 11,95 kod karfiola)...
AB  - Plants are continuously exposed to various types of stress from the environment that adversely affect their growth and development. The increased salinity of the soil is one of the environmetal factors that greatly limits the productivity of the cultivated plants. Cauliflower (Brassica oleracea L. var. botrytis), is an important vegetable crop, which, like other Brassica vegetable, is grown under the influence of various biotic and abiotic stresses, including the increased salinity due to intensive watering. In addition to classical breeding methods, biotechnology methods may be an effective approach to increase plants stress tolerance. The mitogen-activated protein kinases (MAPK) play a key role in the growth and development of plants and their response to biotic and abiotic stresses. Introduction of genes for components of MAPK cascade may lead to the development of transgenic plants with the increased tolerance to stress, wherein MAPKKK are most suitable for genetic engineering, because they act at the beginning of the cascade that leads to activation of several transcription factors and stress inducible genes. A prerequisite for the application of these methods in plant breeding is establishing an efficient plant regeneration system in vitro. A regenerative system via organogenesis was established in four B. oleracea varieties (cauliflower, cabbage, broccoli, kale). The hypocotyl explants in almost all the tested varieties have the best regenerative potential (from 75.0-91.2% with the production of 3.5-7.4 shoots per explant). The media containing N6-benzyladenine (BA) were optimal both for shoot regeneration and their subsequent multiplication. Regenerated shoots were succesfully rooted and acclimatized. In cabbage and cauliflower a simple protocol was established for the direct somatic embryogenesis from immature zigotic embryos grown on B5 medium without growth regulators (B5-0), which enabled the efficient transformation of cauliflower, in this regard, as a problematic variety. In both varieties, zygotic embryos at the cotyledonary stage of development had the highest embryo-forming capacity (11.84 in cabbage and 11.95 in cauliflower)...
PB  - Univerzitet u Beogradu, Biološki fakultet
T1  - Efekat ekspresije gena za protein-kinazu 1 duvana (NPK1) kod transformisanih biljaka karfiola (Brassica oleracea L. var. botrytis) na razvoj tolerancije prema povišenim koncentracijama NaCI in vitro
T1  - Effect of tobacco protein kinase 1 (NPK1) gene expression in transformed cauliflower (Brassica oleracea L. var. botrytis) plants on the development of tolerance to increased concentrations of NaCl in vitro
UR  - t-492
UR  - https://hdl.handle.net/21.15107/rcub_nardus_4622
ER  - 
@phdthesis{
author = "Pavlović, Suzana",
year = "2015",
abstract = "Biljke su neprekidno izloţene razliĉitim vrstama stresa iz spoljašnje sredine, koji nepovoljno utiĉu na njihov rast i razviće. Povećan salinitet zemljišta je jedan od faktora spoljašnje sredine, koji u velikoj meri ograniĉava produktivnost gajenih biljaka. Karfiol (Brassica oleracea L. var. botrytis), je znaĉajna povrtarska kultura koja je, kao i ostale povrtarske kupusnjaĉe, podloţna uticaju razliĉitih stresnih faktora, pa i povećanom salinitetu usled intenzivnog zalivanja. Uz klasiĉne metode oplemenjivanja, biotehnološke metode mogu biti efikasan pristup za povećanje tolerancije biljaka prema stresu. Mitogenom aktivirane protein-kinaze (MAPK) igraju kljuĉnu ulogu u rastu i razviću biljaka i njihovom odgovoru na biotiĉke i abiotiĉke faktore spoljašnje sredine. Introdukcija gena za komponente MAPK kaskada moţe dovesti do razvoja transgenih biljaka sa povećanom tolerancijom prema stresu, pri ĉemu su MAPKKK najpogodnije za genetiĉki inţenjering jer funkcionišu na poĉetku kaskade koja vodi aktivaciji više transkripcionih faktora i stres inducibilnih gena. Preduslov za primenu ovih metoda u oplemenjivanju biljaka jeste uspostavljanje efikasnog sistema za regeneraciju biljaka in vitro. Kod ĉetiri varijeteta B. oleracea (karfiola, kupusa, brokolija i kelja) uspostavljen je sistem za regeneraciju pupoljaka in vitro putem organogeneze. Eksplantati hipokotila su kod skoro svih testiranih varijeteta imali najbolji regenerativni potencijal (od 75,0-91,2% sa produkcijom 3,5-7,4 pupoljaka po eksplantatu). Podloge koje su sadrţale N6-benziladenin (BA) su bile optimalne, kako za regeneraciju pupoljaka, tako i za njihovu kasniju multiplikaciju. Regenerisani izdanci su uspešno oţiljeni i aklimatizovani. Kod kupusa i karfiola uspostavljen je i jednostavan protokol za direktnu somatsku embriogenezu iz nezrelih zigotskih embriona gajenih na B5 podlozi bez regulatora rastenja (B5-0), koji je omogućio efikasnu transformaciju karfiola kao, u tom pogledu, problematiĉnog varijeteta. Kod oba varijeteta, zigotski embrioni u kotiledonarnom stadijumu razvoja imali su najveći kapacitet za formiranje somatskih embriona (11,84 kod kupusa i 11,95 kod karfiola)..., Plants are continuously exposed to various types of stress from the environment that adversely affect their growth and development. The increased salinity of the soil is one of the environmetal factors that greatly limits the productivity of the cultivated plants. Cauliflower (Brassica oleracea L. var. botrytis), is an important vegetable crop, which, like other Brassica vegetable, is grown under the influence of various biotic and abiotic stresses, including the increased salinity due to intensive watering. In addition to classical breeding methods, biotechnology methods may be an effective approach to increase plants stress tolerance. The mitogen-activated protein kinases (MAPK) play a key role in the growth and development of plants and their response to biotic and abiotic stresses. Introduction of genes for components of MAPK cascade may lead to the development of transgenic plants with the increased tolerance to stress, wherein MAPKKK are most suitable for genetic engineering, because they act at the beginning of the cascade that leads to activation of several transcription factors and stress inducible genes. A prerequisite for the application of these methods in plant breeding is establishing an efficient plant regeneration system in vitro. A regenerative system via organogenesis was established in four B. oleracea varieties (cauliflower, cabbage, broccoli, kale). The hypocotyl explants in almost all the tested varieties have the best regenerative potential (from 75.0-91.2% with the production of 3.5-7.4 shoots per explant). The media containing N6-benzyladenine (BA) were optimal both for shoot regeneration and their subsequent multiplication. Regenerated shoots were succesfully rooted and acclimatized. In cabbage and cauliflower a simple protocol was established for the direct somatic embryogenesis from immature zigotic embryos grown on B5 medium without growth regulators (B5-0), which enabled the efficient transformation of cauliflower, in this regard, as a problematic variety. In both varieties, zygotic embryos at the cotyledonary stage of development had the highest embryo-forming capacity (11.84 in cabbage and 11.95 in cauliflower)...",
publisher = "Univerzitet u Beogradu, Biološki fakultet",
title = "Efekat ekspresije gena za protein-kinazu 1 duvana (NPK1) kod transformisanih biljaka karfiola (Brassica oleracea L. var. botrytis) na razvoj tolerancije prema povišenim koncentracijama NaCI in vitro, Effect of tobacco protein kinase 1 (NPK1) gene expression in transformed cauliflower (Brassica oleracea L. var. botrytis) plants on the development of tolerance to increased concentrations of NaCl in vitro",
url = "t-492, https://hdl.handle.net/21.15107/rcub_nardus_4622"
}
Pavlović, S.. (2015). Efekat ekspresije gena za protein-kinazu 1 duvana (NPK1) kod transformisanih biljaka karfiola (Brassica oleracea L. var. botrytis) na razvoj tolerancije prema povišenim koncentracijama NaCI in vitro. 
Univerzitet u Beogradu, Biološki fakultet..
t-492
Pavlović S. Efekat ekspresije gena za protein-kinazu 1 duvana (NPK1) kod transformisanih biljaka karfiola (Brassica oleracea L. var. botrytis) na razvoj tolerancije prema povišenim koncentracijama NaCI in vitro. 2015;.
t-492 .
Pavlović, Suzana, "Efekat ekspresije gena za protein-kinazu 1 duvana (NPK1) kod transformisanih biljaka karfiola (Brassica oleracea L. var. botrytis) na razvoj tolerancije prema povišenim koncentracijama NaCI in vitro" (2015),
t-492 .

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