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Introduction of the Nicotiana protein kinase (NPK1) gene by combiningAgrobacterium-mediated transformation and recurrent somatic embryogenesis to enhance salt tolerance in cauliflower

Authorized Users Only
2020
Authors
Pavlović, Suzana
Savić, Jelena
Milojević, Jelena
Vinterhalter, Branka
Girek, Zdenka
Adžić, Slađan
Zečević, Bogoljub
Banjac, Nevena
Article (Published version)
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Abstract
Key message The results underlined that constitutively expressingNPK1can significantly contribute to enhanced salt stress tolerance in cauliflower, suggesting that this could be a promising basis for the creation of new stress tolerance cruciferous vegetable lines. Cauliflower is exposed to various biotic and abiotic stresses, including increased salinity due to the intensive irrigation of crops. Mitogen-activated protein kinase (MAPK) cascades are universal signal transduction modules that play important roles in regulating innate immune responses in plants. Based on involvement of tobacco MAP kinase kinase kinase (NPK1) in stress response, the effect of the expression ofNPK1transgene to NaCl salt stress tolerance in cauliflower KFRM4 lines was studied. TheAgrobacterium tumefaciens-mediated transformation protocol, using EHA101(pSHX004) vector harbouring theNPK1and phosphinothricinN-acetyltransferase (bar) genes, the cyclic somatic embryogenesis regeneration pathway, the application o...f acetosyringone (AS) during co-cultivation and a delayed phosphinothricine (PPT) selection procedure provided sufficient transformation efficiency of 7.33% without escapes. PCR analysis indicated the integration of bothNPK1andbartransgenes in regenerated cauliflower lines. Transgenic cauliflower lines, exposed to NaCl stress in vitro, showed higher growth rates, greater ability to retain chlorophyll and carotenoids, and increased osmotic regulation capacity compared with non-transformed control plants. The tolerance level of transformed lines correlated with the level ofNPK1gene expression estimated by RT-qPCR, and the L2 line with the highestNPK1expression displayed the greatest tolerance to NaCl stress. None of the obtained cauliflower transformed lines grown in greenhouses showed any morphological or yield differences compared with non-transformed plants. Furthermore, the expression of thebargene facilitated the tolerance of transformed lines to the total herbicide PPT, applied at concentrations 2-3 times higher than those routinely used for weed control in the crop field.

Keywords:
Brassica oleraceavar / botrytisl / Cyclic somatic embryogenesis / Agrobacterium tumefaciens-mediated genetic transformation / NPK1gene / Herbicide resistance / Salt stress
Source:
Plant Cell Tissue and Organ Culture, 2020, 143, 3, 635-651
Publisher:
  • Springer, Dordrecht
Funding / projects:
  • Biotechnology in vitro - crop, medicinal and endangered plant species (RS-173015)
  • Integrating biotechnology approach in breeding vegetable crops for sustainable agricultural systems (RS-31059)

DOI: 10.1007/s11240-020-01948-6

ISSN: 0167-6857

WoS: 000577961700001

Scopus: 2-s2.0-85096238113
[ Google Scholar ]
URI
http://RIVeC.institut-palanka.rs/handle/123456789/215
Collections
  • Radovi sa afilijacijom Instituta za povrtarstvo / Papers with the Institute for Vegetable Crops affiliation
Institution/Community
Institut za povrtarstvo
TY  - JOUR
AU  - Pavlović, Suzana
AU  - Savić, Jelena
AU  - Milojević, Jelena
AU  - Vinterhalter, Branka
AU  - Girek, Zdenka
AU  - Adžić, Slađan
AU  - Zečević, Bogoljub
AU  - Banjac, Nevena
PY  - 2020
UR  - http://RIVeC.institut-palanka.rs/handle/123456789/215
AB  - Key message The results underlined that constitutively expressingNPK1can significantly contribute to enhanced salt stress tolerance in cauliflower, suggesting that this could be a promising basis for the creation of new stress tolerance cruciferous vegetable lines. Cauliflower is exposed to various biotic and abiotic stresses, including increased salinity due to the intensive irrigation of crops. Mitogen-activated protein kinase (MAPK) cascades are universal signal transduction modules that play important roles in regulating innate immune responses in plants. Based on involvement of tobacco MAP kinase kinase kinase (NPK1) in stress response, the effect of the expression ofNPK1transgene to NaCl salt stress tolerance in cauliflower KFRM4 lines was studied. TheAgrobacterium tumefaciens-mediated transformation protocol, using EHA101(pSHX004) vector harbouring theNPK1and phosphinothricinN-acetyltransferase (bar) genes, the cyclic somatic embryogenesis regeneration pathway, the application of acetosyringone (AS) during co-cultivation and a delayed phosphinothricine (PPT) selection procedure provided sufficient transformation efficiency of 7.33% without escapes. PCR analysis indicated the integration of bothNPK1andbartransgenes in regenerated cauliflower lines. Transgenic cauliflower lines, exposed to NaCl stress in vitro, showed higher growth rates, greater ability to retain chlorophyll and carotenoids, and increased osmotic regulation capacity compared with non-transformed control plants. The tolerance level of transformed lines correlated with the level ofNPK1gene expression estimated by RT-qPCR, and the L2 line with the highestNPK1expression displayed the greatest tolerance to NaCl stress. None of the obtained cauliflower transformed lines grown in greenhouses showed any morphological or yield differences compared with non-transformed plants. Furthermore, the expression of thebargene facilitated the tolerance of transformed lines to the total herbicide PPT, applied at concentrations 2-3 times higher than those routinely used for weed control in the crop field.
PB  - Springer, Dordrecht
T2  - Plant Cell Tissue and Organ Culture
T1  - Introduction of the Nicotiana protein kinase (NPK1) gene by combiningAgrobacterium-mediated transformation and recurrent somatic embryogenesis to enhance salt tolerance in cauliflower
EP  - 651
IS  - 3
SP  - 635
VL  - 143
DO  - 10.1007/s11240-020-01948-6
UR  - conv_220
ER  - 
@article{
author = "Pavlović, Suzana and Savić, Jelena and Milojević, Jelena and Vinterhalter, Branka and Girek, Zdenka and Adžić, Slađan and Zečević, Bogoljub and Banjac, Nevena",
year = "2020",
abstract = "Key message The results underlined that constitutively expressingNPK1can significantly contribute to enhanced salt stress tolerance in cauliflower, suggesting that this could be a promising basis for the creation of new stress tolerance cruciferous vegetable lines. Cauliflower is exposed to various biotic and abiotic stresses, including increased salinity due to the intensive irrigation of crops. Mitogen-activated protein kinase (MAPK) cascades are universal signal transduction modules that play important roles in regulating innate immune responses in plants. Based on involvement of tobacco MAP kinase kinase kinase (NPK1) in stress response, the effect of the expression ofNPK1transgene to NaCl salt stress tolerance in cauliflower KFRM4 lines was studied. TheAgrobacterium tumefaciens-mediated transformation protocol, using EHA101(pSHX004) vector harbouring theNPK1and phosphinothricinN-acetyltransferase (bar) genes, the cyclic somatic embryogenesis regeneration pathway, the application of acetosyringone (AS) during co-cultivation and a delayed phosphinothricine (PPT) selection procedure provided sufficient transformation efficiency of 7.33% without escapes. PCR analysis indicated the integration of bothNPK1andbartransgenes in regenerated cauliflower lines. Transgenic cauliflower lines, exposed to NaCl stress in vitro, showed higher growth rates, greater ability to retain chlorophyll and carotenoids, and increased osmotic regulation capacity compared with non-transformed control plants. The tolerance level of transformed lines correlated with the level ofNPK1gene expression estimated by RT-qPCR, and the L2 line with the highestNPK1expression displayed the greatest tolerance to NaCl stress. None of the obtained cauliflower transformed lines grown in greenhouses showed any morphological or yield differences compared with non-transformed plants. Furthermore, the expression of thebargene facilitated the tolerance of transformed lines to the total herbicide PPT, applied at concentrations 2-3 times higher than those routinely used for weed control in the crop field.",
publisher = "Springer, Dordrecht",
journal = "Plant Cell Tissue and Organ Culture",
title = "Introduction of the Nicotiana protein kinase (NPK1) gene by combiningAgrobacterium-mediated transformation and recurrent somatic embryogenesis to enhance salt tolerance in cauliflower",
pages = "651-635",
number = "3",
volume = "143",
doi = "10.1007/s11240-020-01948-6",
url = "conv_220"
}
Pavlović, S., Savić, J., Milojević, J., Vinterhalter, B., Girek, Z., Adžić, S., Zečević, B.,& Banjac, N.. (2020). Introduction of the Nicotiana protein kinase (NPK1) gene by combiningAgrobacterium-mediated transformation and recurrent somatic embryogenesis to enhance salt tolerance in cauliflower. in Plant Cell Tissue and Organ Culture
Springer, Dordrecht., 143(3), 635-651.
https://doi.org/10.1007/s11240-020-01948-6
conv_220
Pavlović S, Savić J, Milojević J, Vinterhalter B, Girek Z, Adžić S, Zečević B, Banjac N. Introduction of the Nicotiana protein kinase (NPK1) gene by combiningAgrobacterium-mediated transformation and recurrent somatic embryogenesis to enhance salt tolerance in cauliflower. in Plant Cell Tissue and Organ Culture. 2020;143(3):635-651.
doi:10.1007/s11240-020-01948-6
conv_220 .
Pavlović, Suzana, Savić, Jelena, Milojević, Jelena, Vinterhalter, Branka, Girek, Zdenka, Adžić, Slađan, Zečević, Bogoljub, Banjac, Nevena, "Introduction of the Nicotiana protein kinase (NPK1) gene by combiningAgrobacterium-mediated transformation and recurrent somatic embryogenesis to enhance salt tolerance in cauliflower" in Plant Cell Tissue and Organ Culture, 143, no. 3 (2020):635-651,
https://doi.org/10.1007/s11240-020-01948-6 .,
conv_220 .

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