Cargando…

A positive experience in applying the biolistic approach to potato varieties Aksor and Nevskiy

The method of biological ballistics (biolistic transformation, genetic bombardment) of plants is one of the most modern methods used for direct gene transfer into plant cells. The main advantages of this method include the ability to simultaneously incorporate several target genes into the plant gen...

Descripción completa

Detalles Bibliográficos
Autores principales: Malakhova, N.P., Skiba, Y.A., Iskakova, G.A., Naizabayeva, D.A., Tezekbaeva, B.K., Ismagulova, G.A., Maltseva, E.R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Federal Research Center Institute of Cytology and Genetics of Siberian Branch of the Russian Academy of Sciences 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8627886/
https://www.ncbi.nlm.nih.gov/pubmed/34901713
http://dx.doi.org/10.18699/VJ21.019
_version_ 1784606906183057408
author Malakhova, N.P.
Skiba, Y.A.
Iskakova, G.A.
Naizabayeva, D.A.
Tezekbaeva, B.K.
Ismagulova, G.A.
Maltseva, E.R.
author_facet Malakhova, N.P.
Skiba, Y.A.
Iskakova, G.A.
Naizabayeva, D.A.
Tezekbaeva, B.K.
Ismagulova, G.A.
Maltseva, E.R.
author_sort Malakhova, N.P.
collection PubMed
description The method of biological ballistics (biolistic transformation, genetic bombardment) of plants is one of the most modern methods used for direct gene transfer into plant cells. The main advantages of this method include the ability to simultaneously incorporate several target genes into the plant genome, carry out transfer without unnecessary agrobacterial parts and plasmid DNA sequences, and the short time needed to produce transgenic cells. For different plant objects, the efficiency of obtaining transgenic plants by the ballistic method varies from 1 to 3 %. For potato plants, the transformation efficiency is quite low at the moment and the selection of optimal conditions for biolistics is one of the pressing issues of practical biotechnology. This article presents a successful experience of introducing two genes of interest into two potato varieties using the biolistic approach. The results of biolistic transformation experiments are presented for two types of explants: potato internodes and calli of the varieties Aksor and Nevskiy. Of the 862 explants used for transformation, 56 regenerated plants were obtained. PCR screening of transformants revealed one plant with the insertion of the chitinase gene, one with the insertion of the endo-β-1,3-glucanase gene, and co-transformation by both genes was confirmed in four regenerants. The average transformation efficiency for potato explants was 0.7 %. A high number of regenerants (56) as opposed to a low number of transformants (6) reflects an attempt to increase the number of regenerants by using a lower concentration of the selective agent (kanamycin). Although this approach requires more effort, it can be used to produce potato lines with integrated genes of interest for further use in crop breeding. The lines of potato obtained in the current study by introducing two genes associated with the plant response to fungal pathogens will be further assessed for their resistance to fungal diseases and, if successful, will be used in potato crop breeding.
format Online
Article
Text
id pubmed-8627886
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher The Federal Research Center Institute of Cytology and Genetics of Siberian Branch of the Russian Academy of Sciences
record_format MEDLINE/PubMed
spelling pubmed-86278862021-12-10 A positive experience in applying the biolistic approach to potato varieties Aksor and Nevskiy Malakhova, N.P. Skiba, Y.A. Iskakova, G.A. Naizabayeva, D.A. Tezekbaeva, B.K. Ismagulova, G.A. Maltseva, E.R. Vavilovskii Zhurnal Genet Selektsii Original Article The method of biological ballistics (biolistic transformation, genetic bombardment) of plants is one of the most modern methods used for direct gene transfer into plant cells. The main advantages of this method include the ability to simultaneously incorporate several target genes into the plant genome, carry out transfer without unnecessary agrobacterial parts and plasmid DNA sequences, and the short time needed to produce transgenic cells. For different plant objects, the efficiency of obtaining transgenic plants by the ballistic method varies from 1 to 3 %. For potato plants, the transformation efficiency is quite low at the moment and the selection of optimal conditions for biolistics is one of the pressing issues of practical biotechnology. This article presents a successful experience of introducing two genes of interest into two potato varieties using the biolistic approach. The results of biolistic transformation experiments are presented for two types of explants: potato internodes and calli of the varieties Aksor and Nevskiy. Of the 862 explants used for transformation, 56 regenerated plants were obtained. PCR screening of transformants revealed one plant with the insertion of the chitinase gene, one with the insertion of the endo-β-1,3-glucanase gene, and co-transformation by both genes was confirmed in four regenerants. The average transformation efficiency for potato explants was 0.7 %. A high number of regenerants (56) as opposed to a low number of transformants (6) reflects an attempt to increase the number of regenerants by using a lower concentration of the selective agent (kanamycin). Although this approach requires more effort, it can be used to produce potato lines with integrated genes of interest for further use in crop breeding. The lines of potato obtained in the current study by introducing two genes associated with the plant response to fungal pathogens will be further assessed for their resistance to fungal diseases and, if successful, will be used in potato crop breeding. The Federal Research Center Institute of Cytology and Genetics of Siberian Branch of the Russian Academy of Sciences 2021-03 /pmc/articles/PMC8627886/ /pubmed/34901713 http://dx.doi.org/10.18699/VJ21.019 Text en Copyright © AUTHORS https://creativecommons.org/licenses/by/2.5/This work is licensed under a Creative Commons Attribution 4.0 License
spellingShingle Original Article
Malakhova, N.P.
Skiba, Y.A.
Iskakova, G.A.
Naizabayeva, D.A.
Tezekbaeva, B.K.
Ismagulova, G.A.
Maltseva, E.R.
A positive experience in applying the biolistic approach to potato varieties Aksor and Nevskiy
title A positive experience in applying the biolistic approach to potato varieties Aksor and Nevskiy
title_full A positive experience in applying the biolistic approach to potato varieties Aksor and Nevskiy
title_fullStr A positive experience in applying the biolistic approach to potato varieties Aksor and Nevskiy
title_full_unstemmed A positive experience in applying the biolistic approach to potato varieties Aksor and Nevskiy
title_short A positive experience in applying the biolistic approach to potato varieties Aksor and Nevskiy
title_sort positive experience in applying the biolistic approach to potato varieties aksor and nevskiy
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8627886/
https://www.ncbi.nlm.nih.gov/pubmed/34901713
http://dx.doi.org/10.18699/VJ21.019
work_keys_str_mv AT malakhovanp apositiveexperienceinapplyingthebiolisticapproachtopotatovarietiesaksorandnevskiy
AT skibaya apositiveexperienceinapplyingthebiolisticapproachtopotatovarietiesaksorandnevskiy
AT iskakovaga apositiveexperienceinapplyingthebiolisticapproachtopotatovarietiesaksorandnevskiy
AT naizabayevada apositiveexperienceinapplyingthebiolisticapproachtopotatovarietiesaksorandnevskiy
AT tezekbaevabk apositiveexperienceinapplyingthebiolisticapproachtopotatovarietiesaksorandnevskiy
AT ismagulovaga apositiveexperienceinapplyingthebiolisticapproachtopotatovarietiesaksorandnevskiy
AT maltsevaer apositiveexperienceinapplyingthebiolisticapproachtopotatovarietiesaksorandnevskiy
AT malakhovanp positiveexperienceinapplyingthebiolisticapproachtopotatovarietiesaksorandnevskiy
AT skibaya positiveexperienceinapplyingthebiolisticapproachtopotatovarietiesaksorandnevskiy
AT iskakovaga positiveexperienceinapplyingthebiolisticapproachtopotatovarietiesaksorandnevskiy
AT naizabayevada positiveexperienceinapplyingthebiolisticapproachtopotatovarietiesaksorandnevskiy
AT tezekbaevabk positiveexperienceinapplyingthebiolisticapproachtopotatovarietiesaksorandnevskiy
AT ismagulovaga positiveexperienceinapplyingthebiolisticapproachtopotatovarietiesaksorandnevskiy
AT maltsevaer positiveexperienceinapplyingthebiolisticapproachtopotatovarietiesaksorandnevskiy