5-Azacytidine mediated reactivation of silenced transgenes in potato (Solanum tuberosum) at the whole plant level

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216208%3A11310%2F17%3A10370613" target="_blank" >RIV/00216208:11310/17:10370613 - isvavai.cz</a>

Result on the web

<a href="http://dx.doi.org/10.1007/s00299-017-2155-7" target="_blank" >http://dx.doi.org/10.1007/s00299-017-2155-7</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1007/s00299-017-2155-7" target="_blank" >10.1007/s00299-017-2155-7</a>

Result language

angličtina

Original language name

5-Azacytidine mediated reactivation of silenced transgenes in potato (Solanum tuberosum) at the whole plant level

Original language description

Transgenes introduced into plant genomes frequently become silenced either at the transcriptional or the posttranscriptional level. Transcriptional silencing is usually associated with DNA methylation in the promoter region. Treatments with inhibitors of maintenance DNA methylation were previously shown to allow reactivation of transcriptionally silenced transgenes in single cells or tissues, but not at the whole plant level. Here we analyzed the effect of DNA methylation inhibitor 5-azacytidine (AzaC) on the expression of two silenced reporter genes encoding green fluorescent protein (GFP) and neomycin phosphotransferase (NPTII) in potato plants. Whereas no obvious reactivation was observed in AzaC-treated stem cuttings, transient treatment of leaf segments with 10 mu M AzaC and subsequent de novo regeneration of shoots on the selective medium with kanamycin resulted in the production of whole plants with clearly reactivated expression of previously silenced transgenes. Reactivation of nptII expression was accompanied by a decrease in cytosine methylation in the promoter region of the gene. Using the plants with reactivated GFP expression, we found that re-silencing of this transgene can be accidentally triggered by de novo regeneration. Thus, testing the incidence of transgene silencing during de novo regeneration could be a suitable procedure for negative selection of transgenic lines (insertion events) which have an inclination to be silenced. Based on our analysis of non-specific inhibitory effects of AzaC on growth of potato shoots in vitro, we estimated that AzaC half-life in the culture media is approximately 2 days.

Czech name

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Czech description

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Type

J_imp - Article in a specialist periodical, which is included in the Web of Science database

CEP classification

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OECD FORD branch

10611 - Plant sciences, botany

Project

<a href="/en/project/LO1417" target="_blank" >LO1417: Centre of Experimental Plant Biology of CU</a><br>

Continuities

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Publication year

2017

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Name of the periodical

Plant Cell Reports

ISSN

0721-7714

e-ISSN

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Volume of the periodical

36

Issue of the periodical within the volume

8

Country of publishing house

DE - GERMANY

Number of pages

12

Pages from-to

1311-1322

UT code for WoS article

000405680300010

EID of the result in the Scopus database

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