Articles
Evaluation of methylation epigenetics changes in micropropagated hop plants
Article number
1328_2
Pages
17 – 22
Language
English
Abstract
In vitro tissue cultures have been used for propagation of plants, due to their totipotency and multiplication efficacy, for more than 50 years.
In vitro tissue cultures have also been used for initiation and formation of somaclonal variability by genetic and epigenetic changes caused by development and stress conditions.
In vitro tissue culture procedures from meristems have been routinely used to produce hop (Humulus lupulus L.) virus-free rootstocks in the Czech Republic.
In this study we investigated the genetic and epigenetic variability of in vitro tissue cultures by molecular methods.
We assessed DNA methylation levels and variability of Saaz Osvalds clones, Sládek and Premiant cultivars by methylation-sensitive amplification polymorphism (MSAP). In vitro plants, regenerated rootstocks, regenerated and control plants in field conditions were used for analyses.
A total 346 clear and repetitive amplified products were detected in MSAP analyses within studied hop plants.
We found 16 polymorphic products (4.6% of products) and 64 products with methylation changes (18.5% of products) in the analyses.
Demethylation events were 14.5 times more often found than de novo methylation events.
The most demethylation changes were found in in vitro plants but only few of them were found in same mericlones under field conditions.
A hierarchical cluster analysis was used for the evaluation of molecular genetic variability within individual samples.
The resulting dendrogram showed that individual samples of the same cultivar were clustered together.
Because of detected demethylation changes for control plants within all studied cultivars, we suggest that certain degree of demethylation occurs in all plants thus it is a natural tool of plant epigenetics and evolution.
Even though, micropropagation increased methylation epigenetics, we did not find any phenotypical variation within plants under field conditions.
In vitro tissue cultures have also been used for initiation and formation of somaclonal variability by genetic and epigenetic changes caused by development and stress conditions.
In vitro tissue culture procedures from meristems have been routinely used to produce hop (Humulus lupulus L.) virus-free rootstocks in the Czech Republic.
In this study we investigated the genetic and epigenetic variability of in vitro tissue cultures by molecular methods.
We assessed DNA methylation levels and variability of Saaz Osvalds clones, Sládek and Premiant cultivars by methylation-sensitive amplification polymorphism (MSAP). In vitro plants, regenerated rootstocks, regenerated and control plants in field conditions were used for analyses.
A total 346 clear and repetitive amplified products were detected in MSAP analyses within studied hop plants.
We found 16 polymorphic products (4.6% of products) and 64 products with methylation changes (18.5% of products) in the analyses.
Demethylation events were 14.5 times more often found than de novo methylation events.
The most demethylation changes were found in in vitro plants but only few of them were found in same mericlones under field conditions.
A hierarchical cluster analysis was used for the evaluation of molecular genetic variability within individual samples.
The resulting dendrogram showed that individual samples of the same cultivar were clustered together.
Because of detected demethylation changes for control plants within all studied cultivars, we suggest that certain degree of demethylation occurs in all plants thus it is a natural tool of plant epigenetics and evolution.
Even though, micropropagation increased methylation epigenetics, we did not find any phenotypical variation within plants under field conditions.
Publication
Authors
J. Patzak, A. Henychová, P. Svoboda, I. Malířová
Keywords
Humulus lupulus L., meristem in vitro tissue cultures, mericlones, methylation-sensitive amplification polymorphism (MSAP), demethylation and de novo methylation change, epigenetic variability, hierarchical cluster analysis
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