Abstract
Chalcone synthase (CHS) has been recognized as an essential enzyme in the phenylpropanoid biosynthesis pathway. Apart from the leading role in the production of phenolic compounds with many valuable biological activities beneficial to biomedicine, CHS is well appreciated in science. Genetic engineering greatly facilitates expanding knowledge on the function and genetics of CHS in plants. The CHS gene is one of the most intensively studied genes in flax. In our study, we investigated engineering of the CHS gene through genetic and epigenetic approaches. Considering the numerous restrictions concerning the application of genetically modified (GM) crops, the main purpose of this research was optimization of the plant's modulation via epigenetics. In our study, plants modified through two methods were compared: a widely popular agrotransformation and a relatively recent oligodeoxynucleotide (ODN) strategy. It was recently highlighted that the ODN technique can be a rapid and time-serving antecedent in quick analysis of gene function before taking vector-mediated transformation. In order to understand the molecular background of epigenetic variation in more detail and evaluate the use of ODNs as a tool for predictable and stable gene engineering, we concentrated on the integration of gene expression and gene-body methylation. The treatment of flax with a series of short oligonucleotides homologous to a different part of CHS gene isoforms revealed that those directed to regulatory gene regions (5'- and 3'-UTR) activated gene expression, directed to non-coding region (introns) caused gen activity reduction, while those homologous to a coding region may have a variable influence on its activity. Gene expression changes were accompanied by changes in its methylation status. However, only certain (CCGG) motifs along the gene sequence were affected. The analyzed DNA motifs of the CHS flax gene are more accessible for methylation when located within a CpG island. The methylation motifs also led to rearrangement of the nucleosome location. The obtained results suggest high specificity of ODN action and establish a potential valuable alternative for improvement of crops.
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Claims 8
Claim · model-written summary
ODNs targeting regulatory regions activated CHS expression.
- Taxon
- flax
Linum usitatissimumt0 - Treatment
- those directed to regulatory gene regions (5'- and 3'-UTR)e0
- Gene
- CHSt0
- Outcome
- gene expressione0
- Direction
- activatede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“those directed to regulatory gene regions (5'- and 3'-UTR) activated gene expression”
t0“CHS Gene Expression in Flax”
Claim · model-written summary
Intron-targeting ODNs reduced gene activity.
- Taxon
- flaxLinum usitatissimumt0
- Treatment
- directed to non-coding region (introns)e0
- Outcome
- gen activitye0
- Direction
- reductione0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“directed to non-coding region (introns) caused gen activity reduction”
t0“CHS Gene Expression in Flax”
Claim · model-written summary
Coding-region-targeting ODNs had variable effects on gene activity.
- Taxon
- flaxLinum usitatissimumt0
- Treatment
- those homologous to a coding regione0
- Outcome
- its activitye0
Not reportedComparatorDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“those homologous to a coding region may have a variable influence on its activity”
t0“CHS Gene Expression in Flax”
Claim · model-written summary
Gene expression changes accompanied methylation changes.
- Taxon
- flaxLinum usitatissimumt0
- Gene
- CHSt0
- Outcome
- methylation statuse0
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Gene expression changes were accompanied by changes in its methylation status”
t0“CHS Gene Expression in Flax”
Claim · model-written summary
Only certain CCGG motifs were affected.
- Taxon
- flaxLinum usitatissimumt0
- Outcome
- (CCGG) motifs along the gene sequencee0
- Qualifier
- only certaine0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“only certain (CCGG) motifs along the gene sequence were affected”
t0“CHS Gene Expression in Flax”
Claim · model-written summary
CHS DNA motifs were more accessible for methylation within CpG islands.
- Taxon
- flaxLinum usitatissimume0
- Gene
- CHS flax genee0
- Outcome
- accessible for methylatione0
- Direction
- more accessiblee0
- Qualifier
- within a CpG islande0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“DNA motifs of the CHS flax gene are more accessible for methylation when located within a CpG island”
Claim · model-written summary
Methylation motifs led to nucleosome rearrangement.
- Taxon
- flaxLinum usitatissimumt0
- Outcome
- rearrangement of the nucleosome locatione0
- Qualifier
- methylation motifse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The methylation motifs also led to rearrangement of the nucleosome location”
t0“CHS Gene Expression in Flax”
Claim · model-written summary
The authors regard ODNs as a potentially valuable crop-improvement alternative.
- Taxon
- flaxLinum usitatissimumt0
- Outcome
- potential valuable alternative for improvement of cropse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“high specificity of ODN action and establish a potential valuable alternative for improvement of crops”
t0“CHS Gene Expression in Flax”