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Oligonucleotide treatment causes flax β-glucanase up-regulation via changes in gene-body methylation.

Wojtasik W, Kulma A, Boba A, Szopa J - BMC Plant Biol. (2014)

Bottom Line: However, the strong social resistance to genetically modified organisms and restrictive regulations in European Union countries necessitated the development of a new technology for new plant types generation which uses the knowledge resulting from analysis of genetically modified plants to generate favourably altered plants while omitting the introduction of heterologous genes to their genome.In addition, EMO-βGlu flax obtained by this technology showed similar features as those obtained by genetic engineering.To our best knowledge, this is the first report on plant gene activation by treatment with oligodeoxynucleotides homologous to the coding region of the gene.

View Article: PubMed Central - PubMed

ABSTRACT

Background: Nowadays, the challenge for biotechnology is to develop tools for agriculture and industry to provide plants characterized by productivity and quality that will satisfy the growing demand for different kinds of natural products. To meet the challenge, the generation and application of genetically modified plants is justified. However, the strong social resistance to genetically modified organisms and restrictive regulations in European Union countries necessitated the development of a new technology for new plant types generation which uses the knowledge resulting from analysis of genetically modified plants to generate favourably altered plants while omitting the introduction of heterologous genes to their genome. Four-year experiments led to the development of a technology inducing heritable epigenetic gene activation without transgenesis.

Results: The method comprises the induction of changes in methylation/demethylation of the endogenous gene by the plant's treatment with short oligodeoxynucleotides antisense to the coding region. In vitro cultured plants and F3 generation flax plants overproducing the β-1,3-glucanase gene (EMO-βGlu flax) were characterized by up-regulation of β-glucanase and chitinase genes, decreases in the methylation of CCGG sequences in the β-glucanase gene and in total DNA methylation and, more importantly, reasonable resistance against Fusarium infection. In addition, EMO-βGlu flax obtained by this technology showed similar features as those obtained by genetic engineering.

Conclusion: To our best knowledge, this is the first report on plant gene activation by treatment with oligodeoxynucleotides homologous to the coding region of the gene. Apart from the evident effectiveness, the most important issue is that the EMO method allows generation of favourably altered plants, whose cultivation makes the plant producer independent from the complicated procedure of obtaining an agreement on GMO release into the environment and whose products might be more easily introduced to the global market.

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Predicted secondary structure of OLIGOs and β-1,3-glucanase mRNA and correlation coefficients between secondary structure of OLIGOs and expression level of β-1,3-glucanase gene. (A) Nucleotide sequences of OLIGOs (B1, B2, B3 and B4) and their predicted secondary structures. (B) The predicted secondary structure of β-1,3-glucanase mRNA and the binding sites of OLIGOs (B1, B2, B3 and B4). (C) Correlation coefficients between secondary structure of OLIGOs and expression level of β-1,3-glucanase gene.
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Fig6: Predicted secondary structure of OLIGOs and β-1,3-glucanase mRNA and correlation coefficients between secondary structure of OLIGOs and expression level of β-1,3-glucanase gene. (A) Nucleotide sequences of OLIGOs (B1, B2, B3 and B4) and their predicted secondary structures. (B) The predicted secondary structure of β-1,3-glucanase mRNA and the binding sites of OLIGOs (B1, B2, B3 and B4). (C) Correlation coefficients between secondary structure of OLIGOs and expression level of β-1,3-glucanase gene.

Mentions: Among several designed and tested OLIGOs the four most effective in gene (β-glucanase) activation were chosen for further analysis. The nucleotide sequences of selected OLIGOs (B1, B2, B3 and B4) and their predicted secondary structures were presented in Figure 6A. The predicted secondary structure of mRNA from the β-1,3-glucanase gene and OLIGO binding sites were shown in Figure 6B.Figure 6


Oligonucleotide treatment causes flax β-glucanase up-regulation via changes in gene-body methylation.

Wojtasik W, Kulma A, Boba A, Szopa J - BMC Plant Biol. (2014)

Predicted secondary structure of OLIGOs and β-1,3-glucanase mRNA and correlation coefficients between secondary structure of OLIGOs and expression level of β-1,3-glucanase gene. (A) Nucleotide sequences of OLIGOs (B1, B2, B3 and B4) and their predicted secondary structures. (B) The predicted secondary structure of β-1,3-glucanase mRNA and the binding sites of OLIGOs (B1, B2, B3 and B4). (C) Correlation coefficients between secondary structure of OLIGOs and expression level of β-1,3-glucanase gene.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4209061&req=5

Fig6: Predicted secondary structure of OLIGOs and β-1,3-glucanase mRNA and correlation coefficients between secondary structure of OLIGOs and expression level of β-1,3-glucanase gene. (A) Nucleotide sequences of OLIGOs (B1, B2, B3 and B4) and their predicted secondary structures. (B) The predicted secondary structure of β-1,3-glucanase mRNA and the binding sites of OLIGOs (B1, B2, B3 and B4). (C) Correlation coefficients between secondary structure of OLIGOs and expression level of β-1,3-glucanase gene.
Mentions: Among several designed and tested OLIGOs the four most effective in gene (β-glucanase) activation were chosen for further analysis. The nucleotide sequences of selected OLIGOs (B1, B2, B3 and B4) and their predicted secondary structures were presented in Figure 6A. The predicted secondary structure of mRNA from the β-1,3-glucanase gene and OLIGO binding sites were shown in Figure 6B.Figure 6

Bottom Line: However, the strong social resistance to genetically modified organisms and restrictive regulations in European Union countries necessitated the development of a new technology for new plant types generation which uses the knowledge resulting from analysis of genetically modified plants to generate favourably altered plants while omitting the introduction of heterologous genes to their genome.In addition, EMO-βGlu flax obtained by this technology showed similar features as those obtained by genetic engineering.To our best knowledge, this is the first report on plant gene activation by treatment with oligodeoxynucleotides homologous to the coding region of the gene.

View Article: PubMed Central - PubMed

ABSTRACT

Background: Nowadays, the challenge for biotechnology is to develop tools for agriculture and industry to provide plants characterized by productivity and quality that will satisfy the growing demand for different kinds of natural products. To meet the challenge, the generation and application of genetically modified plants is justified. However, the strong social resistance to genetically modified organisms and restrictive regulations in European Union countries necessitated the development of a new technology for new plant types generation which uses the knowledge resulting from analysis of genetically modified plants to generate favourably altered plants while omitting the introduction of heterologous genes to their genome. Four-year experiments led to the development of a technology inducing heritable epigenetic gene activation without transgenesis.

Results: The method comprises the induction of changes in methylation/demethylation of the endogenous gene by the plant's treatment with short oligodeoxynucleotides antisense to the coding region. In vitro cultured plants and F3 generation flax plants overproducing the β-1,3-glucanase gene (EMO-βGlu flax) were characterized by up-regulation of β-glucanase and chitinase genes, decreases in the methylation of CCGG sequences in the β-glucanase gene and in total DNA methylation and, more importantly, reasonable resistance against Fusarium infection. In addition, EMO-βGlu flax obtained by this technology showed similar features as those obtained by genetic engineering.

Conclusion: To our best knowledge, this is the first report on plant gene activation by treatment with oligodeoxynucleotides homologous to the coding region of the gene. Apart from the evident effectiveness, the most important issue is that the EMO method allows generation of favourably altered plants, whose cultivation makes the plant producer independent from the complicated procedure of obtaining an agreement on GMO release into the environment and whose products might be more easily introduced to the global market.

Show MeSH
Related in: MedlinePlus