*4.10. Statistical Analysis*

Presented experiments were performed at least three times, independently (three technical repeats of each of three biological samples). Data represent the mean value ± standard deviation (SD) from at least three independent experiments. The significance of the differences between each mean and control was determined by Student's t-test. An asterisk above each bar indicates *p* < 0.05. The statistical analysis was performed using STATISTICA (ver. 12, StatSoft Inc.—Dell, Round Rock, TX, USA).

## **5. Conclusions**

In this article, we presented the latest research regarding ODN technology in flax, considering two genes: *LuCHS6* and *LuCHS7*. The main purpose was to demonstrate the potential maintenance of induced epigenetic changes in CHS gene expression and methylation via short oligodeoxynucleotides complementary to the gene of interest. Induced changes in the methylation status (5-methylcytosine) and CHS gene expression can be perpetuated in the next generation. Modified ODNs (especially, by methylation) were assumed to be successful in prolonging the ODN-triggered effect. Although the mechanism of ODNs' action and heredity of ODN-induced changes are not fully explained yet, the processes involving the sRNAs (small non-coding RNAs) and RdDM (RNA-directed DNA methylation) along with the rearrangement of nucleosome positioning are suspected to play a substantial role in defining the epigenome [8,9]. Thus, we suggested the great potential of ODN technology in plant research as a significant tool not only for genetic and epigenetic studies, but also crop improvement.

**Supplementary Materials:** Supplementary materials can be found at http://www.mdpi.com/1422-0067/20/16/ 3983/s1.

**Author Contributions:** M.D. and M.Z. conceived and designed the experiments and analyzed the data; M.D. performed the experiments and wrote the manuscript; J.S. and M.Z. supervised the work and revised the manuscript; A.H. performed the bioinformatic in silico analysis. All authors reviewed and approved the manuscript.

**Funding:** The research was supported by grants no. 2016/23/N/NZ9/01619 and 2013/11/B/NZ9/00150 from the National Science Centre (Available online: http://www.ncn.gov.pl).

**Acknowledgments:** We would like to give special thanks to Aleksandra Boba for sharing the primers sequences of same previously identified flax genes analyzed in this work.

**Conflicts of Interest:** The author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
