Research Article

Novel methylation specific bisulfite primer pairs for epigenetic studies of Capsicum spp.

Volume: 7 Number: 4 December 29, 2023
EN

Novel methylation specific bisulfite primer pairs for epigenetic studies of Capsicum spp.

Abstract

Over the past ten years, interest in epigenetic has rapidly increased. Heritable and stable changes in gene expression without any change in DNA sequence is in the field of epigenetics. Plants have a well-preserved epigenetic signature called DNA methylation. It is an essential epigenetic mark that protects genomic stability, silences harmful transposon insertions, and controls global gene expression in all developmental stages and environmental circumstances. All three sequence contexts, the asymmetric CpHpH context and the symmetric CpG and CpHpG contexts (where H is C, A, or T), are among DNA methylation sites in plants. Particularly, DNA cytosine methylation affects a wide range of biological processes, such as gene expression, chromatin structure, DNA packing, recombination, genomic imprinting, and DNA replication. The choice of primer pairs that flank cytosine methylation contexts is critical when designing for the detection of DNA cytosine methylation using bisulfite sequencing. We have developed and synthesized 26 bisulfite specific primer pairs suitable for DNA cytosine methylation investigations in peppers. These primers are specific to certain promoters, intergenic regions, and gene bodies (exons, introns, and UTRs). DNA samples taken from various tissues and developmental stages of Capsicum annuum L. Demre Sivrisi were analyzed by these primer pairs to confirm their utilization.

Keywords

Cytosine methylation, Epigenetics, Exon, Intron, Promoter

Supporting Institution

TÜBİTAK

Project Number

113O935

Ethical Statement

Ethics committee approval is not required.

Thanks

This study was financially supported by a grant from The Scientific and Technological Research Council of Turkey (Project No: 113O935).

References

  1. Araz, O., Ekinci, M., Yüce, M., Shams, M., Ağar, G., & Yildirim, E., (2022). Low-temperature modified DNA methylation level, genome template stability, enzyme activity, and proline content in pepper (Capsicum annuum L.) genotypes. Scientia Horticulturae, 294, 110761. https://doi.org/10.1016/j.scienta.2021.110761
  2. Cai, Y., Xu, M., Liu, J., Zeng, H., Song, J., Sun, B., Chen, S., Deng, Q., Lei, J., Cao, B., Chen, C., Chen, M., Chen, K., Chen, G., & Zhu, Z. (2022) Genome-wide analysis of histone acetyltransferase and histone deacetylase families and their expression in fruit development and ripening stage of pepper (Capsicum annuum). Frontiers in Plant Science, 13, 971230. https://doi.org/10.3389/fpls.2022.971230
  3. Dhingra, T., Mittal, K., & Sarma, G. S. (2014). Analytical techniques for DNA methylation - An Overview. Current Pharmaceutical Analysis, 10: 71-85.
  4. Gallusci, P., Hodgman, C., Teyssier, E., & Seymour, G. B. (2016). DNA methylation and chromatin regulation during fleshy fruit development and ripening. Frontiers Plant Science, 7, 807.
  5. Gruntman, E., Qi, Y., Slotkin, R. K., Roeder, T., Martienssen R. A., & Sachidanandam, R. (2008). Kismeth: analyzer of plant methylation states through bisulfite sequencing. BMC Bioinformatics, 9: 371.
  6. Hulse-Kemp, A. M., Maheshwari, S., Stoffel, K., Hill, T. A., Jaffe, D., Williams, S. R., et al. (2018). Reference quality assembly of the 3.5-Gb genome of Capsicum annuum from a single linked-read library. Horticulture Research, 5(4), 10. https://doi.org/1038/s41438-017-0011-0
  7. Ince, A. G., Karaca, M., & Onus, A. N. (2010a). Genetic relationships within and between Capsicum species. Biochemical Genetics. 48, 83-95.
  8. Ince, A. G., Karaca, M., & Onus, A. N. (2010b). Differential expression patterns of genes containing microsatellites in Capsicum annuum L. Molecular Breeding, 25, 645-658.
  9. Ince, A. G., & Karaca, M. (2017). Development and utilization of bisulfite specific primer pairs for epigenetic studies in Solanum lycopersicum L. Journal of Scientific and Engineering Research, 4(11), 145-153.
  10. Ince, A. G., & Karaca, M. (2021). Tissue and/or developmental stage specific methylation of nrDNA in Capsicum annuum. Journal of Plant Research, 134(4), 841-855. https://doi.org/10.1007/s10265-021-01287-3
APA
Ince, A. G., & Karaca, M. (2023). Novel methylation specific bisulfite primer pairs for epigenetic studies of Capsicum spp. International Journal of Agriculture Environment and Food Sciences, 7(4), 918-925. https://doi.org/10.31015/jaefs.2024.4.23
AMA
1.Ince AG, Karaca M. Novel methylation specific bisulfite primer pairs for epigenetic studies of Capsicum spp. int. j. agric. environ. food sci. 2023;7(4):918-925. doi:10.31015/jaefs.2024.4.23
Chicago
Ince, Ayse Gul, and Mehmet Karaca. 2023. “Novel Methylation Specific Bisulfite Primer Pairs for Epigenetic Studies of Capsicum Spp”. International Journal of Agriculture Environment and Food Sciences 7 (4): 918-25. https://doi.org/10.31015/jaefs.2024.4.23.
EndNote
Ince AG, Karaca M (December 1, 2023) Novel methylation specific bisulfite primer pairs for epigenetic studies of Capsicum spp. International Journal of Agriculture Environment and Food Sciences 7 4 918–925.
IEEE
[1]A. G. Ince and M. Karaca, “Novel methylation specific bisulfite primer pairs for epigenetic studies of Capsicum spp”., int. j. agric. environ. food sci., vol. 7, no. 4, pp. 918–925, Dec. 2023, doi: 10.31015/jaefs.2024.4.23.
ISNAD
Ince, Ayse Gul - Karaca, Mehmet. “Novel Methylation Specific Bisulfite Primer Pairs for Epigenetic Studies of Capsicum Spp”. International Journal of Agriculture Environment and Food Sciences 7/4 (December 1, 2023): 918-925. https://doi.org/10.31015/jaefs.2024.4.23.
JAMA
1.Ince AG, Karaca M. Novel methylation specific bisulfite primer pairs for epigenetic studies of Capsicum spp. int. j. agric. environ. food sci. 2023;7:918–925.
MLA
Ince, Ayse Gul, and Mehmet Karaca. “Novel Methylation Specific Bisulfite Primer Pairs for Epigenetic Studies of Capsicum Spp”. International Journal of Agriculture Environment and Food Sciences, vol. 7, no. 4, Dec. 2023, pp. 918-25, doi:10.31015/jaefs.2024.4.23.
Vancouver
1.Ayse Gul Ince, Mehmet Karaca. Novel methylation specific bisulfite primer pairs for epigenetic studies of Capsicum spp. int. j. agric. environ. food sci. 2023 Dec. 1;7(4):918-25. doi:10.31015/jaefs.2024.4.23