Improved demethylation in ecological epigenetic experiments: Testing a simple and harmless foliar demethylation application
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| Formato: | Recurso digital |
| Lenguaje: | inglés |
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2017
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| _version_ | 1866901273965494272 |
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| author | Puy, Javier Dvořáková, Hana Carmona, Carlos P de Bello, Francesco Hiiesalu, Inga Latzel, Vít |
| author_facet | Puy, Javier Dvořáková, Hana Carmona, Carlos P de Bello, Francesco Hiiesalu, Inga Latzel, Vít |
| contents | <p>Experimental demethylation of plant DNA enables testing for epigenetic effects ina simple and straightforward way without the use of expensive and laborious DNAsequencing. Plants are commonly demethylated during their germination with theapplication of agents such as 5-azacytidine (5-azaC). However, this approach cancause unwanted effects such as underdeveloped root systems and high mortality oftreated plants, hindering a full comparison with untreated plants, and can be ap-plied only on plant reproducing by seeds. Here we test a simple alternative methodof plant demethylation designed to overcome the shortcomings of the germinatingmethod. We compared a novel method of demethylating plants, based on periodical spray-ing of 5-azaC aqueous solution on established seedlings, with the previous methodin which seeds were germinated directly in 5-azaC solution. We quantified theamount of methylated DNA and measured various aspects of plant performance.Also, we demonstrated its applicability in ecological epigenetic experiments bytesting transgenerational effects of plant–plant competition. We found that the spray application had similar DNA-demethylating efficiencythan the germination method, particularly in the earlier phases of plant develop-ment, but without unwanted effects. The spray application method did not reduceplant growth and performance compared to untreated plants, as opposed to thetraditional method which showed reduced growth. Also, the spray applicationmethod equalized the epigenetically modified plant features of seedlings comingfrom plants grown under competition and plants growing without competition,demonstrating its application in ecological epigenetic experiments. We conclude that regular spraying of 5-azaC solution onto established seedlingssurpassed the germination-in-solution method in terms of vigour and fitness oftreated plants. This novel method could thus be better suited for experimentalstudies seeking valuable insights into ecological epigenetics. Furthermore, thespray method can be suitable for clonal species reproducing asexually, and mostimportantly, it opens the possibility of community-level experimental demethylation of plants.</p> |
| format | Recurso digital |
| id | zenodo_https___doi_org_10_1111_2041-210X_12903 |
| institution | Zenodo |
| language | eng |
| publishDate | 2017 |
| publisher | Zenodo |
| record_format | zenodo |
| spellingShingle | Improved demethylation in ecological epigenetic experiments: Testing a simple and harmless foliar demethylation application Puy, Javier Dvořáková, Hana Carmona, Carlos P de Bello, Francesco Hiiesalu, Inga Latzel, Vít 5-azacytidine competition Demethylation DNA Methylation ecological epigenetics Ecology plant memory plant performance Toxicity transgenerational effects <p>Experimental demethylation of plant DNA enables testing for epigenetic effects ina simple and straightforward way without the use of expensive and laborious DNAsequencing. Plants are commonly demethylated during their germination with theapplication of agents such as 5-azacytidine (5-azaC). However, this approach cancause unwanted effects such as underdeveloped root systems and high mortality oftreated plants, hindering a full comparison with untreated plants, and can be ap-plied only on plant reproducing by seeds. Here we test a simple alternative methodof plant demethylation designed to overcome the shortcomings of the germinatingmethod. We compared a novel method of demethylating plants, based on periodical spray-ing of 5-azaC aqueous solution on established seedlings, with the previous methodin which seeds were germinated directly in 5-azaC solution. We quantified theamount of methylated DNA and measured various aspects of plant performance.Also, we demonstrated its applicability in ecological epigenetic experiments bytesting transgenerational effects of plant–plant competition. We found that the spray application had similar DNA-demethylating efficiencythan the germination method, particularly in the earlier phases of plant develop-ment, but without unwanted effects. The spray application method did not reduceplant growth and performance compared to untreated plants, as opposed to thetraditional method which showed reduced growth. Also, the spray applicationmethod equalized the epigenetically modified plant features of seedlings comingfrom plants grown under competition and plants growing without competition,demonstrating its application in ecological epigenetic experiments. We conclude that regular spraying of 5-azaC solution onto established seedlingssurpassed the germination-in-solution method in terms of vigour and fitness oftreated plants. This novel method could thus be better suited for experimentalstudies seeking valuable insights into ecological epigenetics. Furthermore, thespray method can be suitable for clonal species reproducing asexually, and mostimportantly, it opens the possibility of community-level experimental demethylation of plants.</p> |
| title | Improved demethylation in ecological epigenetic experiments: Testing a simple and harmless foliar demethylation application |
| topic | 5-azacytidine competition Demethylation DNA Methylation ecological epigenetics Ecology plant memory plant performance Toxicity transgenerational effects |
| url | https://doi.org/10.1111/2041-210X.12903 |