Influence of cryopreservation on the cytosine methlation state of potato genomic DNA
Shoot tips of Solanum tuberosum 'Desiree' were successfully cryopreserved by the DMSO droplet method and stored for almost 7 years, while control material was maintained in vitro for the same period of time. To analyse potential epigenetic changes, the DNA methylation status was assayed by...
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| Format: | Journal Article |
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CryoLetters C/- Royal Veterinary College
2010
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| Online Access: | http://www.ingentaconnect.com/content/cryo/cryo/2010/00000031/00000005/art00003 http://hdl.handle.net/20.500.11937/25312 |
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| author | Kaczmarczyk, Anja Houben, A. Keller, E.R. Joachim Mette, M. |
| author_facet | Kaczmarczyk, Anja Houben, A. Keller, E.R. Joachim Mette, M. |
| author_sort | Kaczmarczyk, Anja |
| building | Curtin Institutional Repository |
| collection | Online Access |
| description | Shoot tips of Solanum tuberosum 'Desiree' were successfully cryopreserved by the DMSO droplet method and stored for almost 7 years, while control material was maintained in vitro for the same period of time. To analyse potential epigenetic changes, the DNA methylation status was assayed by methylation-sensitive amplified polymorphism (MSAP) analysis using restriction endonucleases MspI and HpaII. An amount of 93.6% of the analysed MSAP signals were stable among all cryopreserved and in vitro maintained samples tested, indicating extensive stability of DNA methylation. Only 0.9% of MSAP signals showed results that differed between the two treatments and at the same time matched for all three biological replications within each treatment. These can be seen as indicating directed effects of the two treatments on the DNA methylation. Cryopreserved samples displayed in comparison to in vitro stored samples consistent hypomethylation for 0.6% (3 of 469) of MSAP signals (Table 4, pattern 4) and consistent hypermethylation for 0.2% (1 of 469), respectively. For 5.6% of all MSAP signals, inconsistent results were observed among the three biological replications at least for one of the two treatments. These were interpreted as resulting from stochastic DNA methylation changes in individual samples. As results for two biological replications were identical and different from the result for the third biological replication, the direction of methylation change could be determined in those cases. Cases of stochastic loss of CG methylation in cryopreserved samples were most frequent among them, adding up to 3.4% of MSAP signals. Stochastic loss of CG methylation was also found inmaterial maintained in vitro, only for 0.6% of all MSAP signals. In conclusion, methylation changes occurred in long-term cryopreservation of potato, in a random rather than directed fashion. Hence, cryopreservation and long-term in vitro maintenance both induce limited changes of DNA methylation status. The order of magnitude of methylation changes observed was consistent with other studies, where similar rates of DNA methylation changes have been found. |
| first_indexed | 2025-11-14T07:56:28Z |
| format | Journal Article |
| id | curtin-20.500.11937-25312 |
| institution | Curtin University Malaysia |
| institution_category | Local University |
| last_indexed | 2025-11-14T07:56:28Z |
| publishDate | 2010 |
| publisher | CryoLetters C/- Royal Veterinary College |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | curtin-20.500.11937-253122017-01-30T12:47:49Z Influence of cryopreservation on the cytosine methlation state of potato genomic DNA Kaczmarczyk, Anja Houben, A. Keller, E.R. Joachim Mette, M. DMSO droplet method Solanum tuberosum DNA methylation genetic stability Shoot tips of Solanum tuberosum 'Desiree' were successfully cryopreserved by the DMSO droplet method and stored for almost 7 years, while control material was maintained in vitro for the same period of time. To analyse potential epigenetic changes, the DNA methylation status was assayed by methylation-sensitive amplified polymorphism (MSAP) analysis using restriction endonucleases MspI and HpaII. An amount of 93.6% of the analysed MSAP signals were stable among all cryopreserved and in vitro maintained samples tested, indicating extensive stability of DNA methylation. Only 0.9% of MSAP signals showed results that differed between the two treatments and at the same time matched for all three biological replications within each treatment. These can be seen as indicating directed effects of the two treatments on the DNA methylation. Cryopreserved samples displayed in comparison to in vitro stored samples consistent hypomethylation for 0.6% (3 of 469) of MSAP signals (Table 4, pattern 4) and consistent hypermethylation for 0.2% (1 of 469), respectively. For 5.6% of all MSAP signals, inconsistent results were observed among the three biological replications at least for one of the two treatments. These were interpreted as resulting from stochastic DNA methylation changes in individual samples. As results for two biological replications were identical and different from the result for the third biological replication, the direction of methylation change could be determined in those cases. Cases of stochastic loss of CG methylation in cryopreserved samples were most frequent among them, adding up to 3.4% of MSAP signals. Stochastic loss of CG methylation was also found inmaterial maintained in vitro, only for 0.6% of all MSAP signals. In conclusion, methylation changes occurred in long-term cryopreservation of potato, in a random rather than directed fashion. Hence, cryopreservation and long-term in vitro maintenance both induce limited changes of DNA methylation status. The order of magnitude of methylation changes observed was consistent with other studies, where similar rates of DNA methylation changes have been found. 2010 Journal Article http://hdl.handle.net/20.500.11937/25312 http://www.ingentaconnect.com/content/cryo/cryo/2010/00000031/00000005/art00003 http://www.cryoletters.org/ CryoLetters C/- Royal Veterinary College fulltext |
| spellingShingle | DMSO droplet method Solanum tuberosum DNA methylation genetic stability Kaczmarczyk, Anja Houben, A. Keller, E.R. Joachim Mette, M. Influence of cryopreservation on the cytosine methlation state of potato genomic DNA |
| title | Influence of cryopreservation on the cytosine methlation state of potato genomic DNA |
| title_full | Influence of cryopreservation on the cytosine methlation state of potato genomic DNA |
| title_fullStr | Influence of cryopreservation on the cytosine methlation state of potato genomic DNA |
| title_full_unstemmed | Influence of cryopreservation on the cytosine methlation state of potato genomic DNA |
| title_short | Influence of cryopreservation on the cytosine methlation state of potato genomic DNA |
| title_sort | influence of cryopreservation on the cytosine methlation state of potato genomic dna |
| topic | DMSO droplet method Solanum tuberosum DNA methylation genetic stability |
| url | http://www.ingentaconnect.com/content/cryo/cryo/2010/00000031/00000005/art00003 http://www.ingentaconnect.com/content/cryo/cryo/2010/00000031/00000005/art00003 http://hdl.handle.net/20.500.11937/25312 |