Influence of cryopreservation on the cytosine methlation state of potato genomic DNA
MetadataShow full item record
Copyright © 2010 CryoLetters
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.
Showing items related by title, author, creator and subject.
Berwick, Lyndon (2009)The analytical capacity of MSSV pyrolysis has been used to extend the structural characterisation of aquatic natural organic matter (NOM). NOM can contribute to various potable water issues and is present in high ...
A cryopreservation protocol for ex situ conservation of terrestrial orchids using asymbiotic primary and secondary (adventitious) protocormsBustam, B.; Dixon, Kingsley; Bunn, E. (2016)© 2015, The Society for In Vitro Biology. In a bid to better conserve endangered terrestrial orchids, we detail cryogenic research using a widely distributed terrestrial orchid, Caladenia latifolia, as a model species for ...
Acclimation-induced changes in cell membrane composition and influence on cryotolerance of in vitro shoots of native plant speciesFunnekotter, Bryn; Kaczmarczyk, Anja; Turner, S.; Bunn, E.; Zhou, W.; Smith, S.; Flematti, G.; Mancera, Ricardo (2013)Cell membranes are the primary sites of cryopreservation injury and measuring changes to membrane composition arising from cold acclimation may assist with providing a rationale for optimising cryopreservation methods. ...