Cryopreservation changes the DNA methylation of embryonic axes of Quercus robur and Fagus sylvatica seeds during in vitro culture

Nuc, Katarzyna; Marszałek, Małgorzata; Pukacki, Paweł M.

doi:10.1007/s00468-016-1416-3

Cited by 12 publications

(7 citation statements)

References 58 publications

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“…In desiccation-sensitive recalcitrant seeds, the use of dry seeds for storage is not possible because of the loss of seed viability following drying. Thus, the use of cryoprotectants is required to enabled cryopreservation of tissues isolated from recalcitrant seeds (Chmielarz et al 2011;Nuc et al 2016;Gaidamashvili et al, 2019). The present study revealed that seeds of M. boninensis have high desiccation tolerance, which is common among orthodox seeds, including Morus species (Kew Royal Botanic Gardens, 2008; https://data.…”

Section: Discussionmentioning

confidence: 83%

Seed production and storage for endangered Morus boninensis using an ex‐situ living collection

et al. 2021

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Meaningful conservation techniques for the endangered Morus boninensis require seed production and storage, since in situ purebred seed production is infrequent in Ogasawara Islands (World Natural Heritage Site) in Japan.• Ex situ living M. boninensis specimens produced seeds by open-and artificialpollination in a greenhouse. Seed desiccation tolerance and storage characteristics were examined. Open-pollinated seeds were dried at 75, 33 and 8% relative humidity (RH) for 5 days then cryopreserved at À170 °C. Artificially pollinated seeds were dried at 8% RH then stored cryogenically at À170 °C and in freezers at À80 or À20 °C for 6 months. Germination rates were compared among seeds before and after drying and storage.• Germination rates of undried seeds were 83% for open-pollinated seeds and 65% for artificially pollinated seeds. M. boninensis seeds exhibited the characteristic high desiccation tolerance of orthodox seeds and maintained a high germination rate after drying to a moisture content of approximately 4%. Cryopreservation of open-pollinated dry seeds with moisture contents of ca. 12, 7 and 4% maintained germination rates, while cryopreservation of undried seeds (moisture content ca. 44%) resulted in no germinability. Cryogenic and freezer storage of artificially pollinated dry seeds for 6 months resulted in maintenance of germination rates comparable to undried seeds before storage.• Seed production was enabled by ex situ M. boninensis collection. The present study also developed a method for low temperature seed storage. Ex situ living tree collection represents a feasible seed resource for reintroduction and restoration programmes for M. boninensis.

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Section: Discussionmentioning

confidence: 83%

Seed production and storage for endangered Morus boninensis using an ex‐situ living collection

et al. 2021

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“…Hao et al (2001) associated cryopreservation-induced demethylation events to the increased root capacity of apple shoots observed after cryopreservation. Furthermore, Nuc et al (2016) studied DNA methylation after cryopreservation of embryonic axes isolated from seeds of pedunculate oak ( Quercus robur ) and European beech ( Fagus sylvatica ), and reported different methylation status depending on their tolerance (beech) or recalcitrance (oak) to dehydration. The loss of embryogenic potential observed in long-term subcultures of Quercus alba was associated by Corredoira et al (2017) to an increase in DNA methylation, and therefore might account for the lower maturation rates observed in aged maritime pine embryogenic lines.…”

Section: Discussionmentioning

confidence: 99%

Assessment of dynamics of global DNA methylation during the cryopreservation process of Pinus pinaster embryogenic lines

Mendoza-Poudereux

Cano

Solís

et al. 2022

Preprint

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Long-term in vitro maintenance of embryogenic lines of Pinus species has been associated with lower maturation capacity, because of this, cryopreservation protocols for the embryogenic lines are needed to maintain valuable genotypes. Since cryopreservation may induce epigenetic variations, we evaluate changes in DNA methylation levels through the course of the cryopreservation of maritime pine embryogenic lines, as compared to those lines maintained by repeated subcultures. Six maritime pine embryogenic lines were cryopreserved following a protocol that includes pre-treatments inducing osmotic stress in liquid media. The percentage of methylated cytosines (%5-mC) in total DNA was determined by using a colorimetric assay. Reactive oxygen species (ROS) accumulation in cell lines was also determined by quantifying dihydroethidium (DHE) fluorescence under a confocal laser-scanning microscope. In the first experiment, we found that global DNA methylation was significantly reduced during the cryopreservation protocol. Subsequently, we evaluated the methylation status of both cryopreserved and no cryopreserved lines (maintained by subcultures) and found differences among embryogenic lines but overall, cryopreservation did not alter %5-mC of the recovered lines while periodical subcultures increased methylation rates. In addition, maltose pretreatment did not increase significantly ROS production in embryogenic lines. Our results demonstrate that the genetic stability during cryopreservation highly depends on the embryogenic line studied, but the protocol allows maintaining methylation DNA rates in most of the recovered lines.

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“…For those species with the shortest longevity predictions, such as F. sylvatica and U. glabra, we recommend retest intervals at least every 5 years and recollection where necessary as well as further research into alternative storage methods such as cryo-storage. Successful cryo-storage has already been reported for seeds of a number of temperate tree species, including A. glutinosa (Chmielarz, 2010a), C. betulus (Chmielarz, 2010b), B. pendula (Chmielarz, 2010c), U. glabra (Chmielarz, 2010d), F. excelsior (Chmielarz, 2009), M. sylvestris (Michalak et al, 2015) and seeds and embryonic axes of F. sylvatica (Chmielarz, 2007;Nuc et al, 2016).…”

Section: Discussionmentioning

confidence: 99%

Exploring seed longevity of UK native trees: implications for ex situ conservation

et al. 2020

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UK trees require increased conservation efforts due to sparse and fragmented populations. Ex situ conservation, including seed banking, can be used to better manage these issues. We conducted accelerated ageing tests on seeds of 22 UK native woody species, in order to assess their likely longevity and optimize their conservation in a seed bank. Germination at four ageing time points was determined to construct survival curves, and it was concluded that multiple samples within a species showed comparable responses for most species tested, except for Fraxinus excelsior. Of all species studied, one could be classified as very short-lived, four as short-lived and 17 as medium, with none exceeding the medium category. The most important finding of this manuscript is that although some taxonomic trends were observed, the results indicate the need for caution when making broad conclusions on potential seed storage life at a species, genus or family level. Longevity predictions were compared to actual performance of older collections held in long-term storage at the Millennium Seed Bank, Kew. Although most collections remain high in viability in storage after more than 20 years, for short-lived species at least, there is some indication that accelerated ageing predicts longevity in seed bank conditions. For species with reduced potential longevity, such as Fagus sylvatica and Ulmus glabra, additional storage options are recommended for long-term gene banking.

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Cryopreservation changes the DNA methylation of embryonic axes of Quercus robur and Fagus sylvatica seeds during in vitro culture

Cited by 12 publications

References 58 publications

Seed production and storage for endangered Morus boninensis using an ex‐situ living collection

Seed production and storage for endangered Morus boninensis using an ex‐situ living collection

Assessment of dynamics of global DNA methylation during the cryopreservation process of Pinus pinaster embryogenic lines

Exploring seed longevity of UK native trees: implications for ex situ conservation

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