Abstract

Epigenetic variation, and particularly DNA methylation, is involved in plasticity and responses to changes in the environment. Conservation biology studies have focused on the measurement of this variation to establish demographic parameters, diversity levels and population structure to design the appropriate conservation strategies. However, in ex situ conservation approaches, the main objective is to guarantee the characteristics of the conserved material (phenotype and epi-genetic). We review the use of the Methylation Sensitive Amplified Polymorphism (MSAP) technique to detect changes in the DNA methylation patterns of plant material conserved by the main ex situ plant conservation methods: seed banks, in vitro slow growth and cryopreservation. Comparison of DNA methylation patterns before and after conservation is a useful tool to check the fidelity of the regenerated plants, and, at the same time, may be related with other genetic variations that might appear during the conservation process (i.e., somaclonal variation). Analyses of MSAP profiles can be useful in the management of ex situ plant conservation but differs in the approach used in the in situ conservation. Likewise, an easy-to-use methodology is necessary for a rapid interpretation of data, in order to be readily implemented by conservation managers.

Highlights

  • Human activity in recent centuries, and in recent decades, has led to overexploitation and a significant degradation of habitats, with a consequent loss of natural populations and even species

  • We review the use of the Methylation Sensitive Amplified Polymorphism (MSAP) technique to detect changes in the DNA methylation patterns of plant material conserved by the main ex situ plant conservation methods: seed banks, in vitro slow growth and cryopreservation

  • Still far from being a reality, it would be compelling and useful to find epigenetic changes associated to specific stresses imposed by storage conditions that could be used as biomarkers

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Summary

Epigenetic Variation in Ex Situ Plant Conservation

Human activity in recent centuries, and in recent decades, has led to overexploitation and a significant degradation of habitats, with a consequent loss of natural populations and even species. Slow-growth and cryopreservation are the main ex situ conservation techniques that can guarantee a higher control of the samples compared to field collections Environmental conditions for these approaches may cause an important stress to the conserved plant material. The stability of epimutations over generations is expected to be higher in plants than in animals [22] All these considerations make ex situ conservation an especially sensitive scenario in which it is important to control the state of epigenetic markers such as DNA methylation. There is a further technical shortcoming regarding the MSAP procedure: the banding pattern observed when both MspI and HpaII fail to cut Such conditions can be generated by both genetic (point mutation to the restriction site, or changes to adjacent restriction sites) and epigenetic (hypermethylation, methylation of all cytosines in the restriction site) causes. Acc65I and KpnI are isoschizomers, which differ in their sensitivity to template methylation, and, together with MseI are used for the initial digestion of genomic DNA [47]

Seed Conservation
In Vitro Plant Conservation
Cryopreservation
DNA demethylation markers out of 314
Statistical Methods for MSAP Analysis in Plant Conservation
Findings
Conclusions
Full Text
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