engleski

Methylation-Sensitive Amplification Polymorphism (MSAP) of White Stork (Ciconia ciconia L.) DNA in Eastern Croatia

White stork (Ciconia ciconia L.) is a migratory bird species that is widespread in Europe and inhabits open arias, often wet grasslands, and agriculture habitats. It is an apex bird species and its diet mostly comprise of various amphibians, fish, small mammals and invertebrates. Nestlings are fed exclusively on local food sources and the food is provided by their parents. For that reason, nestlings are suitable bioindicators of pollution in surrounding habitats. Epigenetic modification leads to changes in gene expression without changes in DNA sequence and can be passed to the next generation. One of the epigenetic modifications is DNA methylation, where cytosines, mainly in CpG dinucleotides, are covalently modified. Stress or changes in the environment, such as pollution can cause changes in DNA methylation which can change phenotype, behaviour, fitness, and survival of the animal. For determination of differences in DNA methylation pattern we used MSAP assay, where restriction enzymes with different sensitivity cut DNA into fragments. Afterwards, adapters are ligated to the fragments and amplified with specific primers. DNA was isolated from blood of the nestlings located in the eastern Croatia during bird ringing in 2016, 2017 and 2020. PCR products were analysed on 3 % agarose gel, and level of DNA methylation was analysed using msap package in R. Results showed differences in DNA methylation between nestlings hatched in different years. We assume that these differences in DNA methylation pattern could be attributed to the factors such as environment, food and brood size.

MSAP ; methylation ; DNA ; white stork

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