Analysis of DNA methylation during the germination of wheat seeds

Abstract

DNA methylation is known to play a crucial role in regulating plant development and organ or tissue differentiation. Here, we focused on the DNA methylation dynamics during the germination of wheat seeds using the adapted AFLP technique so called methylation-sensitive amplified polymorphism (MSAP). The MSAP profiles of genomic DNA in embryo and endosperm tissues of germinating seeds, as well as dry seeds were characterized and notable changes of cytosine methylation were detected. Comparisons of MSAP profiles in different tissues tested showed that the methylation level in dry seeds is the highest. The alteration analysis of cytosine methylation displayed that the number of demethylation events were three times higher than that of de novo methylation, which indicated that the demethylation was predominant in germinating wheat seeds, though the methylation events occurred as well. Sixteen differentially displayed DNA fragments in MSAP profiles were cloned and the sequencing analysis confirmed that nine of them contained CCGG sites. The further BLAST search showed that four of the cloned sequences were located in coding regions. Interestingly, three of the sixteen candidates were homologous to retrotransposons, which indicated that switches between DNA methylation and demethylation occurred in retrotransposon elements along with the germination of wheat seeds.

Scoring and analysis of methylation‐sensitive amplification polymorphisms for epigenetic population studies

“…Ever since, the approach was adopted in more than 100 publications, focusing mainly on developmental biology (e.g. Portis et al 2004;Hanai et al 2010;Moran & Perez-Figueroa 2011;Meng et al 2012), hybridization and polyploidization (e.g. Salmon et al 2005;Zhao et al 2008;Hegarty et al 2011;Rodriguez et al 2012) and plant breeding (e.g.…”

Analysis of DNA methylation of perennial ryegrass under drought using the methylation-sensitive amplification polymorphism (MSAP) technique

“…Most importantly, using this method, the cytosine methylation level within the global genome and specific loci can be analyzed without advance knowledge of the DNA sequence. MSAP has been widely employed to detect cytosine methylation in various plants, such as maize (Shan et al 2013), wheat (Meng et al 2012), banana (Baurens et al 2003), and Leymus chinensis (Yu et al 2013b).…”

Genomics: A Hallmark to Monitor Molecular and Biochemical Processes Leading Toward a Better Perceptive of Seed Aging and ex-situ Conservation

“…The involvement of DNA methylation in the seed germination process was c o n f i r m e d t h r o u g h m e t h y l a t i o n -s e n s i t i v e amplification polymorphisms (Portis et al, 2004;Meng et al, 2012).…”

“…This strategy could also be used to monitor seed aging in the future, especially for the mitochondrial genome (Lu et al, 2006;Birtic et al, 2011). DNA methylation has been implicated inthe seed germination process, using methylation sensitive amplification polymorphisms (Portis et al, 2004;Meng et al, 2012). Therefore, to characterize DNA methylation during seed aging, methylationsensitive amplification polymorphisms may serve as a new tool.…”