Differential Transgeneration Methylation of Exogenous Promoters in T1 Transgenic Wheat ( Triticum aestivum )
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ifferential Transgeneration Methylation of Exogenous Promoters in T1 Transgenic Wheat (Triticum aestivum) Mona Mohamed Elseehy* Department of Genetics, Faculty of Agriculture, University of Alexandria, Elshatby, Alexandria, Egypt *e-mail: [email protected] Received June 18, 2019; revised October 16, 2019; accepted September 18, 2020
Abstract—DNA methylation has become an essential molecular approach to regulate gene expression through the regulation of methyl group addition/removal at the 5th position of Cytosine. The T1 progeny of T0 transgenic wheat plants were used to study the transgeneration methylation of promoter proximal regions of two exogenous promoters using bisulfite sequencing. The progeny of low and high T0 IFS expressers with high and low methylation levels respectively were used. Results of this study revealed that T1 plants that have 35S promoter driving the IFS inherited the methylation status of 35S promoter and IFS expression, especially methylation at the –56 and –88 CpG islands. On the other hand, the high IFS expresser of T0 plants with OL promoter driving the IFS expression passed the same pattern of IFS expression and methylation to their T1 progeny, whereas the low IFS expresser of T0 changed the pattern of expression and methylation to the high T0 expresser in their T1 progeny. This indicates that T1 wheat plants were able to demethylates DNA of the OL promoter proximal region, especially at –106 and –151 and reconstitute the IFS expression from low to high expresser through one generation. This also could indicate that plant promoters are more suitable for driving transgene in plant biotechnology. Results will improve our understanding of regulation of gene expression by DNA methylation and their application in plant biotechnology. Keywords: transgenic, wheat, methylation, transgeneration, epigenetic, DNA methyltransferases DOI: 10.3103/S0095452720050151
INTRODUCTION Epigenetics has made it difficult to understand how genetic information is stored, expressed, and shape the phenotype. Currently, there is intensive research to better understand the epigenetic regulation of development, phenotype characteristics, and adaptation of higher organisms to changing environments [1, 2]. The best studied mechanism of epigenetics is the DNA methylation which has become a important route for crop development via applied epigenetics [1]. Addition of methyl group to carbon 5 of cytosine results in DNA methylation. DNA methylation in plants has been reported at three specific nucleotide sequences: CpG, CpNpG, and CpNpN, where N indicates A, T, or C [3]. Cytosine methylation at these sites is achieved by different DNA methyltransferases. The CpG methylation is achieved and maintained by DNA methyltransferase I (MET1) [4, 5], while CpNpG methylation is maintained by Chromomethyltransferase 3 (CMT3) [6]. The methylation of CpG and CpNpG is inherited via meiosis [7]. On the other hand, methylation of the CpNpN is not inherited and maintained by Domains Rearranged Methyltransferase 2 (DRM2) through de no
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