We applied an integrative approach using multiple methods to verify cytosine methylation in the chloroplast DNA of the multicellular brown alga Saccharina japonica.
Cytosine DNA methylation is a heritable process which plays important roles in regulating development throughout the life cycle of an organism. Although methylation of nuclear DNA has been studied extensively, little is known about the state and role of DNA methylation in chloroplast genomes, especially in marine algae. Here, we have applied an integrated approach encompassing whole-genome bisulfite sequencing, methylated DNA immunoprecipitation, gene co-expression networks and photophysiological analyses to provide evidence for the role of chloroplast DNA methylation in a marine alga, the multicellular brown alga Saccharina japonica. Although the overall methylation level was relatively low in the chloroplast genome of S. japonica, gametophytes exhibited higher methylation levels than sporophytes. Gene-specific bisulfite-cloning sequencing provided additional evidence for the methylation of key photosynthetic genes. Many of them were highly expressed in sporophytes whereas genes involved in transcription, translation and biosynthesis were strongly expressed in gametophytes. Nucleus-encoded photosynthesis genes were co-expressed with their chloroplast-encoded counterparts potentially contributing to the higher photosynthetic performance in sporophytes compared to gametophytes where these co-expression networks were less pronounced. A nucleus-encoded DNA methyltransferase of the DNMT2 family is assumed to be responsible for the methylation of the chloroplast genome because it is predicted to possess a plastid transit peptide.
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The Illumina data for whole genome bisulfite sequencing were deposited at NCBI with the GEO accession number GSE117191 (https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSM3273187). The Illumina data of the MeDIP experiments were deposited at CNGB Nucleotide Sequence Archive with the accession number CNP0000364 (https://db.cngb.org/search/project/CNP0000364/). The Illumina raw data for transcriptomes were deposited at NCBI database with the SRA accession numbers SRR5860560–SRR5860568.
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TM acknowledges partial support from the School of Environmental Sciences, University of East Anglia, Norwich Research Park, UK
This work was supported by National Key Research and Development Program of China (2018YFD0900703); National Natural Science Foundation of China (32000402, 41976110); Fund for Doctor Research of Dezhou University, China (2019xgrc26); Marine S&T Fund of Shandong Province for Pilot National Laboratory for Marine Science and Technology (Qingdao) (2018SDKJ0406-3); Major Scientific and Technological Innovation Project of Shandong Provincial Key Research and Development Program (2019JZZY020706); Central Public-interest Scientific Institution Basal Research Fund, CAFS (2020TD27); China Agriculture Research System (CARS-50); Financial Fund of the Ministry of Agriculture and Rural Affairs, P.R. China (NFZX2018); Taishan Scholars Funding and Talent Projects of Distinguished Scientific Scholars in Agriculture; A Project of Shandong Province Higher Educational Science and Technology (J16LC59).
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Teng, L., Han, W., Fan, X. et al. Integrative analysis of chloroplast DNA methylation in a marine alga—Saccharina japonica. Plant Mol Biol (2021). https://doi.org/10.1007/s11103-020-01113-9
- Saccharina japonica
- Chloroplast genome
- DNA methylation