Arteaga-Vazquez, M. A. | bioRxiv Science

bioRxiv2018

Loss of CG methylation in Marchantia polymorpha causes disorganization of cell division and reveals unique DNA methylation regulatory mechanisms of non-CG methylation.

DNA methylation is an epigenetic mark that ensures silencing of transposable elements (TEs) and affects gene expression in many organisms. The function of different DNA methylation regulatory pathways has been largely characterized in the model plant Arabidopsis thaliana. However, far less is known about DNA methylation regulation and functions in basal land plants. Here we focus on the liverwort Marchantia polymorpha, an emerging model species that represents a basal lineage of land plants. We identified MpMET, the M. polymorpha orthologue of the METHYLTRANSFERASE 1 (MET1) gene required for maintenance of methylation at CG sites in angiosperms. We generated Mpmet mutants using the CRISPR/Cas9 system, which showed a significant loss of CG methylation and severe morphological changes and developmental defects. The mutants developed many adventitious shoot-like structures, suggesting that MpMET is required for maintaining differentiated cellular identities in the gametophyte. Numerous TEs were up-regulated, even though non-CG methylation was highly increased at TEs in the Mpmet mutants. Closer inspection of CHG methylation revealed features unique to M. polymorpha. Methylation of CCG sites in M. polymorpha does not depend on MET1, unlike in A. thaliana and Physcomitrella patens. Furthermore, unlike A. thaliana, M. polymorpha shows higher methylation level at CAG sites than at other CHG contexts and CAG/CTG sites are mostly methylated asymmetrically. Interestingly, CAG and CTG methylation reached comparable levels and symmetry upon loss of CG methylation. Our results highlight the diversity of non-CG methylation regulatory mechanisms in plants.

plant biology↗

bioRxiv2017

Marchantia liverworts as a proxy to plants basal microbiomes.

Microbiomes influence plant development, establishment, nutrient acquisition, pathogen defense, and the myriad of roles that ultimately impacts plant health. Plants microbiome are shaped through interactions between the microbes (ranging from cooperative functions to chemical warfare) and a selection process entailed by the host plants that distinguishes between pathogens, commensals, symbionts and transient bacteria. The soil is a primary source for microbes colonizing plants, along with other environmental sources including rain and interactions with other organisms. In this work, we explore the microbiomes through massive sequencing of the 16S rRNA gene in the eldest terrestrial plants: Marchantia liverworts. We compared microbiomes from M. polymorpha, and M. paleacea plants collected in the wild and their soils, all together luckily in the same geographical location (sympatric) thus reducing geographic effects; and also from plants grown in vitro and established from gemmae obtained from the same population of wild plants. Qualitative and quantitative microbiome analysis allowed us to identify microbes conserved in both native and in vitro Marchantia species. While M. polymorpha native plants microbiomes richness is reduced about M. paleacea, containing almost half of the Operative Taxonomic Units (OTUs) observed in M. paleacea, M. polymorpha grown in vitro exhibits larger OTUs. This diversity differences might be the result of impairment to recognize their microbial partners and being an open niche for opportunistic bacteria. The main OTUs in Marchantia microbiomes were assigned to the genera: Methylobacterium, Rhizobium, Paenibacillus, Lysobacter, Pirellula, Steroidobacter, and Bryobacter. The assigned genera correspond to bacteria capable of plant-growth promotion, complex exudates degradation, nitrogen fixation, methylotrophs, and disease-suppressive bacteria, all hosted in the relatively simple anatomy of the plant that provides refuge on their surfaces, rhizoids, and multiple gas chambers that work as specialized niches for different bacterial groups. Marchantia is a promising model to study not only long-term relationships between plants and their microbes but also the transgenerational impact of microbiomes because of Marchantia long 450 million years under climate change conditions testing microbiome configurations.

microbiology↗