Silene latifolia Model Dioecious Plant Transformed

Silene latifolia is a model plant for study of plant sex chromosomes evolution. It has XY chromosomes of middle age (~5–10 million years old) which allow us to study processes sex chromosomes undergone in this time period. It has large genome with many repetitive sequences which prevents it from being succesfuly sequenced and assembeled. Until very recently it wasn't possible to transform S. latifolia or any of dioecious species which severly limited our means to study many aspects of dioecy such as evolution of sex chromososomes and sex determination. Our team has devised protocol for transformation of S. latifolia. The main obstacles (low susceptibility of S. latifolia to Agrobacterium and low to none regeneration rate) were overcome by using very agresive strains of A. rhisogenes and by adding hormone metatopolin to the regeneration media. We are now using this transformation protocol to study sex determinig genes and epigenetic regulation of transposable elements by targeted mutagenesis via CRISPR/Cas9 system. We also study transgenerational proliferation of transposable elements which are unevenly distributed on sex chromosomes. We transformed Silene with artificial nonautonomous element carrying reporter GUS gene surounded by LTRs. We follow this element through crosses and assess if its copy number increases only after passing through female reproductive lineage.

Funding: This work was supported by the grant 15-21523Y from Czech Science Foundation.