DNA content varies significantly over land plants, and is known to correlate with various aspects of plant form and function. In the present study, two measures of DNA content were examined in taxa across the land plant phylogeny: genome size (or C-value) and endopolyploidy (or endoreduplication index, EI). Additionally, the relationships between DNA content and various morphological and ecological traits were assessed. DNA content was determined for 64 liverwort species from 33 families. There was a large range in 1C-values from 0.27 to 20.46 pg, but no endopolyploidy was observed. There was no correlation between genome size and breeding system (monoecy vs. dioecy). Genome size and degree of endopolyploidy were determined for 74 moss species from 21 families. Genome sizes were constrained in this group, with 1C-values ranging from 0.25 to 1.21 pg. Endopolyploidy was high in all species except from the Sphagnaceae. Additionally, 1C-value was negatively correlated with desiccation tolerance but was not correlated with breeding system. DNA content variation was determined in 31 monilophyte (fern) species (including three horsetails) and 6 lycophyte (clubmoss) species. There was a wide range in 1C-values from 2.79 to 26.90 pg, and there was no indication of endopolyploidy in any of the species. Multivariate analyses were used to explore the relative contribution of traits and phylogenetic placement to DNA content varation in 41 angiosperm species. Six measures of DNA content (2C-value, 1Cx-value, leaf EI, stem EI, petal EI and root EI) were assessed. Phylogeny explained more of the variation observed in the six measures of DNA content than 21 ecological and morphological traits. However, many of the traits were able to explain some of the variation in DNA content, both with and without phylogeny included as a covaraite. One trait that was consistently correlated with DNA content was the association with arbuscular mycorrhizal (AM) fungi. In a controlled experiment, the EI in root cells colonized by AM fungi significantly increased compared to non-mycorrhizal plants. This thesis increases our knowledge regarding the extent and significance of variation in DNA content in land plants.

The colonization of the arid continental surface by plants was one of the milestones in Earth's history. Morphological innovations, such as the origin of complex 3D tissues, allowed the successful colonization and radiation of plants on land. The epidermis is the outermost plant tissue that constitutes the interface between the plant and the environment. Thus, the evolution of epidermal cells was crucial for the adaptation of plants on the terrestrial arid environment. I undertook a combined approach that aims to understand the evolutionary trends that drove land plant colonization and the genetic mechanisms that underlie the development of the epidermis. This approach includes: 1) analyses of plant transcription factors (TFs) families distribution and diversification, with a particular focus on the basic Helix-Loop-Helix (bHLH) TF family, and 2) functional characterization of a putatively conserved bHLH TF subfamily involved in epidermal cell development in land plants. Here, I showed that there was a stepwise increase in the number of transcription factor (TF) families and bHLH subfamilies that predated the colonization of the terrestrial surface by plants. The subsequent increase in TF number on land was through duplication within pre-existing TF families and subfamilies. Moreover, a similar trend occurred in metazoan bHLH TF, suggesting that the majority of innovation in plant and metazoan TF families occurred in the Precambrian before the Phanerozoic radiation of land plants and metazoans. Furthermore, I demonstrated that the function of IIIf bHLH TFs in controlling the development of the epidermal cell layer is conserved between liverworts and angiosperms. This suggests that IIIf bHLH TFs are ancient and conserved regulators of epidermal cell development since the early colonization of the land by plants. Moreover, these bHLH TFs were recruited during the evolution of land plants to control the development of seemingly unrelated morphological characters in specific lineages of extant land plants. The recruitment of ancient developmental regulators to control distinct and unrelated developmental processes in land plants might underlie the huge morphological and taxonomic radiation of plants on land.

京都大学 / 新制・課程博士 / 博士(生命科学) / 甲第24982号 / 生博第511号 / 新制||生||68(附属図書館) / 京都大学大学院生命科学研究科統合生命科学専攻 / (主査)教授 河内 孝之, 教授 荒木 崇, 教授 中野 雄司 / 学位規則第4条第1項該当 / Doctor of Philosophy in Life Sciences / Kyoto University / DFAM

Plants produce a rich variety of natural products to face environmental constraints. Enzymes of the cytochrome P450 CYP98 family are key actors in the production of phenolic bioactive compounds. They hydroxylate phenolic esters for lignin biosynthesis in angiosperms, but also produce various other bioactive phenolics. We characterized CYP98s from a moss, a lycopod, a fern, a conifer, a basal angiosperm, a monocot and from two eudicots. We found that substrate preference of the enzymes has changed during evolution of land plants with typical lignin-related activities only appearing in angiosperms, suggesting that ferns, similar to lycopods, produce lignin through an alternative route. A moss CYP98 knock-out mutant revealed coumaroyl-threonate as CYP98 substrate in vivo and showed a severe phenotype. Multiple CYP98s per species exist only in the angiosperms, where we generally found one isoform presumably involved in the biosynthesis of monolignols, and additional isoforms, resulting from independent duplications, with a broad range of functions in vitro / Graduate / 2017-08-31