Gamete expression of TALE class HD genes activates the diploid sporophyte program in Marchantia polymorpha

Eukaryotic life cycles alternate between haploid and diploid phases and in phylogenetically diverse unicellular eukaryotes, expression of paralogous homeodomain genes in gametes primes the haploid-to-diploid transition. In the unicellular chlorophyte alga Chlamydomonas, KNOX and BELL TALE-homeodomain genes mediate this transition. We demonstrate that in the liverwort Marchantia polymorpha, paternal (sperm) expression of three of five phylogenetically diverse BELL genes, MpBELL234, and maternal (egg) expression of both MpKNOX1 and MpBELL34 mediate the haploid-to-diploid transition. Loss-of-function alleles of MpKNOX1 result in zygotic arrest, whereas a loss of either maternal or paternal MpBELL234 results in variable zygotic and early embryonic arrest. Expression of MpKNOX1 and MpBELL34 during diploid sporophyte development is consistent with a later role for these genes in patterning the sporophyte. These results indicate that the ancestral mechanism to activate diploid gene expression was retained in early diverging land plants and subsequently co-opted during evolution of the diploid sporophyte body.

Genome-Wide Mapping of Cytosine Methylation Revealed Dynamic DNA Methylation Patterns Associated with Sporophyte Development of Saccharina japonica

Cytosine methylation plays vital roles in regulating gene expression and plant development. However, the function of DNA methylation in the development of macroalgae remains unclear. Through the genome-wide bisulfite sequencing of cytosine methylation in holdfast, stipe and blade, we obtained the complete 5-mC methylation landscape of Saccharina japonica sporophyte. Our results revealed that the total DNA methylation level of sporophyte was less than 0.9%, and the content of CHH contexts was dominant. Moreover, the distribution of CHH methylation within the genes exhibited exon-enriched characteristics. Profiling of DNA methylation in three parts revealed the diverse methylation pattern of sporophyte development. These pivotal DMRs were involved in cell motility, cell cycle and cell wall/membrane biogenesis. In comparison with stipe and blade, hypermethylation of mannuronate C5-epimerase in holdfast decreased the transcript abundance, which affected the synthesis of alginate, the key component of cell walls. Additionally, 5-mC modification participated in the regulation of blade and holdfast development by the glutamate content respectively via glutamine synthetase and amidophosphoribosyl transferase, which may act as the epigenetic regulation signal. Overall, our study revealed the global methylation characteristics of the well-defined holdfast, stipe and blade, and provided evidence for epigenetic regulation of sporophyte development in brown macroalgae.

Assessment of the independent and combined effects of copper and polycyclic aromatic hydrocarbons on gametogenesis and sporophyte development of the kelp Lessonia spicata (Phaeophyceae, Ochrophyta)

Coastal shores near the industrial park of Quintero Bay in central Chile exhibit increasing concentrations of copper (Cu) and polycyclic aromatic hydrocarbons (PAHs), well above international standards. This raises concern about their combined toxic effects on early development stages of kelps. Accordingly, we aimed to assess more accurately the independent and combined effects of Cu and PAHs on gametogenesis and sporophyte development in the kelp Lessonia spicata from central Chile by in vitro cultivation. Independent Cu and PAH trials were performed using increasing nominal concentrations of Cu and PAHs in the ranges 0.8–200 µg L−1 and 0.05–100 µg L−1, respectively. Cu and PAH median effective concentrations (EC50) on gametogenesis and early sporophyte formation were calculated using DRC in the R environment. Then, combined EC50 Cu + PAH trials were conducted to determine their effects on gametogenesis and sporophyte formation. Cu EC50 values on gametogenesis and sporophyte formation were up to three orders of magnitude lower than EC50 reported previously on spore germination in kelps. The gametogenesis (EC50 = 1.39 µg L−1) was more sensitive to Cu than sporophyte formation (EC50 = 11 µg L−1). Inversely, sporophyte formation (EC50 = 0.04 µg L−1) was more sensitive to PAHs (EC50 = 0.11 µg L−1). Considering the entire exposure period, the combined EC50 Cu + PAH exposure was the most harmful and rapid for L. spicata microscopic stages, especially the synergistic effect on early sporophytes. This highlights the need to acquire an integrated knowledge of the seasonal variation of pollutants and their combination on highly intervened coasts.

The Habitat of the Neglected Independent Protonemal Stage of Buxbaumia viridis

Buxbaumia viridis is a well-known species of decaying deadwood, which is protected in Europe. All previous studies dealing with the ecology of B. viridis rely on the sporophyte generation because the gametophyte generation is allegedly undetectable. Recent advances have shown that the protonemal stage, including gemmae, is recognizable in the field, thereby considerably modifying our perception of the species’ range and habitat. In France, we demonstrate the existence of independent protonemal populations, with the implication that the range of B. viridis is widely underestimated. Sporophytes and sterile protonema do not share the same ecological requirements. The sporophyte stage was found in montane zones, almost exclusively in coniferous forests, and on well-decayed wood. The sterile protonemal stage extends to lower elevations, in broad-leaved forests, and on wood in a less advanced state of decay. Our results suggest that the humidity could be one of the most relevant explanatory variables for the occurrence of sporophytes. Opening of the canopy seems to promote sporophyte development. Previous anomalous observations of B. viridis growing on humus or bark might be explained by the presence of a protonemal population that is able to produce sporophytes under rarely occurring but favorable climatic events.

How opposite may differ from opposite: a lesson from the peristome development in the moss Discelium

Previous morphological studies and molecular phylogenetic reconstructions resolved Discelium in the diplolepideous-opposite peristome group of mosses among the early-diverging mosses with arthrodontous peristomes. However, sporophyte development in Discelium differs from that of the other ‘diplolepideous-opposite’ families, Funariaceae and Encalyptaceae, in that the transverse sections of the peristome in the early stages of sporophyte differentiation exhibit diplolepideous-opposite, diplolepideous-alternate and haplolepideous patterns. Although the proportion of diplolepideous-opposite vs. haplolepideous patterns increases as the capsules mature, a haplolepideous peristomial formula persists in about one-third of the peristome sectors, reducing in frequency only in the lower parts of the teeth. This is the first evidence of the presence of the haplolepideous pattern in the ‘diplolepideous-opposite’ lineage; although appearing in the course of development it does not end in a really haplolepideous peristome, as its endostome and exostome elements remain opposite due to adhesion throughout their length. In contrast to Discelium, the peristome in Encalyptaceae maintains a typical diplolepideous-opposite pattern of cell divisions from the earliest stages of development, as determined by the unusually thick cells of the inner peristomial layer. The presence of the haplolepideous pattern in Discelium fills an enigmatic gap between the earliest-diverging arthrodontous lineage Diphysciaceae and the terminal lineages Dicranidae, in which the haplolepideous pattern prevails, and Bryideae, in which it appears only as a transitional stage towards the more complex structure. The diplolepideous-opposite peristome may not represent a synapomorphy for the ‘diplolepideous-opposite’ group of mosses as a whole (including Discelium), thus supporting treating Discelium in its own order.

Gamete-specific expression of TALE class HD genes activates the diploid sporophyte program in Marchantia polymorpha

Eukaryotic life cycles alternate between haploid and diploid phases and in phylogenetically diverse unicellular eukaryotes, expression of paralogous homeodomain genes in the two gametes directs the haploid-to-diploid transition. In the unicellular Chlorophyte alga Chlamydomonas KNOX and BELL TALE-homeodomain genes mediate the transition. Here we demonstrate that in the liverwort Marchantia polymorpha paternal (sperm) expression three of the five phylogenetically diverse BELL genes, MpBELL234, and maternal (egg) expression of MpKNOX1 mediate the haploid-to-diploid transition. Loss-of-function alleles of either result in zygotic or early embryonic arrest. In land plants both the haploid gametophyte and diploid sporophyte are complex multicellular organisms. Expression of MpKNOX1 and two other paralogs, MpBELL1 and MpKNOX2, during sporophyte development is consistent with a later role in patterning the sporophyte. These results indicate that the ancestral mechanism to activate diploid gene expression was retained in early diverging land plants and subsequently co-opted during evolution of the diploid sporophyte body.

Convergent recruitment of TALE homeodomain life cycle regulators to direct sporophyte development in land plants and brown algae

Three amino acid loop extension homeodomain transcription factors (TALE HD TFs) act as life cycle regulators in green algae and land plants. In mosses these regulators are required for the deployment of the sporophyte developmental program. We demonstrate that mutations in either of two TALE HD TF genes, OUROBOROS or SAMSARA, in the brown alga Ectocarpus result in conversion of the sporophyte generation into a gametophyte. The OUROBOROS and SAMSARA proteins heterodimerise in a similar manner to TALE HD TF life cycle regulators in the green lineage. These observations demonstrate that TALE-HD-TF-based life cycle regulation systems have an extremely ancient origin, and that these systems have been independently recruited to regulate sporophyte developmental programs in at least two different complex multicellular eukaryotic supergroups, Archaeplastida and Chromalveolata.