A better understanding of germination and seedling establishment is needed for conservation of orchid populations. Due to the obligate association with a mycobiont, the germination niches in orchid species are extremely complex and varied. Microsites suitable for germination can be small and transient, and direct observation is difficult. An experimental approach using several levels of environmental manipulation/control is recommended.

The Pladias (Plant Diversity Analysis and Synthesis) Database of the Czech Flora and Vegetation was developed by the Pladias project team in 2014-2018 and has been continuously updated since then. The flora section of the database contains critically revised information on the Czech vascular flora, including 13.6 million plant occurrence records, which are dynamically displayed in maps, and data on 120 plant characteristics (traits, environmental associations and other information), divided into the sections: (1) Habitus and growth type, (2) Leaf, (3) Flower, (4) Fruit, seed and dispersal, (5) Belowground organs and clonality, (6) Trophic mode, (7) Karyology, (8) Taxon origin, (9) Ecological indicator values, (10) Habitat and sociology, (11) Distribution and frequency, and (12) Threats and protection. The vegetation section of the database contains information on Czech vegetation types extracted from the monograph Vegetation of the Czech Republic. The data are supplemented by national botanical bibliographies, electronic versions of the standard national flora and vegetation monographs, a database of more than 19,000 pictures of plant taxa and vegetation types, and digital maps (shapefiles) with botanical information. The data from the database are available online on a public portal www.pladias.cz, which also provides download options for various datasets and online identification keys to the species and vegetation types of the Czech Republic. In this paper, we describe the general scope, structure and content of the database, and details of the data on plant characteristics. To illustrate the data and describe the main geographic patterns in selected plant characteristics, we provide maps of mean values of numerical characteristics or proportions of categories for categorical characteristics on the map of the country in a grid of 5 longitudinal × 3 latitudinal minutes (approximately 6.0 km × 5.5 km). We also summarize the main variation patterns in the functional traits in the Czech flora using the principal component analysis.

Plant dependence on fungal carbon (mycoheterotrophy) evolved repeatedly. In orchids, it is connected with a mycorrhizal shift from rhizoctonia to ectomycorrhizal fungi and a high natural (13)C and (15)N abundance. Some green relatives of mycoheterotrophic species show identical trends, but most of these remain unstudied, blurring our understanding of evolution to mycoheterotrophy. We analysed mycorrhizal associations and (13)C and (15)N biomass content in two green species, Neottia ovata and N. cordata (tribe Neottieae), from a genus comprising green and nongreen (mycoheterotrophic) species. Our study covered 41 European sites, including different meadow and forest habitats and orchid developmental stages. Fungal ITS barcoding and electron microscopy showed that both Neottia species associated mainly with nonectomycorrhizal Sebacinales Clade B, a group of rhizoctonia symbionts of green orchids, regardless of the habitat or growth stage. Few additional rhizoctonias from Ceratobasidiaceae and Tulasnellaceae, and ectomycorrhizal fungi were detected. Isotope abundances did not detect carbon gain from the ectomycorrhizal fungi, suggesting a usual nutrition of rhizoctonia-associated green orchids. Considering associations of related partially or fully mycoheterotrophic species such as Neottia camtschatea or N. nidus-avis with ectomycorrhizal Sebacinales Clade A, we propose that the genus Neottia displays a mycorrhizal preference for Sebacinales and that the association with nonectomycorrhizal Sebacinales Clade B is likely ancestral. Such a change in preference for mycorrhizal associates differing in ecology within the same fungal taxon is rare among orchids. Moreover, the existence of rhizoctonia-associated Neottia spp. challenges the shift to ectomycorrhizal fungi as an ancestral pre-adaptation to mycoheterotrophy in the whole Neottieae.

SummaryFor germination and establishment, orchids depend on carbon (C) and nutrients supplied by mycorrhizal fungi. As adults, the majority of orchids then appear to become autotrophic. To compare the proportional C and nitrogen (N) gain from fungi in mycoheterotrophic seedlings and in adults, here we examined in the field C and N stable isotope compositions in seedlings and adults of orchids associated with ectomycorrhizal and saprotrophic fungi.Using a new highly sensitive approach, we measured the isotope compositions of seedlings and adults of four orchid species belonging to different functional groups: fully and partially mycoheterotrophic orchids associated with narrow or broad sets of ectomycorrhizal fungi, and two adult putatively autotrophic orchids associated exclusively with saprotrophic fungi.Seedlings of orchids associated with ectomycorrhizal fungi were enriched in 13 C and 15 N similarly to fully mycoheterotrophic adults. Seedlings of saprotroph-associated orchids were also enriched in 13 C and 15 N, but unexpectedly their enrichment was significantly lower, making them hardly distinguishable from their respective adult stages and neighbouring autotrophic plants.We conclude that partial mycoheterotrophy among saprotroph-associated orchids cannot be identified unequivocally based on C and N isotope compositions alone. Thus, partial mycoheterotrophy may be much more widely distributed among orchids than hitherto assumed.

SummaryPolyploidy is widely recognized as a major mechanism of sympatric speciation in plants, yet little is known about its effects on interactions with other organisms. Mycorrhizal fungi are among the most common plant symbionts and play an important role in plant nutrient supply. It remains to be understood whether mycorrhizal associations of ploidy-variable plants can be ploidy-specific.We examined mycorrhizal associations in three cytotypes (2x, 3x, 4x) of the Gymnadenia conopsea group (Orchidaceae), involving G. conopsea s.s. and G. densiflora, at different spatial scales and during different ontogenetic stages. We analysed: adults from mixed-and single-ploidy populations at a regional scale; closely spaced adults within a mixed-ploidy site; and mycorrhizal seedlings.All Gymnadenia cytotypes associated mainly with saprotrophic Tulasnellaceae (Basidiomycota). Nonetheless, both adults and seedlings of diploids and their autotetraploid derivatives significantly differed in the identity of their mycorrhizal symbionts. Interploidy segregation of mycorrhizal symbionts was most pronounced within a site with closely spaced adults.This study provides the first evidence that polyploidization of a plant species can be associated with a shift in mycorrhizal symbionts. This divergence may contribute to niche partitioning and facilitate establishment and co-existence of different cytotypes.