An Insight into the Biology of the Rare and Peculiar Moss Pterygoneurum sibiricum (Pottiaceae): A Conservation Physiology Approach.

The biological features of the recently described peculiar and rare pottioid moss species Pterygoneurum sibiricum have been studied. A conservation physiology approach through in vitro axenic establishment and laboratory-controlled tests was applied to learn more about its development, physiology, and ecology. Additionally, ex situ collection for this species was established, and a micropropagation methodology was developed. The results obtained clearly document its reaction to salt stress in contrast to its sibling bryo-halophyte species P. kozlovii. The reaction to exogenously applied plant growth regulators, auxin and cytokinin, can be used in the different moss propagation phases of this species or for target structure production and development. Inference to the poorly known ecology of this species should also help in recent species records, and thus improve knowledge about its distribution and conservation.

Need for split: integrative taxonomy reveals unnoticed diversity in the subaquatic species of Pseudohygrohypnum (Pylaisiaceae, Bryophyta).

We present an integrative molecular and morphological study of subaquatic representatives of the genus Pseudohygrohypnum (Pylaisiaceae, Bryophyta), supplemented by distribution modelling of the revealed phylogenetic lineages. Phylogenetic analyses of nuclear and plastid datasets combined with the assemble species by automatic partitioning (ASAP) algorithm revealed eight distinct species within the traditionally circumscribed P. eugyrium and P. subeugyrium. These species are therefore yet another example of seemingly widely distributed taxa that harbour molecularly well-differentiated lineages with narrower distribution ranges. Studied accessions that were previously assigned to P. eugyrium form three clearly allopatric lineages, associated with temperate regions of Europe, eastern North America and eastern Asia. Remarkably, accessions falling under the current morphological concept of P. subeugyrium were shown to be even more diverse, containing five phylogenetic lineages. Three of these lineages occur under harsh Asian continental climates from cool-temperate to Arctic regions, while the remaining two, referred to P. subeugyrium s.str. and P. purpurascens, have more oceanic North Atlantic and East Asian distributions. Niche identity and similarity tests suggested no similarity in the distributions of the phylogenetically related lineages but revealed the identity of two East Asian species and the similarity of two pairs of unrelated species. A morphological survey confirmed the distinctness of all eight phylogenetic lineages, requiring the description of five new species. Pseudohygrohypnum appalachianum and P. orientale are described for North American and East Asian plants of P. eugyrium s.l., while P. sibiricum, P. subarcticum and P. neglectum are described for the three continental, predominantly Asian lineages of P. subeugyrium s.l. Our results highlight the importance of nontropical Asia as a center of bryophyte diversity. Phylogenic dating suggests that the diversification of subaquatic Pseudohygrohypnum lineages appeared in late Miocene, while mesophilous species of the genus split before Miocene cooling, in climatic conditions close to those where the ancestor of Pseudohygrohypnum appeared. We speculate that radiation of the P. subeugyrium complex in temperate Asia might have been driven by progressive cooling, aridification, and increases in seasonality, temperature and humidity gradients. Our results parallel those of several integrative taxonomic studies of North Asian mosses, which have resulted in a number of newly revealed species. These include various endemics from continental areas of Asia suggesting that the so-called Rapoport's rule of low diversity and wide distribution range in subpolar regions might not be applicable to bryophytes. Rather, the strong climatic oscillations in these regions may have served as a driving force of speciation and niche divergence.

MHA Herbarium: Collections of mosses from Yana-Indigirka Region, Yakutia, Russia.

BACKGROUND: The Skvortsov Herbarium of the Tsitsin Main Botanical Garden, Russian Academy of Sciences (MHA) in the 1945-1980s dealt with vascular plants and only scattered occasional collections of bryophytes and lichens were accumulated there without special arrangement. Since the late 1980s, the bryophyte studies in the MHA Herbarium became permanent and several projects were started since then, including the currently conducted "Moss Flora of Russia". There are many white spots on the map of bryophyte exploration of Russia, but one of the most conspicuous was Yakutia, the largest administrative unit of Russia, covering 3,081,000 km2. Yana-Indigirka Region, originally defined as a floristic region, includes Verkhoyansky Range and some smaller adjacent mountain areas. It is the largest amongst the bryofloristic regions in Russia, but exploration of its territory, which is difficult to access, remains far from complete. NEW INFORMATION: Several expeditions of the Institute for Biological Problems of Cryolithozone, Siberian Branch of Russian Academy of Sciences, and the Main Botanical Garden, Russian Academy of Sciences in 2000-2018 yielded in many bryophyte specimens, partly published in a number of papers. This dataset comprehensively represents the diversity of mosses of the Region. It includes 7,738 records of moss specimens preserved in the MHA Herbarium.

Cell Division Patterns in the Peristomial Layers of the Moss Genus Costesia: Two Hypotheses and a Third Solution.

The Chilean endemic genus Costesia belongs to the Gigaspermaceae, one of the most basal groups of arthrodontous mosses. While none of the species in this family has a peristome, earlier stages of sporophyte development often disclose its basic structure. The study of Costesia sporophytes at the early stages of development was conducted to identify possible similarities with Diphyscium, the genus sister to Gigaspermaceae plus all other arthrodontous mosses in the moss phylogenetic tree. Diphyscium shares a strongly unequal cell division pattern with the Dicranidae. In groups more closely related to Diphyscium, as it is the case of Costesia, this pattern is not known. Our study of Costesia found only irregular presence of slightly unequal cell divisions that may then be considered as a plesiomorphic state in peristomate mosses. The most frequently present pattern revealed in Costesia is common with the Polytrichaceae, a more basal moss group with nematodontous peristomes.

Phylogenetic structure in the Sphagnum recurvum complex (Bryophyta) in relation to taxonomy and geography.

PREMISE: The Sphagnum recurvum complex comprises a group of closely related peat mosses that are dominant components of many northern wetland ecosystems. Taxonomic hypotheses for the group range from interpreting the whole complex as one polymorphic species to distinguishing 6-10 species. The complex occurs throughout the Northern Hemisphere, and some of the putative species have intercontinental ranges. Our goals were to delimit the complex and assess its phylogenetic structure in relation to morphologically defined species and intercontinental geography. METHODS: RADseq analyses were applied to a sample of 384 collections from Europe, North America, and Asia. The data were subjected to maximum likelihood phylogenetic analyses and analyses of genetic structure using the software STRUCTURE and multivariate ordination approaches. RESULTS: The S. recurvum complex includes S. angustifolium, S. fallax, S. flexuosum, S. pacificum, and S. recurvum as clades with little evidence of admixture. We also resolved an unnamed clade that is referred to here as S. "pseudopacificum." We confirm that S. balticum and S. obtusum are nested within the complex. Species with bluntly acute to obtuse stem leaf apices are sister to those with acute to apiculate leaves. Most of the species exhibit some differentiation between intraspecific population systems disjunct on different continents. CONCLUSIONS: We recognize seven species in the amended S. recurvum complex, including S. balticum and S. obtusum, in addition to the informal clade S. "pseudopacificum." Although we detected some geographically correlated phylogenetic structure within widespread morphospecies, our RADseq data support the interpretation that these species have intercontinental geographic ranges.

Are All Paraphyllia the Same?

Moss paraphyllia, the trichome-like or foliose structures on moss stem surfaces, are usually treated as epidermal outgrowths. However, in some taxa of the moss families Leskeaceae, Neckeraceae, and Amblystegiaceae their distribution along the stem is consistently correlated with parts of the stem surface near branch primordia. In other moss families, Climaciaceae, Hylocomiaceae, and Pseudoleskeaceae the specific paraphyllia-generating epidermal layer produces paraphyllia evenly all along the stem. These results suggest that there are at least two different types of regulation of paraphyllia development; however, both of them may be involved in the morphogenesis of paraphyllia in some families, for example in the Thuidiaceae. Exogenous abscisic acid treatment consistently increases the number of paraphyllia of the Leskea-type, and it also induces the development of proximal branch leaves that normally do not develop a lamina above the stem surface. This fact supports conclusions regarding the homology of the Leskea-type of paraphyllia with leaves.

Placing the regionally threatened moss Orthodontium gracile in the big picture - Phylogeny, genome incongruence and anthropogenic dispersal in the order Orthodontiales.

The Orthodontiaceae is a small family of predominantly Southern Hemisphere temperate and South East Asian mosses that has a key phylogenetic position for research into the evolution of pleurocarpy. In the United Kingdom it is represented by the rare conservation priority species Orthodontium gracile and the abundant exotic O. lineare, introduced from the Southern Hemisphere around a century ago. Although the two species are superficially very similar and difficult to tell apart in the field, very little is known about how closely they are related or about the phylogeny, biogeography and evolutionary history of the genus Orthodontium as a whole. Phylogenetic inference and divergence time estimation were used to explore relationships within the genus globally, date major lineage splits, detect reticulate evolutionary processes and test monophyly of taxa. It was shown that Orthodontium gracile belongs to a Holarctic and Asian clade that diverged from the exclusively southern temperate lineage of O. lineare approximately 53 Ma and that it is sister to the Himalayan and South Siberian bispecific genus Orthodontopsis, which we now recognise as a single species within Orthodontium, O. lignicola. Orthodontium lignicola is quite distinct from O. gracile morphologically but may have a closely overlapping centre of extant diversity in the Himalaya, in contrast to O. lineare which is morphologically similar but biogeographically dissimilar. The introduced European populations of Orthodontium lineare were shown to share plastid and nuclear haplotypes with four collections from Tasmania and Southern Chile, but to be distinct from other Chilean and South African haplotypes. Finally, well-supported incongruence between nuclear and plastid sequences in some Western North American populations of Orthodontium gracile strongly implies one or more chloroplast capture or horizontal genome transfer events involving this species and the regionally sympatric O. pellucens. An appeal is made for targeting phylogenetic research at the intersection points of practical conservation, taxonomic uncertainty and wider biological questions and for the factoring of historical evolutionary and phylogenetic diversity into conservation assessments.

Correction: Complete mitochondrial genome sequence of the "copper moss" Mielichhoferia elongata reveals independent nad7 gene functionality loss.

[This corrects the article DOI: 10.7717/peerj.4350.].

Staminate flower of Prunus s. l. (Rosaceae) from Eocene Rovno amber (Ukraine).

The late Eocene ambers provide plethora of animal and plant fossils including well-preserved angiosperm flowers from the Baltic amber. The Rovno amber from NW Ukraine resembles in many aspects the Baltic amber; however, only fossilized animals and some bryophytes have yet been studied from the Rovno amber. We provide the first detailed description of an angiosperm flower from Rovno amber. The flower is staminate with conspicuous hypanthium, double pentamerous perianth and whorled androecium of 24 stamens much longer than the petals. Sepals are sparsely pubescent and petals are densely hirsute outside. The fossil shares important features with extant members of Prunus subgen. Padus s. l. (incl. Laurocerasus, Pygeum and Maddenia), especially with its evergreen paleotropical species. It is described here as a new species Prunus hirsutipetala D.D.Sokoloff, Remizowa et Nuraliev. Our study provides the first convincing record of fossil flowers of Rosaceae from Eocene of Europe and the earliest fossil flower of Prunus outside North America. Our record of a plant resembling extant tropical species supports palaeoentomological evidences for warm winters in northwestern Ukraine during the late Eocene, as well as suggesting a more significant role of tropical insects in Rovno amber than inferred from Baltic amber.

Development of the sphagnoid areolation pattern in leaves of Palaeozoic protosphagnalean mosses.

Background and Aims: Protosphagnalean mosses constitute the largest group of extinct mosses of still uncertain affinity. Having the general morphology of the Bryopsida, some have leaves with an areolation pattern characteristic of modern Sphagna. This study describes the structure and variation of these patterns in protosphagnalean mosses and provides a comparison with those of modern Sphagna. Methods: Preparations of fossil mosses showing preserved leaf cell structure were obtained by dissolving rock, photographed, and the resulting images were transformed to graphical format and analysed with Areoana computer software. Key Results: The sphagnoid areolation pattern is identical in its basic structure for both modern Sphagnum and Palaeozoic protosphagnalean mosses. However, in the former group the pattern develops through unequal oblique cell divisions, while in the latter the same pattern is a result of equal cell divisions taking place in a specific order with subsequent uneven cell growth. The protosphagnalean pathway leads to considerable variability in leaf structure. Conclusions: Protosphagnalean mosses had a unique ability to switch the development of leaf areolation between a pathway unique to Sphagnum and another one common to all other mosses. This developmental polyvariancy hinders attempts to classify these mosses, as characters previously considered to be of generic significance can be shown to co-occur in one individual leaf. New understanding of the ontogeny has allowed us to re-evaluate the systematic significance of such diagnostic characters in these Palaeozoic plants, showing that their similarity to Sphagnum is less substantial.

Complete mitochondrial genome sequence of the "copper moss" Mielichhoferia elongata reveals independent nad7 gene functionality loss.

The mitochondrial genome of moss Mielichhoferia elongata has been sequenced and assembled with Spades genome assembler. It consists of 100,342 base pairs and has practically the same gene set and order as in other known bryophyte chondriomes. The genome contains 66 genes including three rRNAs, 24 tRNAs, and 40 conserved mitochondrial proteins genes. Unlike the majority of previously sequenced bryophyte mitogenomes, it lacks the functional nad7 gene. The phylogenetic reconstruction and scrutiny analysis of the primary structure of nad7 gene carried out in this study suggest its independent pseudogenization in different bryophyte lineages. Evaluation of the microsatellite (simple sequence repeat) content of the M. elongata mitochondrial genome indicates that it could be used as a tool in further studies as a phylogenetic marker. The strongly supported phylogenetic tree presented here, derived from 33 protein coding sequences of 40 bryophyte species, is consistent with other reconstructions based on a number of different data sets.

And if Engler was not completely wrong? Evidence for multiple evolutionary origins in the moss flora of Macaronesia.

The Macaronesian endemic flora has traditionally been interpreted as a relict of a subtropical element that spanned across Europe in the Tertiary. This hypothesis is revisited in the moss subfamily Helicodontioideae based on molecular divergence estimates derived from two independent calibration techniques either employing fossil evidence or using an Monte Carlo Markov Chain (MCMC) to sample absolute rates of nucleotide substitution from a prior distribution encompassing a wide range of rates documented across land plants. Both analyses suggest that the monotypic Madeiran endemic genus Hedenasiastrum diverged of other Helicodontioideae about 40 million years, that is, well before Macaronesian archipelagos actually emerged, in agreement with the relict hypothesis. Hedenasiastrum is characterized by a plesiomorphic morphology, which is suggestive of a complete morphological stasis over 40 million years. Macaronesian endemic Rhynchostegiella species, whose polyphyletic origin involves multiple colonization events, evolved much more recently, and yet accumulated many more morphological novelties than H. percurrens. The Macaronesian moss flora thus appears as a complex mix of ancient relicts and more recently dispersed, fast-evolving taxa.

Origin and evolution of the northern hemisphere disjunction in the moss genus Homalothecium (Brachytheciaceae).

Competing hypotheses that rely either on a stepping-stone dispersal via the North Atlantic or the Bering land bridges, or more recent transoceanic dispersal, have been proposed to explain the disjunct distribution of Mediterranean flora in southern Europe and western North America. These hypotheses were tested with molecular dating using a phylogeny of the moss genus Homalothecium based on ITS, atpB-rbcL, and rpl16 sequence data. The monophyly of two main lineages in Western Palearctic (Europe, central Asia and north Africa) and North America is consistent with the ancient vicariance hypothesis. The monophyly of Madeiran H. sericeum accessions supports the recognition of the Macaronesian endemic H. mandonii. A range of absolute rates of molecular evolution documented in land plants was used as probabilistic calibration prior by a Bayesian inference implementing a relaxed-clock model to derive ages for the nodes of interest. Our age estimates for the divergence of the American and Western Palearctic Homalothecium clade (5.7 Ma, IC 3.52-8.26) and the origin of H. mandonii (2.52 Myr IC 0.86-8.25) are not compatible with the ancient vicariance hypothesis. Age estimates suggests that species distributions result from rare instances of dispersal and subsequent sympatric diversification. The calibrated phylogeny indicates that Homalothecium has undergone a fast radiation during the last 4 Myr, which is consistent with the low levels of morphological divergence among sibling species.

Phylogeny of the Brachytheciaceae (Bryophyta) based on morphology and sequence level data.

Brachytheciaceae is often considered a taxonomically difficult group of mosses. For example, morphological variation has led to difficulty in generic delimitation. We used DNA sequence data (chloroplast psbT-H and trnL-F and nuclear ITS2) together with morphology (63 characters) to examine the relationships within this family. The combined unaligned length of the DNA sequences used in the phylogenetic analyses varied between 1277 and 1343 bp. For phylogeny reconstruction we performed direct optimization, as implemented in POY. Analyses were performed with three different gap costs and the morphological data partition was weighted both: (1) equal to gap cost, and (2) with a weight of one. The utility of sensitivity analysis has recently been cast into doubt; hence in this study it was performed only to explore the effects of weighting on homology statements and topologies and to enable more detailed comparisons between earlier studies utilizing the direct optimization method. The wide sequence length variation of non-coding ITS2 sequences resulted in character optimizations (i.e., "alignments") of very different lengths when various gap costs were applied. Despite this variation, the topologies of equally parsimonious trees remained fairly stable. The inclusion of several outgroups, instead of only one, was observed to increase the congruence between data sets and to slightly increase the resolution. An inversion event in the 9 bp loop region in the chloroplast psbT-N spacer in mosses has been postulated to include only uninformative variation, thus possibly negatively impacting the phylogeny reconstruction. Despite this inversion, its variation within Brachytheciaceae was clearly congruent with information from other sources, but inclusion of these 9 bp in the analysis had only a minor effect on the phylogenetic results. In the most parsimonious topology, which was obtained with equal weighting of all data, Meteoriaceae and Brachytheciaceae were resolved as monophyletic sister groups, which had recently been suggested based on a few shared morphological characters. Our study revealed some new generic relationships within the Brachytheciaceae, which are discussed in light of the morphological characters traditionally used for generic delimitation.