Phytohormone Profiling across the Bryophytes

BackgroundBryophytes represent a very diverse group of non-vascular plants such as mosses, liverworts and hornworts and the oldest extant lineage of land plants. Determination of endogenous phytohormone profiles in bryophytes can provide substantial information about early land plant evolution. In this study, we screened thirty bryophyte species including six liverworts and twenty-four mosses for their phytohormone profiles in order to relate the hormonome with phylogeny in the plant kingdom.MethodologySamples belonging to nine orders (Pelliales, Jungermanniales, Porellales, Sphagnales, Tetraphidales, Polytrichales, Dicranales, Bryales, Hypnales) were collected in Central and Northern Bohemia. The phytohormone content was analysed with a high performance liquid chromatography electrospray tandem-mass spectrometry (HPLC-ESI-MS/MS).ConclusionThe apparent differences in conjugation and/or degradation strategies of growth hormones between liverworts and mosses might potentially show a hidden link between vascular plants and liverworts. On the other hand, the complement of stress hormones in bryophytes probably correlate rather with prevailing environmental conditions and plant survival strategy than with plant evolution.     

粤东亚热带森林群落树种组成对苔藓植物分布的影响

为探究森林群落树种组成对苔藓植物分布的影响,利用多元统计方法研究粤东亚热带地区苔藓植物的组成和分布对林分类型的响应。根据森林乔木层的树种组成划分为翻白叶树(Pterospermum heterophyllum)、木荷(Schima superba)+米锥(Castanopsis carlesii)和米锥3种森林类型。结果表明,3种森林类型中苔类(liverworts)和藓类(mosses)植物群落组成特征总体上存在显著差异;双齿裂萼苔(Chiloscyphus latifolius)和细指苔(Kurzia gonyotricha)在3种类型林分中的重要值变化指示了苔类植物群落的差异。3种类型林分中,均以东亚拟鳞叶藓(Pseudotaxiphyllum pohliaecarpum)为优势种。不同林分中藓类植物种类组成主要表现为亚优种分布的不同。这表明森林群落树种组成作为重要的生物因子,对林内苔藓植物的分布和种类组成有重要影响。     

Tree hollows can affect epiphyte species composition

Tree hollows often harbor animals and microorganisms, thereby storing nutritive resources derived from their biological activities. The outflows from tree hollows can create unique microenvironments, which may affect communities of epiphytic organisms on trunk surfaces below the hollows. In this study, we tested whether the species richness and composition of epiphytic bryophytes (liverworts and mosses) and lichens differ above and below tree hollows of Aria japonica and Cercidiphyllum japonicum in a Japanese temperate forest. The species richness of epiphytic bryophytes and lichens did not differ above and below hollows; however, the species composition of bryophytes differed significantly above and below hollows. Indicator species analyses showed that the moss species Anomodon tristis and the liverwort species Porella vernicosa were significantly more common below than above hollows, while the liverwort species Radula japonica and four lichen species, including Leptogium cyanescens, occurred more frequently above than below hollows. Our results highlight that tree hollows can produce unique microenvironments on trunk surfaces that potentially contribute to the maintenance of epiphytic diversity on a local scale.     

Introducing Intron Locus cox1i624 for Phylogenetic Analyses in Bryophytes: On the Issue of Takakia as Sister Genus to All Other Extant Mosses

Liverworts are well supported as the sister group to all other land plants (embryophytes) by molecular data. Observations strongly supporting this earliest dichotomy in embryophyte evolution are the strikingly different introns occurring in the mitochondrial DNAs of liverworts versus non-liverwort embryophytes (NLE), including the mosses. A final conclusion on the most basal lineages of mosses, for which genera such as Sphagnum and Takakia are the most likely candidates, is lacking. We have now investigated cox1i624, a mitochondrial group I intron conserved between the moss Physcomitrella patens and the liverwort Marchantia polymorpha. Focusing on a sampling of liverwort and moss genera, which had previously been identified as early branching taxa in their respective clades, we find that group I intron cox1i624 is universally conserved in all 33 mosses and 11 liverworts investigated. The group I intron core secondary structure is well conserved between the two ancient land plant clades. However, whereas dramatic size reductions are seen in the moss phylogeny, exactly the opposite is observed for liverworts. The cox1i624g1 locus was used for phylogenetic tree reconstruction also in combination with data sets of nad5i753g1 as well as chloroplast loci rbcL and rps4. The phylogenetic analyses revealed (i) very good support for the Treubiopsida as sister clade to all other liverworts, (ii) a sister group relationship of the nematodontous Tetraphidopsida and Polytrichopsida and (iii) two rivalling hypotheses about the basal-most moss genus with mitochondrial loci suggesting an isolated Takakia as sister to all other mosses and chloroplast loci indicating a Takakia–Sphagnum clade.     

Plant Mitochondrial RNA Editing

RNA editing affects messenger RNAs and transfer RNAs in plant mitochondria by site-specific exchange of cytidine and uridine bases in both seed and nonseed plants. Distribution of the phenomenon among bryophytes has been unclear since RNA editing has been detected in some but not all liverworts and mosses. A more detailed understanding of RNA editing in plants required extended data sets for taxa and sequences investigated. Toward this aim an internal region of the mitochondrial nad5 gene (1104 nt) was analyzed in a large collection of bryophytes and green algae (Charales). The genomic nad5 sequences predict editing in 30 mosses, 2 hornworts, and 7 simple thalloid and leafy liverworts (Jungermanniidae). No editing is, however, required in seven species of the complex thalloid liverworts (Marchantiidae) and the algae. RNA editing among the Jungermanniidae, on the other hand, reaches frequencies of up to 6% of codons being modified. Predictability of RNA editing from the genomic sequences was confirmed by cDNA analysis in the mosses Schistostega pennata and Rhodobryum roseum, the hornworts Anthoceros husnotii and A. punctatus, and the liverworts Metzgeria conjugata and Moerckia flotoviana. All C-to-U nucleotide exchanges predicted to reestablish conserved codons were confirmed. Editing in the hornworts includes the removal of genomic stop codons by frequent reverse U-to-C edits. Expectedly, no RNA editing events were identified by cDNA analysis in the marchantiid liverworts Ricciocarpos natans, Corsinia coriandra, and Lunularia cruciata. The findings are discussed in relation to models on the phylogeny of land plants. Received: 2 April 1998 / Accepted: 4 August 1998     

Mosses,Liverworts, and Hornworts Screened for Antitumor Agents

The National Cancer Institute’s record of bryophytes (mosses, liverworts, and hornworts) screened for biologically active chemicals is reviewed. In 1977, an extract of a moss from Maryland, Polytrichum ohioense, showed cytotoxicity in KB cell culture, but antitumor activity was not discovered in bryophytes until 1980. This led to a greater emphasis in collecting and screening bryophytes during 1980–1981. Samples weighing 0.5–2 kg (dried) were collected for each of 208 species and varieties. These included 184 species in 97 genera of mosses, 23 species in 16 genera of liverworts, and 1 hornwort. The methodologies for field work, extraction, and screening are briefly described. The screening results are summarized in a table listing all species alphabetically by family and by genus. For each species, the origin of the sample (country or state), collector’s number, extract number, type of extract, and 3 bioassays employed (including the dose levels used and toxic dose for the P388 lymphocytic leukemia assay) are indicated. Extracts of 75 species were toxic; extracts of 43 species were active. Activity was especially noted in the moss families Thuidiaceae, Mniaceae, Neckeraceae, Hypnaceae, Brachytheciaceae, Polytrichaceae, Dicranaceae, and Grimmiaceae. The authors suggest that bryophytes are a promising source for discovery of novel biologically active compounds. The possibility that biological activity in bryophytes may be the result of allelopathy, or the reputed ability of bryophytes to accumulate toxic substances, is considered.     

The phylogeny of land plants: A cladistic analysis based on male gametogenesis

A cladistic analysis was carried out to resolve phylogenetic pattern among bryophytes and other land plants. The analysis used 22 taxa of land plants and 90 characters relating to male gametogenesis.Coleochaete orChara/Nitella were the outgroups in various analyses using HENNIG86, PAUP, and MacClade, and the land plant phylogeny was unchanged regardless of outgroup utilized. The most parsimonious cladograms from HENNIG86 (7 trees) have treelengths of 243 (C.I. = 0.58, R.I. = 0.82). Bryophytes are monophyletic as are hornworts, liverworts, and mosses, with hornworts identified as the sister group of a liverwort/moss assemblage. In vascular plants, lycophytes are polyphyletic andSelaginella is close to the bryophytes.Lycopodium is the sister group of the remaining vascular plants (minusSelaginella). Longer treelengths (over 250) are required to produce tree topologies in which either lycophytes are monophyletic or to reconstruct the paraphyletic bryophyte phylogeny of recent authors. This analysis challenges existing concepts of bryophyte phylogeny based on more classical data and interpretations, and provides new insight into land plant evolution.     

New national and regional bryophyte records, 37

Abstract

We analyzed the form of the UV-absorption spectra and calculated the bulk UV-absorption capacity of the methanolic extracts (BUVACME) in 15 bryophytes (five liverworts and 10 mosses) from unshaded aquatic habitats of Tierra del Fuego (Argentina). BUVACME was obtained as the area under the absorbance curve (AUC) in several ranges of the UV band to allow comparison with the literature data. Both the spectra form and the AUC values depended on the species considered. The spectra showed either no, one or two defined peaks. The peaks were probably due to phenolic derivatives, which could act as both screening compounds and antioxidants. The different AUCs calculated were highly and significantly correlated, and thus it may be unimportant which of these is used. The AUC values of most liverworts were higher than those of most mosses and in particular, the liverworts Noteroclada confluens and Triandrophyllum subtrifidum showed much higher BUVACMEs than those analyzed in any other bryophyte. Thus, the accumulation of UV-absorbing compounds might often increase protection against UV radiation in liverworts, but rarely in mosses. This is in line with the evolutionary differences between these two groups of bryophytes. Except for the two abovementioned liverwort species, BUVACME of aquatic bryophytes from Tierra del Fuego is not particularly different to that found in bryophytes from other zones of the planet.     

Bryophytes and lichens in 16th-century herbaria

The diversity of bryophyte and lichen collections in 9 of the oldest preserved herbaria (dating from ca 1542 to 1577) was compared, including the first reports of bryophytes and lichens from the ‘En Tibi’ herbarium (possibly 1542–1544) and the herbarium of Leonhard Rauwolf (1560–1563). Bryophytes and lichens formed only a minority in each herbarium compared to the numbers of vascular plant specimens; numbers ranged from representatives of 21 genera in the herbarium of Ulisse Aldrovandi to the single genus Conocephalum in the Rauwolf herbarium. The focus was on large, handsome species of bryophytes and macrolichens, apart from small amounts of additional species collected as ‘by-catch’ in mixed collections. All herbaria together included 34 genera of bryophytes (36 species and 10 specimens identified to genus level) and 13 genera of lichens (24 species and 4 specimens identified to genus level). The diversity of mosses was higher than that of liverworts, and pleurocarpous mosses were more diverse than acrocarpous mosses. The collectors probably aimed at selecting material that was either characteristic of the vegetation in the respective areas of collecting or used for certain purposes (or both). The former hypothesis is supported by the small overlap in taxonomic diversity between the herbaria, and the latter by the fact that several moss, liverwort, and lichen genera are included whose traditional uses are well documented.     

Phylogeny and diversification of bryophytes

The bryophytes comprise three phyla of embryophytes that are well established to occupy the first nodes among extant lineages in the land-plant tree of life. The three bryophyte groups (hornworts, liverworts, mosses) may not form a monophyletic clade, but they share life history features including dominant free-living gametophytes and matrotrophic monosporangiate sporophytes. Because of their unique vegetative and reproductive innovations and their critical position in embryophyte phylogeny, studies of bryophytes are crucial to understanding the evolution of land plant morphology and genomes. This review focuses on phylogenetic relationships within each of the three divisions of bryophytes and relates morphological diversity to new insights about those relationships. Most previous work has been on the mosses, but progress on understanding the phylogeny of hornworts and liverworts is advancing at a rapid pace. Multilocus multigenome studies have been successful at resolving deep relationships within the mosses and liverworts, whereas single-gene analyses have advanced understanding of hornwort evolution.     

Geothermal bryophyte habitats in the South Sandwich Islands,maritime Antarctic

Question: How does geothermal activity influence terrestrial plant colonization, species composition and community development in the Antarctic? Location: South Sandwich Islands, maritime Antarctic. Methods: Bryophytes were documented during a biological survey of the archipelago in January and February 1997. Particular attention was given to sites under current or recent influence of geothermal activity. Temperature profiles obtained across defined areas of activity on several islands were linked with the presence of specific bryophytes. Results: Greatest bryophyte richness was associated with geothermally influenced ground. Of 35 moss and nine liverwort species recorded, only four mosses were never associated with heated ground, while eight of the liverworts and 50% of the mosses were found only on actively or recently heated ground. Some species occur in unheated sites elsewhere in the maritime Antarctic, but were absent from such habitats on the South Sandwich Islands. Several species occurred in distinct zones around fumaroles. Maximum temperatures recorded within the upper 0.5 cm of the vegetation surface were 40 ‐ 47 °C, with only Campylopus introflexus tolerating such temperatures. Maximum temperatures 2.5 or 5 cm below the vegetation surface of this moss reached 75 °C. Other bryophytes regularly present in zoned vegetation included the mosses Dicranella hookeri, Sanionia georgico‐uncinata, Pohlia nutans and Notoligotrichum trichodon, and the liverworts Cryptochila grandiflora and Marchantia berteroana. Surface temperatures of 25 ‐ 35 °C and subsurface temperatures of 50 ‐ 60 °C were recorded in these species. Conclusions: These exceptional plant communities illustrate the transport of viable propagules into the Antarctic. Individually ephemeral in nature, the longer term existence of geothermal habitats on islands along the Scotia Arc may have provided refugia during periods of glacial expansion, facilitating subsequent recolonization of Antarctic terrestrial habitats.     

Distribution and floristics of moss- and lichen-dominated soil crusts in a patterned Callitris glaucophylla woodland in eastern Australia

The distribution and abundance of soil crust lichens and bryophytes was examined in a patterned Callitris glaucophylla woodland in eastern Australia. Twenty-one lichen species and 26 bryophyte species were collected within thirty quadrats along a sequence of runoff, interception and runoff zones. Crust cover was significantly greatest in the interception zones (79.0 %), followed by the runoff zones (24.0 %), and lowest in the groved, runon zones (6.6 %). Lichens and bryophytes were distributed across all geomorphic zones, and, although there were significantly more moss species in the interception zones (mean = 9.1) compared with either the runoff (4.2) or runon (3.2) zones, the number of lichen species did not vary between zones. Ordination of a reduced data set of 32 species revealed a separation of taxa into distinct groups corresponding to the three geomorphic zones. Canonical correspondence analysis (CCA) of the 32 species and thirteen environmental variables revealed that the most important factors associated with the distribution of species were sheet and scarp erosion, soil stability and coherence, litter cover and crust cover. Surface cracking, microtopography and plant cover were of intermediate importance. The CCA biplot revealed that the timbered runon zones (groves) were dominated by `shade-tolerant' mosses Fissidens vittatus and Barbula hornschuchiana, whilst the heavily eroded runoff zones supported sparse populations of `erosion tolerant' lichens (Endocarpon rogersii) and mosses (Bryum argenteum and Didymodon torquatus). Interception zones supported a rich suite of `crust forming' mosses and lichens capable of tolerating moderate inundation by overland flow. Two other groups of taxa were identified by this analysis: the `pioneer' group, comprising mainly nitrogen-fixing lichens which occupy the zone of active erosion at the lower edge of the groves, and the `opportunists' dominated by liverworts, occupying the shallow depressions or bays at the margins of the groves and the interception zones. This study confirms that the non-vascular lichens and bryophytes in these arid soil crusts, are, like the vascular plants, strongly patterned according to geomorphic zone, being most strongly associated with soil surface and erosional features.     

The bryophytes Physcomitrium patens and Marchantia polymorpha as model systems for studying evolutionary cell and developmental biology in plants

Bryophytes are nonvascular spore-forming plants. Unlike in flowering plants, the gametophyte (haploid) generation of bryophytes dominates the sporophyte (diploid) generation. A comparison of bryophytes with flowering plants allows us to answer some fundamental questions raised in evolutionary cell and developmental biology. The moss Physcomitrium patens was the first bryophyte with a sequenced genome. Many cell and developmental studies have been conducted in this species using gene targeting by homologous recombination. The liverwort Marchantia polymorpha has recently emerged as an excellent model system with low genomic redundancy in most of its regulatory pathways. With the development of molecular genetic tools such as efficient genome editing, both P. patens and M. polymorpha have provided many valuable insights. Here, we review these advances with a special focus on polarity formation at the cell and tissue levels. We examine current knowledge regarding the cellular mechanisms of polarized cell elongation and cell division, including symmetric and asymmetric cell division. We also examine the role of polar auxin transport in mosses and liverworts. Finally, we discuss the future of evolutionary cell and developmental biological studies in plants.

A review of the cell biological and developmental mechanisms of bryophytes, including Physcomitrium patens and Marchantia polymorpha.     

Comprehensive phylogenomic time tree of bryophytes reveals deep relationships and uncovers gene incongruences in the last 500 million years of diversification

Premise

Bryophytes form a major component of terrestrial plant biomass, structuring ecological communities in all biomes. Our understanding of the evolutionary history of hornworts, liverworts, and mosses has been significantly reshaped by inferences from molecular data, which have highlighted extensive homoplasy in various traits and repeated bursts of diversification. However, the timing of key events in the phylogeny, patterns, and processes of diversification across bryophytes remain unclear.

Methods

Using the GoFlag probe set, we sequenced 405 exons representing 228 nuclear genes for 531 species from 52 of the 54 orders of bryophytes. We inferred the species phylogeny from gene tree analyses using concatenated and coalescence approaches, assessed gene conflict, and estimated the timing of divergences based on 29 fossil calibrations.

Results

The phylogeny resolves many relationships across the bryophytes, enabling us to resurrect five liverwort orders and recognize three more and propose 10 new orders of mosses. Most orders originated in the Jurassic and diversified in the Cretaceous or later. The phylogenomic data also highlight topological conflict in parts of the tree, suggesting complex processes of diversification that cannot be adequately captured in a single gene-tree topology.

Conclusions

We sampled hundreds of loci across a broad phylogenetic spectrum spanning at least 450 Ma of evolution; these data resolved many of the critical nodes of the diversification of bryophytes. The data also highlight the need to explore the mechanisms underlying the phylogenetic ambiguity at specific nodes. The phylogenomic data provide an expandable framework toward reconstructing a comprehensive phylogeny of this important group of plants.     

Fungal biomass associated with the phyllosphere of bryophytes and vascular plants

Little is known about the amount of fungal biomass in the phyllosphere of bryophytes compared to higher plants. In this study, fungal biomass associated with the phyllosphere of three bryophytes (Hylocomium splendens, Pleurozium schreberi, Polytrichum commune) and three vascular plants (Avenella flexuosa, Gymnocarpium dryopteris, Vaccinium myrtillus) was investigated using ergosterol content as a proxy for fungal biomass. Phyllosphere fungi accounted for 0.2-4.0 % of the dry mass of moss gametophytes, representing the first estimation of fungal biomass associated with bryophytes. Significantly more fungal biomass was associated with the phyllosphere of bryophytes than co-occurring vascular plants. The ergosterol present in moss gametophytic tissues differed significantly between species, while the ergosterol present in vascular plant leaf tissues did not. The photosynthetic tissues of mosses had less associated fungal biomass than their senescent tissues, and the magnitude of this difference varied in a species-specific manner. The fungal biomass associated with the vascular plants studied varied significantly between localities, while that of mosses did not. The observed differences in phyllosphere community biomass suggest their size could be affected by host anatomical and physiological attributes, including micro-niche availability and chemical host defenses, in addition to abiotic factors like moisture and nutrient availability.     

中国与北美苔藓植物区系关系的探讨

关于中国和北美间苔藓植物区系关系的研究是在20世纪90年代开始的。研究表明中国和北美苔类和藓类的种间关系疏远,这一事实说明两个大陆在第三纪后期分离后,由于地质、地理、气候等因素的差异致使苔藓植物区系成分也产生明显分异。虽然它们在科、属方面差异较小,但两个大陆已各自形成地区特有的苔类和藓类的属。已知的中国特有及东亚特有的苔类属有5个,藓类属有22个;而已知北美地区特有的苔类属有4个、藓类属有5个。北美东部和西部形成于不同地质时期,在苔藓植物方面与中国共同分布的科、属和种存在一定差异。从神农架藓类植物研究表明,从共同分布的种来看,北美东部与中国比北美西部与中国的相似性系数高约6％。从两地区间苔类和藓类植物的雌雄同株或雌雄异株及孢子直径大小而论,这两个因素似乎并不是影响苔藓植物在两大地区间分布的主要原因,但对苔藓植物的分布仍然十分重要。本文作者认为结合中国和北美的地史、地理条件及苔藓植物自身的特点,可以认为白令海峡不是阻碍中国和北美苔藓植物交流的绝对自然屏障;中国和北美苔藓植物区系关系目前的状况,是长期多次交流融合和分异的结果。     

Nitrogenase Activity in Relation to Intracellular Organisms in Sphagnum Mosses

Electron microscopic studies of Sphagnum lindbergii (Schimp.) and S. riparium (Ångstr.) have revealed the presence of intracellular organisms such as blue-green algae, green algae, bacteria and fungi. Nitrogenase activities of these Sphagnum mosses were found to be related mainly to the presence of intracellular Nostoc filaments. The appearance of nitrogen-fixing blue-green algae within bryophytes is thus not restricted to liverworts. The association is likely to be of ecological importance as it seems to occur in very acid habitats generally lacking blue-green algae. Possible interrelations between the moss, the blue-green algae and different types of bacteria are discussed.     

Fifty mosses on five trees: comparing phylogenetic information in three types of non-coding mitochondrial DNA and two chloroplast loci

Given the frequent genomic recombinations in plant mitochondrial DNA, intergenic regions of this organelle genome had so far not been considered as loci of potential phylogenetic information. Based on the recent evidence that an evolutionary ancient mitochondrial nad5-nad4 gene continuum is conserved in bryophytes we have compiled a dataset for a phylogenetically wide sampling of 50 mosses covering this intergenic region. The length of the intergenic region was generally in the range of 585 (Diphyscium) to 646 bp (Tomentypnum) with rare exceptions, for example all Polytrichales taxa showing a 200 bp deletion as an apparent synapomorphy of this order. Phylogenetic information in the novel marker sequence was compared with that of a nad5 gene region containing a group I intron and a nad2 gene region containing a group II intron as well as with two widely sampled chloroplast data sets, rbcL and rps4. Indel evolution in the three types of non-coding mitochondrial sequences is obviously more taxon-dependent than locus-dependent, indicating lineage-specific insertion/deletion rates. For example, larger sequence deletions are a general feature in Schistostega and Tetraphis. Although confidence for particular nodes in the phylogeny was found to vary among data sets, gene trees were essentially without conflict with respect to well-supported ones and add up in information towards a reasonably well-resolved moss phylogeny. However, while a consensus on the latter is clearly emerging, sufficient confidence is still lacking for the first dichotomies among the arthrodontous mosses leading into subclasses Bryidae, Dicranidae and Funariidae and the relative placement of nematodontous mosses (Polytrichales and Tetraphidales) on the backbone of early moss phylogeny.     

Cutin acids from bryophytes: An ω-1 hydroxy alkanoic acid in two liverwort species

The cutin acids of two species of Sphagnum moss and two species of liverwort from the order Marchantiales have been analysed. The liverwort species contain as a bound lipid, 15-hydroxyhexadecanoic acid, hitherto unreported in higher plants. The relative abundances of 8,16-, 9,16- and 10,16-dihydroxyhexadecanoic acids may be of value in the chemotaxonomy of Sphagnum mosses. Based on their cutin acid compositions, mosses and liverworts may be less related than generally supposed, the Sphagnum cutins resembling more those of angiosperms and gymnosperms. The n-alkane content of Sphagnum is very low, but there are quantitative differences between species.     

The phylogeny of mosses – Addressing open issues with a new mitochondrial locus: Group I intron cobi420

Mosses are an ancient clade of land plants, set apart from the two other monophyletic groups of bryophytes, the liverworts and the hornworts. Different molecular data sets seem to converge towards a consensus backbone phylogeny of mosses. Nevertheless, for some crucial nodes open questions remain, which obviously require additional phylogenetic information. We here report that a group I intron in the mitochondrial cob gene (cobi420) is universally conserved in the mosses, including basal genera such as Sphagnum and Takakia. Well resolved phylogenetic trees were obtained for 56 mosses of wide phylogenetic sampling, e.g. supporting a placement of Drummondia branching after the genera Timmiella and Catoscopium basal in the Dicranidae. Taxon gaps have been filled in the previously established data sets nad5i753, nad2i156, the nad5-nad4 intergenic region and chloroplast rbcL and rps4 genes. A concatenated 6-loci analysis suggests reconsideration of a Funariidae concept which includes Gigaspermaceae but leaves the isolated genus Timmia at the radiation of arthrodontous mosses unresolved.