We are continuing the chemosystematic study of the liverworts by gas chromatography (GC)-and liquid chromatography (LC)-MS analysis, as resolution of the relationships between species of liverworts is difficult because of their small size, and ephemeral nature of the sporophytic generation, which yields many conventionally critical taxonomic characters. This is particularly true of the genus Radula. This investigation is part of a chemosystematic study devoted to assisting the resolution of infrageneric relationships within Radula. During our investigation of the genus of Radula, we found the cannabinoid type compound, perrottetinene in the Japanese liverwort, Radula perrottetii.1) Radula species are very rich sources of aromatic compounds and are extremely rich in prenylated bibenzyls.2-4) The ether extract of the New Zealand liverwort R. marginata, which have not yet been investigated phytochemically, afforded new cannabinoid type and prenylated bibenzyls.The ether extract of R. marginata was chromatographed on silica gel to give three new aromatic compounds 1, 3 and 4, together with known 2, 5, 6 and d-tocopherol (Fig. 1 Hz, H-5Љ, 7Љ) were observed in the 1 H-NMR spectrum of 1, due to a 1-substituted aromatic ring. A fragment peak, tropylium cation (C 7 H 7 ϩ ) at m/z 91 in the electron impact (El)-MS of 1 provided further evidence for this aromatic ring. The EI-MS spectrum of 1 showed a molecular ion peak at m/z 392, its constitution of C 25 H 28 O 4 was confirmed by its high resolution EI-MS measurement. Analysis of the heteronuclear multiple quantum coherence (HMQC) and heteronuclear multiple bond correlation (HMBC) spectra (important correlations are summarized in Fig. 2) supports the structural assignment. The nuclear Overhauser enhancement spectroscopy (NOESY spectrum of 1 (Fig. 3) showed correlations between H-3 and 4, confirming the stereochemistry of a cis-fused ring. Consideration of these spectral data led to the conclusion that the compound was perrottetinenic acid (1), although its absolute configuration remains to be clarified. It is noteworthy that in this investigation the GC-MS analysis of 1 showed a same retention time and mass spectrum as those of 2 because of decarboxylation of 1 in an injection at 250°C of GC.Compound 3 was isolated as a minor constituent of this species, the coloration of which showed dark blue with spray of FeCl 3 on a TLC. The presence of a phenolic hydroxyl group was anticipated. The structure of 3 was deduced by comparing its spectral data with those of 2. Whereas the car-* To whom correspondence should be addressed. e-mail: toyota@ph.bunri-u.ac. The ether extract of the New Zealand liverwort Radula marginata afforded a new cannabinoid type bibenzyl compound named perrottetinenic acid, and two new bibenzyls, together with a known cannabinoid, perrottetinene. Their structures were established by two dimensional (2D) NMR spectral data. The structure of perrottetinenic acid was a similar to that of D D 1 -tetrahydrocannabinol, a known hallucinogen. Cannabinoid type bibe...

Summary• Shifts in sexual systems are among the most common and important transitions in plants and are correlated with a suite of life-history traits. The evolution of sexual systems and their relationships to gametophyte size, sexual and asexual reproduction, and epiphytism are examined here in the liverwort genus Radula.• The sequence of trait acquisition and the phylogenetic correlations between those traits was investigated using comparative methods.• Shifts in sexual systems recurrently occurred from dioecy to monoecy within facultative epiphyte lineages. Production of specialized asexual gemmae was correlated to neither dioecy nor strict epiphytism.• The significant correlations among life-history traits related to sexual systems and habitat conditions suggest the existence of evolutionary trade-offs. Obligate epiphytes do not produce gemmae more frequently than facultative epiphytes and disperse by whole gametophyte fragments, presumably to avoid the sensitive protonemal stage in a habitat prone to rapid changes in moisture availability. As dispersal ranges correlate with diaspore size, this reinforces the notion that epiphytes experience strong dispersal limitations. Our results thus provide the evolutionary complement to metapopulation, metacommunity and experimental studies demonstrating trade-offs between dispersal distance, establishment ability, and life-history strategy, which may be central to the evolution of reproductive strategies in bryophytes.

Summary Australia is an excellent setting to explore relationships between climate change and diversification dynamics. Aridification since the Eocene has resulted in spectacular radiations within one or more Australian biomes. Acacia is the largest plant genus on the Australian continent, with around 1000 species, and is present in all biomes. We investigated the macroevolutionary dynamics of Acacia within climate space. We analysed phylogenetic and climatic data for 503 Acacia species to estimate a time‐calibrated phylogeny and central climatic tendencies for BioClim layers from 132 000 herbarium specimens. Diversification rate heterogeneity and rates of climate space exploration were tested. We inferred two diversification rate increases, both associated with significantly higher rates of climate space exploration. Observed spikes in climate disparity within the Pleistocene correspond with onset of Pleistocene glacial–interglacial cycling. Positive time dependency in environmental disparity applies in the basal grade of Acacia, though climate space exploration rates were lower. Incongruence between rates of climate space exploration and disparity suggests different Acacia lineages have experienced different macroevolutionary processes. The second diversification rate increase is associated with a south‐east Australian mesic lineage, suggesting adaptations to progressively aridifying environments and ability to transition into mesic environments contributed to Acacia's dominance across Australia.

The number of available liverwort fossils substantially increased within the past decade, which is mainly due to new findings from Cretaceous and Cenozoic amber deposits. Many of them, however, are fragmentary and not predestined for consideration in evolutionary analyses. Here, we list those liverwort fossils that we suggest as suitable for calibrating phylogenetic reconstructions, along with brief descriptions, justification of their use, and age information. Our recommendations are based on thorough microscopic investigation of available fossils from worldwide amber collections including recent findings. We recommend that the following 42 fossil taxa can be used as confident minimum age constraints in phylogenetic reconstructions: Acrolejeunea ucrainica (35 Ma), Anastrophyllum rovnoi (35 Ma), Bazzania polyodus (34 Ma), Blepharolejeunea obovata (15 Ma), the genus Bryopteris with B. bispinosa and B. succinea (15 Ma), Calypogeia stenzeliana (34 Ma), Cephaloziella nadezhdae (35 Ma), the genus Ceratolejeunea with C. antiqua, C. palaeomexicana, and C. sublaetefusca (15 Ma), Cheilolejeunea latiloba (34 Ma), Cheirorhiza brittae (158 Ma), Cololejeunea sp. (15 Ma), Cyclolejeunea archaica (15 Ma), Dibrachiella grollei (15 Ma), Diettertia montanensis (112 Ma), Drepanolejeunea eogena (15 Ma), the genus Frullania with F. baerlocheri, F. cretacea, and F. partita (99 Ma), Frullania subgen. Frullania with F. casparyi (34 Ma) and F. riclefgrollei (35 Ma), F. subgen. Trachycolea with F. rovnoi (35 Ma) and F. schumannii (34 Ma), Gackstroemia cretacea (99 Ma), Geocalyx heinrichsii (34 Ma), the genus Lejeunea with L. hamatiloba, L. miocenica, L. resinata, and L. urbanioides (15 Ma), Lopholejeunea subnigricans (15 Ma), Marchantites cyathodoides (228 Ma), Marchesinia brachiata (15 Ma), Metzgeriothallus sharonae (383 Ma), Microlejeunea nyiahae (52 Ma), Neurolejeunea macrostipula (15 Ma), Nipponolejeunea europaea (34 Ma), Notoscyphus balticus (34 Ma), Odontoschisma (sect. Iwatsukia) dimorpha (34 Ma), Plagiochila groehnii (34 Ma), Porella subgrandiloba (34 Ma), Protolophozia kutscheri (34 Ma), Radula (subgen. Odontoradula) cretacea (99 Ma), R. (subgen. Amentuloradula) heinrichsii (99 Ma), Scapania hoffeinsiana (34 Ma), Solenostoma berendtii (34 Ma), Spruceanthus polonicus (34 Ma), Stictolejeunea squamata (15 Ma), Tetralophozia groehnii (34 Ma), Thysananthus auriculatus (15 Ma), Thysananthus contortus (34 Ma). Furthermore, we transfer Lophozia kutscheri to Protolophozia, Archilejeunea grollei to Dibrachiella, Frullania ucrainica to Acrolejeunea, and Mastigolejeunea extincta to Spruceanthus, based on new fossil evidence and morphological revisions.

Molecular data from three chloroplast markers resolve individuals attributable to Radula buccinifera in six lineages belonging to two subgenera, indicating the species is polyphyletic as currently circumscribed. All lineages are morphologically diagnosable, but one pair exhibits such morphological overlap that they can be considered cryptic. Molecular and morphological data justify the re-instatement of a broadly circumscribed ecologically variable R. strangulata, of R. mittenii, and the description of five new species. Two species Radula mittenii Steph. and R. notabilis sp. nov. are endemic to the Wet Tropics Bioregion of north-east Queensland, suggesting high diversity and high endemism might characterise the bryoflora of this relatively isolated wet-tropical region. Radula demissa sp. nov. is endemic to southern temperate Australasia, and like R. strangulata occurs on both sides of the Tasman Sea. Radula imposita sp. nov. is a twig and leaf epiphyte found in association with waterways in New South Wales and Queensland. Another species, R. pugioniformis sp. nov., has been confused with Radula buccinifera but was not included in the molecular phylogeny. Morphological data suggest it may belong to subg. Odontoradula. Radula buccinifera is endemic to Australia including Western Australia and Tasmania, and to date is known from south of the Clarence River on the north coast of New South Wales. Nested within R. buccinifera is a morphologically distinct plant from Norfolk Island described as R. anisotoma sp. nov. Radula australiana is resolved as monophyletic, sister to a species occurring in east coast Australian rainforests, and nesting among the R. buccinifera lineages with strong support. The molecular phylogeny suggests several long-distance dispersal events may have occurred. These include two east-west dispersal events from New Zealand to Tasmania and south-east Australia in R. strangulata, one east-west dispersal event from Tasmania to Western Australia in R. buccinifera, and at least one west-east dispersal from Australia to New Zealand in R. australiana. Another west-east dispersal event from Australia to Norfolk Island may have led to the budding speciation of R. anisotoma. In contrast, Radula demissa is phylogeographically subdivided into strongly supported clades either side of the Tasman Sea, suggesting long distance dispersal is infrequent in this species.