Retrotransposon distribution and copy number variation in gymnosperm genomes

Retrotransposable elements (REs) and related sequences form a large proportion of conifer genomes. During genome evolution, some RE sequences are degraded or eliminated, but some are evolutionarily stable, and can be identified even in distantly related species. Use of genome sequence information from loblolly pine (Pinus taeda) enables investigation of divergent non-coding RE sequences in other pine and conifer species, including Scots pine (Pinus sylvestris). Non-specific inter-retrotransposon amplified polymorphism technique (IRAP) as well as the amplification polymorphism of 12 RE families were investigated in 80 gymnosperm species. The obtained results were compared with phylogenetic relationships among gymnosperms. Investigation of distantly related gymnosperm species reveals persistent RE sequences, such as IFG and Pineywoods, distributed among a wide range of plant lineages. RE sequence divergence was observed, reflecting periods of inactivity and degradation during speciation of pine lineages, as demonstrated by the delineation of the main pine subgenera. Intraspecific variation of 10 RE copy numbers (CN) between Scots pine individuals ranged from 8.9 to 26.6% of the overall mean estimates. CN analyses were performed in 16 additional gymnosperm species. The analysed pine species contained a similar complement of RE families; however, CN and genome occupation proportions differ. A decrease in RE CN estimates can reflect sequence divergence, associated with independent transposition events. Transposition of some REs can be induced by stress conditions; therefore, even distantly related species inhabiting extreme environments could have similar patterns or distribution of these elements.     

Divergent life history strategies in congeneric hyperparasitoids

Life histories can reveal important information on the performance of individuals within their environment and how that affects evolutionary change. Major trait changes, such as trait decay or loss, may lead to pronounced differences in life history strategies when tight correlations between traits exist. Here, we show that three congeneric hyperparasitoids (Gelis agilis, Gelis acarorum and Gelis areator) that have diverged in wing development and reproductive mode employ markedly different life history strategies. Potential fecundity of Gelis sp. varied, with the wingless G. acarorum maturing a much higher number of eggs throughout life compared with the other two species. Realized lifetime fecundity, in terms of total offspring number was, however, highest for the winged G. areator. The parthenogenic G. agilis invests its resources solely in females, whilst the sexually reproducing species both invested heavily in males to reduce competitive pressures for their female offspring. Longevity also differed between species, as did the direction of the reproduction-longevity trade-off, where reproduction is heavily traded off against longevity only in the asexual G. agilis. Resting metabolic rates also differed between the winged and wingless species, with the highest metabolic rate observed in the winged G. areator. Overall, these geline hyperparasitoids showed considerable divergence in life history strategies, both in terms of timing and investment patterns. Major trait changes observed between closely related species, such as the loss of wings and sexual reproduction, may contribute to the divergence in key life history traits.     

Functional convergence and phenotypic divergence in two specialist species of pine-associated ladybirds

Traditionally, convergent evolution has been considered to produce phenotypic similarity in independently evolved species. By contrast, recent studies have detected morphological divergence between species even in similar selective environments when different morphological traits combine to produce a specific functional output. However, it is still unclear whether a complex combination of non-morphological phenotypic traits, such as behavioural and life-history traits, can produce a similar performance in different species. In this study, I examined prey capture performance and related phenotypes in two sympatric ladybird species, Sospita oblongoguttata and Harmonia yedoensis, which specialize on the giant pine aphid, which is known to be elusive for ladybird hatchlings. In particular, I focused on egg size and proportion of trophic eggs in the clutch, since the amount of maternal investment per offspring can contribute to prey capture performance of ladybird hatchlings. Predation success of hatchlings against the giant pine aphid was higher in both S. oblongoguttata and H. yedoensis than in Harmonia axyridis, a generalist ladybird species that feeds on various kinds of aphid species in nature. Sospita oblongoguttata females, however, produced relatively larger eggs and in most clutches provided no trophic eggs, whereas H. yedoensis females produced smaller eggs and provided more trophic eggs per clutch. Moreover, hatchling morphology in H. yedoensis more closely resembled that of its congener, H. axyridis, than that of the more distantly related S. oblongoguttata, although like H. yedoensis, S. oblongoguttata predates on the elusive pine aphid. These results in two pine-associated specialist ladybirds indicate that divergent phenotypes can nonetheless have similar prey capture performance. In conclusion, this work demonstrates that the general ultimate function can be achieved by various mechanisms through convergence that operates at different level of life.     

Comparative morphological analysis of two parallel mycoheterotrophic transitions reveals divergent and convergent traits in the genus <Emphasis Type="Italic">Pyrola</Emphasis> (Pyroleae,Ericaceae)

The genus Pyrola includes species with different degree of mycoheterotrophy; some species possess individuals that rely on all carbon through their associations with fungi (full mycoheterotrophy, FM), whereas some species obtain carbon through both fungi and photosynthesis by itself (partial mycoheterotrophy, PM). To investigate how plant functional traits of photosynthesis and reproduction are related to the degree of mycoheterotrophy in the initial stage of the transition from PM to FM, we determined morphological traits in FM (or nearly FM) and PM species in two independent lineages, P. picta and P. japonica complexes. We used herbarium specimens and examined leaf number, leaf area, flower number, and scape length in FM or nearly FM species (P. aphylla and P. subaphylla) and PM species (P. picta s.l. and P. japonica). We found a leaf area reduction in FM (or nearly FM) species in both lineages, suggesting that this is a convergent trait. The number of flowers was not significantly different between FM (or nearly FM) and PM species in both lineages. On the other hand, differences in the variation between FM (or nearly FM) and PM species were found in some traits between the two lineages. The FM (or nearly FM) species in one lineage only possessed rudimentary leaves, whereas that in the other linage possessed a few small, ordinary leaves in addition to those with only rudimentary leaves. The scape length of the FM (or nearly FM) species was significantly longer than that of PM species in one lineage, whereas it was shorter in the other lineage. The different and common variations are divergent and convergent traits, respectively, that could be associated with the transition to FM in Pylora. In addition, shoots of both PM species occasionally lacked ordinary leaves, possibly indicating possession of these shoots is preadaptation for the transition to FM in Pyrola.     

Disentangling the effects of isolation-by-distance and isolation-by-environment on genetic differentiation among <Emphasis Type="Italic">Rhododendron</Emphasis> lineages in the subgenus <Emphasis Type="Italic">Tsutsusi</Emphasis>

Ecological speciation has long been noted as a central topic in the field of evolutionary biology, and investigation into the relative importance of ecological and geographical factors is becoming increasingly emphasized. We surveyed genetic variation of 277 samples from 25 populations of nine Rhododendron species within Tsutsusi subgenus in Taiwan using simple sequence repeats of expressed sequence tags. Bayesian clustering revealed four genetic lineages: (1) the Rhododendron simsii, Rhododendron kanehirai, and Rhododendron nakaharae lineage (lineage 1); (2) the Rhododendron longiperulatum, Rhododendron breviperulatum, and Rhododendron noriakianum lineage (lineage 2); (3) the Rhododendron rubropilosum lineage (lineage 3); and (4) the Rhododendron oldhamii lineage (lineage 4). Asymmetric introgressions were found from lineage 3 into lineages 1 and 2 (introgressed lineages). Genetic admixture of non-R. oldhamii species was also revealed by a neighbor-joining tree. Variation partitioning showed that environment explained much larger portions of genetic variation than geography between non-introgressed lineages (i.e., between R. oldhamii and other lineages). However, the Mantel and partial Mantel tests and the multiple matrix regression with randomization found that isolation-by-distance played a more important role than isolation-by-environment (IBE) in contributing to genetic variation in most between lineage comparisons. Nevertheless, strong IBE was found when compared between non-introgressed lineages of R. oldhamii and R. rubropilosum, suggesting post-speciation ecological divergence. Several environmental variables, including annual mean temperature, aspect, isothermality, seasonal precipitation, slope, and soil pH, could be important ecological drivers involved in reproductive isolation between R. oldhamii and non-R. oldhamii species within the Tsutsusi subgenus.     

Genetic drift and uniform selection shape evolution of most traits in <Emphasis Type="Italic">Eugenia dysenterica</Emphasis> DC. (Myrtaceae)

Knowing how microevolutionary processes, such as genetic drift and natural selection, shape variation in adaptive traits is strategic for conservation measures. One way to estimate local adaptation is to compare divergences in quantitative traits (Q_ST) and neutral loci (F_ST). Therefore, we have assessed the pattern of phenotypic and molecular genetic divergence among natural subpopulations of the fruit tree Eugenia dysenterica DC. A provenance and progeny test was performed to assess the quantitative traits of the subpopulations collected in a wide distribution area of the species in the Brazilian Cerrado. The sampled environments are in a biodiversity hotspot with heterogeneous soil and climate conditions. By associating quantitative trait variation in initial seedling development with neutral microsatellite marker variation, we tested the local adaptation of the traits by the Q_ST–F_ST contrast. Genetic drift was prevalent in the phenotypic differentiation among the subpopulations, although the traits seedling emergence time and root green mass, which are relevant for adaptation to the Cerrado climate, showed signs of uniform selection. Our results suggest that E. dysenterica has a spatial genetic structure divided into two large groups, separated by a line that divides the Cerrado biome in a southwestern to northeastern direction. This structure must be taken into account for managing E. dysenterica genetic resources both for conservation and breeding purposes.     

A molecular phylogeny of the Old World cypresses (<Emphasis Type="Italic">Cupressus</Emphasis>: Cupressaceae): evidence from nuclear and chloroplast DNA sequences

Studies of phylogenetic relationships among cypresses of the Old World (Cupressus; Cupressaceae) have been plagued by unresolved relationships, poor branch support, and conflict between data sets and methods of analysis. In this study, we combined 5.4 kb of aligned DNA sequence and 157 binary characters with previously published data in examining phylogenetic relationships among Cupressus species. Bayesian and parsimony analysis of the combined data or of the nuclear data alone always recovered three principal clades of Cupressus; however, tests of phylogenetic incongruence could not distinguish between competing relationships among the three principal Cupressus lineages. In contrast, incongruence tests often found statistically significant conflict between the nuclear and plastid data, particularly with respect to the placement of C. chengiana. Consistent with previous studies and prevailing taxonomic opinion, we find C. darjeelingensis more closely related to cypresses of the New World (Hesperocyparis). In contrast, we placed accessions of C. assamica and C. tonkinensis, two putatively Old World species suggested to be misidentified New World taxa by some authors, within well-supported Old World clades. Statistical analysis of genetic distances suggests instances in which taxa recognized as distinct species by some authors are identical or nearly so and may best be considered a single taxon. Conversely, we identify instances in which infraspecific taxa are more distantly related to one another than those traditionally recognized as distinct species. Factors confounding cypress taxonomies, including poor morphological differentiation, misidentification, and the use of accessions of questionable provenance, are discussed.     

Morphological diversity and phylogeography of the Georgian durmast oak (<Emphasis Type="Italic">Q. petraea</Emphasis> subsp. <Emphasis Type="Italic">iberica</Emphasis>) and related Caucasian oak species in Georgia (South Caucasus)

The Caucasus region is one of the 25 global biodiversity hotspots and constitutes a shelter area for Neogene relict species as well as a center of ongoing radiation. In order to elucidate the taxonomic identity, divergence patterns, and evolutionary history of the largely widespread Georgian durmast oak (Quercus petraea subsp. iberica), we examined leaf morphology and chloroplast DNA (cpDNA) (trnH-psbA, trnK-matK) sequence variation across its South Caucasian range. Six other oak taxa distributed throughout Georgia were included in the dataset and used for comparison. Evidence for differentiation in both sets of traits was found. Populations represented by different taxa from ecologically equivalent areas showed common morphological features and genetic structures. Molecular analysis clearly indicated the presence of two major haplotype lineages (West Caucasian vs. East Caucasian zonation type) and suggested a maternal lineage diversification of Q. petraea subsp. iberica in the Late Miocene, as a likely result of complex patterns associated with major orogenic and climatic changes. The Quaternary glacial oscillations resulted in a number of less common, derived haplotypes. Based on mismatch distribution analysis and neutrality tests, we found no evidence of demographic expansion for the populations from the West and East Caucasian zonation types. The two Caucasian provinces therefore acted as important shelter/diversification areas and as a lineage crossroad for the Georgian oaks. Close intra- and interspecific cpDNA relationships shared with other oaks from bordering countries support the relevant role played by the Colchis region as a primary refugium for the European temperate forest species.     

Historical patterns of niche dynamics in Neotropical species of the <Emphasis Type="Italic">Drosophila</Emphasis> subgenus (Drosophilidae,Diptera)

Niche conservatism (NC) presence is a controversial question in evolutionary ecology. In Drosophila, little is known about which is the preponderant evolutionary pattern, since the adaptive radiation hypothesis first proposed by Throckmorton assumed niche divergence (ND) according to a niche occupancy scenario. Nevertheless, this hypothesis has not yet been straightforwardly tested. Our aim here was to test the role of NC patterns across evolution of American drosophilids belonging to the tripunctata and virilis-repleta lineages of the Drosophila subgenus, through measures of geographical, abiotic and biotic niche overlap and evaluations regarding the presence of phylogenetic signal or niche identity. We recovered phylogenetic signal attributable to phylogenetic niche conservatism when all species were analyzed together, but not in more restricted groups. Identity tests showed that niche equivalency was seldom rejected for the tripunctata lineage species. So, in general, neither the results for the Drosophila subgenus nor those for the tripunctata lineage support the hypothesis of an adaptive radiation. Notwithstanding, there were also several isolated cases supporting a scenario of ND, and ecological speciation was evident in some of the evaluated sister species pairs.     

Tracking the diversity of the flatworm genus <Emphasis Type="Italic">Imbira</Emphasis> (Platyhelminthes) in the Atlantic Forest

The genus Imbira Carbayo et al., 2013 encompasses two species, Imbira guaiana (Leal-Zanchet & Carbayo, 2001) and Imbira marcusi Carbayo et al., 2013, which occur in south Brazil, in areas originally covered by the Atlantic Forest. In the present study, we examine the genetic diversity within the genus, investigate the occurrence of molecular autapomorphies for its species and describe a new species for the genus based on an integrative approach. The Bayesian and maximum likelihood analyses based on DNA barcoding recovered the monophyly of the genus Imbira, but indicate that specimens representing I. marcusi correspond to five distinct lineages. These analyses, as well as sequence divergence data, revealed that the new species herein described is closely related to I. guaiana and that the specific status of specimens of I. marcusi available in GenBank should be reviewed. In addition, sequence analysis revealed 32 molecular autapomorphies for all independent evolutionary units within the genus. The new species described herein seems to be endemic to its type locality, a private area without legal protection.     

Inferring the potential of plastid DNA-based identification of derived ferns: a case study on the <Emphasis Type="Italic">Asplenium trichomanes</Emphasis> aggregate in Europe

The utility of three plastid DNA regions to identify fern species was explored with focus on the European representatives of the Asplenium trichomanes aggregate. The sampling included representatives of the three diploid and the four tetraploid taxa recognized in the European flora plus Macaronesia. Besides European samples, the compiled data set comprised specimens of a putative Hawaiian endemic and one species occurring in Southeast Asia. By combining the sequences of three non-coding plastid regions, 13 haplotypes were recovered of which four were found in more than one taxon. Evidences for four distinct diploid lineages were found that correspond to Asplenium anceps, A. inexpectans, A. trichomanes s.s., and A. tripteropus. The four tetraploids occurring in Europe shared haplotypes with A. inexpectans. Thus, DNA barcoding can successfully identify the diploids, but fail to separate the tetraploids from their diploid ancestors. As a consequence, barcoding analyses of ferns need to take into account the differences of ploidy level measured by evidence independent from the DNA barcode. Evidence for uneven accumulation of intra-species DNA variation was recovered by comparing all species. Furthermore, the study provided evidence that the current taxonomy of these ferns requires to be revised. The two European diploids form well-separated clades and need to be recognized as A. inexpectans and A. trichomanes s.s. To keep name consistency for all European tetraploids, a new name Asplenium jessenii is introduced to replace A. trichomanes subsp. hastatum.     

Intraspecific Variation in Mitogenomes of Five <Emphasis Type="Italic">Crassostrea</Emphasis> Species Provides Insight into Oyster Diversification and Speciation

A large number of Crassostrea oysters are found in Asia-Pacific. While analyses of interspecific variation have helped to establish historical relationships among these species, studies on intraspecific variation are necessary to understand their recent evolutionary history and current forces driving population biology. We resequenced 18 and analyzed 31 mitogenomes of five Crassostrea species from China: Crassostrea gigas, Crassostrea angulata, Crassostrea sikamea, Crassostrea ariakensis, and Crassostrea hongkongensis. Our analysis finds abundant insertions, deletions, and single-nucleotide polymorphisms in all species. Intraspecific variation varies greatly among species with polymorphic sites ranging from 54 to 293 and nucleotide diversity ranging from 0.00106 to 0.00683. In all measurements, C. hongkongensis that has the narrowest geographic distribution exhibits the least sequence diversity; C. ariakensis that has the widest distribution shows the highest diversity, and species with intermediate distribution show intermediate levels of diversity. Low sequence diversity in C. hongkongensis may reflect recent bottlenecks that are probably exacerbated by human transplantation. High diversity in C. ariakensis is likely due to divergence of northern and southern China populations that have been separated without gene flow. The significant differences in mitogenome diversity suggest that the five sister species of Crassostrea have experienced different evolutionary forces since their divergence. The recent divergence of two C. ariakensis populations and the C. gigas/angulata species complex provides evidence for continued diversification and speciation of Crassostrea species along China’s coast, which are shaped by unknown mechanisms in a north–south divide.     

<Emphasis Type="Italic">Shackletonia cryodesertorum</Emphasis> (<Emphasis Type="Italic">Teloschistaceae</Emphasis>, Ascomycota), a new species from the McMurdo Dry Valleys (Antarctica) with notes on the biogeography of the genus <Emphasis Type="Italic">Shackletonia</Emphasis>

A new species of Shackletonia (Teloschistaceae) is described from the McMurdo Dry Valleys in Antarctica, one of the regions with the harshest conditions on Earth. Distinctive traits of the new taxon are the grey thallus, its black lecideine apothecia with a dark greenish blue exterior side of the exciple, Lecidea green pigment present at the cortex and exciple, emodin-dominated anthraquinones only in epithecium, and spores on average 11.2?×?6.0 μm with 3.6 μm wide septum. New chemical data from HPLC analyses further supports the uniqueness of the genus Shackletonia regarding secondary metabolite production within subfamily Xanthorioideae. Using three molecular markers (nrITS, nuLSU, and mtSSU) we found the new species sister to S. sauronii, a species so far known only from Livingston Island (Antarctica). Using secondary calibrations we inferred a long-time evolution of Shackletonia in the Southern Hemisphere, which separated from the remaining lineages of Xanthorioideae between the late Cretaceous and the early Paleogene, and diversified during the late Paleocene and early Oligocene.     

A preliminary molecular phylogeny shows Japanese and Austrian populations of the red mite <Emphasis Type="Italic">Balaustium murorum</Emphasis> (Acari: Trombidiformes: Erythraeidae) to be closely related

The red mite Balaustium murorum (Hermann) inhabits the Western Palaearctic realm and is well adapted to man-made structures. In Japan, B. murorum had been reported more frequently after the 1980s. A molecular phylogeny based on the nuclear 18S rRNA and mitochondrial COI genes, and including B. murorum individuals from Japan and Austria and representatives of related species from Japan showed four Balaustium species-level lineages in Japan (B. murorum, Balaustium sp. 1, Balaustium sp. 2, Balaustium sp. 3). The B. murorum lineage shared identical 18S sequence and COI haplotype with the Austrian population. Balaustium sp. 1 was detected from the Tokyo and Misaki area (Honshu Island) and was the sister group to B. murorum; the other two lineages inhabited coastal environments of Erimo, Hokkaido Island (Balaustium sp. 2) and Ainan, Shikoku Island (Balaustium sp. 3). The high genetic distances among these four lineages indicate that each lineage is a distinct species, with three of the lineages representing undescribed species. Our results are compatible with the conclusion that B. murorum was introduced to Japan from Europe, although our study did not resolve the polarity or timing of migration events.     

Intraspecific functional trait variability across different spatial scales: a case study of two dominant trees in Korean pine broadleaved forest

Intraspecific functional trait variability plays an important role in the response of plants to environmental changes. However, it is still unclear how the variability differs across three nested spatial scales (individual, plot, and site) and which determinants (climatic, soil, and ontogenetic variables) shape the trait variability. Along a latitudinal gradient in Korean pine broadleaved forest of northeast China, we quantified the extent of intraspecific variability of four functional traits in two dominant trees Pinus koraiensis and Fraxinus mandshurica at eight sites, including specific leaf area, leaf dry matter content (morphological traits) and leaf nitrogen content, leaf phosphorus content (physiological traits). Results showed a large trait variation within and between species (coefficient variation: 6.07–23.3%). The leaf physiological traits of F. mandshurica and morphological traits of P. koraiensis were more responsive at site scale, while the morphological traits of F. mandshurica and physiological traits of P. koraiensis were more responsive at individual scale. In addition, abiotic and biotic factors explaining functional trait variation differ markedly between the two tree species, with physiological trait of F. mandshurica being more associated with climate and soil, while traits variability in P. koraiensis was not affected by climate, soil, and ontogeny, except for leaf phosphorus content. Overall, we can predict that the physiological traits of broadleaved species tend to be more sensitive to environmental changes, while pines are more sensitive to competition. It is critical to determine which spatial scale and trait type should be taken into account in predictive models of vegetation dynamics.     

Response of floodplain understorey species to environmental gradients and tree invasion: a functional trait perspective

Plants are connected to habitats by functional traits which are filtered by environmental gradients. Since tree species composition in the forest canopy can influence ecosystem processes by changing resource availability, litter accumulation, and soil nutrient content, we hypothesised that non-native invasive trees can establish new environmental filters on the understorey communities. In the hardwood floodplain forests in Northern Italy, the invasive trees Robinia pseudoacacia L. and Prunus serotina Ehrh. are the dominant canopy species. We used trait data assembled from databases and iterative RLQ analysis to identify a parsimonious set of functional traits responding to environmental variables (soil, light availability, disturbance, and stand structure) and the dominant native and invasive canopy species. Then, RLQ and fourth-corner analysis was conducted to investigate the joint structure between macro-environmental variables and species traits and functional groups were identified. The trait composition of the herb-layer was significantly related to the main environmental gradients and the presence of the invaders in the canopy showed significant relationships with several traits. In particular, the presence of P. serotina may mitigate or even erase the effect of disturbances, maintaining a stable forest microclimate and thus favouring ‘true’ forest species, while R. pseudoacacia may slow down forest succession and regeneration by establishing new stable associations with a graminoid-dominated understorey. The impact of the two invasive trees on herb layer composition appears to differ, indicating that different management and control strategies may be needed.     

Specific leaf area variations drive acclimation of <Emphasis Type="Italic">Cistus salvifolius</Emphasis> in different light environments

Cistus salvifolius L. is the most widely spread Cistus species around the Mediterranean basin. It colonizes a wide range of habitats growing from sea level to 1,800 m a.s.l., on silicolous and calcicolous soils, in sun areas as well as in the understory of wooded areas. Nevertheless, this species has been mainly investigated in term of its responsiveness to drought. Our aim was to understand which leaf traits allow C. salvifolius to cope with low-light environments. We questioned if biochemical and physiological leaf trait variations in response to a reduced photosynthetic photon flux density were related to leaf morphological plasticity, expressed by variations of specific leaf area (SLA) and its anatomical components (leaf tissue density and thickness). C. salvifolius shrubs growing along the Latium coast (41°43'N,12°18'E, 14 m a.s.l., Italy) in the open and in the understory of a Pinus pinea forest, were selected and the relationships between anatomical, gas exchange, chlorophyll (Chl) fluorescence, and biochemical parameters with SLA and PPFD variations were tested. The obtained results suggested long-term acclimation of the selected shrubs to contrasting light environments. In high-light conditions, leaf nitrogen and Chl contents per leaf area unit, leaf thickness, and Chl a/b ratio increased, thus maximizing net photosynthesis, while in shade photosynthesis, it was downregulated by a significant reduction in the electron transport rate. Nevertheless, the increased pigment-protein complexes and the decreased Chl a/b in shade drove to an increased light-harvesting capacity (i.e. higher actual quantum efficiency of PSII). Moreover, the measured vitality index highlighted the photosynthetic acclimation of C. salvifolius to contrasting light environments. Overall, our results demonstrated the morphological, anatomical, and physiological acclimation of C. salvifolius to a reduced light environment.     

Phylogeography of the Macaronesian Lettuce Species <Emphasis Type="Italic">Lactuca watsoniana</Emphasis> and <Emphasis Type="Italic">L. palmensis</Emphasis> (Asteraceae)

The phylogenetic relationships and phylogeography of two relatively rare Macaronesian Lactuca species, Lactuca watsoniana (Azores) and L. palmensis (Canary Islands), were, until this date, unclear. Karyological information of the Azorean species was also unknown. For this study, a chromosome count was performed and L. watsoniana showed 2n = 34. A phylogenetic approach was used to clarify the relationships of the Azorean endemic L. watsoniana and the La Palma endemic L. palmensis within the subtribe Lactucinae. Maximum parsimony, Maximum likelihood and Bayesian analysis of a combined molecular dataset (ITS and four chloroplast DNA regions) and molecular clock analyses were performed with the Macaronesian Lactuca species, as well as a TCS haplotype network. The analyses revealed that L. watsoniana and L. palmensis belong to different subclades of the Lactuca clade. Lactuca watsoniana showed a strongly supported phylogenetic relationship with North American species, while L. palmensis was closely related to L. tenerrima and L. inermis, from Europe and Africa. Lactuca watsoniana showed four single-island haplotypes. A divergence time estimation of the Macaronesian lineages was used to examine island colonization pathways. Results obtained with BEAST suggest a divergence of L. palmensis and L. watsoniana clades c. 11 million years ago, L. watsoniana diverged from its North American sister species c. 3.8 million years ago and L. palmensis diverged from its sister L. tenerrima, c. 1.3 million years ago, probably originating from an African ancestral lineage which colonized the Canary Islands. Divergence analyses with *BEAST indicate a more recent divergence of the L. watsoniana crown, c. 0.9 million years ago. In the Azores colonization, in a stepping stone, east-to-west dispersal pattern, associated with geological events might explain the current distribution range of L. watsoniana.     

<Emphasis Type="Italic">Calcarisporium xylariicola</Emphasis> sp. nov. and introduction of <Emphasis Type="Italic">Calcarisporiaceae</Emphasis> fam. nov. in Hypocreales

During a survey of fungicolous fungi, a novel taxon from the surface of stroma of an unidentified Xylaria species was collected. Phylogenetic analyses showed that this taxon clustered with Calcarisporium sp. and C. arbuscula isolates, but was resolved as a distinct species. A detailed morphological examination coupled with phylogenetic analysis indicated that the taxon represented a new species. Calcarisporium xylariicola sp. nov. is thus introduced. The new taxon is characterized by short conidiophores with swollen bases and less length/width ratio of conidia that distinguish it from other Calcarisporium species. Calcarisporium is presently placed in Hypocreales genera, incertae sedis genus. Species in the genus are largely fungicolous, or occasionally caulicolous or foliicolous, and have hyaline, erect, verticillate conidiophores and sympodial, polyblastic conidiation. A phylogenetic analysis of combined SSU, ITS, LSU, TEF and RPB2 sequence data from Calcarisporium species and other taxa in Hypocreales indicate that Calcarisporium is a distinct lineage from other families. Therefore, a new family, Calcarisporiaceae, in Hypocreales is introduced.