Ruth Jaén-Molina

A local dormancy cline is related to the seed maturation environment, population genetic composition and climate

BACKGROUND AND AIMS Seed dormancy varies within species in response to climate, both in the long term (through ecotypes or clines) and in the short term (through the influence of the seed maturation environment). Disentangling both processes is crucial to understand plant adaptation to environmental changes. In this study, the local patterns of seed dormancy were investigated in a narrow endemic species, Centaurium somedanum, in order to determine the influence of the seed maturation environment, population genetic composition and climate. METHODS Laboratory germination experiments were performed to measure dormancy in (1) seeds collected from different wild populations along a local altitudinal gradient and (2) seeds of a subsequent generation produced in a common garden. The genetic composition of the original populations was characterized using intersimple sequence repeat (ISSR) PCR and principal co-ordinate analysis (PCoA), and its correlation with the dormancy patterns of both generations was analysed. The effect of the local climate on dormancy was also modelled. KEY RESULTS An altitudinal dormancy cline was found in the wild populations, which was maintained by the plants grown in the common garden. However, seeds from the common garden responded better to stratification, and their release from dormancy was more intense. The patterns of dormancy variation were correlated with genetic composition, whereas lower temperature and summer precipitation at the population sites predicted higher dormancy in the seeds of both generations. CONCLUSIONS The dormancy cline in C. somedanum is related to a local climatic gradient and also corresponds to genetic differentiation among populations. This cline is further affected by the weather conditions during seed maturation, which influence the receptiveness to dormancy-breaking factors. These results show that dormancy is influenced by both long-and short-term climatic variation. Such processes at such a reduced spatial scale highlight the potential of plants to adapt to fast environmental changes.

The colonization history of Juniperus brevifolia (Cupressaceae) in the Azores Islands.

Background A central aim of island biogeography is to understand the colonization history of insular species using current distributions, fossil records and genetic diversity. Here, we analyze five plastid DNA regions of the endangered Juniperus brevifolia, which is endemic to the Azores archipelago. Methodology/Principal Findings The phylogeny of the section Juniperus and the phylogeographic analyses of J. brevifolia based on the coalescence theory of allele (plastid) diversity suggest that: (1) a single introduction event likely occurred from Europe; (2) genetic diversification and inter-island dispersal postdated the emergence of the oldest island (Santa Maria, 8.12 Ma); (3) the genetic differentiation found in populations on the islands with higher age and smaller distance to the continent is significantly higher than that on the younger, more remote ones; (4) the high number of haplotypes observed (16), and the widespread distribution of the most frequent and ancestral ones across the archipelago, are indicating early diversification, demographic expansion, and recurrent dispersal. In contrast, restriction of six of the seven derived haplotypes to single islands is construed as reflecting significant isolation time prior to colonization. Conclusions/Significance Our phylogeographic reconstruction points to the sequence of island emergence as the key factor to explain the distribution of plastid DNA variation. The reproductive traits of this juniper species (anemophily, ornithochory, multi-seeded cones), together with its broad ecological range, appear to be largely responsible for recurrent inter-island colonization of ancestral haplotypes. In contrast, certain delay in colonization of new haplotypes may reflect intraspecific habitat competition on islands where this juniper was already present.

Comparative Micromorphology of Petals in Macaronesian Lotus (Leguminosae) Reveals a Loss of Papillose Conical Cells during the Evolution of Bird Pollination

Bird pollination has evolved in four species of Macaronesian Lotus from a bee-pollinated ancestor. The transition is associated with the modification of several floral traits, including flower color and size, relative size and orientation of the petals, and nectar composition and quantity. Here, we examine petal surface micromorphology in relation to pollination type, using SEM and LM. In the bee-pollinated Lotus (the majority of the genus), papillose conical cells (PCS) are the most abundant epidermal type on dorsal and lateral petals. However, bird-pollinated species completely lack PCS on their dorsal petals and have only a small patch of PCS in a highly localized region of the lateral petal. In the bee-pollinated species (including those most closely related to the bird-pollinated species), PCS develop early in floral development. In contrast, the small amount of residual PCS in bird-pollinated species forms later in development, after the other two major epidermal types have been formed. The almost complete elimination of PCS during the shift of pollination syndrome from bee to birds may be adaptively driven as a both probird and antibee trait.

The molecular phylogeny of Matthiola R. Br. (Brassicaceae) inferred from ITS sequences, with special emphasis on the Macaronesian endemics.

Matthiola (Brassicaceae) is a genus that is widespread in the Mediterranean and Irano-Turanian regions and includes two species that are endemic to the archipelagos of Madeira and the Canaries in Macaronesia, which is an insular oceanic hotspot of biodiversity harboring many radiating endemic plant lineages. Sequence analyses of the nuclear ITS-1 and ITS-2 regions in a comprehensive geographical sample of Matthiola, encompassing all the endemic Macaronesian populations known to date, suggest independent Mediterranean and NW African origins of the taxa in Madeira and the Canaries, respectively. These molecular data reveal a complex evolutionary landscape that converges with morphological analyses in the recognition of two new Madeiran species. The data also suggest that the Canarian infra-specific endemic taxa described thus far have high (but non-diagnostic) levels of morphological and genetic diversity, and should be included in the single endemic Matthiola bolleana. In agreement with earlier investigations that revealed a high genetic differentiation between the populations of Matthiola in Fuerteventura and Lanzarote, our phylogeny supports independent founder events from the same mainland congener to either island. The consistently derived position of the Moroccan populations within a mostly Canarian clade suggests a further back-colonization of the continent. Notably, the ITS sequence resolution offered by Matthiola is higher than that found in many of the radiating Canarian endemic lineages for which molecular phylogenetic studies abound. Hence, our research discovers largely unexplored pathways to understand plant diversification in this oceanic insular hotspot through the investigation of non-speciose endemics.

The origin of bird pollination in Macaronesian Lotus (Loteae, Leguminosae)

The four red-flowered, apparently bird-pollinated Lotus species from the Canary Islands have previously been classified in their own genus, Rhyncholotus. Currently, they are considered as a separate section within genus Lotus, distinct from other herbaceous Canarian congeners which are yellow-flowered and bee-pollinated. A combined analysis of four nuclear regions (including ITS and three homologues of CYCLOIDEA) and three plastid regions (CYB6, matK and trnH-psbA) nests the four bird-pollinated species within a single extant species of bee-pollinated Lotus (L. sessilifolius), in a very extreme example of species paraphyly. Therefore, our data compellingly support the hypothesis that the Macaronesian Lotus species with a bird pollination syndrome are recently derived from entomophilous ancestors. Calibration of the phylogenetic trees using geological age estimates of the most recent islands (La Palma and El Hierro) together with oldest ages of Fuerteventura indicates that bird pollination evolved ca. 1.7 Ma in the Canarian Lotus. These four bird-pollinated species share a most recent common ancestor (MRCA) with L. sessilifolius that dates to about 2.2 Ma. Our analyses further suggest that the evolution of the bird pollination syndrome was likely triggered by the availability of new niches in La Palma and Tenerife as a result of recent volcanic activity.

Phylogenetic position of Dunaliella acidophila (Chlorophyceae) based on ITS and rbcL sequences

Dunaliella acidophila is one of the most extreme acidophiles on earth and is able to survive in highly acidic habitats. This characteristic has made this organism the universal model for the study of abiotic stress. Although D. acidophila is currently circumscribed to the subgenus Pascheria within Dunaliella Teodoresco (Chlorophyceae), its taxonomic position has stirred controversy. The comparison of D. acidophila CCAP19/35 internal transcribed spacers (including ITS2 secondary structure analysis) and RuBisCo large subunit (rbcL) sequences with other Dunaliella species confirms that D. acidophila should maintain its phylogenetic position within the genus Dunaliella, suggesting its inclusion within the subgenus Dunaliella. Furthermore, the ITS1 and ITS2 data revealed that D. acidophila was highly divergent from the other freshwater species assessed, D. lateralis, with which it barely shares a 56.8% similarity.

Phylogenetic analysis of ITS2 sequences suggests the taxonomic re-structuring of Dunaliella viridis (Chlorophyceae, Dunaliellales)

We analyzed the ITS2 primary and secondary structure (including Compensatory Base Changes (CBCs)) of 17 new Dunaliella strains (11 D. viridis, two D. tertiolecta, and four Dunaliella sp.), and compared these with other Dunaliella sequences available from the ITS2 database to circumscribe their taxonomic position. The ITS2 primary and secondary structure analysis positioned the majority of D. viridis strains in four main clades, showing that D. viridis is polyphyletic. The detection of at least one CBC among these clades strongly suggests that they could correspond to different biological species. Unexpectedly, while D. viridis var. euchlora (CCAP19/21) was positioned within the subgenus Dunaliella, D. viridis var. palmelloides (CCAP11/34) was positioned clearly outside this subgenus, suggesting that this taxon may not be properly placed in Dunaliella. Furthermore, the detection of at least three compensatory base changes (CBCs) between D. viridis var. palmelloides (CCAP11/34) and the other strains analyzed, confirm that this strain is a different species. For these reasons we propose re‐naming D. viridis var. palmelloides (CCAP11/34) to incertae sedis, and D. viridis var. euchlora (CCAP19/21) to Dunaliella sp. Therefore, the ITS2 primary and secondary structure data suggest a taxonomic re‐structuring of D. viridis.

Molecular taxonomic identification in the absence of a ‘barcoding gap’: a test with the endemic flora of the Canarian oceanic hotspot

We use a comprehensive subset of Canarian angiosperms corresponding to 23 families, 35 genera and 60 Canarian endemic taxa to test whether this flora is suitable to taxonomic identification with the two proposed plant DNA barcode sequences and whether these sequences may reveal the existence of cryptic species overlooked by morphology. The rate of discrimination success between the insular congeneric samples using the rbcL+matK combination and a ‘character‐based’ approach (where we use only the combination of nucleotide positions in an alignment that allows unambiguous species identification) is higher (82.29%) than that obtained with the ‘distance‐based’ approach (80.20%) used by the CBOL Plant Working Group in 2009 and also when compared with tests conducted in other floras. This suggests that the molecular identification of the Canarian endemic flora can be achieved as successfully as in other floras where the incidence of radiation is not as relevant. The facts that (i) a distance‐based criterion was unable to discriminate between congeneric and conspecific comparisons and (ii) only the character‐based discrimination criterion resolved cases that the distance‐based criterion did not, further support the use of a character discrimination approach for a more efficient DNA barcoding of floras from oceanic islands like the Canaries. Thus, a barcoding gap seems not to be necessary for the correct molecular characterization of the Canarian flora. DNA barcodes also suggest the possible existence of cryptic taxa to be further investigated by morphology and that the current taxonomic status of some of the taxa analysed may need revision.

DNA barcodes successfully identified Macaronesian Lotus (Leguminosae) species within early diverged lineages of Cape Verde and mainland Africa

Plant barcoding uses short DNA sequences to identify unknown samples at species level. This technique relies on the universality of these gene regions and the existence of enough variation among species to allow discrimination. Island radiations pose one challenging scenario where insufficient variation has accumulated in recently diverged groups to allow species identification. In this work we tested whether six gene regions are suitable for barcoding such a radiation in the Macaronesian Lotus. We found high levels of species discrimination in lineages of 3.5 Mya old or older and that the efficiency drastically reduces for younger radiations.

A tale of two forests: ongoing aridification drives population decline and genetic diversity loss at continental scale in Afro-Macaronesian evergreen-forest archipelago endemics

Background and Aims Various studies and conservationist reports have warned about the contraction of the last subtropical Afro-Macaronesian forests. These relict vegetation zones have been restricted to a few oceanic and continental islands around the edges of Africa, due to aridification. Previous studies on relict species have generally focused on glacial effects on narrow endemics; however, little is known about the effects of aridification on the fates of previously widespread subtropical lineages. Methods Nuclear microsatellites and ecological niche modelling were used to understand observed patterns of genetic diversity in two emblematic species, widely distributed in these ecosystems: Canarina eminii (a palaeoendemic of the eastern Afromontane forests) and Canarina canariensis (a palaeoendemic of the Canarian laurel forests). The software DIYABC was used to test alternative demographic scenarios and an ensemble method was employed to model potential distributions of the selected plants from the end of the deglaciation to the present. Key Results All the populations assessed experienced a strong and recent population decline, revealing that locally widespread endemisms may also be alarmingly threatened. Conclusions The detected extinction debt, as well as the extinction spiral to which these populations are subjected, demands urgent conservation measures for the unique, biodiversity-rich ecosystems that they inhabit.