Cristina Pardo

Molecular phylogeny and systematics of Genista (Leguminosae) and related genera based on nucleotide sequences of nrDNA (ITS region) and cpDNA (trnL-trnF intergenic spacer)

Phylogenetic relationships of Genista and related genera (Teline, Chamaespartium, Pterospartum, Echinospartum, Ulex, Stauracanthus and Retama) were assessed by the analysis of sequences of the nrDNA internal transcribed spacer (ITS region), and the cpDNA trnL-trnF intergenic spacer. The tree obtained by combining both sets of data indicates the existence of three lines of diversification within Genista, that correspond to three subgenera: Genista, Phyllobotrys and Spartocarpus, however, each of these lineages encompass also species of the related genera Echinospartum, Teline, Retama, Chamaespartium, Pterospartum, Ulex, Stauracanthus. The molecular data do not support division of these subgenera into taxonomical units at the sectional level; only sections Genista and Spartocarpus are monophyletic groups. The sequences of both regions are also informative at the specific level, grouping morphologically related species (e.g. the G. cinerea aggregate). The molecular data have also helped to clarify the position of taxa whose relationships were not well established (e.g. G. valdes-bermejoi). The relationships of related genera that belong to the Genista lines of diversification have also been investigated. Echinospartum splits into two separate clades matching the separation of two ecological and caryological differentiated groups. Teline also forms two groups, both placed near to Genista subgenus Genista, but that separated from the main core of the group. Retama, morphologically well differentiated from Genista, is close to Genista subgenus Spartocarpus. Chamaespartium and Pterospartum do not form a monophyletic group. Chamaespartium is closer to Genista subgenus Genista, whereas Pterospartum stands close to: 1) Genista subgenus Spartocarpus (particularly, sect. Cephalospartum); and 2) the Ulex-Stauracanthus clade (a terminal derivative of Genista subgenus Spartocarpus). Cases of incongruence (e.g. Echinospartum, Chamaespartium, Teline) between the trees obtained from the two molecular markers, may be indicating hybridisation and/or introgression between different lines of Genisteae.

Genetic variation and phylogeography of Stauracanthus (Fabaceae, Genisteae) from the Iberian Peninsula and northern Morocco assessed by chloroplast microsatellite (cpSSR) markers

The tribe Genisteae includes genera of great ecological importance in Mediterranean countries because they are dominant elements of many plant communities. Genetic variation and diversification patterns in Stauracanthus (Genisteae) provide information relevant for the study of the processes of diversification in relation to the environmental history of the western Mediterranean. Nineteen populations of S. boivinii and S. genistoides were assessed by 11 chloroplast microsatellite markers, revealing 44 haplotypes. Both species had different haplotypes and contrasting patterns of karyological, morphological, and genetic variation. In the minimum spanning tree of the haplotypes, AMOVA analysis, and nested clade analysis, S. boivinii had high levels of differentiation and restricted gene flow among populations. Allopatric differentiation occurred between the Moroccan and Iberian populations of S. genistoides, although S. genistoides subsp. spectabilis and subsp. vicentinus had high levels of differentiation among populations (F(ST)), whereas S. genistoides subsp. genistoides had a low F(ST). Genetic patterns are discussed in relation to the Messinian salinity crisis (MSC): hard conditions drove plants to refuge habitats along the Atlantic coast and higher altitude areas in the Moroccan mountains (S. genistoides subsp. spectabilis and S. boivinii). After the MSC, S. boivinii underwent polyploidization and expansion, whereas S. genistoides expanded and continued diversifying into S. genistoides subspp. genistoides and vicentinus.

Molecular approach to the phylogeny and systematics of Cytisus (Leguminosae) and related genera based on nucleotide sequences of nrDNA (ITS region) and cpDNA (trnL-trnF intergenic spacer)

Abstract. Phylogenetic relationships of Cytisus and allied genera (Argyrocytisus, Calicotome, Chamaecytisus, Cytisophyllum, and Spartocytisus) were assessed by analysis of sequences of the nrDNA internal transcribed spacer (ITS) and the cpDNA trnL-trnF intergenic spacer. Genera of the Genista-group (Chamaespartium, Echinospartum, Genista, Pterospartum, Spartium, Teline and Ulex) were included to check the position of Cytisus species transferred to Teline. The tree obtained by combining both sets of data indicates that the Genista and Cytisus groups form two separate clades. Cytisus heterochrous and C. tribracteolatus are more closely related to the Cytisus-group, thus their transfer to Teline is not supported by molecular data. Cytisus fontanesii (syn. Chronanthos biflorus) groups with Cytisophyllum sessilifolium and Cytisus heterochrous within the Cytisus-group. Similarly, Argyrocytisus battandieri falls within the Cytisus-group as a well differentiated taxon. All these taxa seem to have early diverged from the Cytisus-group. Their taxonomic rank should be reconsidered to better reflect their phylogenetic separation from Cytisus. On the contrary, Chamaecytisus proliferus and Spartocytisus supranubius enter in the main core of Cytisus, and they should better be included in sections of Cytisus (sect. Tubocytisus and Oreosparton, respectively). Sect. Spartopsis is not monophyletic and the position of several species, currently included in this section, deserves reevaluation: C. arboreus aggregate is closely related to C. villosus (sect. Cytisus) and to Calicotome; C. striatus is closely related to Cytisus sect. Alburnoides; and the position of C. commutatus (incl. C. ingramii) remains unclear. The relationships and positioning of several minor taxa (C. transiens, C. megalanthus, and C. maurus) are also discussed.

Genetic variation and relationships among Ulex (Fabaceae) species in southern Spain and northern Morocco assessed by chloroplast microsatellite (cpSSR) markers

Genetic variation in 27 populations of Ulex species from southern Spain and northern Morocco (Betic-Rif arc) was assessed using 11 chloroplast microsatellite (cpSSR) markers, which revealed 47 different haplotypes. These nonrecombinant, haploid markers allow measurement of genetic variation in closely related species of Ulex where molecular phylogenetic analyses have not provided a clear view of interspecific relationships. Discriminant analysis indicates that the haplotypes are useful to differentiate among species, and analysis of molecular variance (AMOVA) shows high levels of differentiation among populations. The minimum spanning tree (MST), that represents the connections between the haplotypes, suggests that the eastern Rifean U. africanus haplotypes are more genetically related than those from southern Spain. The latter may have lost genetic diversity while colonizing new habitats, eventually differentiating into U. baeticus and U. scaber. Hybridization between these populations, followed by polyploidization, may have originated the tetraploids (U. congestus and U. borgiae) that colonized new habitats associated with acidic rocks. Separate groupings of U. scaber populations may indicate multiple origins from different stocks. Diversification in this group of Ulex species could be related to the opening of the Alboran Sea by Middle Miocene, when the populations from Morocco and Spain became isolated from each other.

Automated pollen identification using microscopic imaging and texture analysis

Pollen identification is required in different scenarios such as prevention of allergic reactions, climate analysis or apiculture. However, it is a time-consuming task since experts are required to recognize each pollen grain through the microscope. In this study, we performed an exhaustive assessment on the utility of texture analysis for automated characterisation of pollen samples. A database composed of 1800 brightfield microscopy images of pollen grains from 15 different taxa was used for this purpose. A pattern recognition-based methodology was adopted to perform pollen classification. Four different methods were evaluated for texture feature extraction from the pollen image: Haralicks gray-level co-occurrence matrices (GLCM), log-Gabor filters (LGF), local binary patterns (LBP) and discrete Tchebichef moments (DTM). Fishers discriminant analysis and k-nearest neighbour were subsequently applied to perform dimensionality reduction and multivariate classification, respectively. Our results reveal that LGF and DTM, which are based on the spectral properties of the image, outperformed GLCM and LBP in the proposed classification problem. Furthermore, we found that the combination of all the texture features resulted in the highest performance, yielding an accuracy of 95%. Therefore, thorough texture characterisation could be considered in further implementations of automatic pollen recognition systems based on image processing techniques.

Aquatic Ranunculus communities in the northern hemisphere: A global review

Abstract This study offers a general review of aquatic ranunculi communities in the northern hemisphere, and includes aspects of their ecology, syntaxonomy and distribution. White-flowered ranunculi communities (Ranunculus subgenus Batrachium) grow in a wide range of habitats, including different types of water regime (still and flowing waters; permanent and seasonal waters) and nutrient states. They are mainly found in Eurasia. High richness is detected in Western Europe and the Mediterranean area of heterophyllous and laminar-leaved species, and in Asia of species with only dissected leaves. The western centre of the heterophyllous species has been related to waters with low mineralization and oscillation events occurring between the Mediterranean and Temperate climates. Yellow-flowered ranunculi communities (R. Xanthobatrachium) include heterophyllous species with a tendency to helophytism. They also are found in a wide range of aquatic habitats. There is an uneven degree of knowledge on the ecology of aquatic ranunculi communities, and further research is required into aquatic ranunculi habitats in Asia and North America.

Habitat Variation in Vernal Pool Ecosystems on Both Sides of the Strait of Gibraltar

Abstract Lumbreras, A.; Tahiri, H.; Pinto-Cruz, C.; Pardo, C., and Molina, J.A., 2012. Habitat variation in vernal pool ecosystems on both sides of the Strait of Gibraltar. We studied vernal pool (VP) ecosystems along a latitudinal gradient crossing the Strait of Gibraltar in order to determine its role in the distribution of VP plant communities. We analyzed flora, vegetation, physical–chemical water parameters, and climatic data from two vernal pool areas on both the European (Iberian) and African (Moroccan) sides of the Strait. Despite the minor distance between both territories, the pools clearly differed in species composition and ecology. However, they showed a similar vegetation zonation in growth forms, including isoetid, batrachiid, and helophytic vegetation. The distribution of the plant communities was related to nutrient load, temperature, and precipitation. Water nitrate concentration was higher in Morocco, where VPs are characterized by Isoetes velata subsp. adspersa and Ranunculus saniculifolius communities. Iberian VPs had lower water nitrate content, and were characterized by Isoetes velata subsp. velata and Ranunculus peltatus communities. We think this nutrient difference is likely to be caused by the different land management regime on each side of the Strait, with more intensive agriculture in Morocco. Long-term (historical) and present-day (ecological) processes have been proposed to account for the habitat variation in vernal pool ecosystems on both sides of the Strait of Gibraltar.

Pollen segmentation and feature evaluation for automatic classification in bright-field microscopy

Pollen collection: 15 types - 120 samples/type.Proposal of contour-inner pollen segmentation: 50% accuracy rates.New contour profile descriptor.LogGabor descriptors firstly tested for pollen classification.Experiments of descriptors state of the art combination: rates above 99%. Besides the well-established healthy properties of pollen, palynology and apiculture are of extreme importance to avoid hard and fast unbalances in our ecosystems. To support such disciplines computer vision comes to alleviate tedious recognition tasks. In this paper we present an applied study of the state of the art in pattern recognition techniques to describe, analyze, and classify pollen grains in an extensive dataset specifically collected (15 types, 120 samples/type). We also propose a novel contour-inner segmentation of grains, improving 50% of accuracy. In addition to published morphological, statistical, and textural descriptors, we introduce a new descriptor to measure the grains contour profile and a logGabor implementation not tested before for this purpose. We found a significant improvement for certain combinations of descriptors, providing an overall accuracy above 99%. Finally, some palynological features that are still difficult to be integrated in computer systems are discussed.

Pollen morphology in Cytisus (Papilionoideae, Leguminosae) from Morocco and the Iberian Peninsula

Although it is generally accepted that the tribe Genisteae has rather homogeneous pollen, we have found considerable variation in pollen size and morphology in Cytisus and related genera (Argyrocytisus and Chamaecytisus), which match taxonomic groups defined on morphological grounds. The results of Principal Component Analysis of pollen characteristics show the following: Cytisus Sect. Alburnoides is well-delimited, with the smallest pollen grains and the simplest pattern in the tectum, fossulate-perforate to perforate; No separation of C. villosus (Sect. Cytisus) from Sect. Alburnoides is observed; Cytisus Sect. Spartopsis, with the largest pollen grains and reticulate to reticulate-fossulate ornamentation, forms an isolated group; C. fontanesii (Sect. Heterocytisus) stands apart from other Cytisus species by its homogeneous ornamentation and the perforate instead of psilate-punctate margo. Thus, the pollen data supports its transfer to the separated genus Chronanthos; Pollen ornamentation and morphology also support the separation of Argyrocytisus battandieri and Chamaecytisus mollis from Cytisus.

Morphological convergence or lineage sorting? The case of Cytisus purgans auct. (Leguminosae)

Genetic distances among populations and phylogenetic hypothesis concerning origin and diversification of Cytisus purgans s.1. are inferred by analysis of the ITS and ETS regions (nrDNA) and the trnL-trnF intergenic spacer (cpDNA). Cytisus purgans s.1., a shrub of western Mediterranean mountains, consists of two evolutionary units: C. balansae (Boiss.) Ball (Morocco, Algeria, southern Spain) and C. oromediterraneus Rivas Mart. & al. (France, Portugal, Andorra, central and northern Spain). The two units are related to different Cytisus groups, in Bayesian and Maximum Parsimony analyses, and may be genetically unrelated, their morphological similarities being the result of convergence. However, the incongruence in the position of both species in the ITS tree and the ETS and trnL-trnF trees suggests an alternative explanation. Genetic differentiation could be due to lineage sorting of allelic polymorphism in the ITS region of the common ancestor, and the morphological similarities the result of morphological parallelism. Cytisus balansae conserves ITS sequences related to old Cytisus taxa; rapid radiation may have led to a western Mediterranean lineage in which C. oromediterraneus originated.