Morphological and molecular analysis of natural hybrids between the diploid Centaurea aspera L. and the tetraploid C. seridis L. (Compositae)

[EN] Polyploidy and hybridisation are the basis of the evolution of Centaurea (Compositae). At the El Saler dune field (eastern Spain), the diploid Centaurea aspera ssp. stenophylla and the tetraploid C. seridis ssp. maritima form a polyploid complex in which C. x subdecurrens individuals occur. This polyploid complex was analysed morphologically and genetically, using random amplified polymorphic DNA (RAPD) and tubulin-based polymorphism (TBP) markers. Flow cytometry showed that the hybrids are triploid, which is a rare finding in Centaurea. Morphologically, in contrast to leaf characters, flowering characters clearly discriminated the three taxa. The genetic analyses confirm that C. x subdecurrens is a result of the hybridisation between Centaurea aspera ssp. stenophylla and C. seridis ssp. maritima, and suggest that backcrossing events and gene flow are very rare or absent. Although the hybrids likely represent true F1 offspring, they displayed some genetic diversity that is probably due to the combination of alleles. Genetic diversity was higher in diploid than in tetraploid individuals. This fact, and the high degree of sterility of the triploid hybrids, may reflect a cytotype minority exclusion effect. This may cause spatial segregation, which effectively takes place in the study area. Dune disturbance may lead to an overlapping of the parents' distribution areas, facilitating hybridisation.This work is posthumously dedicated to Antonio Samo Lumbreras, to whom we are very grateful for all his help. This study was sponsored by the Valencian Government (Research Project GVPRE/2008/130) and the Universitat Politecnica de Valencia (Research Project Ref. 3241).Ferriol Molina, M.; Garmendia, A.; Ruiz, J.; Merle Farinós, HB.; Boira Tortajada, H. (2012). Morphological and molecular analysis of natural hybrids between the diploid Centaurea aspera L. and the tetraploid C. seridis L. (Compositae). Plant Biosystems. 146(1):86-100. https://doi.org/10.1080/11263504.2012.727878S86100146

Molecular and morphological data resurrect the long neglected <em>Carex laxula</em> (Cyperaceae) and expand its range in the western Mediterranean

Carex sylvatica subsp. paui is a poorly studied taxon considered endemic from a few places in the western Mediterranean. It has been frequently misidentified as C. sylvatica subsp. sylvatica. To date, it has been reported only from the NE Iberian Peninsula and the NW Africa. We use molecular —nuclear ribosomal and plastid sequences— and morphological data to shed light on the taxonomic circumscription and distribution of this taxon, especially regarding its distinction from the type subspecies. The genetic data support the recognition of C. sylvatica subsp. paui as an independent taxon, and confirm new records from the Balearic and Tuscan archipelagos. It implies a considerable increase in its range and a new taxon for the Italian flora. Strikingly, the morphometric analyses revealed that the Sicilian type specimen ofC. laxula identifies this species with C. sylvatica subsp. paui. We consider that the taxon should be ranked at the species level. Based on the priority of the name C. laxula over C. paui, we subsume C. sylvatica subsp. paui in C. laxula. We also provide notes on the ecology of the species

Palynological and chemical volatile components of tipically autumnal honeys of the western Mediterranean

[EN] Twenty-five samples of autumnal honeys from the western Mediterranean (Mallorca and Eivissa, Balearic Islands) were examined for pollen content (qualitative and quantitative melissopalynological analysis), moisture, electrical conductivity, colour, sensorial qualities and volatile components. Quantitative analysis showed that the honey contained Maurizio's Class II: 64%, Class III: 28%, Class IV: 4% and Class V: 4%. Fifty-four pollen types, with an average number of 16.68 per sample, were identified, belonging to 29 botanical families. Only two taxa (Ceratonia siliqua and Erica multiflora) were found in all samples. Seventeen samples were unifloral (68%) - ten (40%) of C. siliqua, six (24%) of E. multiflora and one (4%) of Hedera helix. All honeys have a low honeydew index (<?0.09%), while the values for electrical conductivity and water content were high. The major honey volatile components are: cis- and trans-linalool oxides (64.2%) and hotrienol (10.4%) for the carob (C. siliqua) and trans-linalool oxide (13.4%), p-menthane-1,8-diol (11.1%), safranal (9.7%), limonene (5,4%), -pinene (3.7%) and oxoisophorone (3.4%) for the winter heather (E. multiflora).The authors would like to extend their gratitude to the Mallorca Rural 'Leader plus' programme and the beekeepers of Mallorca and Eivissa for their support and friendly collaboration. The authors also thank an anonymous reviewer for useful comments and suggestions on an earlier version of the manuscript.Boi, M.; Llorens Molina, JA.; Cortés, L.; Lladó, G.; Llorens, L. (2013). Palynological and chemical volatile components of tipically autumnal honeys of the western Mediterranean. Grana. 52(2):93-105. doi:10.1080/00173134.2012.744774S93105522Andrade, P. B., Amaral, M. T., Isabel, P., Carvalho, J. C. M. F., Seabra, R. M., & Proença da Cunha, A. (1999). Physicochemical attributes and pollen spectrum of Portuguese heather honeys. Food Chemistry, 66(4), 503-510. doi:10.1016/s0308-8146(99)00100-4Anklam, E. (1998). A review of the analytical methods to determine the geographical and botanical origin of honey. Food Chemistry, 63(4), 549-562. doi:10.1016/s0308-8146(98)00057-0Bosch, J., Del Pino, F. G., Ramoneda, J., & Retana, J. (1996). FRUITING PHENOLOGY AND FRUIT SET OF CAROB, CERATONIA SILIQUA L. (CESALPINACEAE). Israel Journal of Plant Sciences, 44(4), 359-368. doi:10.1080/07929978.1996.10676657Bouseta, A., Collin, S., & Dufour, J.-P. (1992). Characteristic aroma profiles of unifloral honeys obtained with a dynamic headspace GC-MS system. Journal of Apicultural Research, 31(2), 96-109. doi:10.1080/00218839.1992.11101268Cajka, T., Hajslova, J., Pudil, F., & Riddellova, K. (2009). Traceability of honey origin based on volatiles pattern processing by artificial neural networks. Journal of Chromatography A, 1216(9), 1458-1462. doi:10.1016/j.chroma.2008.12.066Castro-Vázquez, L., Díaz-Maroto, M. C., González-Viñas, M. A., & Pérez-Coello, M. S. (2009). Differentiation of monofloral citrus, rosemary, eucalyptus, lavender, thyme and heather honeys based on volatile composition and sensory descriptive analysis. Food Chemistry, 112(4), 1022-1030. doi:10.1016/j.foodchem.2008.06.036Conti, M. E., Stripeikis, J., Campanella, L., Cucina, D., & Tudino, M. B. (2007). Characterization of Italian honeys (Marche Region) on the basis of their mineral content and some typical quality parameters. Chemistry Central Journal, 1(1). doi:10.1186/1752-153x-1-14Custódio, L., Serra, H., Nogueira, J. M. F., Gonçalves, S., & Romano, A. (2006). Analysis of the Volatiles Emitted by Whole Flowers and Isolated Flower Organs of the Carob Tree Using HS-SPME-GC/MS. Journal of Chemical Ecology, 32(5), 929-942. doi:10.1007/s10886-006-9044-9Cuevas-Glory, L., Ortiz-Vázquez, E., Pino, J. A., & Sauri-Duch, E. (2012). Floral classification of Yucatan Peninsula honeys by PCA & HS-SPME/GC-MS of volatile compounds. International Journal of Food Science & Technology, 47(7), 1378-1383. doi:10.1111/j.1365-2621.2012.02983.xDe Bolòs, O., & Molinier, R. (1984). Vegetation of the Pityusic Islands. Biogeography and Ecology of the Pityusic Islands, 185-221. doi:10.1007/978-94-009-6539-3_9De Maria, C. A. B., & Moreira, R. F. A. (2003). Compostos voláteis em méis florais. Química Nova, 26(1), 90-96. doi:10.1590/s0100-40422003000100016Guyot, C., Scheirman, V., & Collin, S. (1999). Floral origin markers of heather honeys: Calluna vulgaris and Erica arborea. Food Chemistry, 64(1), 3-11. doi:10.1016/s0308-8146(98)00122-8Herrera, J. (1988). Pollination Relationships in Southern Spanish Mediterranean Shrublands. The Journal of Ecology, 76(1), 274. doi:10.2307/2260469Jerković, I., & Marijanović, Z. (2010). Volatile Composition Screening of Salix spp. Nectar Honey: Benzenecarboxylic Acids, Norisoprenoids, Terpenes, and Others. Chemistry & Biodiversity, 7(9), 2309-2325. doi:10.1002/cbdv.201000021Jones, G. D., & Bryant, Jr, V. M. (2004). The use of ETOH for the dilution of honey. Grana, 43(3), 174-182. doi:10.1080/00173130410019497Kummerow, J. (1983). Comparative Phenology of Mediterranean-Type Plant Communities. Ecological Studies, 300-317. doi:10.1007/978-3-642-68935-2_17La‐Serna Ramos, I. E., & GÓmez Ferreras, C. (2006). Pollen and sensorial characterization of different honeys from El Hierro (Canary Islands). Grana, 45(2), 146-159. doi:10.1080/00173130600578658Del Carmen Llasat, M., Ramis, C., & Barrantes, J. (1996). The meteorology of high‐intensity rainfall events over the west Mediterranean region. Remote Sensing Reviews, 14(1-3), 51-90. doi:10.1080/02757259609532313Louveaux, J., Maurizio, A., & Vorwohl, G. (1978). Methods of Melissopalynology. Bee World, 59(4), 139-157. doi:10.1080/0005772x.1978.11097714Martins, R. C., Lopes, V. V., Valentão, P., Carvalho, J. C. M. F., Isabel, P., Amaral, M. T., … Silva, B. M. (2008). Relevant principal component analysis applied to the characterisation of Portuguese heather honey. Natural Product Research, 22(17), 1560-1582. doi:10.1080/14786410701825004Melliou, E., & Chinou, I. (2011). Chemical constituents of selected unifloral Greek bee-honeys with antimicrobial activity. Food Chemistry, 129(2), 284-290. doi:10.1016/j.foodchem.2011.04.047Pendleton, M. (2006). Descriptions of melissopalynological methods involving centrifugation should include data for calculating Relative Centrifugal Force (RCF) or should express data in units of RCF or gravities (g). Grana, 45(1), 71-72. doi:10.1080/00173130500520479Pérez, R. A., Sánchez-Brunete, C., Calvo, R. M., & Tadeo, J. L. (2002). Analysis of Volatiles from Spanish Honeys by Solid-Phase Microextraction and Gas Chromatography−Mass Spectrometry. Journal of Agricultural and Food Chemistry, 50(9), 2633-2637. doi:10.1021/jf011551rPersano Oddo, L., Piana, L., Bogdanov, S., Bentabol, A., Gotsiou, P., Kerkvliet, J., … von der Ohe, K. (2004). Botanical species giving unifloral honey in Europe. Apidologie, 35(Suppl. 1), S82-S93. doi:10.1051/apido:2004045Persano Oddo, L., & Piro, R. (2004). Main European unifloral honeys: descriptive sheets. Apidologie, 35(Suppl. 1), S38-S81. doi:10.1051/apido:2004049Piana, M. L., Persano Oddo, L., Bentabol, A., Bruneau, E., Bogdanov, S., & Guyot Declerck, C. (2004). Sensory analysis applied to honey: state of the art. Apidologie, 35(Suppl. 1), S26-S37. doi:10.1051/apido:2004048Piasenzotto, L., Gracco, L., & Conte, L. (2003). Solid phase microextraction (SPME) applied to honey quality control. Journal of the Science of Food and Agriculture, 83(10), 1037-1044. doi:10.1002/jsfa.1502Radovic, B. S., Careri, M., Mangia, A., Musci, M., Gerboles, M., & Anklam, E. (2001). Contribution of dynamic headspace GC–MS analysis of aroma compounds to authenticity testing of honey. Food Chemistry, 72(4), 511-520. doi:10.1016/s0308-8146(00)00263-6RAMÓN-LACA, L., & MABBERLEY, D. J. (2004). The ecological status of the carob-tree (Ceratonia siliqua, Leguminosae) in the Mediterranean. Botanical Journal of the Linnean Society, 144(4), 431-436. doi:10.1111/j.1095-8339.2003.00254.xRetana, J., Ramoneda, J., Garcia Del Pino, F., & Bosch, J. (1994). Flowering phenology of carob,Ceratonia siliquaL. (Cesalpinaceae). Journal of Horticultural Science, 69(1), 97-103. doi:10.1080/14620316.1994.11515254Ricciardelli d’Albore, G. & Vorwohl, G. (1979). Mieles monoflorales en el Mediterráneo documentado con ayuda del análisis microscópico de mieles. Actas de XXVII Congreso Internacional de Apicultura, Athens, Greece, 14–20 September 1979, 201–208.Pilar de Sá‐Otero, M., Armesto‐Baztan, S., & DÍaz‐Losada, E. (2006). A study of variation in the pollen spectra of honeys sampled from the Baixa Limia‐Serra do Xurés Nature Reserve in north‐west Spain. Grana, 45(2), 137-145. doi:10.1080/00173130600708537Seijo, M. C., Jato, M. V., Aira, M. J., & Iglesias, I. (1997). Unifloral honeys of Galicia (north-west Spain). Journal of Apicultural Research, 36(3-4), 133-140. doi:10.1080/00218839.1997.11100939Terrab, A., Diez, M. J., & Heredia, F. J. (2003). Palynological, physico-chemical and colour characterization of Moroccan honeys: III. Other unifloral honey types. International Journal of Food Science and Technology, 38(4), 395-402. doi:10.1046/j.1365-2621.2003.00713.xTERRAB, A., PONTES, A., HEREDIA, F. J., & DÍEZ, M. J. (2004). A preliminary palynological characterization of Spanish thyme honeys. Botanical Journal of the Linnean Society, 146(3), 323-330. doi:10.1111/j.1095-8339.2004.00335.xTerrab, A., Valdés, B., & Josefa Díez, M. (2003). Pollen analysis of honeys from the Mamora forest region (NW Morocco). 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A global view on the riparian forests with Salix neotricha and Populus alba in the Iberian Peninsula (Portugal and Spain)

Forests dominated by Salix neotricha, and Populus alba found along the mesoeutrophic rivers in the Iberian Peninsula, were studied. We discuss the floristic circumscription, chorology, and community segregation based on the available releve´s of all Iberian riparian communities included in Populenion albae. Eleven formerly described communities were analyzed and due to original floristic combination, habitat features, and biogeographic scope, a new willow and poplar forest type is proposed within a well-defined biogeographical unit (Sadensean-Dividing Portuguese Subprovince): Clematido campaniflorae- Salicetum neotrichae. This syntaxon is found under a semi-hyperoceanic thermomediterranean to lower mesomediterranean, subhumid to humid bioclimate. Cluster analysis including all Iberian communities of Populenion albae shows a clear floristic segregation within the suballiance and confirms the originality of the new association. Furthermore, chemical characteristics of the water along some of the Portuguese watercourses with Populenion albae were studied and compared to the oligotrophic rivers occupied by Osmundo-Alnion communities. This study suggests that floristic separation between the communities of Populenion and Osmundo-Alnion is accompanied by a differentiation of the water trophic level

Influence of phenological barriers and habitat differentiation on the population genetic structure of the balearic endemic Rhamnus ludovici-salvatoris Chodat and R. alaternus L

[EN] Rhamnus ludovici-salvatoris, endemic to the Gymnesian Islands, coexists with the related and widespread R. alaternus in Mallorca and Menorca. In both species, the population genetic structure using RAPD, and flowering during a 3-year period to check for possible phenological barriers, were analyzed. Rhamnus ludovici-salvatoris showed lower genetic diversity and stronger population structure than R. alaternus, the Cabrera population being less diverse and the most differentiated. Rhamnus ludovici-salvatoris flowered one month later, although flowering of both species coincided sporadically. These congeners seem to have diverged through isolation by time and differentiation in habitat. The population genetic structure of R. ludovici-salvatoris could mainly be due to the existence of small populations on the one hand, and a gene flow caused by rare hybridization events on the other, which may also explain the presence of morphologically intermediate individuals in Menorca. The conservation of R. ludovici-salvatoris populations may include population reinforcements and other in situ interventions.Ferriol Molina, M.; Llorens García, L.; Gil, L.; Boira Tortajada, H. (2009). Influence of phenological barriers and habitat differentiation on the population genetic structure of the balearic endemic Rhamnus ludovici-salvatoris Chodat and R. alaternus L. Plant Systematics and Evolution. 277(1-2):105-116. doi:10.1007/s00606-008-0110-3S1051162771-2Affre L, Thompson JD, Debussche M (1997) Genetic structure of continental and island populations of the Mediterranean endemic Cyclamen balearicum (Primulaceae). Amer J Bot 84(4): 437–451BOIB (2005) Decreto 75/2005. BOIB 106: 29–32Bolmgren K, Oxelman B (2004) Generic limits in Rhamnus L. s.l. (Rhamnaceae) inferred from nuclear and chloroplast DNA sequence phylogenies. Taxon 53(2):383–390Bolòs O, Molinier R (1958) Recherches phytosociologiques dans l’île de Majorque. 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Effects of Salt Stress on Three Ecologically Distinct Plantago Species

Comparative studies on the responses to salt stress of taxonomically related taxa should help to elucidate relevant mechanisms of stress tolerance in plants. We have applied this strategy to three Plantago species adapted to different natural habitats, P. crassifolia and P. coronopus both halophytes and P. major, considered as salt-sensitive since it is never found in natural saline habitats. Growth inhibition measurements in controlled salt treatments indicated, however, that P. major is quite resistant to salt stress, although less than its halophytic congeners. The contents of monovalent ions and specific osmolytes were determined in plant leaves after four-week salt treatments. Salt-treated plants of the three taxa accumulated Na+ and Cl- in response to increasing external NaCl concentrations, to a lesser extent in P. major than in the halophytes; the latter species also showed higher ion contents in the non-stressed plants. In the halophytes, K+ concentration decreased at moderate salinity levels, to increase again under high salt conditions, whereas in P. major K+ contents were reduced only above 400 mM NaCl. Sorbitol contents augmented in all plants, roughly in parallel with increasing salinity, but the relative increments and the absolute values reached did not differ much in the three taxa. On the contrary, a strong (relative) accumulation of proline in response to high salt concentrations (600 800 mM NaCl) was observed in the halophytes, but not in P. major. These results indicate that the responses to salt stress triggered specifically in the halophytes, and therefore the most relevant for tolerance in the genus Plantago are: a higher efficiency in the transport of toxic ions to the leaves, the capacity to use inorganic ions as osmotica, even under low salinity conditions, and the activation, in response to very high salt concentrations, of proline accumulation and K+ transport to the leaves of the plants.MAH was a recipient of an Erasmus Mundus pre-doctoral scholarship financed by the European Commission (Welcome Consortium). AP acknowledges the Erasmus mobility programme for funding her stay in Valencia to carry out her Master Thesis.Al Hassan, M.; Pacurar, AM.; López Gresa, MP.; Donat Torres, MDP.; Llinares Palacios, JV.; Boscaiu Neagu, MT.; Vicente Meana, Ó. (2016). Effects of Salt Stress on Three Ecologically Distinct Plantago Species. PLoS ONE. 11(8):1-21. doi:10.1371/journal.pone.0160236S12111

α2,3-Sialyltransferase ST3Gal III Modulates Pancreatic Cancer Cell Motility and Adhesion In Vitro and Enhances Its Metastatic Potential In Vivo

Background: Cell surface sialylation is emerging as an important feature of cancer cell metastasis. Sialyltransferase expression has been reported to be altered in tumours and may account for the formation of sialylated tumour antigens. We have focused on the influence of alpha-2,3-sialyltransferase ST3Gal III in key steps of the pancreatic tumorigenic process. Methodology/Principal Findings: ST3Gal III overexpressing pancreatic adenocarcinoma cell lines Capan-1 and MDAPanc-28 were generated. They showed an increase of the tumour associated antigen sialyl-Lewis x. The transfectants ’ E-selectin binding capacity was proportional to cell surface sialyl-Lewis x levels. Cellular migration positively correlated with ST3Gal III and sialyl-Lewis x levels. Moreover, intrasplenic injection of the ST3Gal III transfected cells into athymic nude mice showed a decrease in survival and higher metastasis formation when compared to the mock cells. Conclusion: In summary, the overexpression of ST3Gal III in these pancreatic adenocarcinoma cell lines underlines the rol

Allogamy-Autogamy Switch Enhance Assortative Mating in the Allotetraploid Centaurea seridis L. Coexisting with the Diploid Centaurea aspera L. and Triggers the Asymmetrical Formation of Triploid Hybrids

[EN] Hybridization between tetraploids and its related diploids is generally unsuccessful in Centaurea, hence natural formation of triploid hybrids is rare. In contrast, the diploid Centaurea aspera and the allotetraploid C. seridis coexist in several contact zones where a high frequency of triploid hybrids is found. We analyzed the floral biology of the three taxa to identify reproductive isolation mechanisms that allow their coexistence. Flowering phenology was recorded, and controlled pollinations within and between the three taxa were performed in the field. Ploidy level and germination of progeny were also assessed. There was a 50% flowering overlap which indicated a phenological shift. Diploids were strictly allogamous and did not display mentor effects, while tetraploids were found to be highly autogamous. This breakdown of self-incompatibility by polyploids is first described in Centaurea. The asymmetrical formation of the hybrid was also found: all the triploid intact cypselae came from the diploid mothers pollinated by the pollen of tetraploids. Pollen and eggs from triploids were totally sterile, acting as a strong triploid block. These prezygotic isolation mechanisms ensured higher assortative mating in tetraploids than in diploids, improving its persistence in the contact zones. However these mechanisms can also be the cause of the low genetic diversity and high genetic structure observed in C. seridis.Ferriol Molina, M.; Garmendia, A.; Ana Gonzalez; Merle Farinós, HB. (2015). Allogamy-Autogamy Switch Enhance Assortative Mating in the Allotetraploid Centaurea seridis L. Coexisting with the Diploid Centaurea aspera L. and Triggers the Asymmetrical Formation of Triploid Hybrids. PLoS ONE. 10(10):1-13. doi:10.1371/journal.pone.0140465S1131010Jiao, Y., Wickett, N. J., Ayyampalayam, S., Chanderbali, A. S., Landherr, L., Ralph, P. E., … dePamphilis, C. W. (2011). 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