Plasticity to drought and ecotypic differentiation in populations of a crop wild relative.

Populations of widely distributed species often exhibit geographic variation in functional traits in response to environmental heterogeneity. Such trait variation may be the result of different adaptive mechanisms, including genetically based differentiation, phenotypic plasticity or a combination of both. Disentangling the genetic and environmental components of trait variation may be particularly interesting in crop wild relatives, since they may provide unique reservoirs of genetic diversity for crop improvement. In this study, we assessed ecotypic differentiation and patterns of plasticity to drought in populations of Lupinus angustifolius, a Mediterranean crop wild relative, from two climatically distinct regions in the Iberian Peninsula. Using an outdoor common garden, we compared phenotypic responses of inbred maternal families to two ecologically meaningful water availability treatments (drought and high-moisture). We measured 18 different functional traits related to growth, morphology, phenology and reproduction. Plants in the drought treatment grew less, had lower leaf chlorophyll content and photochemical efficiency, but also reproduced faster, produced larger seeds and altered leaflet morphology through increased leaflet thickness, higher leaflet dry matter content and lower specific leaf area. We also found significant differences between regions that likely reflect adaptation to climatically distinct environments, with populations from the south showing a faster onset of reproduction, higher leaf thickness and higher seed size, consistent with the drier conditions experienced in southern sites. Plasticity to drought was in most cases in the same direction as quantitative genetic differentiation (i.e. cogradient variation), providing evidence of the adaptive value of the plastic change. Our results show that both genetic differentiation and plasticity can generate adaptive phenotypic variation in L. angustifolius, and help to identify potentially valuable genetic resources to incorporate into breeding programmes.

Ecotypic differentiation reveals seed colour-related alkaloid content in a crop wild relative.

Crop wild relatives can be a useful source of genotypes that maximise crop survival and yield in specific habitats. Lupinus angustifolius is an annual forb with crop varieties derived from a narrow genetic basis but that are cultivated worldwide. Its seeds have high nutritional value, but they naturally contain alkaloids with anti-nutritive factors. The study of its wild populations can be useful to find genotypes that contribute to higher climate resilience and greater yield under stressing environmental conditions. Using a common garden, we evaluated ecotypic differentiation in four natural populations from two contrasting latitudes in terms of plant biomass, seed mass and number, alkaloid content in seeds for the three main alkaloids present in the plant and seed colour, including its possible influence on post-dispersal predation. Correlations among traits were also assessed. We found differences among populations for all traits except final biomass. Northern populations had lighter seeds and a tendency to yield more seeds when they produced white seeds, compared to southern populations and variegated seeds. Regardless of latitude, populations showed differences in alkaloid concentration, with all three alkaloids found generally in high or low concentrations in each population. Proportion of white seed morphs varied in each population. Seed colour did not influence predator preference. In addition, white seed colour was related to a low alkaloid content. Our results evidence the existence of natural ecotypic differentiation in L. angustifolius not only due to latitudinal range, but also to local environmental factors. White seed coat colour could be used as a visual clue for identification of low-alkaloid genotypes, a priority trait in L. angustifolius breeding programmes.

Reproductive traits and evolutionary divergence between Mediterranean crops and their wild relatives.

Changes in reproductive traits associated with domestication critically determine the evolutionary divergence between crops and their wild relatives, as well as the potential of crop plants to become feral. In this review, we examine the genetic mechanisms of plant domestication and the different types of selection involved, and describe the particularities of domestication of Mediterranean field crops with regard to their reproductive traits, showing illustrative examples. We also explore gene flow patterns between Mediterranean field crops and their wild relatives, along with their ecological, evolutionary and economic implications. Domestication entails multiple selective processes, including direct selection, environmental adaptation and developmental constraints. In contrast to clonal propagation in perennials, sexual reproduction and seed propagation in annuals and biennials have led to a distinct pathway of evolution of reproductive traits. Thus, the initial domestication and further breeding of Mediterranean field crops has brought about changes in reproductive traits, such as higher mean values and variance of seed and fruit sizes, reduced fruit and seed toxicity, non-shattering seeds and loss of seed dormancy. Evolution under domestication is not a linear process, and bi-directional gene flow between wild and crop taxa is a frequent phenomenon. Thus, hybridisation and introgression have played a very important role in determining the genetics of current cultivars. In turn, gene flow from crops to wild relatives can lead to introgression of crop genes into wild populations and potentially alter the characteristics of natural communities. In conclusion, plant evolution under domestication has not only changed the reproductive biology of cultivated taxa, its effects are multifaceted and have implications beyond agriculture.

How does climate change affect regeneration of Mediterranean high-mountain plants? An integration and synthesis of current knowledge.

Mediterranean mountains are extraordinarily diverse and hold a high proportion of endemic plants, but they are particularly vulnerable to climate change, and most species distribution models project drastic changes in community composition. Retrospective studies and long-term monitoring also highlight that Mediterranean high-mountain plants are suffering severe range contractions. The aim of this work is to review the current knowledge of climate change impacts on the process of plant regeneration by seed in Mediterranean high-mountain plants, by combining available information from observational and experimental studies. We also discuss some processes that may provide resilience against changing environmental conditions and suggest some research priorities for the future. With some exceptions, there is still little evidence of the direct effects of climate change on pollination and reproductive success of Mediterranean high-mountain plants, and most works are observational and/or centred only in the post-dispersal stages (germination and establishment). The great majority of studies agree that the characteristic summer drought and the extreme heatwaves, which are projected to be more intense in the future, are the most limiting factors for the regeneration process. However, there is an urgent need for studies combining elevational gradient approaches with experimental manipulations of temperature and drought to confirm the magnitude and variability of species' responses. There is also limited knowledge about the ability of Mediterranean high-mountain plants to cope with climate change through phenotypic plasticity and local adaptation processes. This could be achieved by performing common garden and reciprocal translocation experiments with species differing in life history traits.

Individual spatial aggregation correlates with between-population variation in fine-scale genetic structure of Silene ciliata (Caryophyllaceae).

Fine-scale genetic structure (FSGS) can vary among populations within species depending on multiple demographic and environmental factors. Theoretical models predict that FSGS should decrease in high-density populations and increase in populations where individuals are spatially aggregated. However, few empirical studies have compared FSGS between populations with different degrees of individual spatial aggregation and microhabitat heterogeneity. In this work, we studied the relationship between spatial and genetic structure in five populations of alpine specialist Silene ciliata Poiret (Caryophyllaceae). We mapped all individuals in each population and genotyped 96 of them using 10 microsatellite markers. We found significant FSGS consistent with an isolation-by-distance process in three of the five populations. The intensity of FSGS was positively associated with individual spatial aggregation. However, no association was found between FSGS and global population density or microhabitat heterogeneity. Overall, our results support theoretical studies indicating that stronger spatial aggregation tends to increase the magnitude of FSGS. They also highlight the relevance of characterizing local plant distribution and microhabitat to better understand the mechanisms that generate intraspecific variation in FSGS across landscapes.

Effects of the duration of cold stratification on early life stages of the Mediterranean alpine plant Silene ciliata.

Cold stratification provided by snow cover is essential to break seed dormancy in many alpine plant species. The forecast reduction in snow precipitation and snow cover duration in most temperate mountains as a result of global warming could threaten alpine plant populations, especially those at the edge of their species distribution, by altering the dynamics of early life stages. We simulated some effects of a reduction in the snow cover period by manipulating the duration of cold stratification in seeds of Silene ciliata, a Mediterranean alpine specialist. Seeds from three populations distributed along an altitudinal gradient were exposed to different periods of cold stratification (2, 4 and 6 months) in the laboratory and then moved to common garden conditions in a greenhouse. The duration of the cold stratification treatment and population origin significantly affected seed emergence percentage, emergence rate and seedling size, but not the number of seedling leaves. The 6-month and 4-month cold stratification treatments produced higher emergence percentages and faster emergence rates than seeds without cold stratification treatment. No significant cold stratification duration x seed population origin interactions were found, thus differential sensitivity to cold stratification along elevation is not supported.

Reproductive limits of a late-flowering high-mountain Mediterranean plant along an elevational climate gradient.

Mountain plants are particularly sensitive to climate warming because snowmelt timing exerts a direct control on their reproduction. Current warming is leading to earlier snowmelt dates and longer snow-free periods. Our hypothesis is that high-mountain Mediterranean plants are not able to take advantage of a lengthened snow-free period because this leads to longer drought that truncates the growing season. However, reproductive timing may somewhat mitigate these negative effects through temporal shifts. We assessed the effects of flowering phenology on the reproductive success of Silene ciliata, a Mediterranean high-mountain plant, across an altitudinal gradient during two climatically contrasting years. The species showed a late-flowering pattern hampering the use of snowmelt water. Plant fitness was largely explained by the elapsed time from snowmelt to onset of flowering, suggesting a selective pressure towards early flowering caused by soil moisture depletion. The proportion of flowering plants decreased at the lowest population, especially in the drier year. Plants produced more flowers, fruits and seeds at the highest population and in the mild year. Our results indicate that water deficit in dry years could threaten the lowland populations of this mountainous species, while high-altitude environments are more stable over time.

[Auricular chondritis. Experimental model in rats]. / Condritis auricular. Modelo experimental en rata.

The general pattern of local inflammation in the rat is developed in the pad of the paw. In otorhinolaryngology a model of local inflammation can be reached in the rat ear by means of the production of an auricular chondritis. For that, we have protocolized the methodology. The induction of inflammation is carried out by the substance vegetable carrageenan. It has been verified by histological studies the inflammation generated, as well as studying the action of antiinflammatory, proinflammatory and inflammatory compounds. The inflammation has been measured by nonius and by determination of the activity of the myeloperoxidase enzyme. After these studies we can validate the auricular chondritis in rat as an experimental model of local inflammation in otorhinolaryngology.

Factors affecting establishment of a gypsophyte: the case of Lepidium subulatum (Brassicaceae).

The restriction of vascular plants to gypsum-rich soils under arid or semiarid climates has been reported by many authors in different parts of the world. However, factors controlling the presence of gypsophytes on these soils are far from understood. We investigated the establishment of Lepidium subulatum, a gypsophyte, in a nondisturbed semiarid gypsum-soil landscape in central Spain, both from spatial and temporal perspectives. Over 1400 seedlings were tagged, and their growth and survival were monitored for a 2-yr period. Several biotic and abiotic variables were measured to determine the factors controlling the emergence and early survival. These variables included the cover of annual plants, bryophytes, lichens, litter, gypsum crystals, bare fraction and cover of each perennial plant, and several soil properties (gravel, fine gravel, and fine-earth fraction, conductivity, pH, gypsum content, organic matter and penetrometer soil resistance). Our results support the linkage of gypsophily with some physical properties of the surface crust. Seedlings tended to establish on the gypsum surface crust, and their survival was size dependent, probably as a consequence of the necessity of rooting below the surface crust before summer drought arrives. However, once seedlings emerged, a higher survival rate occurred on the alluvial soils of the piedmont-slope boundary where soil crusts are absent or thinner. We conclude that Lepidium subulatum may be considered a refuge model endemic with a distribution range that occupies a reduced fraction of a wider habitat from which it is probably excluded by competition.

Organ-preserving management of rhabdomyosarcoma of the prostate and bladder in children.

Between November 1978 and July 1991, ten children between one and eight years of age with Group III pelvic rhabdomyosarcoma (IRS classification) and considered inoperable at diagnosis were treated primarily with intensive poly-chemotherapy. Complementary radiotherapy and conservative surgery were added as needed. Eight of the ten survive free of disease with functioning bladders for periods ranging from 5.7-18.4 years.

Micropropagation of Elaeagnus angustifolia from mature trees.

Multiple shoots were obtained from nodal segments of mature trees of Elaeagnus angustifolia L. cultured on MS medium (Murashige and Skoog 1962) supplemented with 0, 0.88 or 2.22 micro M N(6)-benzyladenine. When nodal segments taken from the in vitro proliferated shoots were cultured under the same conditions, additional multiple shoots were obtained. Rooting of the in vitro propagated shoots was achieved on full strength MS medium or on MS supplemented with 2.46 micro M indole-3-butyric acid. Regenerated plantlets were acclimatized and successfully transplanted to soil.

Effects of Seed Cryopreservation and Priming on Germination in Several Cultivars of Apium graveolens.

Seed germination of seven celery cultivars was studied after storage in liquid nitrogen for 1 or 30 d. Cryopreservation was also carried out on pelleted and primed seeds. None of the treatments applied reduced germination percentages. T(50) (time for germination to reach 50%) significantly decreased in Florida, Utah and Istar cultivars when priming, alone or in combination with cryopreservation, was used.

Micropropagation of an endangered plant species: Coronopus navasii (Brassicaceae).

Plantlets of Coronopus navasii, an endangered species from SE Spain, were successfully regenerated from shoot and root segments excised from young seedlings. Initiation of multiple buds and development of leaves were obtained in MS modified medium plus l mg/l BAP and 0.1 mg/l NAA. Rooting was achieved by transfer of the isolated shoots to fresh MS medium without plant growth regulators. Plant survival of 47% was obtained six weeks after removal from in vitro culture conditions.

Malignant fibrous histiocytoma of bone. A clinico-pathological and electronmicroscopical study.

The clinicopathological and ultrastructural features of seven Malignant Fibrous Histiocytomas reported to the Spanish Registry of Bone Tumors were studied. Two patients were females (20 and 60 years old) and the other five males (15, 37, 67, 38 and 17 years old). The tumors were located in the lower femur (3 cases), upper tibia, upper humerus, rib and iliac bone. The past medical history revealed previous fracture in one patient and previous radiation of the affected bone in another; in a third patient the tumor associated to a bone infarct. Microscopically all tumors presented a storiform pattern composed of neoplastic fibroblasts, histiocytes, and malignant multinucleated giant cells. There was no evidence of osteoid or bone tissue formation by the tumor cells. Five cases were studied with electronmicroscopy; the tumor cells had features of fibroblasts, myofibroblasts, histiocytes and multinucleated histiocytes. Primary and secondary lysosomes, along with lipid vacuoles were common findings in the tumor cells. The value of the electron-microscopy in the differential diagnosis of this tumor is emphasized.