Integrated conservation of important plant taxa through the improvement of the original plant micro-reserve (PMR) approach: The intensive PMR monitoring case of Ophrys kotschyi.

Developing a monitoring system and a conservation strategy against the negative impact of global change on threatened plant species, is nowadays the challenge for conservation experts. The Plant Micro-Reserve (PMR) approach became a highly effective approach in protecting plant species, since mild active management of vegetation plots and protection of plant populations takes place. The PMR has greatly evolved since its initial concept of managing a large network of PMRs, to having fewer protected areas subject to intensive scientific monitoring (e.g. Intensive Monitoring PMR; IM-PMR). This study further improved the IM-PMR approach by focusing on the threatened plant species of Ophrys kotschyi in Cyprus. The proposed IM-PMR enhances the available knowledge on the biology, physiology and ecology of the targeted plant species, through implementing an intensive monitoring system and assessing its genetic diversity. Within the framework of IM-PMR, the population size of O. kotschyi recorded statistically significant differentiation during the monitoring period, most likely due to the vegetative dormancy of the species. The subpopulation size and dormancy in O. kotschyi was correlated with precipitation and air temperature for specific months. In addition, the different local climatic conditions and the species dormancy between years seem to influence the flowers production among individuals, in the four monitoring years. Nevertheless, the low natural fecundity compared to the artificially pollinated plants and the absent correlation with any of the climatic parameters, might be closely related to the lack of pollinators at this site. The genetic diversity (HT = 0.456) is higher compared to other endemic and short-lived perennial species, while the genetic differentiation among the subpopulations of O. kotschyi showed significant substructure (FSTFIS=0.5 = 0.097*). The subpopulation within IM-PMR showed relatively lower genetic diversity among the largest subpopulations of O. kotschyi, and the highest percentage of linked loci. Such observations support the non-random association of different loci in this subpopulation, and the ineffective pollen flow within this single subpopulation. The improvement of the original PMR approach in the current IM-PMR proposal denotes that different ecological aspects are taken into account towards gaining a holistic knowledge on a target species. The IM-PMR approach as implemented for O. kotschyi, could lead to the development of an integrated conservation approach for rare, threatened, or endangered species.

Postfire biodiversity database for eastern Iberia.

In the summer of 2012, two fires affected Mediterranean ecosystems in the eastern Iberian Peninsula. The size of these fires was at the extreme of the historical variability (megafires). Animals are traditionally assumed to recolonize from source populations outside of the burned area (exogenous regeneration) while plants recover from endogenous regeneration (resprouting and seeding). However, there is increasing evidence of in situ fire survival in animals. To evaluate the effect of large-scale fires on biodiversity and the mechanism of recovery, in 2013, we set up 12 plots per fire, covering burned vegetation at different distances from the fire perimeter and unburned vegetation. In each plot, we followed the postfire recovery of arthropods, reptiles (including some of their parasites), and plants for 2 to 5 years. Here we present the resulting database (POSTDIV) of taxon abundance. POSTDIV totals 19,906 records for 457 arthropod taxa (113,681 individuals), 12 reptile taxa (503 individuals), 4 reptile parasites (234 individuals), and 518 plant taxa (cover-abundance). We provide examples in the R language to query the database.

Constitutive and Adaptive Traits of Environmental Stress Tolerance in the Threatened Halophyte Limonium angustebracteatum Erben (Plumbaginaceae).

Limonium angustebracteatum is a halophyte endemic to the E and SE Iberian Peninsula with interest in conservation. Salt glands represent an important adaptive trait in recretohalophytes like this and other Limonium species, as they allow the excretion of excess salts, reducing the concentration of toxic ions in foliar tissues. This study included the analysis of the salt gland structure, composed of 12 cells, 4 secretory and 8 accessory. Several anatomical, physiological and biochemical responses to stress were also analysed in adult plants subjected to one month of water stress, complete lack of irrigation, and salt stress, by watering with aqueous solutions of 200, 400, 600 and 800 mM NaCl. Plant growth was inhibited by the severe water deficit and, to a lesser extent, by high NaCl concentrations. A variation in the anatomical structure of the leaves was detected under conditions of salt and water stress; plants from the salt stress treatment showed salt glands sunken between epidermal cells, bordered by very large epidermal cells, whereas in those from the water stress treatment, the epidermal cells were heterogeneous in shape and size. In both, the palisade structure of the leaves was altered. Salt excretion is usually accompanied by the accumulation of salts in the foliar tissue. This was also found in L. angustebracteatum, in which the concentration of all ions analysed was higher in the leaves than in the roots. The increase of K+ in the roots of plants subjected to water stress was also remarkable. The multivariate analysis indicated differences in water and salt stress responses, such as the accumulation of Na and Cl, or proline, but K+ homeostasis played a relevant role in the mechanism of tolerance to both stressful conditions.

Seventeen 'extinct' plant species back to conservation attention in Europe.

Seventeen European endemic plant species were considered extinct, but improved taxonomic and distribution knowledge as well as ex situ collecting activities brought them out of the extinct status. These species have now been reported into a conservation framework that may promote legal protection and in situ and ex situ conservation.

Responses to Increased Salinity and Severe Drought in the Eastern Iberian Endemic Species Thalictrum maritimum (Ranunculaceae), Threatened by Climate Change.

Thalictrum maritimum is an endangered, endemic species in East Spain, growing in areas of relatively low salinity in littoral salt marshes. A regression of its populations and the number of individuals has been registered in the last decade. This study aimed at establishing the causes of this reduction using a multidisciplinary approach, including climatic, ecological, physiological and biochemical analyses. The climatic data indicated that there was a direct negative correlation between increased drought, especially during autumn, and the number of individuals censused in the area of study. The susceptibility of this species to water deficit was confirmed by the analysis of growth parameters upon a water deficit treatment applied under controlled greenhouse conditions, with the plants withstanding only 23 days of complete absence of irrigation. On the other hand, increased salinity does not seem to be a risk factor for this species, which behaves as a halophyte, tolerating in controlled treatments salinities much higher than those registered in its natural habitat. The most relevant mechanisms of salt tolerance in T. maritimum appear to be based on the control of ion transport, by (i) the active transport of toxic ions to the aerial parts of the plants at high external salinity-where they are presumably stored in the leaf vacuoles to avoid their deleterious effects in the cytosol, (ii) the maintenance of K+ concentrations in belowground and aboveground organs, despite the increase of Na+ levels, and (iii) the salt-induced accumulation of Ca2+, particularly in stems and leaves. This study provides useful information for the management of the conservation plans of this rare and endangered species.

Ethnopharmacology in the Upper Guadiana River area (Castile-La Mancha, Spain).

ETHNOPHARMACOLOGICAL RELEVANCE: Determining traditional remedies for human pathologies is relevant, when compared with the standard materia medica of the pharmacopoeias and dietary supplement databases, because we can assess the species and uses that have been previously studied and target understudied species for further pharmacological investigation. BACKGROUND: The aim of this study was to systematically record and analyze medicinal uses of natural resources (Plantae, Animalia, Fungi and minerals), mostly local, in the territories adjoining the upper Guadiana River and its tributaries. We were particularly interested in recording resources and pathologies linked to wetland areas, especially in the National Parks Las Tablas de Daimiel and Cabañeros. Wetlands are interesting because they present a double face in relation with human health: Wetlands furnish hydration, safe water, nutrition, and medicinal resources; are places from which people derive their livelihood. However wetlands are also sites of exposure to pollution or toxicants, and infectious diseases; and sites of physical hazards. We wanted to identify procedures for preparation of medicinal formulae and routes of administration. We also intended to detect whether a geographical pattern exists or not in our records in relation to the use of local resources. METHODS: We used semi-structured interviews with one-to-one informants or groups, from 1998 to 2018. Raw data were introduced in a Firebird database and analyzed. To identify ingredients and pathologies we consulted local floras and epidemiological literature. Finally, we compared documented pathologies, remedies and ingredients in the historical context of medicinal uses of natural resources in Castile-La Mancha and especially in Ciudad Real. RESULTS: 126 pathologies and 220 species furnishing ingredients have been recorded from the interviews. In total, 188 are plants and 20 animals. The most commonly used species include Malva sylvestris, Phlomis lychnitis, Genista tridentata and Thymus mastichina. Most records refer to flowers, or fruits, of locally available plant species, classified as Mediterranean, European or widespread that belong to the Lamiaceae, Compositae or Leguminosae. Ingredients which are collected in open shrublands, known as "garrigue", and dry grasslands furnish a relevant proportion of records while the imported ingredients remain marginal. The contribution of wetlands, riverine habitats and irrigated fields and gardens as a source of medicinal resources is 36% of the records. It is relatively high considering its limited presence in terms of total extension within the study area. The most frequently reported diseases are respiratory, gastrointestinal, dermatological and infectious or parasitic. CONCLUSIONS: Along the Guadiana River in the Ciudad Real province exists a wide and deep knowledge of traditional remedies for the treatment of common pathologies, based fundamentally on the use of local flora, fauna and mineral resources. The uses and ingredients documented are useful for further pharmacological investigation to improve health care for a wide range of pathologies.

Insights on Salt Tolerance of Two Endemic Limonium Species from Spain.

We have analysed the salt tolerance of two endemic halophytes of the genus Limonium, with high conservation value. In the present study, seed germination and growth parameters as well as different biomarkers-photosynthetic pigments, mono and divalent ion contents-associated to salt stress were evaluated in response to high levels of NaCl. The study was completed with an untargeted metabolomics analysis of the primary compounds including carbohydrates, phosphoric and organic acids, and amino acids, identified by using a gas chromatography and mass spectrometry platform. Limonium albuferae proved to be more salt-tolerant than L. doufourii, both at the germination stage and during vegetative growth. The degradation of photosynthetic pigments and the increase of Na+/K+ ratio under salt stress were more accentuated in the less tolerant second species. The metabolomics analysis unravelled several differences between the two species. The higher salt tolerance of L. albuferae may rely on its specific accumulation of fructose and glucose under high salinity conditions, the first considered as a major osmolyte in this genus. In addition, L. albuferae showed steady levels of citric and malic acids, whereas the glutamate family pathway was strongly activated under stress in both species, leading to the accumulation of proline (Pro) and γ-aminobutyric acid (GABA).

Plant micro-reserves in Valencia (E. Spain): A model to preserve threatened flora in China?

The Valencian Community (eastern Spain) was the pioneer territory establishing plant micro-reserves (PMRs). Its model to protect small sites for endemic and endangered plants has been exported to several countries around the globe. This paper highlights 1) the role of PMRs to complement the protection provided by large protected areas, 2) how the establishment of PMRs fosters the increase of floristic knowledge, and 3) the fact that continuous monitoring of PMRs also yields new records of endangered species found within the same PMRs. The flexibility of the PMR approach -it can be adapted to other national and regional legislations- allows its transfer to other rich-biodiversity regions and countries such as China.

Conservation of endangered Lupinus mariae-josephae in its natural habitat by inoculation with selected, native Bradyrhizobium strains.

Lupinus mariae-josephae is a recently discovered endemism that is only found in alkaline-limed soils, a unique habitat for lupines, from a small area in Valencia region (Spain). In these soils, L. mariae-josephae grows in just a few defined patches, and previous conservation efforts directed towards controlled plant reproduction have been unsuccessful. We have previously shown that L. mariae-josephae plants establish a specific root nodule symbiosis with bradyrhizobia present in those soils, and we reasoned that the paucity of these bacteria in soils might contribute to the lack of success in reproducing plants for conservation purposes. Greenhouse experiments using L. mariae-josephae trap-plants showed the absence or near absence of L. mariae-josephae-nodulating bacteria in "terra rossa" soils of Valencia outside of L. mariae-josephae plant patches, and in other "terra rossa" or alkaline red soils of the Iberian Peninsula and Balearic Islands outside of the Valencia L. mariae-josephae endemism region. Among the bradyrhizobia able to establish an efficient symbiosis with L. mariae-josephae plants, two strains, LmjC and LmjM3 were selected as inoculum for seed coating. Two planting experiments were carried out in consecutive years under natural conditions in areas with edapho-climatic characteristics identical to those sustaining natural L. mariae-josephae populations, and successful reproduction of the plant was achieved. Interestingly, the successful reproductive cycle was absolutely dependent on seedling inoculation with effective bradyrhizobia, and optimal performance was observed in plants inoculated with LmjC, a strain that had previously shown the most efficient behavior under controlled conditions. Our results define conditions for L. mariae-josephae conservation and for extension to alkaline-limed soil habitats, where no other known lupine can thrive.