Fire and summer temperatures work together breaking physical seed dormancy.

Fire and high summer soil temperatures can break physical seed dormancy in Mediterranean fire-prone ecosystems. Their independent effect is somewhat recognized but both factors may act together with a synergistic effect yet unknown. This study aims to determine the isolated and combined effects of fire and summer temperatures on the release of physical seed dormancy in Cistaceae species. Fire and summer temperature treatments were applied in a factorial experiment to seeds of 12 species of Cistaceae. Seeds previously exposed or not to a heat shock (fire simulation) were kept for 1 or 2 months at constant or alternating temperatures (summer temperatures simulation). Additionally, I compared the effect of exposing the seeds to a heat shock before or after they had been subjected to the summer temperatures. Heat shock increased germination of all species, but summer temperatures produced different results. When seeds were exposed to summer temperatures after heat shock, germination decreased. This negative effect disappeared when heat shock was simulated at the end of the summer temperatures. Fire and summer temperatures modulate timing of germination in Cistaceae with a joint control on post-fire regeneration. Cycling of sensitivity to physical dormancy release may be the mechanism to explain this fine-tuning, which would ensure germination when environmental conditions are suitable for growth. These results contribute to our understanding of vegetation dynamics and postfire regeneration in Mediterranean ecosystems.

Purification and characterization of Terfezia claveryi TcCAT-1, a desert truffle catalase upregulated in mycorrhizal symbiosis.

Terfezia claveryi Chatin is a mycorrhizal fungus that forms ectendomycorrhizal associations with plants of Helianthemum genus. Its appreciated edibility and drought resistance make this fungus a potential alternative crop in arid and semiarid areas of the Mediterranean region. In order to increase the knowledge about the biology of this fungus in terms of mycorrhiza formation and response to drought stress, a catalase from T. claveryi (TcCAT-1) has been purified to apparent homogeneity and biochemically characterized; in addition, the expression pattern of this enzyme during different stages of T. claveryi biological cycle and under drought stress conditions are reported. The results obtained, together with the phylogenetic analysis and homology modeling, indicate that TcCAT-1 is a homotetramer large subunit size monofunctional-heme catalase belonging to Clade 2. The highest expression of this enzyme occurs in mature mycorrhiza, revealing a possible role in mycorrhiza colonization, but it is not upregulated under drought stress. However, the H2O2 content of mycorrhizal plants submitted to drought stress is lower than in well watered treatments, suggesting that mycorrhization improves the plant's oxidative stress response, although not via TcCAT-1 upregulation.

The demography of climate-driven and density-regulated population dynamics in a perennial plant.

Identifying the internal and external drivers of population dynamics is a key objective in ecology, currently accentuated by the need to forecast the effects of climate change on species distributions and abundances. The interplay between environmental and density effects is one particularly important aspect of such forecasts. We examined the simultaneous impact of climate and intraspecific density on vital rates of the dwarf shrub Fumana procumbens over 20 yr, using generalized additive mixed models. We then analyzed effects on population dynamics using integral projection models. The population projection models accurately captured observed fluctuations in population size. Our analyses suggested the population was intrinsically regulated but with annual fluctuations in response to variation in weather. Simulations showed that implicitly assuming variation in demographic rates to be driven solely by the environment can overestimate extinction risks if there is density dependence. We conclude that density regulation can dampen effects of climate change on Fumana population size, and discuss the need to quantify density dependence in predictions of population responses to environmental changes.

Morphological characterization of mycorrhizae formed between three Terfezia species (desert truffles) and several Cistaceae and Aleppo pine.

Six Cistaceae species, Helianthemum ledifolium, Helianthemum lippii, Fumana procumbens, Cistus albidus, Cistus incanus, Cistus salvifolius, and Pinus halepensis (Aleppo pine) were inoculated with three mycorrhizal desert truffles, Terfezia leptoderma, Terfezia boudieri, and Terfezia claveryi under greenhouse conditions, on soil originating from desert truffle natural habitat in Algeria. The syntheses have led to the formation of typical endomycorrhizae in annual Cistaceae (H. ledifolium) and perennial ones (H. lippii and F. procumbens) and an ectomycorrhiza with a less developed sheath in Cistus species and Aleppo pine. These results demonstrate the plasticity of Terfezia species to form different mycorrhizal types. The formation of an endomycorrhiza with H. ledifolium and F. procumbens and a sheathing ectomycorrhiza with P. halepensis inoculated by T. leptoderma in in vivo culture conditions was obtained for the first time.

Expression analysis of aquaporins from desert truffle mycorrhizal symbiosis reveals a fine-tuned regulation under drought.

We have performed the isolation, functional characterization, and expression analysis of aquaporins in roots and leaves of Helianthemum almeriense, in order to evaluate their roles in tolerance to water deficit. Five cDNAs, named HaPIP1;1, HaPIP1;2, HaPIP2;1, HaPIP2;2, and HaTIP1;1, were isolated from H. almeriense. A phylogenetic analysis of deduced proteins confirmed that they belong to the water channel proteins family. The HaPIP1;1, HaPIP2;1, and HaTIP1;1 genes encode functional water channel proteins, as indicated by expression assays in Saccharomyces cerevisiae, showing divergent roles in the transport of water, CO2, and NH3. The expression patterns of the genes isolated from H. almeriense and of a previously described gene from Terfezia claveryi (TcAQP1) were analyzed in mycorrhizal and nonmycorrhizal plants cultivated under well-watered or drought-stress conditions. Some of the studied aquaporins were subjected to fine-tuned expression only under drought-stress conditions. A beneficial effect on plant physiological parameters was observed in mycorrhizal plants with respect to nonmycorrhizal ones. Moreover, stress induced a change in the mycorrhizal type formed, which was more intracellular under drought stress. The combination of a high intracellular colonization, together with the fine-tuned expression of aquaporins could result in a morphophysiological adaptation of this symbiosis to drought conditions.

The role of phosphorus in the ectendomycorrhiza continuum of desert truffle mycorrhizal plants.

The influence of inorganic and organic phosphorus (P) and the absence of P in the culture medium on the type of mycorrhizal colonization formed (ecto-, ectendo-, or endomycorrhiza) during Helianthemum almeriense x Terfezia claveryi symbiosis in in vitro conditions was analyzed. This is the first time that the relative proportions of the different mycorrhizal types in mycorrhizal roots of H. almeriense have been quantified and statistically analyzed. The relative proportions of the mycorrhizal types depended on the P source in the medium, suggesting that it is the organic P form that induces the formation of intracellular colonization. The above association should be considered as a continuum between intra- and intercellular colonizations, the most appropriate term for defining it being ectendomycorrhiza. The influence of the endogenous concentration of P on plant growth was also analyzed. P translocation was observed from shoot to roots, especially in mycorrhizal plants because mycorrhizal roots showed higher growth than non-mycorrhizal roots and/or because of an extra P demand from mycelium inside the roots. Soluble and cell wall acid phosphatases activities from H. almeriense roots were kinetically characterized at optimum pH (5.0), using p-nitrophenyl phosphate as substrate, with K (m) values of 3.4 and 1.8 mM, respectively. Moreover, the plant acid phosphatase and fungal alkaline phosphatases activities were histochemically localised in mycorrhizal H. almeriense roots by fluorescence with enzyme-labelled fluorescence substrate.

Site effect is stronger than species identity in driving demographic responses of Helianthemum (Cistaceae) shrubs in gypsum environments.

PREMISE OF THE STUDY: Regional climatic patterns result in the synchrony of biological processes along large spatial areas. These patterns may be critical for effective plant recruitment in (semi)arid environments. Nevertheless, recruitment patterns of plant species within their range of distribution are still poorly known. Moreover, this response may be species-dependent, particularly between coexisting congenerics, which are thought to vary in demographic responses to climatic conditions as a coexistence-promoting mechanism. In this context, we investigated whether two congeneric plant species show synchronic age structures at varying spatial scales and whether they display demographic differences. METHODS: We sampled 600 Helianthemum squamatum and H. syriacum individuals at six localities in Spain. We used dendrochronological techniques to estimate age and radial growth. We compared the age structure among populations and between species and assessed the effect of site and species on different demographic parameters. Correlations between age structure and climatic factors were also determined. KEY RESULTS: We detected a very low intraspecific synchrony across sites but a high interspecific coupling in age structure within localities. Secondary growth, mean age, and flowering showed large intersite differences and small interspecific differences. Summer rainfall was a good predictor of age structure. CONCLUSIONS: Fine-scale climatic variability plays a major role in determining age structure of the studied species. Climatic variability is more relevant than interspecific differences. Our results illustrate the relevance of including spatial variation in summer rainfall patterns when modeling the impact of climate change on Mediterranean plant demography.

Mycorrhizal association between the desert truffle Terfezia boudieri and Helianthemum sessiliflorum alters plant physiology and fitness to arid conditions.

The host plant Helianthemum sessiliflorum was inoculated with the mycorrhizal desert truffle Terfezia boudieri Chatin, and the subsequent effects of the ectomycorrhizal relationship on host physiology were determined. Diurnal measurements revealed that mycorrhizal (M) plants had higher rates of photosynthesis (35%), transpiration (18%), and night respiration (49%) than non-mycorrhizal (NM) plants. Consequently, M plants exhibited higher biomass accumulation, higher shoot-to-root ratios, and improved water use efficiency compared to NM plants. Total chlorophyll content was higher in M plants, and the ratio between chlorophyll a to chlorophyll b was altered in M plants. The increase in chlorophyll b content was significantly higher than the increase in chlorophyll a content (2.58- and 1.52-fold, respectively) compared to control. Calculation of the photosynthetic activation energy indicated lower energy requirements for CO(2) assimilation in M plants than in NM plants (48.62 and 61.56 kJ mol(-1), respectively). Continuous measurements of CO(2) exchange and transpiration in M plants versus NM plants provided a complete picture of the daily physiological differences brought on by the ectomycorrhizal relationships. The enhanced competence of M plants to withstand the harsh environmental conditions of the desert is discussed in view of the mycorrhizal-derived alterations in host physiology.

The endophytic system of mediterranean Cytinus (cytinaceae) developing on five host Cistaceae species.

BACKGROUND AND AIMS: One of the most extreme manifestations of parasitism is found in the genus Cytinus, a holoparasite whose vegetative body is reduced to an endophytic system living within its host root. There are two species of Cytinus in the Mediterranean, C. hypocistis and C. ruber, which parasitize various genera of Cistaceae, one of the most characteristic families of the Mediterranean scrublands. The aim of this work is to describe the endophytic systems of C. hypocistis and C. ruber, and their tissue relationships with their host. METHODS: Roots from five different hosts infected with C. hypocistis and C. ruber were harvested, and examined by anatomical techniques under light microscopy to elucidate the characteristics of the endophytic system of Cytinus, and to determine if differences in endophytic systems occur between the two species and in response to different hosts. KEY RESULTS: The endophyte structure is similar in both Cytinus species irrespective of the host species. In the initial stages of the endophyte, rows of parenchymal cells spread through the host pericyclic derivatives and phloem, and begin to generate small nodules in the outermost region of the host xylem. Later the nodules anastomose, and bands of parasitic tissue are formed. The host cambium continues to develop xylem tissue, and consequently the endophyte becomes enclosed within the xylem. The bands of parasitic tissue fuse to form a continuous sheath. This mature endophyte has well-developed vascular system with xylem and phloem, and forms sinkers with transfer cells that grow through the host xylem. CONCLUSIONS: The endophytic system of Cytinus develops in all host root tissues and reaches its most mature stages in the host xylem. It is more complex than previously reported, showing parenchyma, xylem and phloem tissues. This is the first report of well-developed phloem in a holoparasitic endophytic species.

Environmental scales on the reproduction of a gypsophyte: a hierarchical approach.

BACKGROUND AND AIMS: Environmental variability at several scales can determine plant reproductive success. The main goal of this work was to model the reproductive flexibility of a semi-arid specialist considering different scales of environmental variability. METHODS: A 2-year field study was performed on the determinants of the female reproductive success of Helianthemum squamatum, an Iberian gypsophyte, considering two scales of environmental variability: differences between two contrasting slope aspects; and, on individual scale, the neighbouring microenvironment. Generalized linear mixed models were used to evaluate simultaneously the potential effects of environmental variability at both scales, together with flowering phenology and plant size on the reproductive output of H. squamatum. The following reproductive response variables were considered: number of flowers, fruit-set, number of viable and aborted seeds per fruit, and number of seeds per plant. KEY RESULTS: Contrary to expectations, environmental variability exerted a weak or even absent effect on the reproductive variables considered, while flowering phenology and plant size, which did not vary between slopes, played a major role. Surprisingly, the absolute reproductive variables were even higher in the extremely dry year of 2003, although only on the south-facing slope. The relatively milder conditions of the north-facing slope did not involve any advantage to this species in terms of reproductive output. CONCLUSIONS: The species seemed to be considerably well adapted to the environmental unpredictability characteristic of Mediterranean systems, considering its ability to maintain reproduction across contrasting environments and contrasting climatic conditions. These findings make us face the question of what must be considered stressful conditions in the case of a stress-tolerant specialist.

[Tissue culture and plant regeneration of Helianthemum songaricum Schrenk].

The cotyledonary segments of sterile seedlings of Helianthemum Songaricum Schrenk were cultured on different media containing different phytohormons. It was found that the calli could be induced efficiently on MS basal medium supplemented with 10.0 mg/L NAA and 0.2 mg/L 6-BA. When calli were transferred on MS medium with 2.0 mg/L 6-BA and 0.2 mg/L NAA, shoots were produced. The frequency of shoot differentiation reached about 85%. The regenerated shoots were rooted on 1/2MS medium added with 0.5 mg/L NAA. The rooting rate was about 76%. Regenerated plantlets were successfully transplanted in soil, with a success rate of 67%.

Effect of smoke, charred wood, and nitrogenous compounds on seed germination of ten species from woodland in central-western Spain.

The effect of smoke, charred wood, and nitrogenous compounds on germination was tested on 10 species of the Cistaceae, Poaceae, Fabaceae, and Asteraceae, from fire-prone, shrubby woodlands in central-western Spain. Dry seeds were exposed to smoke, by watering with distilled water-charred wood suspensions, or NaNO2, KNO3, NH4Cl, and NH4NO3. Smoke enhanced germination in 9 of 10 of the species. In species of Poaceae, germination was stimulated by 20 min of smoke exposure. In Asteraceae and Fabaceae species, 10 min of smoke exposure was the most effective treatment for enhancing germination. Three species--Cistus ladanifer, Cistus crispus, and Cistus monspeliensis--had a positive response to 20 min of smoke exposure; germination of Cistus salviifolius L. was also enhanced after 10 min. The effect of charred wood was variable, with no consistent germination pattern within the families. Trifolium angustifolium and Retama sphaerocarpa showed no stimulation of germination under most of the charred wood concentrations. Similarly, germination of Senecio jacobea under the charred wood treatment did not surpass that of the control. NaNO2 promoted seed germination in Dactylis glomerata (10 mM), Cistus ladanifer (1, 10, and 25 mM), and Cistus crispus (1 and 10 mM). KNO3 enhanced germination in Dactylis glomerata (1 and 25 mM), Dittrichia viscosa (10 and 25 mM), C. ladanifer (1, 10, and 25 mM), Cistus crispus (1 and 25 mM), and C. salviifolius aud C. monspeliensis (25 mM). NH4Cl induced germination of Dactylis glomerata and Dittrichia viscosa (1 mM), and Cistus species germinated best in 25 mM of this salt. NH4NO3 induced germination only in Cistus species. Holcus lanatus had the highest level of germination regardless of treatment.