The effects of nutrient pulsing on the threatened,floodplain species,Boltonia decurrens

Prior to anthropogenic disturbance, the floodplains of the Illinois River (USA) experienced moderate, cyclical flooding; a phenomenon termed a 'flood pulse'. Significant changes in the hydrology of the Illinois River caused by human alteration of the river channel resulted in a decline in populations of Boltonia decurrens, a native, herbaceous floodplain species of the Illinois River Valley and the demography of the species and underlying mechanisms causing the severe decline of B. decurrens are currently being studied. One component of the river flood pulse is the increased availability of nutrients in floodplain soils. We studied the effects of increased nutrient availability on growth and reproduction of B. decurrens and Aster pilosus, a species replacing B. decurrens in the floodplain. In this study, we examined the effects of a spring nutrient pulse and a late summer nutrient pulse, as compared to a control treatment, on rosettes of B. decurrens and A. pilosus grown in three soil types: sandy loam, silty loam and clay. Biomass, plant height, inflorescences per plant, seed production, and seed mass were measured and compared. Individuals of B. decurrens grown in sandy loam, the predominant soil type of the Illinois River Valley, had thegreatest increase in growth and fecundity in response to nutrient pulsing. Total biomass and inflorescences per plant were significantly greater than the control regardless of the timing of the nutrient pulse. Aster pilosus did not exhibit a clear pattern of increased growth and fecundity in a specific soil type or pulse treatment. The early pulse treatment appeared to have a greater effect across the range of soil types than the late pulse treatment; however, it did not seem to influence any particular variable(s) that we measured. A pulse of nutrients benefits B. decurrens more than A. pilosus, particularly in sandy loam. The increased vigor and fecundity of B. decurrens with a nutrient pulse could be a factor contributing to population survival during the period between disturbance events.     

Dynamics of patchy stomatal movements, and their contribution to steady-state and oscillating stomatal conductance calculated using gas-exchange techniques

Patchy stomatal movements were induced in leaves of Helianthus annuus L. and Xanthium strumarium L. by increasing Δw and decreasing light in a gas-exchange cuvette. The dynamics of the patchy movements were recorded and analysed using images of chlorophyll fluorescence, and the influence of heterogeneous stomatal activity on gas-exchange measurements of whole-leaf stomatal conductance was explored. Image series and gas-exchange measurements from two contrasting 100 min experiments are presented. One series of images, taken using Helianthus annuus, was characterized by strongly oscillating stomatal conductance induced by a decrease in light at high Δw. Fluorescence analysis revealed that individual patches of the leaf displayed a variety of behaviours (from static to strongly oscillating fluorescence), which, when averaged, matched the time dependence of the oscillating stomatal conductance measured by gas-exchange techniques. During the second series of images, taken using Xanthium strumarium, stomatal conductance (measured with gas exchange) declined slightly after an increase in Δw, and then maintained a steady state. Again, some patches in this leaf showed highly dynamic q_NP, although on the whole q_NP varied without any obvious pattern or frequency. When all patch activity in this series was averaged, it paralleled the steady whole-leaf stomatal conductance determined by gas-exchange measurements. It is clear from this work that coordinated patchy stomatal movements can contribute significantly to the dynamics of whole-leaf stomatal conductance, and, in contrast, that dynamic but uncoordinated patchy movements can average to produce a steady gas-exchange trace.     

Measurement of cystic fibrosis transmembrane conductance regulator activity using fluorescence spectrophotometry

Cystic fibrosis (CF) is a frequent autosomal recessive disease caused by mutations that impair the CF transmembrane conductance regulator (CFTR) protein function. CFTR is a chloride channel activated by cyclic AMP (cAMP) via protein kinase A (PKA) and ATP hydrolysis. We describe here a method to measure CFTR activity in a monolayer of cultured cells using a fluorescence spectrophotometer and the chloride-sensitive probe 6-methoxy-N-(3-sulfopropyl)quinolinium (SPQ). Modifying a slice holder, the spectrophotometer quartz cuvette was converted in a perfusion chamber, allowing measurement of CFTR activity in real time, in a monolayer of T84 colon carcinoma cells. The SPQ Stern–Volmer constant (K_Cl^-) for chloride in water solution was 115.0 ± 2.8 M⁻¹, whereas the intracellular K_Cl^- was 17.8 ± 0.8 M⁻¹, for T84 cells. A functional analysis was performed by measuring CFTR activity in T84 cells. The CFTR transport inhibitors CFTR(inh)-172 (5 μM) and glibenclamide (100 μM) showed a significant reduction (P < 0.05) in CFTR activity. This simple method allows measuring CFTR activity in a very simple, reproducible, and sensitive way.     

Diurnal patterns of chlorophyll a fluorescence and stomatal conductance in species of two types of coastal tree vegetation in southeastern Brazil

 Diurnal measurements of chlorophyll fluorescence parameters and stomatal conductance to water vapour were carried out in five woody species and a bromeliad from two coastal vegetation types in Brazil, the rock outcrop of Pao de Acúcar and the sand dunes of Macaé. The environmental conditions of both study sites are characterized by high temperatures and light levels. The studied species comprised C₃ plants with different degrees of stomatal closure during the day, overall daily stomatal conductance and a plant with a typical CAM pattern. Plants on Pao de Acúcar exhibited only a small decline in potential quantum yield throughout the day. The non-photochemical quenching and the approximate photosynthetic electron transport rates were maximal during the peak of irradiance. In Macaé, light response curves of fluorescence parameters in the CAM-tree Clusia hilariana showed a clear differentiation between phases III and IV of CAM. In phase III, decarboxylation of organic acids probably maintained high internal CO₂ levels and there was only a small decrease in photochemical quenching with saturating light levels. In phase IV, the depletion of the organic acid pool and low stomatal conductances resulted in much lower levels of effective quantum yield and a substantial increase in both 1-q_P and non-photochemical quenching with increasing light levels. This behaviour during phase IV in the CAM-tree C. hilariana was comparable to the performance of the C₃-Clusia C. aff parviflora at Pao de Acúcar. It is seen that both the C₃ species and the CAM tree occurring in the two coastal communities effectively adjust their photochemical efficiency to environmental conditions and especially to diurnal variations of stress. Received: 29 April 1996 / Accepted: 14 November 1996     

Evolutionary rescue and the coexistence of generalist and specialist competitors: an experimental test

Competition for resources is thought to play a critical role in both the origins and maintenance of biodiversity. Although numerous laboratory evolution experiments have confirmed that competition can be a key driver of adaptive diversification, few have demonstrated its role in the maintenance of the resulting diversity. We investigate the conditions that favour the origin and maintenance of alternative generalist and specialist resource-use phenotypes within the same population. Previously, we confirmed that competition for hosts among φ6 bacteriophage in a mixed novel (non-permissive) and ancestral (permissive) host microcosm triggered the evolution of a generalist phenotype capable of infecting both hosts. However, because the newly evolved generalists tended to competitively exclude the ancestral specialists, coexistence between the two phenotypes was rare. Here, we show that reducing the relative abundance of the novel host slowed the increase in frequency of the generalist phenotype, allowing sufficient time for the specialist to further adapt to the ancestral host. This adaptation resulted in ‘evolutionary rescue’ of the specialists, preventing their competitive exclusion by the generalists. Thus, our results suggest that competition promotes both the origin and maintenance of biodiversity when it is strong enough to favour a novel resource-use phenotype, but weak enough to allow adaptation of both the novel and ancestral phenotypes to their respective niches.     

Assessing photosynthetic efficiency in an experimental mangrove canopy using remote sensing and chlorophyll fluorescence

This study examined the ability of the photochemical reflectance index (PRI) to track changes in effective quantum yield (Δ F/F _m ′), non-photochemical quenching (NPQ), and the xanthophyll cycle de-epoxidation (DPS) in an experimental mangrove canopy. PRI was correlated with (Δ F/F _m ′) and NPQ over the 4-week measurement period and over the diurnal cycle. The normalised difference vegetation index (NDVI) was not correlated with any aspect of photochemical efficiency measured using chlorophyll fluorescence or xanthophyll pigments. This study demonstrated that photochemical adjustments were responsible for controlling the flow of energy through the photosynthetic apparatus in this mangrove forest canopy rather than canopy structural or chlorophyll adjustments.     

Epidermal conductance, stomatal density and stomatal size among genotypes of Sorghum bicolor (L.) Moench

Abstract. The ability of a plant to survive severe water deficits depends on its ability to restrict water loss through the leaf epidermis after stomata attain minimum aperture. At this stage, the rate of water loss is regulated by the epidermal conductance (g_c). Low g_c would be a useful selection criterion to identify genotypes with enhanced survival capability. Consequently, variation in g_c among Sorghum bicolor (L.) Moench genotypes was evaluated. Since there is little conclusive evidence linking g _c with leaf waxiness, alternative hypotheses relating g _c to stomatal trails were also examined. Epidermal conductance varied from 6.3 to 17.6mmol m⁻² s⁻¹ among sorghum genotypes. It was unrelated to stomatal pore length which varied with genotype and to pore depth which was similar for all genotypes measured. However, g _c, increased with increasing stomatal density. This indicates that stomatal density plays a direct role in water loss even at very low conductances. The association of low stomatal density with low g _c is consistent with the hypothesis that at the smallest stomata aperture, water loss from the epidermis above guard cell teichodes becomes a significant source of leaf water loss. Since low g _c is directly related to crop survival under severe water deficits, it is recommended that genotypes with low g _c. be selected using the selection criterion of stomatal density.     

Simultaneous measurement of stomatal conductance, non-photochemical quenching, and photochemical yield of photosystem II in intact leaves by thermal and chlorophyll fluorescence imaging

A new imaging system capable of simultaneously measuring stomatal conductance and fluorescence parameters, non-photochemical quenching (NPQ) and photochemical yield of photosystem II (Phi(PSII)), in intact leaves under aerobic conditions by both thermal imaging and chlorophyll fluorescence imaging was developed. Changes in distributions of stomatal conductance and fluorescence parameters across Phaseolus vulgaris L. leaves induced by abscisic acid treatment were analyzed. A decrease in stomatal conductance expanded in all directions from the treatment site, then mainly spread along the lateral vein toward the leaf edge, depending on the ABA concentration gradient and the transpiration stream. The relationships between stomatal conductance and fluorescence parameters depended on the actinic light intensity, i.e. NPQ was greater and Phi(PSII) was lower at high light intensity. The fluorescence parameters did not change, regardless of stomatal closure levels at a photosynthetically active photon flux (PPF) of 270 micro mol m(-2) s(-1); however, they drastically changed at PPF values of 350 and 700 micro mol m(-2) s(-1), when the total stomatal conductance decreased to less than 80 and 200 mmol m(-2) s(-1), respectively. This study has, for the first time, quantitatively analyzed relationships between spatiotemporal variations in stomatal conductance and fluorescence parameters in intact leaves under aerobic conditions.     

Development of tetranucleotide microsatellite markers for the cushion star, Asterina gibbosa, and cross-species amplification

We describe nine polymorphic tetranucleotide microsatellite loci from the starfish, Asterina gibbosa. Loci were isolated from a partial genomic library that had been enriched for AAAC repeat sequences. Number of alleles per locus ranged from two to 14 in a sample of 85 individuals from three populations (two from Spain and one from the UK). Observed and expected heterozygosities per population ranged from 0.000 to 0.400 and from 0.040 to 0.784, respectively. All loci presented significant heterozygote deficits in one or more populations. Eight of these loci were amplified and variable in A. pancerii and A. phylactica. These loci will be used to study population structure in A. gibbosa.     

Habitat distribution modelling,under the present climatic scenario,of the threatened endemic Iberian Delphinium fissum subsp. sordidum (Ranunculaceae) and implications for its conservation

We modelled the potential habitat of a threatened species D. fissum subsp. sordidum, an endemic hemicryptophyte with a disjunct distribution in the Iberian Peninsula. Maxent was used to predict the subspecies habitat suitability by relating field sample-based distributional information with environmental and topographic variables. Our results suggest that the model performed well, predicting with high accuracy the current distribution of the species. The variables that most contributed to the model were Mean Temperature of Wettest Quarter (MTWtQ), Precipitation of Warmest Quarter (PWmQ), Temperature Annual Range (TAR) and Slope (Slo). These variables are biological significant for the taxon, as they have decisive influence in the critical stages of germination and fruiting. The current and potential distributional areas identified by the model fall mainly in regions with some degree of environmental protection, with some exceptions. A recovery plan for the species should be considered. Species Distribution Modelling cannot substitute long-term monitoring programmes, yet it is a useful tool for identifying appropriate areas of taxon occurrence, and thus allow for efficient use of the economic and human resources.     

Evaluation of the potential to measure photosynthetic rates in C3 plants (Flaveria pringlei and Oryza sativa) by combining chlorophyll fluorescence analysis and a stomatal conductance model

The response curves of leaf photosynthesis to varying light, temperature and leaf-to-air vapour pressure deficit were measured in the C₃ plants Flaveria pringlei and Oryza sativa in normal air with a computerized open infrared gas analysis (IRGA) system, and the photochemical efficiency of photosystem II, described as (1–F,/F′_m) after Genty. Briantais & Baker (1989, Biochimica et Biophysica Acta 990, 87–92), was simultaneously measured with a modulated fluorometer. A model was written for rates of CO₂ fixation as a function of the true rate of O₂ evolution measured by fluorescene analysis (Jo₂), mesophyll conductance and intercellular CO₂ partial pressure. A second model was developed for rates of CO₂ fixation as a function of Jo₂, mesophyll conductance and stomatal conductance. In the latter case, leaf stomatal conductance was simulated using the stomatal model proposed by Leuning (1995, Plant, Cell and Environment 18 , 339–355). The rates of CO₂ fixation predicted from the models were similar to rates measured by IRGA. The results indicate that there is potential to measure CO₂ fixation in C₃ plants by combining the non-invasive measurement of Jo₂ by chlorophyll fluorescence analysis with the stomatal conductance model.     

An experimental removal of a territorial pomacentrid: effects on the occurrence and behavior of competitors

Synopsis Stegastes fasciolatus is the most common territorial damselfish in the shallow waters of Hawaii. Territorial defense was observed against other herbivorous fishes, especially acanthorids, scarids and one omnivorous chaetodontid. One acanthurid,Acanthurus nigrofuscus was found to differ in abundance and social behavior in areas whereS. fasciolatus was present, compared to areas where it was absent. The chaetodontid,Chaetodon quadrimaculatus was sheltered during the day in areas where the pomacentrid was abundant, apparently feeding at night. In other areas it fed during the day and at night, depending on the phase of the moon.S. fasciolatus were then experimentally removed from one study site, to test whether the differences in abundance and behavior of the other species were due to the presence of the damselfish. There was a significant increase in numbers of the surgeonfishAcanthurus nigrofuscus in the removal area, as well as changes in social behavior from schooling to defense of small territories. The butterflyfish,C. quadrimaculatus, was observed to forage during the day in the removal area. There were no significant changes in the control sites. The presence of the interspecifically territorial damselfish,S. fasciolatus, thus appears to be an important determinant of the behavior of these potential food competitors.     

Lack of genetic variability in the rare and endangered Limonium cavanillesii (Plumbaginaceae) using RAPD markers

Limonium cavanillesii is an extremely endangered plant species endemic to the east Mediterranean region of Spain. Regarded as extinct for several years, the recent discovery of a small population (only 29 individuals) has prompted the adoption of measures for its conservation by official agencies. As part of this effort, we have analysed genetic variation in this population by means of random amplified polymorphic DNA (RAPDs). The analysis of 29 individuals with 11 different primers produced 131 monomorphic bands. To our knowledge, this is the lowest level of genetic variation detected in plants using RAPD markers. This result could be explained both by the apomictic reproductive system of this species and by the passage through a severe bottleneck in recent times, after which there has been no chance for mutation to restore detectable genetic variation.     

Floristic diversity and the richness of locally endangered plant species of semi-natural grasslands under different management practices,southern Kyushu,Japan

Background: The rapid decline of semi-natural grasslands in Japan threatens many relic and endemic plant species. There is insufficient knowledge on how the impacts of land-use changes and management of grasslands have been affecting grassland ecosystems and what conservation measures may be taken to conserve as much of the existing plant diversity as possible.

Aim: We assessed the existing management regimes for their suitability for conserving Red Data Book (RDB) species.

Methods: We conducted our study in four districts of Kushima, Kyushu, south-west Japan, with different land-use histories. We compared species richness, plant density and abundance in six grassland types: regularly burnt, regularly mown, paddy levee, roadside, landslip and wetland communities in a total of 289 1 m x 1 m quadrats, recorded in172 grassland patches. Species richness plant density and abundance were analysed with special reference to RDB species under different land use history.

Results: Species richness of grasslands did not differ across different land use histories, yet our analysis showed that the reduced area of grasslands markedly affected the density of RDB species. Grassland types differed in their ability to support RDB species: regularly burnt grasslands were the richest in RDB species and poorest in alien species, followed by regularly mown grasslands, paddy levees, landslip, wetland and roadside communities.

Conclusions: Traditional management regimes, such as regular burning or mowing of grasslands have the best potential for conserving RDB species, and thus should be part of conservation management practices of semi-natural grasslands.     

Desert dogma revisited: coupling of stomatal conductance and photosynthesis in the desert shrub, Larrea tridentata

The success of the desert shrub Larrea tridentata (creosotebush) has been largely attributed to temperature acclimation and stomatal control of photosynthesis (A) under drought stress. However, there is a paucity of field data on these relationships. To address this void, we conducted a joint field and modelling study that encompassed a diverse set of environmental conditions. At a Larrea‐dominated site in southern New Mexico we manipulated soil moisture during the growing season over a 2‐year period and measured plant pre‐dawn water potential (Ψ_pd), stomatal conductance (g) and A of individual shrubs. We used these data to develop a semi‐mechanistic photosynthesis model (A–Season) that explicitly couples internal CO₂ (C_i) and g. Vapour pressure deficit (VPD) and Ψ_pd affect instantaneous g in a manner that is consistent with a biophysical model of stomatal regulation of leaf water potential. C_i is modelled as a function of g, derived from a simplification of a typical A–C_i curve. After incorporating the effects of growing temperature on stomatal behaviour, the model was able to capture the large diurnal fluctuations in A, g and C_i and the observed hysteresis in g versus C_i dynamics. Our field data and application of the A–Season model suggest that dogma attributed to Larrea's success is supported with regard to stomatal responses to VPD and Ψ_pd, but not for mechanisms of temperature acclimation and CO₂ demand.     

Time resolved fluorescence of bacteriophage Pfl DNA binding protein and its complex with DNA

The DNA binding protein of the filamentous bacteriophage Pfl exhibits fluorescence from a single tryptophan residue. The location of the emission maximum at 340 nm ist quite common for proteins, but the single lifetime of 7.8 ns is one of the longest yet reported. Protein fluorescence is quenched more efficiently by Cs⁺ than by I^-; the Trp is located in a partially exposed pocket, in the vicinity of a negative charge.In the native complex of the binding protein with Pfl DNA the fluorescence emission maximum is at 330 nm, indicating a more apolar environment for Trp 14. The native nucleoprotein complex exhibits a similar fluorescence lifetime (6.5 ns) and an approximately equal fluorescence yield, indicating the absence of Trp-DNA stacking. The tryptophan in the complex is virtually inaccessible to ionic quenchers, and thus appears to be buried.Fluorescence depolarisation measurements have been used to examine the rotational mobility of the tryptophan in the protein and in the nucleoprotein complex. In the protein alone a single rotational correlation time () of 19 ns is observed, corresponding to rotation of the entire dimeric molecule; in the native nucleoprotein complex with Pfl DNA, a of 500 ns is observed, corresponding to a rigid unit of at least 50 subunits. In neither case does the tryptophan exhibit any detectable flexibility on the subnanosecond time scale.