Sun-induced fluorescence and gross primary productivity during a heat wave.

Remote sensing of sun-induced chlorophyll fluorescence (SIF) has been suggested as a promising approach for probing changes in global terrestrial gross primary productivity (GPP). To date, however, most studies were conducted in situations when/where changes in both SIF and GPP were driven by large changes in the absorbed photosynthetically active radiation (APAR) and phenology. Here we quantified SIF and GPP during a short-term intense heat wave at a Mediterranean pine forest, during which changes in APAR were negligible. GPP decreased linearly during the course of the heat wave, while SIF declined slightly initially and then dropped dramatically during the peak of the heat wave, temporally coinciding with a biochemical impairment of photosynthesis inferred from the increase in the uptake ratio of carbonyl sulfide to carbon dioxide. SIF thus accounted for less than 35% of the variability in GPP and, even though it responded to the impairment of photosynthesis, appears to offer limited potential for quantitatively monitoring GPP during heat waves in the absence of large changes in APAR.

Slowing of nitrogen cycling and increasing nitrogen use efficiency following afforestation of semi-arid shrubland.

Nitrogen (N) and water availability are important factors affecting ecosystem productivity that can be influenced by land-use change. We hypothesized that the observed increase in carbon (C) sequestration associated with afforestation of semi-arid sparse shrubland must also be associated with an increase in N input. We tested this hypothesis by reconstructing the ecosystem N budget of two ecosystems, a semi-arid shrubland and a nearby planted pine forest, using measurements augmented with literature-based estimates. Our findings demonstrate that, contrary to our hypothesis, massive C sequestration by the pine forest could be accounted for without a change in the net N budget (i.e., neither elevated N inputs nor reduced N losses). However, in comparison to the shrubland, the forest showed an almost tripling in aboveground N use efficiency (NUE; 235 vs. 83 kg dry mass kg(-1) N) and a doubling in ecosystem level C/N ratio (16 vs. 8, for the forest and shrubland, respectively). Nitrogen cycling slowed in the forest compared to the shrubland: net N mineralization rates in soils decreased by approximately 50%, decomposition rates decreased by approximately 20%, and NO(x) loss decreased by approximately 64%. These adjustments in N cycling provide a possible basis for increased NUE and subsequent C sequestration without net change in the overall N budget, which should be addressed in future investigations.

A direct and rapid leaf water extraction method for isotopic analysis.

Isotopic measurements of leaf water have provided insights into a range of ecophysiological and biogeochemical processes, but require an extraction step which often constitutes the major analytical bottleneck in large-scale studies. Current standard procedures for leaf water analysis are based on cryogenic vacuum or azeotrophic distillation, and are laborious, require sophisticated distillation lines and the use of toxic materials. We report a rapid technique based on centrifugation/filtration of leaf samples pulverised in their original sampling tubes, using a specifically adapted, simple apparatus. The leaf water extracts produced are suitable for isotopic analysis via pyrolysis gas chromatography isotope ratio mass spectrometry (PYR/GC/IRMS). The new method was validated against cryogenic vacuum distillation and showed an overall accuracy of +/-0.5 per thousand (nine grouped comparisons, n = 110) over a range of 21 per thousand. Effects due to the presence of soluble carbohydrates were near the detection limits for most samples analysed, and these effects could be corrected for (the extracted soluble organics could also be used for isotopic analysis). The extraction time for a routine eight-sample subset was reduced from 4 h (cryogenic distillation) to 45 min, limited only by the size of the centrifuge(s) used. This method provides a rapid, low-cost and reliable alternative to conventional vacuum and other distillation methods that can alleviate current restrictions on ecosystem- and global-scale studies that require high-throughput leaf water isotopic analysis.

Increased growth of young citrus trees under reduced radiation load in a semi-arid climate.

This study investigated the effects of radiation heat-load reduction by shading on the growth and development of citrus trees in a warm subtropical region. The experiment was conducted from mid-June until late October when daily maximal air temperature averaged 29.3 degrees C. Two-year-old de-fruited Murcott tangor (Citrus reticulata BlancoxCitrus sinensis (L.) Osb.) trees were grown under 30% or 60% shade tunnels, or 60% flat shade (providing midday shade only), using highly reflective aluminized nets. Non-shaded trees were used as the control. Shading reduced direct more than diffuse radiation. Daily radiation was reduced by 35% for the 30% Tunnel and 60% Flat treatments, and by 55% for the 60% Tunnel. Two days of intensive measurement showed that shading increased average sunlit leaf conductance by 44% and photosynthesis by 29%. Shading did not significantly influence root and stem dry weight growth, but it increased the increment in leaf dry weight during the three month period by an average of 28% relative to the control, while final tree height in the 30% Tunnel treatment exceeded the control by 35%. Shoot to root and shoot mass ratios increased and root mass ratio decreased due to shading because of the increase in leaf dry weight. Shading increased starch concentration in leaves while the shadiest treatment, 60% Tunnel, decreased starch concentration in the roots. Carbon isotope ratio (delta(13)C) of exposed leaves that developed under shading was significantly reduced by 1.9 per thousand in the 60% Tunnel, indicating that shading increased CO(2) concentrations at the chloroplasts (C(c)), as would be expected from increased conductance. Substomatal CO(2) concentrations, C(i), computed from leaf net CO(2) assimilation rate and conductance values, also indicate that shading increases internal CO(2) concentrations. Based on tree dry mass, tree height, and total carbohydrates fractions, the 30% Tunnel and the 60% Flat were the optimal shade treatments.

Genomic dissection of genotype x environment interactions conferring adaptation of cotton to arid conditions.

The interaction of genotype with environment is of primary importance in many aspects of genomic research and is a special priority in the study of major crops grown in a wide range of environments. Water deficit, the major factor limiting plant growth and crop productivity worldwide, is expected to increase with the spread of arid lands. In genetically equivalent cotton populations grown under well-watered and water-limited conditions (the latter is responsible for yield reduction of approximately 50% relative to well-watered conditions), productivity and quality were shown to be partly accounted for by different quantitative trait loci (QTLs), indicating that adaptation to both arid and favorable conditions can be combined in the same genotype. QTL mapping was also used to test the association between productivity and quality under water deficit with a suite of traits often found to differ between genotypes adapted to arid versus well-watered conditions. In this study, only reduced plant osmotic potential was clearly implicated in improved cotton productivity under arid conditions. Genomic tools and approaches may expedite breeding of genotypes that respond favorably to specific environments, help test roles of additional physiological factors, and guide the isolation of genes that protect crop performance under arid conditions toward improved adaptation of crops to arid cultivation.

Influence of carbonic anhydrase activity in terrestrial vegetation on the 18O content of atmospheric CO2.

The oxygen-18 (18O) content of atmospheric carbon dioxide (CO2) is an important indicator of CO2 uptake on land. It has generally been assumed that during photosynthesis, oxygen in CO2 reaches isotopic equilibrium with oxygen in 18O-enriched water in leaves. We show, however, large differences in the activity of carbonic anhydrase (which catalyzes CO2 hydration and 18O exchange in leaves) among major plant groups that cause variations in the extent of 18O equilibrium (theta(eq)). A clear distinction in theta(eq) between C3 trees and shrubs, and C4 grasses makes atmospheric C18OO a potentially sensitive indicator to changes in C3 and C4 productivity. We estimate a global mean theta(eq) value of approximately 0.8, which reasonably reconciles inconsistencies between 18O budgets of atmospheric O2 (Dole effect) and CO2.

Internal conductance to CO(2) diffusion and C(18)OO discrimination in C(3) leaves.

(18)O discrimination in CO(2) stems from the oxygen exchange between (18)O-enriched water and CO(2) in the chloroplast, a process catalyzed by carbonic anhydrase (CA). A proportion of this (18)O-labeled CO(2) escapes back to the atmosphere, resulting in an effective discrimination against C(18)OO during photosynthesis (Delta(18)O). By constraining the delta(18)O of chloroplast water (delta(e)) by analysis of transpired water and the extent of CO(2)-H(2)O isotopic equilibrium (theta(eq)) by measurements of CA activity (theta(eq) = 0.75-1.0 for tobacco, soybean, and oak), we could apply measured Delta(18)O in a leaf cuvette attached to a mass spectrometer to derive the CO(2) concentration at the physical limit of CA activity, i.e. the chloroplast surface (c(cs)). From the CO(2) drawdown sequence between stomatal cavities from gas exchange (c(i)), from Delta(18)O (c(cs)), and at Rubisco sites from Delta(13)C (c(c)), the internal CO(2) conductance (g(i)) was partitioned into cell wall (g(w)) and chloroplast (g(ch)) components. The results indicated that g(ch) is variable (0.42-1.13 mol m(-2) s(-1)) and proportional to CA activity. We suggest that the influence of CA activity on the CO(2) assimilation rate should be important mainly in plants with low internal conductances.

The use of stable isotopes to study ecosystem gas exchange.

Stable isotopes are a powerful research tool in environmental sciences and their use in ecosystem research is increasing. In this review we introduce and discuss the relevant details underlying the use of carbon and oxygen isotopic compositions in ecosystem gas exchange research. The current use and potential developments of stable isotope measurements together with concentration and flux measurements of CO2 and water vapor are emphasized. For these applications it is critical to know the isotopic identity of specific ecosystem components such as the isotopic composition of CO2, organic matter, liquid water, and water vapor, as well as the associated isotopic fractionations, in the soil-plant- atmosphere system. Combining stable isotopes and concentration measurements is very effective through the use of "Keeling plots." This approach allows the identification of the isotopic composition and the contribution of ecosystem, or ecosystem components, to the exchange fluxes with the atmosphere. It also allows the estimation of net ecosystem discrimination and soil disequilibrium effects. Recent modifications of the Keeling plot approach permit examination of CO2 recycling in ecosystems. Combining stable isotopes with dynamic flux measurements requires precision in isotopic sampling and analysis, which is currently at the limit of detection. Combined with the micrometeorological gradient approach (applicable mostly in grasslands and crop fields), stable isotope measurements allow separation of net CO2 exchange into photosynthetic and soil respiration components, and the evapotranspiration flux into soil evaporation and leaf transpiration. Similar applications in conjunction with eddy correlation techniques (applicable to forests, in addition to grasslands and crop fields) are more demanding, but can potentially be applied in combination with the Keeling plot relationship. The advance and potential in using stable isotope measurements should make their use a standard component in the limited arsenal of ecosystem-scale research tools.

Immunization of rabbits with faecal extract of Pediculus humanus, the human body louse: effects on louse development and reproduction.

Immunization of rabbits with a faecal extract of the human body louse (Pediculus humanus) induced a high titre of specific IgG. The mean weight of blood taken by females fed on the immunized rabbits was significantly lower (29%) than taken by females fed on the control rabbits. The mean number of eggs per female fed on the immunized rabbits was significantly lower than for females fed on the control rabbits. The hatchability of the eggs laid by lice fed on immunized rabbits (91%) was significantly lower than of those fed on control rabbits (94%). The rate of development of nymphs fed on control rabbits was significantly higher than those fed on the immunized rabbits. There was no difference in survival rates of lice fed on immunized and control rabbits.

Characterization of body louse midgut proteins recognized by resistant hosts.

The human body louse, Pediculus humanus, showed eighteen midgut proteins ranging between 12 and 117 kDa, when analysed by SDS-PAGE electrophoresis. Seven of them (12 kDa, 17 kDa, 29 kDa, 35 kDa, 40 kDa, 55 kDa and 97 kDa) were major bands based on their intensity of staining. The immunization of rabbits with a midgut extract elicited the production of protective polyclonal antibodies. These antibodies reacted strongly with all major midgut proteins as well as with 63 kDa and 117 kDa proteins when tested by the Western blot technique. The analysis of the proteins revealed that the 12 kDa, 25 kDa, 29 kDa, 35 kDa, 45 kDa, 87 kDa and 97 kDa proteins are glycosylated and none of them contained a lipid moiety. By electroelution, the proteins of 35 kDa and 63 kDa were purified. On trypsinization, the proteins of 35 kDa and 63 kDa produced four major fragments (F1, F2, F3, and F4) when resolved on a 18% SDS-PAGE. The F1 fragment of the 35 kDa protein reacted with the polyclonal antibodies by the immunoblot technique.

Localization of immunogenic antigens on midgut of the human body louse Pediculus humanus humanus (Anoplura: Pediculidae).

Immunogenic midgut antigens of the human body louse, Pediculus humanus humanus L., were localized using rabbit antisera against a louse-midgut extract followed by a 2nd antibody conjugated to either fluorescein or colloidal gold. Strong fluorescence was observed on the outer membrane of the epithelial cell of the midgut. The immunogold technique revealed that most of the antigens were localized on the microvilli of the midgut cells. Small numbers of gold particles were also seen in the gut lumen and within the cell cytoplasm. Only a few gold particles were seen in the lumen of the gut sections incubated with control sera.

Electron Partitioning between the Cytochrome and Alternative Pathways in Plant Mitochondria.

The contribution of the cyanide-resistant, alternative pathway to plant mitochondrial electron transport has been studied using a modified aqueous phase on-line mass spectrometry-gas chromatography system. This technique permits direct measurement of the partitioning of electrons between the cytochrome and alternative pathways in the absence of added inhibitors. We demonstrate that in mitochondria isolated from soybean (Glycine max L. cv Ransom) cotyledons, the alternative pathway contributes significantly to oxygen uptake under state 4 conditions, when succinate is used as a substrate. However, when NADH is the substrate, addition of pyruvate, an allosteric activator of the alternative pathway, is required to achieve the same level of alternative pathway activity. Under state 3 conditions, when the reduction state of the ubiquinone pool is low, the addition of pyruvate allows the alternative pathway to compete with the cytochrome pathway for electrons from the ubiquinone pool when the cytochrome pathway is not saturated. These results provide direct experimental verification of the kinetics consequences of pyruvate addition on the partitioning of electron flow between the two respiratory pathways. This distribution of electrons between the two unsaturated pathways could not be measured using conventional oxygen electrode methods and illustrates a clear advantage of the mass spectrometry technique. These results have significant ramifications for studies of plant respiration using the oxygen electrode, particularly those studies involving intact tissues.

Immunization of rabbits with a midgut extract of the human body louse Pediculus humanus humanus: the effect of induced resistance on the louse population.

Resistance to human body lice, Pediculus humanus humanus L, induced by feeding on rabbits immunized with an extract of louse gut was studied. The mortality of lice fed on immunized rabbits was 73%, significantly higher than that of lice fed on control rabbits (52%) (P < 0.01). The proportion of dead nymphs and female lice with ruptured guts was significantly higher in lice fed on immunized rabbits (P < 0.01). The size of the bloodmeal was 35% greater in female lice fed on control rabbits than on immunized rabbits. Lice fed on immunized rabbits laid 40% less eggs than those fed on the controls, they also demonstrated a significant decrease in the number of eggs per female over time (P < 0.01). 86% of the eggs laid by lice fed on immunized animals hatched, compared with 92% hatching of eggs laid by the lice fed on control animals (P < 0.01). With the exception of the first bloodmeal the percentage of hatched eggs which were laid between any two bloodmeals was significantly smaller (P < 0.01) in the lice fed on immunized rabbits than in the control group. The first nymphal stage of lice fed on immunized rabbits took an average of 5.2 days to moult to the second stage, compared with 4 days for those fed on control rabbits.

Mechanism of aggregation behavior inMaladera matrida Argaman (Coleoptera: Scarabaeidae).

AdultMaladera matrida Argaman (Coleoptera: Scarabaeidae: Melolonthinae) males emerge from soil for an active period at dusk, a few minutes before the females. Adults are found during most of the active hours on the foliage in aggregations composed of an equal sex ratio. The mechanism of aggregation behavior ofM. matrida beetles was studied in a Y-shaped olfactometer. No evidence was found for the existence of an aggregation pheromone released either by males or by females, but behavior tests indicate that adultM. matrida beetles, males as well as females, are attracted to volatiles of an injured host plant. The following scenario is suggested: Males emerge daily from soil at dusk, a few minutes before the females, and immediately start feeding. Additional males are attracted to the injured host's volatiles and form aggregations. When females emerge from soil, the attractant volatiles are concentrated in spots, and the females join the aggregations, forming an equal sex ratio.

Measurements of the Engagement of Cyanide-Resistant Respiration in the Crassulacean Acid Metabolism Plant Kalanchoë daigremontiana with the Use of On-Line Oxygen Isotope Discrimination.

Discrimination against (18)O during dark respiration in tissues of Kalanchoë daigremontiana, Medicago sativa, and Glycine max was measured using an on-line system that enabled direct measurements of the oxygen fractionation of samples in a gas-phase leaf disk electrode unit. Discrimination factors for cytochrome pathway respiration were 18.6 to 19.8%(o) for all tissues. However, discrimination in cyanide-resistant respiration was significantly higher in green tissues (30.4-31.2%(o)) compared with nongreen tissues (25.3-25.9%(o)). Using these discrimination factors, the partitioning of electron transport to these pathways was calculated from measurements of discrimination in the absence of inhibitors. Changes in flux through the alternative pathway were measured during the light and dark phases of Crassulacean acid metabolism in leaf disks of K. daigremontiana. The flux of electrons through the alternative pathway was higher during deacidification than during the other phases of Crassulacean acid metabolism. The increase in alternative pathway electron flux accounted for all of the increased respiration in the light phase. Despite this increase, simultaneous measurements of malate concentration and respiratory flux confirm that only a small proportion of the total malate decarboxylation occurs in the mitochondria.

Autotrophy in maize husk leaves : evaluation using natural abundance of stable isotopes.

The natural abundance of carbon and hydrogen isotopic composition, expressed as a delta(13)C value of plant dry matter and cellulose in the hypsophylls (husk leaves) of maize (Zea mays L.) was measured and compared with that of leaves and cobs. The delta(13)C values of outer hypsophylls were usually 2 to 3% per thousand more negative than leaves or other tissues, and became more negative with increasing chlorophyll content, indicating significant local C(3) pathway fixation of CO(2) in the outer hypsophylls. The deltaD values indicated a significant part of hypsophyll cellulose was derived from heterotrophic sources (sucrose from C(4) photosynthesis in other tissues). Isotopic mass balance calculations allowed quantitative estimation of these carbon sources and, in the samples examined, about 16% of hypsophyll cellulose was derived from local C(3) photosynthesis, about 62% from local C(4) photosynthesis, and about 22% from sucrose imported from other leaves.

Oxygen and Hydrogen Isotope Fractionation during Cellulose Metabolism in Lemna gibba L.

Lemna gibba L. B3 was grown under heterotrophic, photoheterotrophic, and autotrophic conditions in water having a variety of hydrogen and oxygen isotopic compositions. The slopes of the linear regression lines between the isotopic composition of water and leaf cellulose indicated that under the three growth conditions about 40, 70, and 100% of oxygens and carbon-bound hydrogens of cellulose exchanged with those of water prior to cellulose formation. Using the equations of the linear relationships, we estimated the overall fractionation factors between water and the exchanged oxygen and carbon bound-hydrogen of cellulose. At least two very different isotope effects must determine the hydrogen isotopic composition of Lemna cellulose. One reflects the photosynthetic reduction of NADP, while the second reflects exchange reactions that occur subsequent to NADP reduction. Oxygen isotopic composition of cellulose apparently is determined by a single type of exchange reaction with water. Under different growth conditions, variations in metabolic fluxes affect the hydrogen isotopic composition of cellulose by influencing the extent to which the two isotope effects mentioned above are recorded. The oxygen isotopic composition of cellulose is not affected by such changes in growth conditions.

Movement of newly imported light-harvesting chlorophyll-binding protein from unstacked to stacked thylakoid membranes is not affected by light treatment or absence of amino-terminal threonines.

In higher plants and algae, the transduction of captured light energy is highly regulated as excess excitation of photosystem II (PSII) reaction centers can be redirected to photosystem I (PSI) reaction centers. Models that attempt to explain this phenomenon involve light-harvesting chlorophyll-protein complexes (LHCII) that capture light energy and migrate between PSII and PSI. This report shows that in pea chloroplasts, the major protein component of LHCII, light-harvesting chlorophyll-binding protein (LHCP), can indeed migrate within the thylakoid membrane. We show, however, that although newly imported LHCP inserts into both stacked and unstacked thylakoid membranes, it then moves only from the unstacked, PSI-rich membranes to the stacked, PSII-rich membranes. The observed migration is not affected by light treatment that induces a redistribution of captured light energy (state I-state II transition) that previously was thought to induce LHCP to migrate in the opposite direction, from stacked to unstacked membranes. A mutation that removes the site of LHCP phosphorylation, the proposed trigger of state transitions, also has no effect on the integration and movement of LHCP, but does render LHCP more susceptible to proteolytic degradation. These results are not consistent with current models that deal with the short-term change in the distribution of light energy.

Quantitative studies of host immunoglobulin G in the hemolymph of ticks (Acari).

A radioimmunoassay was used to measure the concentration of host immunoglobulin G (IgG) in the hemolymph of female hard and soft ticks. Hyalomma excavatum Koch, Rhipicephalus sanguineus Latreille, Ornithodoros tholozani (Laboulbene and Megnin), and O. moubata (Murry) were fed on rabbits immunized with ovalbumin; Argas persicus (Oken) was fed on chickens immunized with cytochrome 'C.' At 24 h after feeding, the concentration of antiovalbumin IgG in the hemolymph was 7 micrograms/ml for H. excavatum, 5 micrograms/ml for R. sanguineus, and 0.15 micrograms/ml for O. moubata; the percentage of intact IgG molecules was 30, 44, and 100%, respectively. Host IgG was not detected in the hemolymph of O. tholozani and A. persicus. There was no increase in the concentrations of host IgG in the hemolymph of the soft ticks during the first week following the blood meal. The potential contribution to the resistance of hosts against ticks by host antibodies that cross into the tick hemocoel is discussed.