Metabolism of Abscisic Acid in Guard Cells of Vicia faba L. and Commelina communis L

Metabolism of abscisic acid (ABA) was investigated in isolated guard cells and in mesophyll tissue of Vicia faba L. and Commelina communis L. After incubation in buffer containing [G-³H]±ABA, the tissue was extracted by grinding and the metabolites separated by thin layer chromatography. Guard cells of Commelina metabolized ABA to phaseic acid (PA), dihydrophaseic acid (DPA), and alkali labile conjugates. Guard cells of Vicia formed only the conjugates. Mesophyll cells of Commelina accumulated DPA while mesophyll cells of Vicia accumulated PA. Controls showed that the observed metabolism was not due to extracellular enzyme contaminants nor to bacterial action.

Metabolism of ABA in guard cells suggests a mechanism for removal of ABA, which causes stomatal closure of both species, from the stomatal complex. Conversion to metabolites which are inactive in stomatal regulation, within the cells controlling stomatal opening, might precede detectable changes in levels of ABA in bulk leaf tissue. The differences observed between Commelina and Vicia in metabolism of ABA in guard cells, and in the accumulation product in the mesophyll, may be related to differences in stomatal sensitivity to PA which have been reported for these species.

     

Genotypic variability in vulnerability of leaf xylem to cavitation in water-stressed and well-irrigated sugarcane

Genotypic variability in vulnerability of leaf xylem to water-stress-induced cavitation was determined in four sugarcane (Saccharum sp.) clones using detached leaf segments in a hydraulic conductivity apparatus. Vulnerability curves were constructed by plotting the percentage of maximum conductivity versus leaf water potential (ψ_I) and fitting curves using a Weibull function. The ψ_I at which each clone lost 10, 50, and 80% of maximum conductivity was determined. Maximum conductivity per unit of leaf width was positively associated with metaxylem vessel diameter. The commercial clone H65-7052 exhibited the highest and the nondomesticated S. spontaneum exhibited the lowest conductivity. All four clones lost substantial conductivity at values of ψ_I less negative than −1.4 MPa, but H65-7052 was able to maintain 50% conductivity to lower ψ_I than the other clones. S. spontaneum sustained the most negative ψ_I (−1.99 MPa) before reaching the 80% conductivity loss point. Clone H69-8235 was consistently the most vulnerable to initial loss of conductivity. These vulnerability functions were used in conjunction with field measurements of ψ_I to estimate diurnal losses in leaf hydraulic conductivity under irrigated and droughted conditions. H69-8235 lost up to 50% of its conductivity during the day, even when well irrigated, and more than 80% when subjected to drought. The other clones exhibited lower conductivity losses. These losses are apparently reversed overnight by root pressure. Despite their close genetic relationships, these clones exhibited large differences in conductivity, in the vulnerability of their xylem to cavitation, and in gas exchange behavior. The potential for altering water relations by selecting for particular hydraulic characteristics is discussed.     

Offsetting high water demands with high productivity: Sorghum as a biofuel crop in a high irradiance arid ecosystem

High irradiance arid environments are promising, yet understudied, areas for biofuel production. We investigated the productivity and environmental trade‐offs of growing sorghum (Sorghum bicolor) as a biofuel feedstock in the low deserts of California (CA). Using a 5.3 ha experimental field in the Imperial Valley, CA, we measured aboveground biomass production and net ecosystem exchange of CO₂ and H₂O via eddy covariance over three growing periods between February and November 2012. Environmental conditions were extreme, with high irradiance, vapor pressure deficit (VPD), and air temperature throughout the growing season. Air temperature peaked in August with a maximum of 45.7 °C. Sorghum produced an annual aboveground biomass yield of 43.7 Mg per hectare. Net ecosystem exchange (NEE) was highest during the summer growth period and reached a maximum of ?68 μmol CO₂ m^?2 s^?1. Water use efficiency, or biomass water ratio (BWR), was high (4.0 g dry biomass kg^?1 H₂O) despite high seasonal evapotranspiration (1094 kg H₂O m^?2). The BWR of sorghum surpassed that of many C4 biofuel candidate crops in the United States, as well as that of alfalfa which is currently widely grown in the Imperial Valley. Sorghum also outperformed many US biofuel crops in terms of radiation use efficiency (RUE), achieving 1.5 g dry biomass MJ^?1. We found no evidence of saturation of NEE at high levels of photosynthetically active radiation (PAR) (up to 2250 μmol m^?2 s^?1). In addition, we found no evidence that NEE was inhibited by either high VPD or air temperature during peak photosynthetic phases. The combination of high productivity, high BWR, and high RUE suggests that sorghum is well adapted to this extreme environment. The biomass production rates and efficiency metrics spanning three growing periods provide fundamental data for future Life Cycle Assessments (LCA), which are needed to assess the sustainability of this sorghum biofuel feedstock system.     

The effect of periphyton stoichiometry and light on biological phosphorus immobilization and release in streams

Periphyton stoichiometry can vary substantially as a result of differences in stream nutrient availability. A decrease in the periphyton carbon to phosphorus (C:P) ratio should decrease the demand for new P to be immobilized from stream water, but no studies to our knowledge have explored the relationship between periphyton stoichiometry and net P immobilization and release by periphyton. We sought to model biological P immobilization and release (flux) in streams by measuring periphyton stoichiometry and light availability. We measured P flux to and from intact periphyton on stream cobbles (20–100 mm diameter) in 1 L microcosms incubated with streamwater under variable light conditions. Net P immobilization occurred in 75% of microcosms, net P release occurred in only 5% of microcosms, and 20% of microcosms had neither net immobilization nor net release. When normalized to stream conditions, net P immobilization was highest when light availability was high (<60% canopy attenuation) and the periphyton C:P ratio was also high. In contrast, net P release occurred only when light availability was low (>60% canopy attenuation) and the periphyton C:P ratio was also low. A multiple regression model that included both periphyton stoichiometry and light availability from the growing season only, and the interaction term of these two variables, explained 99% of the variation in daily periphyton P flux observed in the study. These results indicate that in order to predict periphyton P immobilization, periphyton stoichiometry and light availability should be considered together. Furthermore, the results indicate that net P immobilization occurs even in very P-rich periphyton, which can act as a P sink when light availability is high.     

GS-8374, a Prototype Phosphonate-Containing Inhibitor of HIV-1 Protease,Effectively Inhibits Protease Mutants with Amino Acid Insertions

Insertions in the protease (PR) region of human immunodeficiency virus (HIV) represent an interesting mechanism of antiviral resistance against HIV PR inhibitors (PIs). Here, we demonstrate the improved ability of a phosphonate-containing experimental HIV PI, GS-8374, relative to that of other PIs, to effectively inhibit patient-derived recombinant HIV strains bearing PR insertions and numerous other mutations. We correlate enzyme inhibition with the catalytic activities of corresponding recombinant PRs in vitro and provide a biochemical and structural analysis of the PR-inhibitor complex.     

Carbon Isotope Discrimination in Coffee Genotypes Grown under Limited Water Supply

Photosynthetic gas exchange, plant-water relations characteristics, and stable carbon isotope discrimination (Δ) were evaluated for five Coffea arabica L. genotypes growing under two soil moisture regimes in the field. The Δ of leaf tissue was strongly correlated (r = −0.95) with inherent water use efficiency (ratio of assimilation to stomatal conductance; A/g). The variation in inherent water use efficiency (WUE) among genotypes was 30% for plants irrigated weekly. The higher WUE exhibited by some of these plants resulted from reduced g rather than increased photosynthetic capacity at a given g. Withholding irrigation for 1 month caused Δ to decline substantially in expanding leaf tissue of all genotypes. A strong correlation (r = 0.92) was found between Δ and plant hydraulic efficiency estimated as the ratio of g to the diurnal range in leaf water potential (Ψ_l). The Δ values for plants irrigated weekly adequately predicted drought-induced changes in Δ (r = 0.99) and midday Ψ_l (r = 0.95). The results indicated that Δ might be used to evaluate several aspects of plant performance and response to specific environmental conditions, once suitable background physiological data have been gathered.     

Stomatal responses to light and leaf-air water vapor pressure difference show similar kinetics in sugarcane and soybean

Stomatal responses to light and humidity (vapor pressure difference, VPD) are important determinants of stomatal conductance. Stomatal movements induced by light are the result of a transduction of the light stimulus into modulated ion fluxes in guard cells and concomitant osmotic adjustments and turgor changes. It is generally assumed that this transduction process is a general stomatal property, with different environmental stimuli integrated into guard cell metabolism through their modulation of ion fluxes. In contrast with this notion, the VPD response, which is unique because both its triggering signal and the turgor changes required for aperture modulations involve water molecules, has been considered to be hydropassive and thus independent of guard cell metabolism. We used a kinetic approach to compare the light and VPD responses in order to test the hypothesis that hydropassive changes in guard cell turgor could be faster than the metabolism-dependent light responses. Changes in stomatal conductance in intact leaves of sugarcane and soybean were measured after application of step changes in VPD and in light. In spite of a 5-fold difference in overall rates between the two species, the response rates following light or VPD steps were similar. Although a coincidental kinetic similarity between two mechanistically different responses cannot be ruled out, the data suggest a common mechanism controlling stomatal movements, with the VPD stimulus inducing metabolic modulations of ion fluxes analogous to other stomatal responses.     

Guard cells of Commelina communis L. do not respond metabolically to osmotic stress in isolated epidermis: Implications for stomatal responses to drought and humidity

We investigated the hypothesis that stomatal aperture is regulated by epidermal water status. Detached epidermal peels of Commelina communis L. or leaf disks with epidermis attached were incubated in graded solutions of mannitol (0–1.2 M) containing KCl. In isolated epidermis, guard-cell solute content of open stomata did not decrease in response to desiccation. Guard cells of closed stomata accumulated solutes to the same extent in all levels of mannitol tested. There was no evidence of stress-induced hydroactive closure nor of inhibition of hydroactive opening, even when guard cells of closed stomata were initially plasmolyzed. Hydropassive, osmometer-like, changes in stomatal aperture in the isolated epidermis were induced by addition or removal of mannitol, but these did not involve changes in guard-cell solute content. In leaf disks, stomata exhibited clear hydroactive stomatal responses. Steady-state guard-cell solute content of initially open and initially closed stomata decreased substantially with increasing mannitol. Stomata were completely closed above approx. 0.4 M mannitol, near the turgor-loss point for the bulk leaf tissue. Stomata of Commelina did not exhibit direct hydroactive responses to environmental or epidermal water status. Stomatal responses to water deficit and low humidity may be indirect, mediated by abscisic acid or other signal metabolite(s) from the mesophyll.Abbreviations ABA abscisic acid - EGTA ethyleneglycol-bis-(-aminoethyl ether)-N,N,N,N-tetraacetic acid - Mes 2-(N-morpholino)ethanesulfonic acid     

Root and shoot gas exchange respond additively to moderate ozone and methyl jasmonate without induction of ethylene: ethylene is induced at higher O3 concentrations

The available literature is conflicting on the potential protection of plants against ozone (O(3)) injury by exogenous jasmonates, including methyl jasmonate (MeJA). Protective antagonistic interactions of O(3) and MeJA have been observed in some systems and purely additive effects in others. Here it is shown that chronic exposure to low to moderate O(3) concentrations (4-114 ppb; 12 h mean) and to MeJA induced additive reductions in carbon assimilation (A (n)) and root respiration (R (r)), and in calculated whole plant carbon balance. Neither this chronic O(3) regime nor MeJA induced emission of ethylene (ET) from the youngest fully expanded leaves. ET emission was induced by acute 3 h pulse exposure to much higher O(3) concentrations (685 ppb). ET emission was further enhanced in plants treated with MeJA. Responses of growth, allocation, photosynthesis, and respiration to moderate O(3) concentrations and to MeJA appear to be independent and additive, and not associated with emission of ET. These results suggest that responses of Pima cotton to environmentally relevant O(3) are not mediated by signalling pathways associated with ET and MeJA, though these pathways are inducible in this species and exhibit a synergistic O(3)×MeJA interaction at very high O(3) concentrations.