Modeling dynamic understory photosynthesis of contrasting species in ambient and elevated carbon dioxide

Dynamic responses of understory plants to sunflecks have been extensively studied, but how much differences in dynamic light responses affect daily photosynthesis (A_day) is still the subject of active research. Recent models of dynamic photosynthesis have provided a quantitative tool that allows the critical assessment of the importance of these sunfleck responses on A_day. Here we used a dynamic photosynthesis model to assess differences in four species that were growing in ambient and elevated CO₂. We hypothesized that Liriodendron tulipifera, a species with rapid photosynthetic induction gain and slow induction loss, would have the least limitations to sunfleck photosynthesis relative to the other three species (Acer rubrum, Cornus florida, Liquidambar styraciflua). As a consequence, L. tulipifera should have the highest A_day in an understory environment, despite being the least shade tolerant of the species tested. We further hypothesized that daily photosynthetic enhancement by elevated CO₂ would differ from enhancement levels observed during light-saturated, steady-state measurements. Both hypotheses were supported by the model results under conditions of low daily photosynthetic photon flux density (PFD; <3% of the above-canopy PFD). However, under moderate PFD (10-20% of the above-canopy PFD), differences in dynamic sunfleck responses had no direct impact on A_day for any of the species, since stomatal and photosynthetic induction limitations to sunfleck photosynthesis were small. Thus, the relative species ranking in A_day under moderate PFD closely matched their rankings in steady-state measurements of light-saturated photosynthesis. Similarly, under elevated CO₂, enhancement of modeled A_day over A_day at ambient CO₂ matched the enhancement measured under light saturation. Thus, the effects of species-specific differences in dynamic sunfleck responses, and differences in elevated CO₂ responses of daily photosynthesis, are most important in marginal light environments.     

Structural basis of stereoselectivity in Candida rugosa lipase-catalyzed hydrolysis of secondary alcohols

Lipases are widely used catalysts for highly enantioselective resolution of chiral secondary alcohols. While stereopreference is determined predominantly by the substrate structure, stereoselectivity (enantioselectivity and diastereoselectivity) depends on the atomic details of interactions between substrate and lipase. Experimentally obtained stereoselectivity and activity in the hydrolysis of butanoic acid esters of two secondary alcohols with two neighboring stereocenters by Candida rugosa lipase have been investigated by computer-aided molecular modeling of tetrahedral substrate intermediates in complex with the lipase. Breakdown of these intermediates is considered to be the rate-limiting step. Steric interactions of stereoisomers with the side chain of catalytic histidine led to different orientations of the imidazole. The distance d(H_N)-O_alc) between H_N) of the imidazole side chain of catalytic histidine and the alcohol oxygen of the substrate was identified to correlate with the experimentally determined reactivity order of the four stereoisomers. Modeled distances d(H_N)-O_alc) were short (=1.8 Å) for RR stereoisomers, which were also found to be hydrolyzed most rapidly experimentally; distances d(H_N)-O_alc) were about 2 Å for SS and SR stereoisomers, which were converted at similar rates but at a lower rate than RR stereoisomers; finally, distances d(H_N)-O_alc) for SR stereoisomers were greater than 4 Å, in accordance with very slow conversion of SR stereoisomers.     

Proton affinity of para-substituted acetophenones in gas phase and in solution: a theoretical study

The gas phase proton affinities PA and basicities GB for a series of para-substituted acetophenones weak bases (B) p.X???C₆H₄CO*CH₃ with X?=?H, F, Cl, Br, I, Me, CF₃, CN, NO₂, OCH₃, NH₂, CH₂OH, N(CH₃)₂, OH, $ {\text{NH}}_3^{+} $ , … have been calculated at 298.15 K at the density functional theory DFT/B3LYP level with a 6-311++G (2d,2p) basis set. Conformational results lead to only one stable planar conformer for both unprotonated compounds and their O*-protonated forms. Satisfactory accuracy and computational efficiency could be reached if the computed PAs are scaled by a factor 0.983. Protonation at more than one site is discussed and the carbonyl oxygen atom is found to be the preferential protonated site rather than the substituent X. The calculated gas phase PAs show a good agreement with the experimental available data. The electron-donating/electron-withdrawing nature of the substituents has an enormous influence upon the thermochemical and structural properties. The influence of environment on the proton affinity has been studied by means of SCRF solvent effect computations using PCM solvation model for two solvents: water and SO₂CI₂. Confrontation between computed and experimental pK(B) values exhibits better agreement in aqueous solution than in organic solvent.     

Robust multiple-input-multiple-output control of non-linear continuous fermentation processes

In this study, the removal of nitrate (NO₃^m) ions from aqueous streams with liquid membrane technique has been investigated. Among the other parameters (temperature, pH, acceptor phase type and medium concentration), the stirring speed was chosen as process parameter. From the experimental results, it has been determined that the reaction was diffusion controlled. The transport efficiency of nitrate ions increased with increasing stirring speed. The membrane entrance and exit rate constants (k_1d, k_2m and k_2a respectively) were linearly dependent on the stirring speed ratios of 100 to 250 rpm. Coupled transport of nitrate ions through a liquid membrane in 85% n-hexane-15% tricloromethane as diluent, containing tetraoctyl ammonium chloride (TOACl) as a carrier was examined at various stirring speeds. Membrane entrance (k_1d) and exit rates (k_2m and k_2a) increase with increasing the stirring speeds. The diffusion of the nitrate ion-carrier complex through the narrow stagnant layers was found to be rate determining step. The membrane was stable during the transport experiments. There is no leakage of carrier or nitrate ion-carrier complex to both aqueous phases and also, no supplementary water penetration into the membrane. This favours interfacial reaction of nitrate ion and carrier.     

Elevated CO2 influences nutrient availability in young beech-spruce communities on two soil types

The objectives of this study were to investigate how different soil types and elevated N deposition (0.7 vs 7 g N m^-2a^-1) influence the effects of elevated CO₂ (370 vs 570 µmol CO₂ mol^-1) on soil nutrients and net accumulation of N, P, K, S, Ca, Mg, Fe, Mn, and Zn in spruce (Picea abies) and beech (Fagus sylvatica). Model ecosystems were established in large open-top chambers on two different forest soils: a nutrient-poor acidic loam and a nutrient-rich calcareous sand. The response of net nutrient accumulation to elevated atmospheric CO₂ depended upon soil type (interaction soil 2 CO₂, P<0.05 for N, P, K, S, Ca, Mg, Zn) and differed between spruce and beech. On the acidic loam, CO₂ enrichment suppressed net accumulation of all nutrients in beech (P<0.05 for P, S, Zn), but stimulated it for spruce (P<0.05 for Fe, Zn) On the nutrient-rich calcareous sand, increased atmospheric CO₂ enhanced nutrient accumulation in both species significantly. Increasing the N deposition did not influence the CO₂ effects on net nutrient accumulation with either soil. Under elevated atmospheric CO₂, the accumulation of N declined relative to other nutrients, as indicated by decreasing ratios of N to other nutrients in tree biomass (all ratios: P<0.001, except the N to S ratio). In both the soil and soil solution, elevated CO₂ did not influence concentrations of base cations and available P. Under CO₂ enrichment, concentrations of exchangeable NH₄⁺ decreased by 22% in the acidic loam and increased by 50% in the calcareous sand (soil 2 CO₂, P<0.001). NO₃^- concentrations decreased by 10-70% at elevated CO₂ in both soils (P<0.01).     

The effects of elevated CO2 on seed production and seedling recruitment in a sheep-grazed pasture

Seed production and seedling recruitment were measured over 2 years under ambient (360 ppm) and elevated (475 ppm) atmospheric CO₂ in a free air carbon dioxide enrichment (FACE) experiment, carried out in a sheep-grazed pasture on dry, sandy soil in New Zealand. In both years elevated CO₂ led to more dispersed seeds of the grasses Anthoxanthum odoratum, Lolium perenne and Poa pratensis, the legumes Trifolium repens and T. subterraneum and the herbs Hypochaeris radicata and Leontodon saxatilis. The increased seed dispersal in A. odoratum, H. radicata, Leontodon saxatilis and T. repens reflected both more inflorescences per unit area and more seeds per inflorescence under elevated CO₂. The increased seed dispersal in Lolium perenne, P. pratensis and T. subterraneum was due solely to more inflorescences per unit area. The number of seedlings that emerged and survived to at least 7 months of age was increased by elevated CO₂ for H. radicata, Leontodon saxatilis, T. repens and T. subterraneum in both years and for A. odoratum and Lolium perenne in the first year. For species where increased seedling recruitment was noted, there was a significant positive correlation between seed production in summer and seedling emergence in the following autumn and winter, and sowing 200 extra seeds per species m^-2 resulted in more seedlings compared to unsown controls. Elevated CO₂ did not affect seedling survival in any species. There was no measurable effect of elevated CO₂ on canopy and soil surface conditions or soil moisture at the time of seedling emergence. The results suggest the dominant effect of elevated CO₂ on seedling recruitment in this pasture was an indirect one, reflecting effects on the number of seeds produced. The biomass of H. radicata, Leontodon saxatilis, T. repens and T. subterraneum in the above-ground vegetation was greater under elevated than ambient CO₂. However, the size of individual seedlings and mature plants of these four species was unaffected by elevated CO₂. The results indicate an important way elevated CO₂ influenced plant species composition in this pasture was through changes in the pattern of seedling recruitment.     

Wine vinasses treatments by ozone and an activated sludge system in continuous reactors

In this work wine vinasses have been treated separately by means of a chemical ozonation and a biological aerobic degradation in an activated sludge system, and later by means of a combined process which consisted of an aerobic pretreatment followed by an ozonation treatment, in continuous reactors in all cases. In the ozonation experiments, the hydraulic retention time and the ozone partial pressure were varied leading to substrate removals in the range 4.4-16%, with increases in this removal when both operating variables were increased. A kinetic study, which combines mixed flow reactor model for the liquid phase and plug flow reactor model for the gas phase, allows to determine the rate constant for the ozone reaction and the consumption ratio, which are k_O3 = 3.6 l/(g COD · h) and b = 22.5 g COD degraded/mol O₃ consumed. The aerobic degradation experiments were conducted in the activated sludge system with variations in the retention time and influent organic substrate concentration in the wastewater. A modified Contois model applied to the experimental results leads to the determination of the kinetic parameters of that model: K₁ = 5.43 l/g VSS and q_max = 6.29 g COD/(g VSS · h). Finally, the combined process reveals an improvement in the efficiency of the ozonation stage due to the previous aerobic treatment with increases in the substrate removal reached and in the rate constant obtained, the last one being k_O3 = 5.6 l/(g COD · h).     

13C content of ecosystem respiration is linked to precipitation and vapor pressure deficit

Variation in the carbon isotopic composition of ecosystem respiration ('¹³C_R) was studied for 3 years along a precipitation gradient in western Oregon, USA, using the Keeling plot approach. Study sites included six coniferous forests, dominated by Picea sitchensis, Tsuga heterophylla, Pseudotsuga menziesii, Pinus ponderosa, and Juniperus occidentalis, and ranged in location from the Pacific coast to the eastern side of the Cascade Mountains (a 250-km transect). Mean annual precipitation across these sites ranged from 227 to 2,760 mm. Overall '¹³C_R varied from -23.1 to -33.1‰, and within a single forest, it varied in magnitude by 3.5-8.5‰. Mean annual '¹³C_R differed significantly in the forests and was strongly correlated with mean annual precipitation. The carbon isotope ratio of carbon stocks (leaves, fine roots, litter, and soil organic matter) varied similarly with mean precipitation (more positive at the drier sites). There was a strong link between '¹³C_R and the vapor saturation deficit of air (vpd) 5-10 days earlier, both across and within sites. This relationship is consistent with stomatal regulation of gas exchange and associated changes in photosynthetic carbon isotope discrimination. Recent freeze events caused significant deviation from the '¹³C_R versus vpd relationship, resulting in higher than expected '¹³C_R values.     

Gas exchange and chlorophyll fluorescence responses of <Emphasis Type="Italic">Pinus radiata</Emphasis> D. Don seedlings during and after several storage regimes and their effects on post-planting survival

Post-storage gas exchange parameters like CO₂ assimilation, stomatal conductance, transpiration, water use efficiency and intercellular CO₂ concentrations, together with several chlorophyll a fluorescence parameters: F_o, F_v, F_v/F_m, F_m/F_o and F_v/F_o were examined in radiata pine (Pinus radiata D. Don) seedlings that were stored for 1, 8 or 15 days at 4° or 10°C with or without soil around the roots. Results were analysed in relation to post-storage water potential and electrolyte leakage in order to forecast their vitality (root growth potential) following cold storage, and post-planting survival potential under optimal conditions. During storage at 4° and 10°C, photosynthesis was reduced, being more pronounced in bare-root seedlings than in seedlings with soil around the roots. The depletion of CO₂ assimilation seemed not to be solely a stomatal effect as effects on chloroplasts contributed to this photosynthetic inhibition. Thus, the fall in the ratios F_v/F_m, F_v/F_o and F_m/F_o indicated photochemical apparatus damage during storage. Photosynthetic rate was positively correlated with the root growth index and new root length showing that new root growth is dependent primarily on current photosynthesis. Pre-planting exposure of bare-root radiata pine seedlings to temperatures of 10°C for more than 24 h during transportation or storage is not recommended.     

The effect of elevated carbon dioxide and ozone on leaf- and branch-level photosynthesis and potential plant-level carbon gain in aspen

Two aspen (Populus tremuloides Michx.) clones, differing in O₃ tolerance, were grown in a free-air CO₂ enrichment (FACE) facility near Rhinelander, Wisconsin, and exposed to ambient air, elevated CO₂, elevated O₃ and elevated CO₂+O₃. Leaf instantaneous light-saturated photosynthesis (P_S) and leaf areas (A) were measured for all leaves of the current terminal, upper (current year) and the current-year increment of lower (1-year-old) lateral branches. An average, representative branch was chosen from each branch class. In addition, the average photosynthetic rate was estimated for the short-shoot leaves. A summing approach was used to estimate potential whole-plant C gain. The results of this method indicated that treatment differences were more pronounced at the plant- than at the leaf- or branch-level, because minor effects within modules accrued in scaling to plant level. The whole-plant response in C gain was determined by the counteracting changes in P_S and A. For example, in the O₃-sensitive clone (259), inhibition of P_S in elevated O₃ (at both ambient and elevated CO₂) was partially ameliorated by an increase in total A. For the O₃-tolerant clone (216), on the other hand, stimulation of photosynthetic rates in elevated CO₂ was nullified by decreased total A.     

Modelling mixing parameters in concentric-tube airlift bioreactors

The mixing behaviour of the liquid phase in concentric-tube airlift bioreactors of different scale (RIMP: V_L=0.070 m³; RIS-1: V_L=2.50 m³; RIS-2: V_L=5.20 m³) in terms of mixing time was investigated. This mixing parameter was determined from the output curves to an initial Dirac pulse, using the classical tracer response technique, and analyzed in relation to process and geometrical parameters, such as: gas superficial velocity, x_SGR; top clearance, h_S; bottom clearance, h_B, and ratio of the resistances at downcomer entrance, A_d/A_R. A correlation between the mixing time and the specified operating and geometrical parameters was developed, which was particularized for two flow regimes: bubbly and transition (x_SGRА.08 m/s) and churn turbulent flow (x_SGR> 0.08 m/s) respectively. The correlation was applied in bioreactors of different scale with a maximum error of ᆲ%.     

Photosynthesis in an invasive grass and native forb at elevated CO2 during an El Niño year in the Mojave Desert

Annual and short-lived perennial plant performance during wet years is important for long-term persistence in the Mojave Desert. Additionally, the effects of elevated CO₂ on desert plants may be relatively greater during years of high resource availability compared to dry years. Therefore, during an El Niño year at the Nevada Desert FACE Facility (a whole-ecosystem CO₂ manipulation), we characterized photosynthetic investment (by assimilation rate-internal CO₂ concentration relationships) and evaluated the seasonal pattern of net photosynthesis (A_net) and stomatal conductance (g_s) for an invasive annual grass, Bromus madritensis ssp. rubens and a native herbaceous perennial, Eriogonum inflatum. Prior to and following flowering, Bromus showed consistent increases in both the maximum rate of carboxylation by Rubisco (V_Cmax) and the light-saturated rate of electron flow (J_max) at elevated CO₂. This resulted in greater A_net at elevated CO₂ throughout most of the life cycle and a decrease in the seasonal decline of maximum midday A_net upon flowering as compared to ambient CO₂. Eriogonum showed significant photosynthetic down-regulation to elevated CO₂ late in the season, but the overall pattern of maximum midday A_net was not altered with respect to phenology. For Eriogonum, this resulted in similar levels of A_net on a leaf area basis as the season progressed between CO₂ treatments, but greater photosynthetic activity over a typical diurnal period. While g_s did not consistently vary with CO₂ in Bromus, it did decrease in Eriogonum at elevated CO₂ throughout much of the season. Since the biomass of both plants increased significantly at elevated CO₂, these patterns of gas exchange highlight the differential mechanisms for increased plant growth. The species-specific interaction between CO₂ and phenology in different growth forms suggests that important plant strategies may be altered by elevated CO₂ in natural settings. These results indicate the importance of evaluating the effects of elevated CO₂ at all life cycle stages to better understand the effects of elevated CO₂ on whole-plant performance in natural ecosystems.     

Transpiration in response to variation in microclimate and soil moisture in southeastern deciduous forests

Responses of forests to changes in environmental conditions reflect the integrated behavior of their constituent species. We investigated sap flux-scaled transpiration responses of two species prevalent in upland eastern hardwood forests, Quercus alba in the upper canopy and Acer rubrum in the low to mid canopy, to changes in photosynthetically active radiation above the canopy (Q_o), vapor pressure deficit within the canopy (D), and soil moisture depletion during an entire growing season. Water loss before bud break (presumably through the bark) increased linearly with D, reaching 8% of daily stand transpiration (E_C) as measured when leaf area index was at maximum, and accounting for 5% of annual water loss. After leaves were completely expanded and when soil moisture was high, sap flux-scaled daily E_C increased linearly with the daily sum of Q_o. Species differences in this response were observed. Q. alba reached a maximum transpiration at low Q_o, while A. rubrum showed increasing transpiration with Q_o at all light levels. Daily E_C increased in response to daily average D, with an asymptotic response due to the behavior of Q. alba. Transpiration of A. rubrum showed a greater response to soil moisture depletion than did that of Q. alba. When evaluated at a half-hourly scale under high Q_o, mean canopy stomatal conductance (G_S) of individuals decreased with D. The sensitivity of G_S to D was greater in species with higher intrinsic G_S. Regardless of position in the canopy, diffuse-porous species in this and an additional, more mesic stand showed higher G_S and greater stomatal sensitivity to environmental variation than do ring-porous species.     

Growth, photosynthesis and ozone uptake of young beech (Fagus sylvatica L.) in response to different ozone exposures

Beech seedlings (Fagus sylvatica L.) were exposed to episodes of O₃ in environmentally controlled growth chambers during one growing season. Three treatments were applied: charcoal-filtered air, charcoal-filtered air with the addition of 40 ppb O₃ for seven episodes of 5 days' duration (9000-1700 hours), and charcoal-filtered air with the addition of 100 ppb O₃ for seven episodes of 5 days' duration (9000-1700 hours). The accumulated exposure over a threshold of 40 ppb in the last treatment reached 13,911 ppb h. Throughout the growing season we measured growth as well as photosynthetic properties and related effects to external and calculated internal doses of O₃, using stomatal conductance (g_s) data. Growth, measured as diameter increment and biomass, was not significantly affected by the O₃ treatments. In the 100-ppb treatment, light-saturated CO₂ assimilation rates and chlorophyll content were significantly reduced, and the chlorophyll fluorescence parameter F_v/F_m was significantly reduced at times of high uptake rates and coincided with strong reductions of assimilation rates. O₃ uptake was lowered in the 100-ppb treatment due to reduced g_s. There was serious visible damage by the end of the exposure period in the 100-ppb treatment, while the treatment with 40 ppb O₃ did not seem to cause any significant changes.     

Photosynthetic characteristics in canopies of Quercus rubra, Quercus prinus and Acer rubrum differ in response to soil water availability

Photosynthesis and related leaf characteristics were measured in canopies of co-occurring Quercus rubra L. (red oak), Quercus prinus L. (chestnut oak) and Acer rubrum L. (red maple) trees. Mature (20+ m tall) trees were investigated at sites of differing soil water availability within a catchment (a drier upper site and a wetter lower site). Leaf photosynthetic characteristics differed significantly between species and in response to site and position in the canopy. Photosynthetic capacity (A_max) was significantly greater at the wetter site in all canopy strata in A. rubrum but not in Q. rubra or Q. prinus. Our findings for A. rubrum are generally consistent with those predicting that species with higher specific leaf area (SLA) will have higher A_max per unit leaf nitrogen (N) and that species with leaves with lower SLA (e.g. Q. rubra and Q. prinus) will have shallower slopes of the A_max-N relationship. Importantly, the relationships between A_max and N_area (and by implication photosynthetic nitrogen-use efficiency, PNUE) differed in A. rubrum between the sites, with PNUE significantly lower at the drier site. The lower photosynthetic capacity and PNUE must substantially reduce carbon acquisition capacity in A. rubrum under these field conditions. Maximum stomatal conductance (g_smax) differed significantly between species, with g_smax greatest in Q. rubra and Q. prinus. In Q. rubra and Q. prinus, g_smax was significantly lower at the upper site than the lower site. There was no significant response of g_smax to site in A. rubrum. These stomatal responses were consistent with the C_i/C_a ratio, which was significantly lower in leaves of Q. rubra and Q. prinus at the upper site, but did not differ between sites in A. rubrum. Leaf '¹³C was significantly lower in A. rubrum than in either Q. rubra or Q. prinus at both sites. These findings indicate differences in stomatal behaviour in A. rubrum which are likely to contribute to lower water use efficiency at both sites. Our results support the hypothesis that the two Quercus species, in contrast to A. rubrum, maintain photosynthetic capacity at the drier site whilst minimising transpirational water loss. They also suggest, based primarily on physiological evidence, that the ability of A. rubrum to compete with other species of these deciduous forests may be limited, particularly in sites of low moisture availability and during low rainfall years.     

In situ oxygen microelectrode measurements of bottom-ice algal production in McMurdo Sound, Antarctica

In-situ estimates of fast-ice algal productivity at Cape Evans, McMurdo Sound, in 1999 were lower than at the same site in previous years. Under-ice irradiance was between 0 and 8 µmol photons m^-2 s^-1; the ice was between 1.9 and 2.0 m thick and the algal biomass averaged 150 mg chl a m^-2, although values as high as 378 mg chl a m^-2 were recorded. Production on 11 and 12 November was between 0.053 and 1.474 mg C m^-2 h^-1. When the data from 11 November were fitted to a hyperbolic tangent function, a multilinear regression gave estimates for P_max of 0.571 nmol O₂ cm^-2 s^-1, an ! of 0.167 nmol O₂ cm^-2 s^-1 µmol^-1 photons m^-2 s^-1 and an E_k of 3.419 µmol photons m^-2 s^-1. A P_max of 2.674 nmol O₂ cm^-2 s^-1, an ! of 0.275 nmol O₂ cm^-2 s^-1 µmol^-1 photons m^-2 s^-1, r of 0.305 nmol O₂ cm^-2 s^-1 and an E_k of 9.724 µmol^-1 photons m^-2 s^-1 were estimated from the 12 November data. The sea-ice algal community was principally comprised of Nitzschia stellata, Entomoneis kjellmanii and Berkeleya adeliensis. Other taxa present included N. lecointei, Fragilariopsis spp., Navicula glaciei, Pleurosigma spp. and Amphora spp. Variations in the method for estimating the thickness of the diffusive boundary layer were not found to significantly affect the measurements of oxygen flux. However, the inability to accurately measure fine-scale variations in biomass is thought to contribute to the scatter of the P versus E data.     

Effects of elevated CO<Subscript>2</Subscript> on foliar chemistry of saplings of nine species of tropical tree

This study examined the effects of elevated CO₂ on secondary metabolites for saplings of tropical trees. In the first experiment, nine species of trees were grown in the ground in open-top chambers in central Panama at ambient and elevated CO₂ (about twice ambient). On average, leaf phenolic contents were 48% higher under elevated CO₂. Biomass accumulation was not affected by CO₂, but starch, total non-structural carbohydrates and C/N ratios all increased. In a second experiment with Ficus, an early successional species, and Virola, a late successional species, treatments were enriched for both CO₂ and nutrients. For both species, nutrient fertilization increased plant growth and decreased leaf carbohydrates, C/N ratios and phenolic contents, as predicted by the carbon/nutrient balance hypothesis. Changes in leaf C/N levels were correlated with changes in phenolic contents for Virola (r=0.95, P<0.05), but not for Ficus. Thus, elevated CO₂, particularly under conditions of low soil fertility, significantly increased phenolic content as well as the C/N ratio of leaves. The magnitude of the changes is sufficient to negatively affect herbivore growth, survival and fecundity, which should have impacts on plant/herbivore interactions.     

Mycorrhizal associations between Tuber melanosporum mycelia and transformed roots of Cistus incanus

We set out to establish root cultures of a host plant with the aim of obtaining dual cultures of Tuber melanosporum mycorrhiza on transformed roots. Seedlings of Cistus incanus germinated under sterile conditions from seeds collected in the wild were treated with Agrobacterium rhizogenes. Nine hairy roots collected from different seedlings were cultured individually by repeated subculturing. The hairy root clones differed in growth rates and in morphology (branching frequency and distance between side roots). Root growth in a liquid medium exhibited a lag phase of about 2 weeks and an exponential phase lasting about 12 days before the start of the stationary phase. Hairy roots could be kept alive on medium M, a special solid minimal medium (low in Fe²⁺, BO₄^3-, Ca²⁺, Cu²⁺ and Zn²⁺, very low in PO₄^3- and lacking MoO₄^2-, NH₄⁺ and Co²⁺), for more than 7 months. T. melanosporum could be grown on the same medium for long periods only by subculturing the fungus with the roots. A mycorrhizal association developed between the roots and the T. melanosporum mycelium within 3 months. The association consisted of elongated roots with a mantle and a Hartig net surrounding two to three layers of cortical cells. Swollen, club-like root tips were discernible 5 months after inoculation. The mycorrhized roots could be subcultured and propagated on medium M and maintain the mycorrhizal association.     

Cytochrome oxidase sensitivity to carbon monoxide in leaves of various tall and dwarf wheat cultivars and their crosses

In order to investigate the possible role of Rht genes in the regulation of the redox condition of cytochrome a₃ (cytochrome c oxidase) during steady-state respiration, wheat cultivars belonging to one of two groups - NP 710, NP 846 and NP 875 belonging to the tall group and Olesons dwarf, HD 1982 and HD 2122 of the dwarf group - and the reciprocal crosses between the varieties of these two groups were examined for carbon monoxide (CO) sensitivity in terms of the inhibition of mitochondrial electron transport. Leaves of young wheat seedlings were used. Differences in the redox state of cytochrome a₃ were monitored using the in vivo aerobic assay of nitrate reduction after a 1-min exposure to CO. Dwarf cultivars possessing Rht genes responded marginally (᜖%) to CO inhibition, whereas the response of tall cultivars to CO was higher (51-70%). Since CO forms a complex only with reduced cytochrome a₃, the results indicate differences in the redox state of cytochrome a₃ during in situ respiration of leaves from tall and dwarf plants that are likely to be controlled by cytoplasmic factors.