Effects of Long Term Exposure to High-CO2 During Storage at 0°C on Biology and Infectivity of Botrytis cinerea in Red Chicory

The effects on Botrytis cinerea of prolonged exposures to CO₂‐enriched atmospheres were studied in vitro and in vivo at 0°C. Mycelial growth on potato dextrose agar decreased linearly with increasing CO₂ concentrations from 5, 10, 15 and 20% CO₂. The growth reduction was greater after 30–40 days of incubation. A reduced production of sclerotia in air by the colonies formerly exposed to various CO₂ concentrations was also detected. Conidial germination was delayed and the amount of germinated conidia decreased with increased CO₂ and at 20% CO₂ it was inhibited. Germ tube elongation was affected in the same way. In artificially inoculated red chicory, lesion area caused by B. cinerea decreased with increasing concentrations of CO₂ up to 60 days storage, later only 10 and 15% CO₂ were really effective, while in the final inspection after 120 days all the concentration tested showed a low efficacy. Similar results were obtained in naturally infected chicory where the severity of the disease decreased by increasing CO₂ from 5 to 10%, higher values did not improve the suppressive effect or determined, after 150 days of storage, an increased vulnerability of the tissues to disease due to the phytotoxic effects of the gas. An atmosphere enriched with 10% CO₂ is advised to suppress Botrytis rot during storage at 0°C of red chicory.     

Comprehensive optimization of composite cryoprotectant for Saccharomyces boulardii during freeze-drying and evaluation of its storage stability

Abstract

Saccharomyces boulardii (S. boulardii) is widely adopted in the diarrhea treatment for humans or livestock, so guaranteeing the survival rate of S. boulardii is the critical issue during freeze-drying process. In this study, the survival rate of S. boulardii with composite cryoprotectants during freeze-drying procedure and the subsequent storage were investigated. With the aid of response surface method, the composite cryoprotectants were comprehensively optimized to be lactose of 21.24%, trehalose of 22.00%, and sodium glutamate of 4.00%, contributing to the supreme survival rate of S. boulardii of 64.22?±?1.35% with the viable cell number of 9.5?±?0.07?×?10⁹ CFU/g, which was very close to the expected rate of 65.55% with a number of 9.6?×?10⁹ CFU/g. The accelerated storage test demonstrated that the inactivation rate constant of the freeze-dried S. boulardii powder was k_?18?=?8.04?×?10^?6. In addition, the freeze-dried goat milk powder results exhibited that the inactivation rate constants were k₄?=?4.48?×?10^?4 and k₂₅?=?9.72?×?10^?3 under 4 and 25?°C, respectively. This work provides a composite cryoprotectant formulation that has a good protective effect for the probiotic S. boulardii during freeze-drying process, possessing the potential application prospect in food, medicine, and even feed industry.     

Photosynthetic and leaf respiration activity of <Emphasis Type="Italic">Malcolmia littorea</Emphasis> (L.) R. Br. in response to air temperature

Plant traits of Malcolmia littorea growing at the Botanic Garden of Rome and transplanted from the wild population developing along the Latium coast (Italy) were analyzed. The highest photosynthetic rates [P _N, 22.5 ± 0.5 μmol(CO₂) m⁻² s⁻¹], associated to the highest chlorophyll content (Chl, 60 ± 5 SPAD units), and respiration rates [R, 11.1 ± 0.2 μmol(CO₂) m⁻² s⁻¹] were reached in spring, when mean air temperature (T _m) was in the range 17°C to 23°C. P _N, Chl, and R decreased by 86, 38, and 59% in summer when mean maximum air temperature (T _max) was 30.3 ± 2.6°C. Leaf water potential decreased by 34% in summer compared to the spring value, and it was associated to a relative water content (RWC) of 74 ± 4%, and to a water-use efficiency (WUE) of 2.15 ± 0.81 μmol(CO₂) mmol⁻¹(H₂O). Moreover, also low air temperatures determined a significant P _N and R decreases (by 52 and 40% compared to the maximum, respectively). Responsiveness of gross photosynthetic rate (P _g) to R was higher than that to P _N as underlined by the slope of the regression line between the two variables. The results underlined a low tolerance to both high- and low air temperatures of M. littorea. The selected key traits (R, WUE, Chl) by the discriminant analysis might be used to monitor the M. littorea wild population in the long time. The ex situ cultivated plants could be propagated and used to increase the individuals number of the wild population.     

Elevated CO2 stimulates associative N2 fixation in a C3 plant of the Chesapeake Bay wetland

In this study, the response of N₂ fixation to elevated CO₂ was measured in Scirpus olneyi, a C₃ sedge, and Spartina patens, a C₄ grass, using acetylene reduction assay and ¹⁵N₂ gas feeding. Field plants grown in PVC tubes (25 cm long, 10 cm internal diameter) were used. Exposure to elevated CO₂ significantly (P < 0·05) caused a 35% increase in nitrogenase activity and 73% increase in ¹⁵N incorporated by Scirpus olneyi. In Spartina patens, elevated CO₂ (660 ± 1 μ mol mol ^{− 1}) increased nitrogenase activity and ¹⁵N incorporation by 13 and 23%, respectively. Estimates showed that the rate of N₂ fixation in Scirpus olneyi under elevated CO₂ was 611 ± 75 ng ¹⁵N fixed plant ^{− 1} h ^{− 1} compared with 367 ± 46 ng ¹⁵N fixed plant ^{− 1} h ^{− 1} in ambient CO₂ plants. In Spartina patens, however, the rate of N₂ fixation was 12·5 ± 1·1 versus 9·8 ± 1·3 ng ¹⁵N fixed plant ^{− 1} h ^{− 1} for elevated and ambient CO₂, respectively. Heterotrophic non-symbiotic N₂ fixation in plant-free marsh sediment also increased significantly (P < 0·05) with elevated CO₂. The proportional increase in ¹⁵N₂ fixation correlated with the relative stimulation of photosynthesis, in that N₂ fixation was high in the C₃ plant in which photosynthesis was also high, and lower in the C₄ plant in which photosynthesis was relatively less stimulated by growth in elevated CO₂. These results are consistent with the hypothesis that carbon fixation in C₃ species, stimulated by rising CO₂, is likely to provide additional carbon to endophytic and below-ground microbial processes.     

A comparative study on the regulation of C3 and C4 carboxylation processes in the constitutive crassulacean acid metabolism (CAM) plant Kalanchoë daigremontiana and the C3-CAM intermediate Clusia minor

A comparison of carbon metabolism in the constitutive crassulacean acid metabolism (CAM) plant Kalanchoë daigremontiana Hamet et Perr. and the C₃-CAM intermediate Clusia minor L. was undertaken under controlled environmental conditions where plants experience gradual changes in light intensity, temperature and humidity at the start and end of the photoperiod. The magnitude of CAM activity was manipulated by maintaining plants in ambient air or by enclosing leaves overnight in an atmosphere of N₂ to suppress C₄ carboxylation. Measurements of diel changes in carbonisotope discrimination and organic acid content were used to quantify the activities of C₃ and C₄ carboxylases in vivo and to indicate the extent to which the activities of phosphoenolpyruvate carboxylase (PEPCase), ribulose-1,5-bisphosphate carboxylase-oxygenase (Rubisco) and decarboxylation processes overlap at the start and end of the photoperiod. These measurements in vivo were compared with measurements in vitro of changes in the diel sensitivity of PEPCase to malate inhibition. The results demonstrate fundamental differences in the down-regulation of PEPCase during the day in the two species. While PEPCase is inactivated within the first 30 min of the photoperiod in K. daigremontiana, the enzyme is active for 4 h at the start and 3 h at the end of the photoperiod in C. minor. Enclosing leaves in N₂ overnight resulted in a two-to threefold increase in PEPCase-mediated CO₂ uptake during Phase II of CAM in both species. However, futile cycling of CO₂ between malate synthesis and decarboxylation does not occur during Phase II in either species. In terms of overall carbon balance, C₄ carboxylation accounted for ≈ 20% of net daytime assimilation in both species under control conditions, increasing to 30–34% after a night in N₂. Although N₂-treated leaves of K. daigremontiana took up 25% more CO₂ than control leaves during the day this was insufficient to compensate for the loss of CO₂ taken up by CAM the previous night. In contrast, in N₂-treated leaves of C. minor, the twofold increase in daytime PEPCase activity and the increase in net CO₂ uptake by Rubisco during Phase III compensated for the inhibition of C₄ carboxylation at night in terms of diel carbon balance.     

Growth pattern of <Emphasis Type="Italic">Bidens cernua</Emphasis> L.: relationships between relative growth rate and its physiological and morphological components

Seedlings of Bidens cernua L. emerged when mean air temperature was 17.0±1.3 °C. The highest net photosynthetic rate (P _N), 13.8±0.8 μmol(CO₂) m⁻² s⁻¹, was monitored during the vegetative period (May–August), decreasing on an average by 50 % during flowering (August–September) and during fruiting (September–November) phases. The senescence phase (October–November) was characterised by 79, 58, and 18 % decrease of P _N, chlorophyll content, and leaf area (LA), respectively, from the maximum values. The time span from seedling emergence to the end of fruiting phase was 202 d. The total plant biomass was 1.58±0.05 g of which 81 % was aboveground plant portion. The total dry mass relative growth rate averaged over the assimilation period was 0.0804±0.0002 kg kg⁻¹ d⁻¹, and it was correlated to both the net assimilation rate (NAR) and the leaf area ratio (LAR).     

Environmental induced variations in leaf dark respiration and net photosynthesis of <Emphasis Type="Italic">Quercus ilex</Emphasis> L.

The relationships between dark respiration rate (R _D) and net photosynthetic rate (P _N) in Quercus ilex L. shrubs growing at the Botanical Garden in Rome were analysed. Correlation analysis of the data sets collected in the year 2006 confirmed the dependence among the considered leaf traits, in particular, R _D was significantly (p<0.05) correlated with P _N (r = 0.40). R _D and P _N increased from March to May [1.40±0.10 and 10.1±1.8 μmol(CO₂) m⁻² s⁻¹ mean values of the period, respectively], when air temperature was in the range 14.8–25.2 °C, underlining the highest metabolic activity in the period of the maximum vegetative activity that favoured biomass accumulation. On the contrary, the highest R _D [1.60±0.02 μmol(CO₂) m⁻² s⁻¹], associated to the lowest P _N rates (44 % of the maximum) and carbon use efficiency (CUE) in July underlined the mobilization of stored material during drought stress by a higher air temperature (32.7 °C).     

Initial nitrous oxide,carbon dioxide,and methane costs of converting conservation reserve program grassland to row crops under no‐till vs. conventional tillage

Around 4.4 million ha of land in USDA Conservation Reserve Program (CRP) contracts will expire between 2013 and 2018 and some will likely return to crop production. No‐till (NT) management offers the potential to reduce the global warming costs of CO₂, CH₄, and N₂O emissions during CRP conversion, but to date there have been no CRP conversion tillage comparisons. In 2009, we converted portions of three 9–21 ha CRP fields in Michigan to conventional tillage (CT) or NT soybean production and reserved a fourth field for reference. Both CO₂ and N₂O fluxes increased following herbicide application in all converted fields, but in the CT treatment substantial and immediate N₂O and CO₂ fluxes occurred after tillage. For the initial 201‐day conversion period, average daily N₂O fluxes (g N₂O‐N ha^?1 d^?1) were significantly different in the order: CT (47.5 ± 6.31, n = 6) ? NT (16.7 ± 2.45, n = 6) ? reference (2.51 ± 0.73, n = 4). Similarly, soil CO₂ fluxes in CT were 1.2 times those in NT and 3.1 times those in the unconverted CRP reference field. All treatments were minor sinks for CH₄ (?0.69 ± 0.42 to ?1.86 ± 0.37 g CH₄–C ha^?1 d^?1) with no significant differences among treatments. The positive global warming impact (GWI) of converted soybean fields under both CT (11.5 Mg CO₂e ha^?1) and NT (2.87 Mg CO₂e ha^?1) was in contrast to the negative GWI of the unconverted reference field (?3.5 Mg CO₂e ha^?1) with on‐going greenhouse gas (GHG) mitigation. N₂O contributed 39.3% and 55.0% of the GWI under CT and NT systems with the remainder contributed by CO₂ (60.7% and 45.0%, respectively). Including foregone mitigation, we conclude that NT management can reduce GHG costs by ~60% compared to CT during initial CRP conversion.     

Interactive effects of inorganic phosphate nutrition and carbon dioxide enrichment on assimilate partitioning in barley roots

The combined effects of inorganic phosphate (Pi) insufficiency and CO₂ enrichment on metabolite levels and carbon partitioning were studied using roots of 9-, 13- and 17-day-old barley seedlings (Hordeum vulgare L. cv. Brant). Plants were grown from seed in controlled environment chambers providing 36 ± 1 Pa (ambient) or 100 ± 2 Pa (elevated) CO₂ and either 1.0 mM (Pi sufficient) or 0.05 mM (Pi insufficient) Pi. When values were combined for both Pi treatments, plants grown under enhanced CO₂ showed increased root dry matter, adenylates (ATP + ADP), glutamine and non- structural carbohydrates other than starch. In contrast with shoots, enhanced CO₂ partially reversed the inhibition of root dry matter formation imposed by Pi insufficiency. The Pi-insufficient treatment also increased sucrose, glucose and fructose levels in barley roots. The Pi and CO₂ treatments were additive, so that the highest soluble carbohydrate levels were observed in roots of Pi-insufficient plants from the elevated CO₂ treatment. Pi limitation decreased dry matter formation, acid-extractable Pi, nitrate, hexose-phosphates, glutamate, glutamine and acid invertase activity of barley roots in plants grown in both ambient and elevated CO₂. Adenylate levels in roots were unaffected by the moderate Pi insufficiency described here. Thus, the reduced hexose-phosphate levels of Pi-insufficient roots were not likely to be the result of low adenylate concentrations. The above results suggest that the capacity of barley roots to utilize carbohydrates from the shoot is inadequate under both Pi-insufficient and CO₂-enriched treatments. In addition, the Pi and CO₂ treatments used here alter the nitrogen metabolism of barley roots. These findings further emphasize the importance of avoiding nutrient stress during CO₂ enrichment experiments.     

Some characteristics of photosynthetic inorganic carbon uptake of a marine macrophytic red alga

Abstract Some characteristics of photosynthetic inorganic carbon uptake by Palmaria palmata, a marine red macroalga, have been measured under physiological conditions in artificial seawater. The apparent affinity of thallus for CO₂ [K_1/2(CO₂)] at pH 8.0 and 15°C was 21.4±3.0mmol m^?3 CO₂ under air, and 25.7±70mmol m^?3 CO₂ under N₂. The corresponding values of V_max were 2.98 ± 0.42 and 3.65±0.87 mmol O₂ evolved g Chr^?1 s^?l. The apparent K_m(CO₂) of isolated ribulose bisphosphate carboxylase was determined at pH 8.0 and 30 °C to be 30.2 mmol m^?3 CO₂, and the corresponding value of V_max was 19.67 μniol CO₂ g protein^?1 s^?1. The CO₂ compensation points of the thallus were measured in artificial seawater at pH 8.0 under air and N₂, using a gas-chromatographic method. The values were relatively low, rising from 10 cm³ m^?3 at 15°C, to 35 cm³ m^?3 at 25°C, but were not affected by the O₂ concentration. The lack of an effect of O₂ on photosynthesis and on compensation point indicates that there is little photorespiratory CO₂ loss in this macroalga. The high affinity of the thallus for CO₂, and the low CO₂ compensation concentrations, are consistent with the occurrence of bicarbonate uptake in this alga.     

Land-use conversion effects on CO2 emissions: from agricultural to hybrid poplar plantation

Land-use changes such as deforestation have been considered one of the main contributors to increased greenhouse gas emissions, while verifiable C sequestration through afforestation projects is eligible to receive C credits under the Kyoto Protocol. We studied the short-term effects on CO₂ emissions of converting agricultural land-use (planted to barley) to a hybrid poplar (Populus deltoids × Populus × petrowskyana var. Walker) plantation in the Parkland region in northern Alberta, where large areas are being planted to hybrid poplars. CO₂ emissions were measured using a static gas chamber method. No differences were found in soil temperature, volumetric moisture content, or soil respiration rates between the barley and Walker plots. The mean soil respiration rate in 2005 was 1.83 ± 0.19 (mean ± 1 SE) and 1.89 ± 0.13 μmol CO₂ m⁻² s⁻¹ in the barley and Walker plots, respectively. However, biomass production was higher in the barley plots, indicating that the agricultural land-use system had a greater ability to fix atmospheric CO₂. The C balance in the land-use systems were estimated to be a small net gain (before considering straw and grain removal through harvesting) of 0.03 ± 0.187 Mg C ha⁻¹ year⁻¹ in the barley plots and a net loss of 3.35 ± 0.080 Mg C ha⁻¹ year⁻¹ from the Walker poplar plots. Over the long-term, we expect the hybrid poplar plantation to become a net C sink as the trees grow bigger and net primary productivity increases.     

Effects of sustained immobilisation on aphids

Immobilisation of aphids of three species, Schizaphis graminum, Myzus persicae and Dactynotus ambrosiae, was accomplished using low temperature (6 ± 1°C; or nitrogen (N₂), argon (Ar), or carbon dioxide (CO₂) gases at 25°C for 1,3,6, or 24 h. Mortality was recorded and also times until treated aphids first walked or probed. Cold immobilisation caused least mortality and allowed rapid recovery of walking and probing abilities. CO₂ treatment, even when accompanied by cyclic administration of bottled air, caused excessive mortality when used beyond 6 h, and long recovery times. After 24 h immobilisation with CO₂ aphids seldom walked or probed. N₂ or Ar gases, when administered with alternating cycles of bottled air, produced a nontoxic, hypoxic immobilisation from which aphids recovered fairly rapidly to walk and probe, apparently normally.     

Radon fluxes in tropical forest ecosystems of Brazilian Amazonia: night-time CO2 net ecosystem exchange derived from radon and eddy covariance methods

Radon‐222 (Rn‐222) is used as a transport tracer of forest canopy–atmosphere CO₂ exchange in an old‐growth, tropical rain forest site near km 67 of the Tapajós National Forest, Pará, Brazil. Initial results, from month‐long periods at the end of the wet season (June–July) and the end of the dry season (November–December) in 2001, demonstrate the potential of new Rn measurement instruments and methods to quantify mass transport processes between forest canopies and the atmosphere. Gas exchange rates yield mean canopy air residence times ranging from minutes during turbulent daytime hours to greater than 12 h during calm nights. Rn is an effective tracer for net ecosystem exchange of CO₂ (CO₂ NEE) during calm, night‐time hours when eddy covariance‐based NEE measurements are less certain because of low atmospheric turbulence. Rn‐derived night‐time CO₂ NEE (9.00±0.99 μmol m^?2 s^?1 in the wet season, 6.39±0.59 in the dry season) was significantly higher than raw uncorrected, eddy covariance‐derived CO₂ NEE (5.96±0.51 wet season, 5.57±0.53 dry season), but agrees with corrected eddy covariance results (8.65±1.07 wet season, 6.56±0.73 dry season) derived by filtering out lower NEE values obtained during calm periods using independent meteorological criteria. The Rn CO₂ results suggest that uncorrected eddy covariance values underestimate night‐time CO₂ loss at this site. If generalizable to other sites, these observations indicate that previous reports of strong net CO₂ uptake in Amazonian terra firme forest may be overestimated.     

Drought tolerance of photosynthetic electron transport under CO2‐enriched and normal air in cereal species

The quantum yield of photosynthetic electron transport (ΦPSII), evaluated by means of chlorophyll (Chl) fluorescence analysis, has proven to be a useful screening test for drought tolerance in durum wheat (Triticum durum Desf.). To explore the potential of this parameter further in detecting drought-tolerant genotypes, three cereal species were studied; ΦPSII measurements were carried out under two different gas mixtures, at three points of the induction curve (to obtain the maximal ΦPSII and both the transient and steady-state actual ΦPSII), and at three different water stress levels (moderate, severe and drastic). The species investigated were durum and bread wheat (Triticum aestivum L.) and barley (Hordeum vulgare L.); two cultivars per species, characterized by different levels of drought tolerance, were tested. The two gas mixtures used were normal air (21% O₂, 0.035% CO₂ in N₂) to monitor the whole photosynthetic process under physiological conditions, and CO₂ enriched-low O₂ air (1% O₂, 5% CO₂ in N₂) to monitor ΦPSII reduction under stress mainly related to Calvin cycle activity. When ΦPSII related to both assimilatory and non-assimilatory metabolism was evaluated, the cultivar differences observed under normal Air were more representative of the agronomic performance upon drought stress than under high CO₂-low O₂ air. Maximal ΦPSII showed no difference among either cultivars, gas mixtures or stress levels, the efficiency of excitation capture being highly resistant to drought. The ΦPSII evaluated during the transient yielded predictable values in respect of drought tolerance for durum wheat and barley cultivars, highlighting the key role of regulatory processes such as the Mehler peroxidase reaction and possibly also cyclic electron transport, in preventing overreduction under stress. The results clearly show that when Chl fluorescence analysis is used as a parameter in plant breeding, different experimental conditions should be used depending on the physiological mechanism that is bred or selected for.     

Growth of Pseudomonas fluorescens in modified atmosphere packaged tofu

Aims: The objective was to study the growth of Pseudomonas in a food product (tofu) where it typically occurs as a spoilage organism, and when this product is stored under modified atmosphere. Methods and Results: A Pseudomonas strain was isolated from the endogenous microflora of tofu. Tofu was inoculated with the strain, packaged in different gas conditions (air, 100% N₂, 30% CO₂/70% N₂ or 100% CO₂) and stored under refrigerated conditions. Microbial loads and the headspace gas composition were monitored during storage. Conclusions: The strain was capable of growing in atmospheres containing no or limited amounts of oxygen and increased amounts of carbon dioxide. Even when 100% CO₂ was used, growth could not be inhibited completely. Significance and Impact of Study: In contrast to the general characteristics of the genus Pseudomonas (strictly aerobic, highly sensitive to CO₂), it should not be expected in the food industry that removing oxygen from the food package and increasing the carbon dioxide content, combined with cold storage, will easily avoid spoilage by Pseudomonas species. Guarantee of hygienic standards and combination of strategies with other microbial growth inhibiting measures should be implemented.     

CO2 uptake is offset by CH4 and N2O emissions in a poplar short‐rotation coppice

The need for renewable energy sources will lead to a considerable expansion in the planting of dedicated fast‐growing biomass crops across Europe. These are commonly cultivated as short‐rotation coppice (SRC), and currently poplar (Populus spp.) is the most widely planted. In this study, we report the greenhouse gas (GHG) fluxes of carbon dioxide (CO₂), methane (CH₄) and nitrous oxide (N₂O) measured using eddy covariance technique in an SRC plantation for bioenergy production. Measurements were made during the period 2010–2013, that is, during the first two rotations of the SRC. The overall GHG balance of the 4 years of the study was an emission of 1.90 (±1.37) Mg CO₂eq ha^?1; this indicated that soil trace gas emissions offset the CO₂ uptake by the plantation. CH₄ and N₂O contributed almost equally to offset the CO₂ uptake of ?5.28 (±0.67) Mg CO₂eq ha^?1 with an overall emission of 3.56 (±0.35) Mg CO₂eq ha^?1 of N₂O and of 3.53 (±0.85) Mg CO₂eq ha^?1 of CH₄. N₂O emissions mostly occurred during one single peak a few months after the site was converted to SRC; this peak comprised 44% of the total N₂O loss during the two rotations. Accurately capturing emission events proved to be critical for deriving correct estimates of the GHG balance. The nitrogen (N) content of the soil and the water table depth were the two drivers that best explained the variability in N₂O and CH_4, respectively. This study underlines the importance of the ‘non‐CO₂ GHGs’ on the overall balance. Further long‐term investigations of soil trace gas emissions should monitor the N content and the mineralization rate of the soil, as well as the microbial community, as drivers of the trace gas emissions.     

Comparative measurements of chlorophyll a fluorescence, acid accumulation and gas exchange in exposed and shaded plants of Clusia minor L. and Clusia multiflora H. B. K. in the field

 Changes in chlorophyll a fluorescence during the day and diurnal-changes of net CO₂-exchange and organic acid contents were determined in two species of the genus Clusia during the dry season in Venezuela. The investigations included plants of the C₃/CAM intermediate species Clusia minor and the C₃ species C. multiflora growing at exposed and shaded sites. Both species showed a C₃ pattern of net CO₂-exchange at the exposed site. In the shade under extreme drought stress C. minor showed a weak expression of CAM without CO₂-uptake during the afternoon (phase IV of CAM). C. multiflora growing in the shade exhibited a C₃-pattern of net CO₂-exchange and a small but significant nocturnal accumulation of citrate. Shaded plants of C. minor were able to double their light utilisation for electron transport and to reduce non-photochemical quenching during phase III compared to phase II of CAM. Furthermore, increase of electron transport rate through photosystem II in phase III of CAM is correlated to decarboxylation of malate. At the exposed site C. multiflora was less negatively affected by high PPFD than C. minor. This was shown by a lower reduction of potential electron quantum yield (F_v/F_m) and higher light utilisation of electron transport of C. multiflora compared to C. minor. At the exposed site C. minor did not make use of the CAM option to increase light utilisation of electron transport and to reduce non-photochemical quenching as did the plants growing in the shade. Received: 20 March 1996 / Accepted: 24 June 1996     

Similarity of PO2 and PCO2 values in the air cell of eggs of birds and in the alveolar gas of humans at sea level and at high altitude

At the end of incubation, the partial pressures of oxygen and carbon dioxide in the air cell of sea-level avian eggs are similar to those in the expiratory air of adult birds. At high altitude, changes in the permeability of the shell and probably in the embryo metabolism partially compensates the increase in the gas diffusion constant resulting from the low barometric pressure. The aim of this study was to test whether-despite of the adaptive responses of the high altitude avian embryo-the air cell values would be similar to those of the alveolar air of high altitude human natives. Air cell O₂ (48.3±1.6 torr) and CO₂ (20.9±0.85 torr) pressure values were obtained by studying naturally incubated eggs of the Andean gull (Larus serranus)_at 4650m. Sea-level chicken (Gallus gallus) air cell pressure values of O₂ (102.3±2.7 torr) and of CO₂ (43.3±1.3 torr) were obtained from the literature for comparison. Both these values were similar to those found in the alveolar air of humans at sea level (O₂: 104.4±0.4 torr, CO₂:40.1±0.25 torr) and at high altitude (4540 m) (O₂:50.5±0.53 torr, CO₂: 29.1±0.37 torr). Despite very large evolutionary changes in morphology and physiology of the respiratory organs, the head pressure of O₂ that oxygenates the blood keeps a constant value in the pre-pipping avian embryo and in the alveolar air of adult mammals. This constancy holds valid at high altitude.     

Biomass and compositional changes occur in chalk grassland turves exposed to elevated CO2 for two seasons in FACE

Artificial turves composed of 7 chalk grassland species (Festuca ovina L.; Briza media L.; Bromopsis erecta (Hudson) Fourr.; Plantago media L.; Sanguisorba minor Scop.; Anthyllis vulneraria L. and Lotus corniculatus L.) were grown from seed and exposed to two seasons of elevated (600 μmol mol^–1) and ambient (340 μmol mol^–1) CO₂ concentrations in free air CO₂ enrichment (ETH-FACE, Zurich). The turves were clipped regularly to a height of 5 cm and assessed for above ground biomass production and relative abundance based on accumulated clipped dry biomass as well as by point quadrat recording. Below ground biomass production was assessed with root in-growth bags during the second season of growth. Increases in total biomass (> 30%) were noted in elevated CO₂, but the differences did not become significant until the second season of growth. Individual species’ biomass varied in response to elevated CO₂, with significant increases in biomass in elevated CO₂ turves for both legume species, and no significant CO₂ effect on S. minor or P. media. An initial positive CO₂ effect on biomass of combined grass species was reversed by the end of the experiment with less biomass and a significantly smaller proportion of total biomass present in elevated CO₂, which was attributed primarily to changes in proportion of F. ovina. Species relative abundance was significantly affected by elevated CO₂ in the final 4 of the 6 clip events, with the legume species increasing in proportion at the expense of the other species, particularly the grasses. Root length and dry weight were both significantly increased in elevated CO₂ (77% and 89%, respectively), and these increases were greater than increases in shoot biomass (36%) from the same period. Species responses to elevated CO₂, within the model community, were not consistent with predictions made from data on individual species, leading to the conclusion that responses to elevated CO₂, at the community level, and species within the community level, are the result of direct physiological effects and indirect competitive effects. These conclusions are discussed with respect to the ecological responses of natural communities, and the chalk grassland community in particular, to elevated CO₂.     

Effect of Modified Atmospheres on the Growth and Extracellular Enzyme Activities of Psychrotrophs in Raw Milk

The conservation of food products within a controlled atmosphere is efficient in packaging. To extend the cold storage of raw milk, the effects of five gas atmospheres enriched with carbon dioxide and nitrogen were investigated. Treated and control milk were stored at 7 °C for 10 days and analyzed for microbial counts, pH, proteolysis and lipolysis. The addition of CO₂, N₂, or their mixture had a significant inhibitory effect on psychrotrophic growth. The generation times of these microorganisms were significantly longer in treated milk, particularly for yeasts where they amounted to 16.63 h. The maximum inhibition was observed when a gas mixture of 50 % CO₂ and 50 % N₂ was used. As a result, psychrotrophic growth was affected to 98 % whereas this inhibition did not exceed 78 % when CO₂ and 41 % N₂ were applied. Milk treatment under the conditions of 50 % CO₂ and 50 % N₂ gave significantly lower counts for all groups of psychrotrophs being more efficient against Enterobacteriaceae with 99.5 % of inhibition. Storage of raw milk under the tested atmospheres had a different effect on extracellular enzyme productions. Significant decreases in protease and lipase activities were observed during the storage at 7 °C. These enzyme activities were not detectable with pure CO₂ and a 50 % CO₂ and 50 % N₂ mixture. N₂ has shown to be the less efficient treatment against lipases (65 %) and proteases (95 %). With regard to growth, the course of the pH and the protease and lipase activities, the tested gas mixture of 50 % CO₂ and 50 % N₂ was more suitable for extending the shelf life of raw milk.