Water stress,temperature, and light effects on the capacity for isoprene emission and photosynthesis of kudzu leaves

Kudzu (Pueraria lobata (Willd) Ohwi.) is a vine which forms large, monospecific stands in disturbed areas of the southeastern United States. Kudzu also emits isoprene, a hydrocarbon which can significantly affect atmospheric chemistry including reactions leading to tropospheric ozone. We have studied physiological aspects of isoprene emission from kudzu so the ecological consequences of isoprene emission can be better understood. We examined: (a) the development of isoprene emission as leaves developed, (b) the interaction between photon flux density and temperature effects on isoprene emission, (c) isoprene emission during and after water stress, and (d) the induction of isoprene emission from leaves grown at low temperature by water stress or elevated temperature. Isoprene emission under standard conditions of 1000 mol photons·m^-2·s^-1 and 30°C developed only after the leaf had reached full expansion, and was not complete until up to two weeks past the point of full expansion of the leaf. The effect of temperature on isoprene emission was much greater than found for other species, with a 10°C increase in temperature causing a eight-fold increase in the rate of isoprene emission. Isoprene emission from kudzu was stimulated by increases in photon flux density up to 3000 mol photons·m^-2·s^-1. In contrast, photosynthesis of kudzu was saturated at less than 1000 mol·m^-2·s^-1 photon flux density and was reduced at high temperature, so that up to 20% of the carbon fixed in photosynthesis was reemitted as isoprene gas at 1000 mol photons·m^-2·s^-1 and 35°C. Withholding water caused photosynthesis to decline nearly to zero after several days but had a much smaller effect on isoprene emission. Following the relief of water stress, photosynthesis recovered to the prestress level but isoprene emission increased to about five times the prestress rate. At 1000 mol photons·m^-2·s^-1 and 35°C as much as 67% of the carbon fixed in photosynthesis was reemitted as isoprene eight days after water stress. Leaves grown at less than 20°C did not make isoprene until an inductive treatment was given. Inductive treatments included growth at 24°C, leaf temperature of 30°C for 5 h, or witholding water from plants. With the new information on temperature and water stress effects on isoprene emission, we speculate that isoprene emission may help plants cope with stressful conditions.     

Fluorescence correlation spectroscopy with high count rate and low background: analysis of translational diffusion

An epi-illuminated microscope configuration for use in fluorescence correlation spectroscopy in bulk solutions has been analyzed. For determining the effective sample dimensions the spatial distribution of the molecule detection efficiency has been computed and conditions for achieving quasi-cylindrical sample shape have been derived. Model experiments on translational diffusion of rhodamine 6G have been carried out using strong focusing of the laser beam, small pinhole size and an avalanche photodiode in single photon counting mode as the detector. A considerable decrease in background light intensity and measurement time has been observed. The background light is 40 times weaker than the fluorescence signal from one molecule of Rh6G, and the correlation function with signal-to-noise ratio of 150 can be collected in 1 second. The effect of the shape of the sample volume on the autocorrelation function has been discussed. Correspondence to: R. Rigler     

Light quality growth promotion of Spiraea nipponica: the influence of a low photon fluence rate and transfer time to a higher fluence rate

Combinations of different light quality and fluence exposure times were investigated for their effects on in vitro growth of the woody plant Spiraea nipponica. An interaction was demonstrated between different levels of benzyladenine (BA) used for in vitro propagation and the specific light regimes investigated. This relationship was affected by the length of exposure to either white or red/FR light and the time of transfer from one fluence rate to another. In all instances exposure to red/FR light resulted in more extensive growth than under white light. Thus explants cultured under 0.25 and 0.4 mg l^-1 of BA exhibited high shoot proliferation rates when transferred, after 4 weeks of low photon fluence red/FR light, to higher fluence white light for a further week. The proliferation rates obtained were higher than any white light treatment including that with the highest BA level of 0.5 mg l^-1. In addition, the combination of red/FR light exposure with a white light stage of higher fluence improved proliferation at lower exogenous BA levels.     

Real-time monitoring of biomass during Escherichia coli high-cell-density cultivations by in-line photon density wave spectroscopy

An efficient monitoring and control strategy is the basis for a reliable production process. Conventional optical density (OD) measurements involve superpositions of light absorption and scattering, and the results are only given in arbitrary units. In contrast, photon density wave (PDW) spectroscopy is a dilution-free method that allows independent quantification of both effects with defined units. For the first time, PDW spectroscopy was evaluated as a novel optical process analytical technology tool for real-time monitoring of biomass formation in Escherichia coli high-cell-density fed-batch cultivations. Inline PDW measurements were compared to a commercially available inline turbidity probe and with offline measurements of OD and cell dry weight (CDW). An accurate correlation of the reduced PDW scattering coefficient µ_s′ with CDW was observed in the range of 5–69 g L⁻¹ (R² = 0.98). The growth rates calculated based on µ_s′ were comparable to the rates determined with all reference methods. Furthermore, quantification of the reduced PDW scattering coefficient µ_s′ as a function of the absorption coefficient µ_a allowed direct detection of unintended process trends caused by overfeeding and subsequent acetate accumulation. Inline PDW spectroscopy can contribute to more robust bioprocess monitoring and consequently improved process performance.     

Floral scent emission is the highest at the second night of anthesis in Lonicera japonica (Caprifoliaceae)

Floral color change in diverse plants has been thought to be a visual signal reflecting changes in floral rewards, promoting pollinator foraging efficiency as well as plant reproductive success. It remains unclear whether olfactory signals co-vary with floral color change. We investigated the production rhythms of floral scent and nectar associated with floral color change in Lonicera japonica. The flowers generally last 2–3 days. They are white on opening at night (N1) and become light yellow the following day (D1), yellow on the second night (N2), and golden on the second day of flowering (D2). Our measurements in the four stages indicated that nectar production decreased significantly from N1 and D1 to N2 and D2, tracking the floral color change. A total of 34 compounds were detected in floral scent and total scent emission was significantly higher in N2 than in the other three stages. The scent emission of three major compounds, Linalool, cis-3-Hexenyl tiglate, and Germacrene D was also significantly higher in N2, but the relative content of Linalool decreased gradually, cis-3-Hexenyl tiglate increased gradually, and the relative content of Germacrene D did not differ among the four measured stages. Greater scent emission by night than by day suggested a strong olfactory signal to attract nocturnal hawkmoths, the effective pollinators. However, floral scent rhythms in the four stages did not match the color change and nectar secretion, suggesting that floral color (visual) and scent (olfactory) in this species may play different roles in attracting or filtering various visitors.     

ACCUMULATION OF ASTAXANTHIN IN HAEMATOCOCCUS LACUSTRIS (CHLOROPHYTA)1

In N-limited continuous chemostat cultures of the green alga Haematococcus lacustris (Gir.) Rostaf. (UTEX 16), the steady-state astaxanthin content of the cells was determined by the specific growth rate of the cultures. The highest, pigment content was obtained at the lowest dilution rate. The specific rate of astaxanthin accumulation was, however, a function of the photon flux density measured at the illuminated culture surface. In nongrowing Haematococcus cultures, the specific rate of astaxanthin accumulation was determined by the growth rate of the culture during growth phase. The highest possible cellular astaxanthin content of all cultures was comparable and independent of the culture parameters.     

Nitrous oxide emissions from silage maize fields under different mineral nitrogen fertilizer and slurry applications

Intensive dairy farming systems are a large source of emission of the greenhouse gas nitrous oxide (N₂O), because of high nitrogen (N) application rates to grasslands and silage maize fields. The objective of this study was to compare measured N₂O emissions from two different soils to default N₂O emission factors, and to look at alternative emission factors based on (i) the N uptake in the crop and (ii) the N surplus of the system, i.e., N applied minus N uptake by the crop. Twelve N fertilization regimes were implemented on a sandy soil (typic endoaquoll) and a clay soil (typic endoaquept) in the Netherlands, and N₂O emissions were measured throughout the growing season. Highest cumulative fluxes of 1.92 and 6.81 kg N₂O-N ha^–1 for the sandy soil and clay soil were measured at the highest slurry application rate of 250 kg N ha^–1. Background emissions from unfertilized soils were 0.14 and 1.52 kg N₂O-N ha^–1 for the sandy soil and the clay soil, respectively. Emission factors for the sandy soil averaged 0.08, 0.51 and 0.26% of the N applied via fertilizer, slurry, and combinations of both. For the clay soil, these numbers were 1.18, 1.21 and 1.69%, respectively. Surplus N was linearly related to N₂O emission for both the sandy soil (R²=0.60) and the clay soil (R²=0.40), indicating a possible alternative emission factor. We concluded that, in our study, N₂O emission was not linearly related to N application rates, and varied with type and application rate of fertilizer. Finally, the relatively high emission from the clay soil indicates that background emissions might have to be taken into account in N₂O budgets.     

Effect of daily photon receipt on the susceptibility of dwarf bean (Phaseolus vulgaris L.) leaves to photoinhibition of photosynthesis

Bean (Phaseolus vulgaris L.) plants were grown at two light periods of 8 and 13 h with a similar photon flux density (PFD) giving a daily photon receipt (DPR) of 17.9 and 38.2 mol · m–2, respectively. Shoot growth and leaf area development were followed at regular intervals and diurnal whole-plant photosynthesis measured. Single mature trifoliate leaves were exposed to photoinhibitory treatments at PFDs of 800 and 1400 mol · m–2 · s–1 and at temperatures of 12 and 20°C. Chlorophyll fluorescence and photon yields were measured at regular intervals throughout each treatment. Plants grown in 13 h had significantly greater leaf areas than those grown in 8 h. There were no differences in maximum rates of photosynthesis, photon yields and only minor but significant differences in F_v/F_m for plants in the two treatments, showing photosynthetic characteristics were dependent on PFD but not DPR. A significant decline in photosynthesis and F_v/F_m occurred over the 13-h but little change in photosynthesis for plants in the 8 h, indicating some feedback inhibition of photosynthesis was occurring. Plants grown in 8 h were consistently more susceptible to photoinhibition of photosynthesis at all treatments than 13-h plants. Nevertheless, photoinhibition was exacerbated by increases in PFD, and by decreases in temperature for leaves from both treatments. However, for plants from the 8-h day, exposing leaves to 12°C and 1400 mol · m–2 · s–1 caused photo-oxidation and severe bleaching but no visible damage on leaves from 13-h-grown plants. Closure of the photosystem II reaction-centre pool was partially correlated with increasing extents of photoinhibition but the relationship was similar for plants from both treatments. There remains no clear explanation for their wide differences in susceptibility to photoinhibition.Abbreviations and Symbols DPR daily photon receipt - F₀ and F_m initial and maximal fluorescence - F_v/F_m fluorescence ratio in dark-treated leaves - F/F_m intrinsic efficiency of PSII during illumination - PFD photon flux density - _i photon yield (incident basis) - _psi quantum yield of PSII electron transport - P_max maximum rate of photosynthesis - q_N non-photochemical quenching coefficient - q_P photochemical quenching coefficient Many thanks to my colleague William Laing who spent a considerable effort in developing the programme to run the photosynthesis apparatus. I am also indebted to one reviewer with whom I corresponded to resolve some issues in the paper. This project was funded by the New Zealand Foundation for Research, Science and Technology.