Evaluation of the Effect of Light Intensity on the Measurement of the Photosynthetic Rate in Plankton Microalgae by the Chlorophyll Fluorescence Method

The use of relative variable fluorescence (RVF) of chlorophyll, as measured in the presence of Diuron, an inhibitor of electron transfer, for the estimation of the photosynthetic activity of plankton microalgae was analyzed under a wide range of light intensities in the PAR region. Oxygen evolution rates (estimated by the method of light and dark bottles and the amperometric method), RVF, and chlorophyll a concentration were measured in parallel in natural algal cenoses and microecosystems. When the previously used regression equation, in the form A = b(F/F _d)C _chl I, where A is O₂ evolution rate (g/(m³ h), F/F _d is RVF (relative units), C _chl is chlorophyll a concentration (mg/m³), and I is light intensity (W/m²), was verified in the PAR region, we observed a nonlinear dependence of the correction coefficient b on I, which can be described by the formula b = 6.227 × 10³I. This result agrees with the hypothesis that chlorophyll a fluorescence quenching comprises photochemical (qQ) and energy (qE) components. On the basis of the energy model, we determined the upper limit b _max = 0.003 for light intensity range I< 4.4 W/m² and the lower limit b _min = 0.0003 for I = 400 W/m².     

Diurnal Changes of Gas Exchange,Chlorophyll Fluorescence,and Stomatal Aperture of Hybrid Poplar Clones Subjected to Midday Light Stress

Diurnal changes in net photosynthetic rate (P _N), chlorophyll (Chl) fluorescence, and stomatal aperture of several hybrid poplar clones subjected to midday light stress were measured in July and August of 1996. Midday depression of P _N, photosystem 2 (PS2) efficiency, stomatal conductance (g _s), and stomatal aperture was observed in all clones, though at differing rates among them. Non-uniform stomatal closure occurred at noon and at other times, requiring a modification of intercellular CO₂ concentration (C ₁). A linear relationship was found between g _s and stomatal aperture. More than half of the photons absorbed by PS2 centre dissipated thermally when subjected to light stress at noon. There was a linear relationship between the rate of PS2 photochemical electron transport (PxPFD) and P _N. There was a consensus for two fluorescence indicators (1 – q_P/q_N and (F_m' – F)/F_m') in assessment of susceptibility of photoinhibition in the clones. According to P _N, Chl fluorescence, and stomatal aperture, we conclude that midday depression of photosynthesis can be attributed to both stomatal and non-stomatal limitations.     

Cloning of a cytosolic ascorbate peroxidase gene from Lycium chinense Mill. and enhanced salt tolerance by overexpressing in tobacco

To evaluate the physiological importance of cytosolic ascorbate peroxidase (APX) in the reactive oxygen species (ROS)-scavenging system, a full-length cDNA clone, named LmAPX, encoding a cytosolic ascorbate peroxidase was isolated from Lycium chinense Mill. using homologous cloning, then the expression of LmAPX under salt stress was investigated. After sequencing and related analysis, the LmAPX cDNA sequence was 965 bp in length and had an open reading frame (ORF) of 750 bp coding for 250 amino acids. Furthermore, the LmAPX sequence was sub-cloned into prokaryotic expression vector pET28a and the recombinant proteins had a high expression level in Escherichia coli. Results from a southern blot analysis indicated that three inserts of this gene existed in the tobacco genome encoding LmAPX. Compared with the control plants (wild-type and empty vector control), the transgenic plants expressing the LmAPX gene exhibited lower amount of hydrogen peroxide (H₂O₂) and relatively higher values of ascorbate peroxidase activity, proline content, and net photosynthetic rate (P_n) under the same salt stress. These results suggested that overexpression of the LmAPX gene could decrease ROS production caused by salt stress and protect plants from oxidative stress.     

Luminescence enhancement of the catalytic 19 kDa protein (KAZ) of Oplophorus luciferase by three amino acid substitutions

To characterize the luminescence properties of nanoKAZ, a 16 amino acid substituted mutant of the catalytic 19 kDa protein (KAZ) of Oplophorus luciferase, the effects of each mutated amino acid were investigated by site-specific mutagenesis. All 16 single substituted KAZ mutants were expressed in Escherichia coli cells and their secretory expressions in CHO-K1 cells were also examined using the signal peptide sequence of Gaussia luciferase. Luminescence activity of KAZ was significantly enhanced by single amino acid substitutions at V44I, A54I, or Y138I. Further, the triple mutant KAZ-V44I/A54I/Y138I, named eKAZ, was prepared and these substitutions synergistically enhanced luminescence activity, showing 66-fold higher activity than wild-KAZ and also 7-fold higher activity than nanoKAZ using coelenterazine as a substrate. Substrate specificity of eKAZ for C2- and/or C6-modified coelenterazine analogues was different from that of nanoKAZ, indicating that three amino acid substitutions may be responsible for the substrate recognition of coelenterazine to increase luminescence activity. In contrast, these substitutions did not stimulate protein secretion from CHO-K1 cells, suggesting that the folded-protein structure of KAZ might be different from that of nanoKAZ.     

Effects of DDT on photosynthetic electron flow in Secale species

Following a survey of a range of varieties of rye, mainly Secale cereale, for reaction to DDT, the mode of action of the pesticide in a susceptible variety was studied. Two sites of interaction of DDT with the photosynthetic electron transport chain were demonstrated. The first site of inhibition was on the oxidizing side of photosystem 2, between the sites of electron donation from diphenylcarbazide at pH 6.0 and pH 8.0 in Tris-washed chloroplasts. The second site of DDT inhibition was in the intermediate electron transport chain, and was demonstrated by using dichlorophenol-indophenol and phenyldiamines as electron donors in chloroplasts where electron flow from photosystem 2 was inhibited by 3-(3,4-dichlorophenyl)-1,1-dimethylurea. The sites are distinct from those characteristic of herbicides which affect photosynthetic electron flow.     

Lemna minor tolerance to metal‐working fluid residues: implications for rhizoremediation

For the first time in the literature, duckweed (Lemna minor) tolerance (alone or in combination with a consortium of bacteria) to spent metal‐working fluid (MWF) was assessed, together with its capacity to reduce the chemical oxygen demand (COD) of this residue. In a preliminary study, L. minor response to pre‐treated MWF residue (ptMWF) and vacuum‐distilled MWF water (MWFw) was tested. Plants were able to grow in both residues at different COD levels tested (up to 2300 mg·l^?1), showing few toxicity symptoms (mainly growth inhibition). Plant response to MWFw was more regular and dose responsive than when exposed to ptMWF. Moreover, COD reduction was less significant in ptMWF. Thus, based on these preliminary results, a second study was conducted using MWFw to test the effectiveness of inoculation with a bacterial consortium isolated from a membrane bioreactor fed with the same residue. After 5 days of exposure, COD in solutions containing inoculated plants was significantly lower than in non‐inoculated ones. Moreover, inoculation reduced β+γ‐tocopherol levels in MWFw‐exposed plants, suggesting pollutant imposed stress was reduced. We therefore conclude from that L. minor is highly tolerant to spent MWF residues and that this species can be very useful, together with the appropriate bacterial consortium, in reducing COD of this residue under local legislation limits and thus minimise its potential environmental impact. Interestingly, the lipophilic antioxidant tocopherol (especially the sum of β+γ isomers) proved to be an effective plant biomarker of pollution.     

Photosynthesis,growth, and decay traits in Sphagnum – a multispecies comparison

Peat mosses (Sphagnum) largely govern carbon sequestration in Northern Hemisphere peatlands. We investigated functional traits related to growth and decomposition in Sphagnum species. We tested the importance of environment and phylogeny in driving species traits and investigated trade‐offs among them. We selected 15 globally important Sphagnum species, representing four sections (subgenera) and a range of peatland habitats. We measured rates of photosynthesis and decomposition in standard laboratory conditions as measures of innate growth and decay potential, and related this to realized growth, production, and decomposition in their natural habitats. In general, we found support for a trade‐off between measures of growth and decomposition. However, the relationships are not strong, with r ranging between 0.24 and 0.45 for different measures of growth versus decomposition. Using photosynthetic rate to predict decomposition in standard conditions yielded R² = 0.20. Habitat and section (phylogeny) affected the traits and the trade‐offs. In a wet year, species from sections Cuspidata and Sphagnum had the highest production, but in a dry year, differences among species, sections, and habitats evened out. Cuspidata species in general produced easily decomposable litter, but their decay in the field was hampered, probably due to near‐surface anoxia in their wet habitats. In a principal components analysis, PCA, photosynthetic capacity, production, and laboratory decomposition acted in the same direction. The species were imperfectly clustered according to vegetation type and phylogeny, so that some species clustered with others in the same section, whereas others clustered more clearly with others from similar vegetation types. Our study includes a wider range of species and habitats than previous trait analyses in Sphagnum and shows that while the previously described growth–decay trade‐off exists, it is far from perfect. We therefore suggest that our species‐specific trait measures offer opportunities for improvements of peatland ecosystem models. Innate qualities measured in laboratory conditions translate differently to field responses. Most dramatically, fast‐growing species could only realize their potential in a wet year. The same species decompose fast in laboratory, but their decomposition was more retarded in the field than that of other species. These relationships are crucial for understanding the long‐term dynamics of peatland communities.     

Potential effects of interaction between CO2 and temperature on forest landscape response to global warming

Projected temperature increases under global warming could benefit southern tree species by providing them the optimal growing temperature and could be detrimental to northern species by exposing them to the supra optimal growing temperatures. This benefit-detriment trade-off could increase the competitive advantage of southern species in the northern species range and cause the increase or even dominance of southern species in the northern domain. However, the optimum temperature for photosynthesis of C3 plants may increase due to CO₂ enrichment. An increase in the optimum temperature could greatly reduce the benefit-detriment effect. In this study, we coupled a forest ecosystem process model (PnET-II) and a forest GAP model (LINKAGES) with a spatially dynamic forest landscape model (LANDIS-II) to study how an optimum temperature increase could affect forest landscape response due to global warming. We simulated 360 years of forest landscape change in the Boundary Water Canoe Area (BWCA) in northern Minnesota, which is transitional between boreal and temperate forest. Our results showed that, under the control scenario of continuing the historic 1984–1993 mean climate (mainly temperature, precipitation and CO₂), the BWCA will become a spruce-fir dominated boreal forest. However, under the scenario of predicted climatic change [the 2000–2099 climates are predicted by Canadian Climate Center (CCC), followed by 200 years of continuing the predicted 2090–2099 mean climate], the BWCA will become a pine-dominated mixed forest. If the optimum temperature increases gradually with [CO₂] (the increase in optimum temperature is assumed to change gradually from 0 °C in year 2000 to 5 °C in year 2099 when [CO₂] reaches 711 ppm and stabilizes at 5 °C after year 2099), the BWCA would remain a fir-dominated boreal forest in areas with relatively high water-holding capacity, but not in areas with relatively low water-holding capacity. Our results suggest that the [CO₂] induced increases in optimum temperature could substantially reduce forest landscape change caused by global warming. However, not all tree species would be able to successfully adapt to future warming as predicted by CCC, regardless of optimum temperature acclimations.     

Novel recombinant human lactoferrin: Differential activation of oxidative stress related gene expression

Lactoferrin, an iron-binding protein found in high concentrations in mammalian exocrine secretions, is an important component of the host defense system. It is also a major protein of the secondary granules of neutrophils from which is released upon activation. Due to its potential clinical utility, recombinant human lactoferrin (rhLF) has been produced in various eukaryotic expression systems; however, none of these are fully compatible with humans. Most of the biopharmaceuticals approved by the FDA for use in humans are produced in mammalian expression systems. The Chinese hamster ovary cells (CHO) have become the system of choice for proteins that require post-translational modifications, such as glycoproteins.