Response of Photosynthetic Apparatus of Spring Barley (Hordeum vulgare L.) to Combined Effect of Elevated CO2 Concentration and Different Growth Irradiance

The response of barley (Hordeum vulgare L. cv. Akcent) to various photosynthetic photon flux densities (PPFDs) and elevated [CO₂] [700 μmol (CO₂) mol⁻¹; EC] was studied by gas exchange, chlorophyll (Chl) a fluorescence, and pigment analysis. In comparison with barley grown under ambient [CO₂] [350 μmol (CO₂) mol⁻¹; AC] the EC acclimation resulted in a decrease in photosynthetic capacity, reduced stomatal conductance, and decreased total Chl content. The extent of acclimation depression of photosynthesis, the most pronounced for the plants grown at 730 μmol m⁻² s⁻¹ (PPFD₇₃₀), may be related to the degree of sink-limitation. The increased non-radiative dissipation of absorbed photon energy for all EC plants corresponded to the higher de-epoxidation state of xanthophylls only for PPFD₇₃₀ barley. Further, a pronounced decrease in photosystem 2 (PS2) photochemical efficiency (given as F_V/F_M) for EC plants grown at 730 and 1 200 μmol m⁻² s⁻¹ in comparison with AC barley was related to the reduced epoxidation of antheraxanthin and zeaxanthin back to violaxanthin in darkness. Thus the EC conditions sensitise the photosynthetic apparatus of high-irradiance acclimated barley plants (particularly PPFD₇₃₀) to the photoinactivation of PS2. This revised version was published online in August 2006 with corrections to the Cover Date.     

Different Responses of Norway Spruce Needles from Shaded and Exposed Crown Layers to the Prolonged Exposure to Elevated CO2 Studied by Various Chlorophyll a Fluorescence Techniques

The acclimation depression of capacity of photon utilisation in photochemical reactions of photosystem 2 (PS2) can develop already after three months of cultivation of the Norway spruces (Picea abies [L.] Karst.) under elevated concentrations of CO₂ (i.e., ambient, AC, + 350 µmol(CO₂) mol^–1 = EC) in glass domes with adjustable windows. To examine the role that duration of EC plays in acclimation response, we determined pigment contents, rate of photosynthesis, and parameters of chlorophyll a fluorescence for sun and shade needles after three seasons of EC exposure. We found responses of shaded and exposed needles to EC. Whereas the shaded needles still profited from the EC and revealed stimulated electron transport, for the exposed needles the stimulation of both electron transport activity and irradiance saturated rate of CO₂ assimilation (P _Nmax) under EC already disappeared. No signs of the PS2 impairment were observed as judged from high values of potential quantum yield of PS2 photochemistry (F_V/F_M) and uniform kinetics of Q_A reoxidation for all variants. Therefore, the long-term acclimation of the sun-exposed needles to EC is not necessarily accompanied with the damage to the PS2 reaction centres. The eco-physiological significance of the reported differentiation between the responses of shaded and sun exposed needles to prolonged EC may be in changed contribution of the upper and lower crown layers to the production activity of the tree. Whereas for the AC spruces, P _Nmax of shaded needles was only less than 25 % compared to exposed ones, for the EC spruces the P _Nmax of shaded needles reached nearly 40 % of that estimated for the exposed ones. Thus, the lower shaded part of the crown may become an effective consumer of CO₂.     

Chlorophyll a Fluorescence Response of Norway Spruce Needles to the Long-Term Effect of Elevated CO2 in Relation to Their Position Within the Canopy

The long-term impact of elevated CO₂ concentration on photosynthetic activity of sun-exposed (E) versus shaded (S) foliage was investigated in a Picea abies stand (age 12 years) after three years of cultivation in adjustable-lamella-domes (ALD). One ALD is supplied with either ambient air [ca. 350 µmol(CO₂) mol^–1; AC-variant) and the second with elevated CO₂ concentration [ambient plus 350 µmol(CO₂) mol^–1; EC-variant). The pronounced vertical profile of the photosynthetically active radiation (PAR) led to the typical differentiation of the photosynthetic apparatus between the S- and E-needles in the AC-variant estimated from the irradiance-responses of various parameters of the room temperature chlorophyll (Chl) a fluorescence parameters. Namely, electron transport rate (ETR), photochemical efficiency of photosystem 2, PS2 (_PS2), irradiance-saturated values of non-photochemical quenching of minimum (SV₀) and maximum (NPQ) fluorescence levels, and photochemical fluorescence quenching (q_p) at higher irradiances were all significantly higher for E-needles as compared with the S-ones. The prolonged exposure to EC did not cause any stimulation of ETR for the E-needles but a strongly positive effect of EC on ETR was observed for the S-needles resulting in more than doubled ETR capacity in comparison with S-needles from the AC-variant. For the E-needles in EC-variant a slightly steeper reduction of the _PS2 and q_p occurred with the increasing irradiance as compared to the E-needles of AC-variant. On the contrary, the S-needles in EC variant revealed a significantly greater capacity to maintain a high _PS2 at irradiances lower than 200 µmol m^–2 s^–1 and to prevent the over-reduction of the PS2 reaction centres. Moreover, compared to the AC-variant the relation between SV₀ and NPQ exhibited a strong decrease (up to 72 %) of the SV₀-NPQ slope for the E-needles and an increase (up to 76 %) of this value for the S-needles. Hence the E- and S-foliage responded differently to the long-term impact of EC. Moreover, this exposure was responsible for the smoothing of the PAR utilisation vertical gradient in PS2 photochemical and non-photochemical reactions within the canopy.     

Response of Photosynthesis to Radiation and Intercellular CO2 Concentration in Sun and Shade Shoots of Norway Spruce

Functional differentiation of assimilation activity of sun versus shade foliage was analysed in a Norway spruce monoculture stand (age 15 years). The investigated stand density (leaf area index 8.6) and crown structure led to variation in the photosynthetically active photon flux density (PPFD) within the crowns of the sampled trees. At the saturating PPFD, the maximum rate of CO₂ uptake (P _Nmax) of exposed shoots (E-shoots) was 1.7 times that of the shaded shoots (S-shoots). The apparent quantum yield (α) of E-shoots was 0.9 times that of the S-shoots. A lower ability to use excess energy at high PPFD in photosynthesis was observed in the S-layer. The CO₂- and PPFD-saturated rate of CO₂ uptake (P _Nsat) of the E-shoots was 1.12 times and the carboxylation efficiency (τ) 1.6 times that of the S-shoots. The CO₂-saturated rate of ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBPCO) carboxylation (V_Cmax) and of actual electron transport (J_amax) in the S-needles amounted to 89 and 95 % of V_Cmax and J_amax in the E-needles. Thus, in addition to the irradiation conditions and thus limitation by low J_a, the important limitation of photosynthesis in shade needles is due to carboxylation. This limitation of photosynthesis is accompanied by lower stomatal conductance. This revised version was published online in June 2006 with corrections to the Cover Date.     

Blood-gas,acid-base and catecholamine responses to lactic acid infusion in larval and adult Ambystoma tigrinum

Larval and adult Ambystoma tigrinum were subjected to acidosis by infusing lactic acid (2 M·g^-1) into the peritoneal cavity. Blood was sampled at intervals to establish the time-course of the ensuing acidosis. Both larvae and adults became significantly acidotic after 1 h. The larval acidosis was more pronounced (-4 pH units versus-2 pH units) than adults due to greater extracellular buffering capacity (higher [HCO₃ ^-]) in adults. Both forms recovered in about 8 h. Larvae showed a typical increase in circulating norepinephrine during the initial stages of the acidosis; adults did not, having significantly lower norepinephrine titer than larvae during the acidosis. Both larvae and adults showed transient increases in PO₂ during the acidosis. The ₁ and ₂ antagonists, timolol and butoxamine respectively, (0.2 g·g^-1) were administered to separate groups of larvae. Butoxamine (₂) delayed the recovery from the acidosis by prolonging the increase in arterial PCO₂ and reversing the recovery of [HCO₃ ^-]. Timolol (₁) did not delay recovery. We conclude that ₂ receptors are involved in the catecholamine responses to acidosis in larvae. Catecholamines appear not to play the same role in adult acid-base disturbances as they seem to in larvae.Abbreviations RBC red blood cell     

Expression of a bacterial carotene hydroxylase gene (crtZ) enhances UV tolerance in tobacco

Carotenoids are essential components of the photosynthetic apparatus involved in plant photoprotection. To investigate the protective role of zeaxanthin under high light and UV stress we have increased the capacity for its biosynthesis in tobacco plants (Nicotiana tabacum L. cv. Samsun) by transformation with a heterologous carotenoid gene encoding -carotene hydroxylase (crtZ) from Erwinia uredovora under constitutive promoter control. This enzyme is responsible for the conversion of -carotene into zeaxanthin. Although the total pigment content of the transgenics was similar to control plants, the transformants synthesized zeaxanthin more rapidly and in larger quantities than controls upon transfer to high-intensity white light. Low-light-adapted tobacco plants were shown to be susceptible to UV exposure and therefore chosen for comparative analysis of wild-type and transgenics. Overall effects of UV irradiation were studied by measuring bioproductivity and pigment content. The UV exposed transformed plants maintained a higher biomass and a greater amount of photosynthetic pigments than controls. For revelation of direct effects, photosynthesis, pigment composition and chlorophyll fluorescence were examined immediately after UV treatment. Low-light-adapted plants of the crtZ transgenics showed less reduction in photosynthetic oxygen evolution and had higher chlorophyll fluorescence levels in comparison to control plants. After 1 h of high-light pre-illumination and subsequent UV exposure a greater amount of xanthophyll cycle pigments was retained in the transformants. In addition, the transgenic plants suffered less lipid peroxidation than the wild-type after treatment with the singlet-oxygen generator rose bengal. Our results indicate that an enhancement of zeaxanthin formation in the presence of a functional xanthophyll cycle contributes to UV stress protection and prevention of UV damage.     

PWP2, a member of the WD-repeat family of proteins, is an essentialSaccharomyces cerevisiae gene involved in cell separation

WD-repeat proteins contain four to eight copies of a conserved motif that usually ends with a tryptophan-aspartate (WD) dipeptide. TheSaccharomyces cerevisiae PWP2 gene, identified by sequencing of chromosome III, is predicted to contain eight so-called WD-repeats, flanked by nonhomologous extensions. This gene is expressed as a 3.2-kb mRNA in all cell types and encodes a protein of 104 kDa. ThePWP2 gene is essential for growth because spores carrying thepwp21::HIS3 disruption germinate before arresting growth with one or two large buds. The growth defect ofpwp21::HIS3 cells was rescued by expression ofPWP2 or epitope-taggedHA-PWP2 using the galactose-inducibleGAL1 promoter. In the absence of galactose, depletion of Pwp2p resulted in multibudded cells with defects in bud site selection, cytokinesis, and hydrolysis of the septal junction between mother and daughter cells. In cell fractionation studies, HA-Pwp2p was localized in the particulate component of cell lysates, from which it would be solubulized by high salt and alkaline buffer but not by nonionic detergents or urea. Indirect immunofluorescence microscopy indicated that HA-Pwp2p was clustered at multiple points in the cytoplasm. These results suggest that Pwp2p exists in a proteinaceous complex, possibly associated with the cytoskeleton, where it functions in control of cell growth and separation.     

The mass culture of Dunaliella viridis (Volvocales,Chlorophyta) for oxygenated carotenoids : laboratory and pilot plant studies

The biology, and hence the mass culture, of Dunaliella viridis closely follows that of Dunaliella salina, which is successfully mass cultured for the production of -carotene. Both algae can grow at extremely high salinities and light intensities. They co-exist in the coastal salt lake, Hutt Lagoon, Western Australia. In contrast to D. salina, D. viridis does not accumulate large amounts of -carotene, producing only up to 0.7% of mixed carotenoids (lutein, zeaxathin, other oxygenated carotenoids and -carotene), compared to D. salina's ca 10% dry wt of mainly -carotene. However, in laboratory experiments, D. viridisgrew much faster and to much higher cell densities than D. salina, and attained levels of mixed carotenoids similar to those of D. salina (ca 13 mg L^–1 carotenoid). Preliminary experiments in outdoor ponds were much less promising. Harvesting by chemical flocculation was as effective as with D. salina, but extraction of carotenoids directly into vegetable oil proved inefficient. When incorporated into feed, caretonoids derived from D. viridis pigmented hen eggs acceptably. Extrapolating from laboratory results, and using costing derived from D. salina technology, the cost of production of mixed oxygenated carotenoids from D. viridis was similar to that for the production of -carotene from D. salina, at ca $A500 kg^–1.     

Transient states of geosmin,pigments, carbohydrates and proteins in continuous cultures of Oscillatoria brevis induced by changes in nitrogen supply

Transitions in the growth limiting factor from light (I) to nitrogen (N) and vice versa caused changes in geosmin production, protein and carbohydrate content, and the synthesis of pigments such as chlorophyll a (Chl a), phycobiliproteins (PBPs), and -carotene of the cyanobacterium Oscillatoria brevis. Following IN transition the first 150h, the decrease in protein content was compensated for by an increase of carbohydrates, and thereby, a constant biomass level was maintained in this period. Thereafter, biimass dropped to 15% of its initial level. A decrease in geosmin and pigment content was observed during transition from IN-limited growth. However, geosmin increased relative to phytol (Chl a) and -carotene which may indicate that a lowered demand for phytol and -carotene during N-limited growth allows isoprenoid precursors to be directed to geosmin rather than to pigment synthesis. Synthesis of Chl a and -carotene at the expense of geosmin was suggested for the observed start of increase in geosmin production only at the time that Chl a and -carotene had reached their I-limited steady state. Transition from nitrogen to light limited growth caused an acceleration of metabolism shown by a rapid decrease in carbohydrate content accompanied by an increase in protein content. The growth rate of the organisms temporarily exceeded the dilution rate of the culture and the biomass level increased 6-fold. Due to the only modest changes in geosmin production (2-fold) compared to changes in biomass level (6-fold) during I-or N-limited growth, environmental factors seem to have limited effect on geosmin production.Abbreviations Chl a chlorophyll a - dry wt dry weight; - I-limited light-limited - N-limited nitrogen-limited - PBP phycobiliprotein This research was performed at the Department of Microbiology, University of Amsterdam, with finacial support provided by the Royal Norwegian Ministry of Foreign Affairs and the Royal Norwegian Council for Scientific and Industrial Research     

Changes in germination and respiratory potential of embryos of dormant Grand Rapids lettuce seeds during long-term imbibed storage,and related changes in the endosperm

Grand Rapids lettuce (Lactuca sativa L.) seeds were stored in an imbibed state for up to two years. Embryos dissected from stored seeds showed a progressive loss with time in their ability to germinate on polyethylene glycol (PEG) solutions. Little germination of dissected embryos from one-month imbibed seeds occurred on-6 bar PEG but only after four months of storage did the dissected embryos fail to germinate on-4 bar PEG. After two years storage 30% of dissected embryos still were able to germinate on-2 bar PEG. This loss of germination potential, which may be a symptom of the development of an embryo dormancy, could be reversed by N⁶-benzyladenine (BA) and red light (R) applied together or separately to dissected embryos. Two weeks of chilling of 12-month imbibed seeds restored sensitivity to R and a 48-h BA pretreatment prior to R resulted in germination rates similar to those of seeds emerging from primary dormancy. There was loss of embryo control of endo--mannanase activity after two weeks of storage even though the endosperms themselves retained their capacity for enzyme synthesis for six more weeks. Eventually, then, endo--mannanase synthesis is not possible because of inherent changes in both the embryo and endosperm, although each tissue undergoes changes at its own rate. Oxygen uptake by embryos dissected from two-month imbibed seeds did not increase to the same extent as embryos dissected from freshly imbibed seeds. In intact seeds germinating from a skotodormant state, oxygen uptake increased at a time coincident with radicle protrusion, but did not achieve the levels of uptake of those seeds germinating from a primary dormant state. The decline in uptake of oxygen by secondary dormant seeds is the result of a lowered respiratory capability of the embryo itself, rather than of changes in permeability of the surrounding structures.Abbreviations BA N⁶-benzyladenine - Pfr active (far-redabsorbing) form of phytochrome - R red light - PEG polyethylene glycol