Microcalorimetric Investigation on Effects of LaIII on Metabolic Activity of Mitochondria Isolated from Hybrid Rice

Rare earth elements (REEs) have beneficial influence on plant growth and are widely used in agriculture practice, but little is known about the behavior of the REEs in mitochondria of plant cell. Thermogenic metabolic curves were determined by the ampoule method at 303 K using a TAM air isothermal microcalorimeter in mitochondria isolated from hybrid rice Liangyoupeijiu (Oryza sativa L.), and the effect of La^III on its mitochondrial metabolism was investigated. From the obtained heat flux curves, the crucial parameters including as activity recovery rate constant (k) and maximum heat power (P_max) were determined. Application of La³⁺ in concentrations ranging from 0 to 120 μg/ml significantly increased k and P_max values with the high point reaching 346 and 222% of the control, respectively. Concentrations from 140–150 μg/ml had the opposite effect. These results were consistent with previous reports on the effects of REEs on plant growth. It was concluded that the La‐induced change of mitochondrial metabolic activity is a possible mechanism by which La^III ions influences hybrid rice growth.     

Photosynthetic and biochemical activities in flag leaves of a newly developed superhigh-yield hybrid rice (<Emphasis Type="Italic">Oryza sativa</Emphasis>) and its parents during the reproductive stage

Responses of net photosynthetic rates to intercellular CO₂ concentration (P _n/C _i curves) and photochemical characteristics were investigated in flag leaves of newly developed superhigh-yield hybrid rice (Oryza sativa L.) LiangYouPeiJiu (LYPJ) and its maternal PeiAi64S (PA64S) and paternal WuMang9311 (WM9311) lines grown in the field during the reproductive stage. The results showed that photosynthetic functions, such as the electron transport activities of photosystems and photophosphorylation, assessed in vivo from P _n/C _i curves under field conditions declined more or earlier than those obtained in vitro. The degradation of polypeptides of thylakoid membranes was slower than those for P _Ca=360 (light-saturated net photosynthetic rate measured at 360 μmol mol⁻¹) and CE (carboxylation efficiency, obtained from the initial slope of the P _n/C _i curve). The initial inhibition of the PSII electron transport and oxygen-evolving activity induced by senescence occurred before the degradation of the oxygen-evolving complex. In comparison, LYPJ had intermediate photosynthetic functions in the early stage of leaf development, but greater photochemical activities in the mid and late stages. WM9311 showed a similar pattern of changes but lower values, and PA64S had higher values in the early stage but showed a faster rate of senescence than LYPJ. These findings implied that the hybrid LYPJ demonstrated intermediate photosynthetic activities between its parents in the early stage of leaf development, whereas it had higher photosynthetic activities than its parents in the mid and late stages, which may be responsible for its high yield.     

Effects of water stress on photosynthetic activity,dry mass partitioning and some associated metabolic changes in four provenances of neem (<Emphasis Type="Italic">Azadirachta indica</Emphasis> A. Juss)

Chlorophyll (Chl) content, dry mass, relative water content (RWC), leaf mass per area (LMA), proline (Pro) content, malondialdehyde (MDA) content, superoxide dismutase (SOD) and peroxidase (POD) activity, P _N-PAR response curves and gas exchange were studied to determine the effects of water stress on photosynthetic activity, dry mass partitioning and metabolic changes in four provenances of neem (Azadirachta indica A. Juss). The results indicated that provenance differences existed in the adaptation response to water stress that included changes to growth strategies coupled with ecophysiological and metabolic adjustments. As water stress increased, stomatal conductance (g _s), net photosynthetic rate (P _N), transpiration rate (E), and leaf RWC decreased while LMA increased in all provenances. Dry mass was reduced in droughted plants and the percentage increased in dry mass allocated to roots, and enzyme activities of SOD and POD were highest in neem originating from Kalyani (KA) provenance and lowest in neem originating from New Dehli (ND) provenance. In contrast, water stress increased MDA content least in KA and most in ND. Furthermore, neem originating from ND also had the greatest decrease in Chl a/b ratio while the ratio was least affected in neem originating from KA. These findings suggest neem originating from KA may have more drought resistance than neem originating from ND. The data from P _N-PAR response curves are less clear. While these curves showed that drought stress increased compensation irradiance (I _c) and dark respiration (R _D) and decreased saturation irradiance (I _s) and maximum net photosynthetic rate (P _max), the extent of decline in P _max was provenance dependent. P _max under non-waterlimiting conditions was higher in neem originating from Jodhpur (MA) (about 14 μmol m⁻² s⁻¹) than in the other three provenances (all about 10 μmol m⁻² s⁻¹), but mild water stress had minimal effect on P _max of these three provenances whereas P _max of MA provenance declined to 10 μmol m⁻² s⁻¹, i.e. a similar value. However, under severe water stress P _max of MA and KA provenances had declined to 40% of non-stressed values (about 6 and 4 μmol m⁻² s⁻¹, respectively) whereas the decline in P _max of neem originating from Kulapachta (KU) and ND provenances was about 50% of nonstressed values (about 5 μmol m⁻² s⁻¹). These data suggest the P _N responses of KU and ND provenances are most tolerant, and KA and MA least tolerant to increasing water stress, but also suggest MA provenance could be the most desired under both non-water-limiting and water-limiting conditions due to highest P _max in all conditions.     

Microcalorimetric study of the effect of Ce(III) on metabolic activity of mitochondria isolated from indice rice 9311

Rare earth elements (REEs) have beneficial influence on plant growth and are widely used in agriculture practice, but little is known about behavior of the REEs on mitochondria in plant cells. Thermogenic metabolic curves were determined by the ampoule method at 303 K using a TAM air isothermal microcalorimeter in mitochondria isolated from indice rice 9311 (Oryza sativa L.), and the effect of Ce(III) on mitochondrial metabolism was investigated. By analyzing the obtained heat flux curves, the crucial parameters such as activity recovery rate constant (k) and maximum heat power (P(m)) were investigated. Application of Ce3+ in concentrations ranging from 0 to 120 microg/ml significantly increased k and P(m) values, with the maximum reaching 261 and 180% of the control, respectively. Concentrations from 140 to 150 microg/ml had the opposite effect. These results were consistent with previous reports on the effects of REEs on plant growth. It was concluded that the Ce(III)-induced change of mitochondrial metabolic activity is a possible mechanism by which Ce(III) influenced indice rice 9311 growth.     

Molecular characterization of marker-free transgenic lines of <Emphasis Type="Italic">indica</Emphasis> rice that accumulate carotenoids in seed endosperm

A single Agrobacterium strain harbouring two binary plasmids was successfully used for the first time to develop a marker-free transgenic rice of improved nutritional value. Sixty-eight T₀ co-transformants were obtained in three indica rice cultivars—two popular high-yielding Bangladeshi varieties (BR28 and BR29), and one high-iron rice cultivar (IR68144). Marker-free lines were obtained from 14 out of 24 selected co-transformants screened in the T₁ generation. The accumulation of total carotenoids in polished T₂ rice seeds of the primary transgenic VPBR29-17-37 reached levels of up to 3.0 μg/g, with the level of β-carotene reaching 1.8 μg/g. In the cultivars BR28 and IR68144, total carotenoid levels in the transformants reached 2.0 μg/g of polished rice seeds. The levels of lutein and other carotenoids in the seeds were also significantly enhanced. T₁ plants obtained from primary transgenics with simple gene-integration patterns tended to have a lower carotenoid content than the original parental lines. This study describes the development of marker-free transgenic rice lines containing high levels of carotenoids, and addresses the relationship between the rearrangement of transgenes and the presence of metabolic end products in transgenic rice.     

Subcellular distribution of selenium during uptake and its influence on mitochondrial oxidations in germinatingVigna radiata L

The metabolic significance of Se in plants is not well documented, though the presence of many selenoenzymes in bacteria and the essentiality of Se in higher animals is established. Since germination is an active process in plant growth and metabolism, the effect of Se was investigated in germinatingVigna radiata L, a nonaccumulating Sedeficient legume. Growth and protein were enhanced in seedlings supplemented with selenium (Se) as sodium selenite in the medium up to 1 μg/mL. The pattern of uptake of⁷⁵Se in the differentiating tissues and the subcellular distribution were investigated. The percentage of incorporation of⁷⁵Se was greater in the mitochondria at the lowest level (0.5 μg/mL) of Se supplementation compared to higher levels of Se exposure. Proteins precipitated from the postmitochondrial supernatant fractions, when separated by means of polyacrylamide gel electrophoresis (PAGE), indicated a major selenoprotein in the seedlings germinated at 2.0 μg/mL Se. In seedlings grown with supplemented Se, enhanced respiratory control ratio and succinate dehydrogenase activity were observed in the mitochondria of tissues, indicative of a role for Se in mitochondrial membrane functions.     

Effect of Rice Straw Incorporation on Phytotoxicity of Isoxaflutole to an Exotic Weed Phalaris minor Retz.

Phalaris minorRetz. is a major exotic annual weed in the wheat (Triticum aestivum L.) crop. Unharvested rice (Oryza sativa L.) straw, unburned and burned, is often incorporated in the field prior to cultivating wheat. Isoxaflutole (Balance), a pre-emergent systemic soil applied herbicide, has potential to control P. minor. Glasshouse experiments were conducted to determine the phytotoxicity of isoxaflutole defined by reductions in relation to shoot length of P. minor when grown in unamended soil or soil amended with unburned or burned rice straw. A 120 g soil was amended with 0, 1, 2 and 4 g of unburned or burned rice straw, and placed in 150 mL styrofoam pots. Appropriate amount of isoxaflutole (75% active ingredient, ai) was added to pots to get final concentration of 0, 7.5, 30, 60 and 120 μg ai/pot. Unamended soil and soil amended with unburned or burned rice straw were analyzed for pH and organic matter; two important determinants of isoxaflutole activity. Results indicate a significant reduction in shoot length of P. minor when grown in soil treated with isoxaflutole at 30, 60 or 120 μg ai/pot. Inhibition in the shoot length of P. minor was observed when soil amended with unburned straw was treated with isoxaflutole at 7.5 and 30 μg ai/pot compared with unamended soil treated with similar amounts of isoxaflutole. No significant change in isoxaflutole toxicity was observed when soil amended with unburned straw was treated with isoxaflutole at 60 and 120 lg ai/pot compared with unamended soil treated with similar amounts of isoxaflutole. Isoxaflutole phytotoxicity to P. minor shoot length was eliminated when soil amended with burned straw was treated with isoxaflutole at 7.5 and 30 μg ai/pot. P. minor shoot length was greater when soil amended with burned straw was treated with isoxaflutole at 60 and 120 μg ai/pot relative to herbicide-treated unamended soils. We conclude that incorporation of burned rice straw greatly reduces the phytotoxicity of isoxaflutole toP. minor.     

Photosynthetic Response of Carrots to Varying Irradiances

Response to irradiance of leaf net photosynthetic rates (P _N) of four carrot cultivars: Cascade, Caro Choice (CC), Oranza, and Red Core Chantenay (RCC) were examined in a controlled environment. Gas exchange measurements were conducted at photosynthetic active radiation (PAR) from 100 to 1 000 μmol m⁻² s⁻¹ at 20 °C and 350 μmol (CO₂) mol⁻¹(air). The values of P _N were fitted to a rectangular hyperbolic nonlinear regression model. P _N for all cultivars increased similarly with increasing PAR but Cascade and Oranza generally had higher P _N than CC. None of the cultivars reached saturation at 1 000 μmol m⁻² s⁻¹. The predicted P _N at saturation (P _Nmax) for Cascade, CC, Oranza, and RCC were 19.78, 16.40, 19.79, and 18.11 μmol (CO₂) m⁻² s⁻¹, respectively. The compensation irradiance (I _c) occurred at 54 μmol m⁻² s⁻¹ for Cascade, 36 μmol m⁻² s⁻¹ for CC, 45 μmol m⁻² s⁻¹ for Oranza, and 25 μmol m⁻² s⁻¹ for RCC. The quantum yield among the cultivars ranged between 0.057–0.033 mol(CO₂) mol⁻¹(PAR) and did not differ. Dark respiration varied from 2.66 μmol m⁻² s⁻¹ for Cascade to 0.85 μmol m⁻² s⁻¹ for RCC. As P _N increased with PAR, intercellular CO₂ decreased in a non-linear manner. Increasing PAR increased stomatal conductance and transpiration rate to a peak between 600 and 800 μmol m⁻² s⁻¹ followed by a steep decline resulting in sharp increases in water use efficiency. This revised version was published online in August 2006 with corrections to the Cover Date.     

Characteristics of Single, Large-Conductance Calcium-Dependent Potassium Channels (BKCa) from Smooth Muscle Cells Isolated from the Rabbit Mesenteric Artery

Smooth muscle cells isolated from the secondary and tertiary branches of the rabbit mesenteric artery contain large Ca²⁺-dependent channels. In excised patches with symmetrical (140 mm) K⁺ solutions, these channels had an average slope conductance of 235 ± 3 pS, and reversed in direction at −6.1 ± 0.4 mV. The channel showed K⁺ selectivity and its open probability (P _o ) was voltage-dependent. Iberiotoxin (50 nm) reversibly decreased P _o , whereas tetraethylammonium (TEA, at 1 mm) reduced the unitary current amplitude. Apamin (200 nm) had no effect. The channel displayed sublevels around 1/3 and 1/2 of the mainstate level. The effect of [Ca²⁺] on P _o was studied and data fitted to Boltzmann relationships. In 0.1, 0.3, 1.0 and 10 μm Ca²⁺, V _1/2 was 77.1 ± 5.3 (n= 18), 71.2 ± 4.8 (n= 16), 47.3 ± 10.1 (n= 11) and −14.9 ± 10.1 mV (n= 6), respectively. Values of k obtained in 1 and 10 μm [Ca²⁺] were significantly larger than that observed in 0.1 μm [Ca²⁺]. With 30 μm NS 1619 (a BK_Ca channel activator), V _1/2 values were shifted by 39 mV to the left (hyperpolarizing direction) and k values were not affected. TEA applied intracellularly, reduced the unitary current amplitude with a K _d of 59 mm. In summary, BK_Ca channels show a particularly weak sensitivity to intracellular TEA and they also display large variation in V _1/2 and k. These findings suggest the possibility that different types (isoforms) of BK_Ca channels may exist in this vascular tissue. Received: 22 December 1997/Revised: 27 March 1998     

Microcalorimetric study of the toxic effect of selenium on the mitochondrial metabolism of cyprinus carpio liver

The metabolic thermograms and heat output of mitochondria isolated from carp liver have been determined by using an LKB bioactivity monitor. The thermogram can be divided into four parts: the lag phase, active recovery phase, stationary phase, and decline phase. The thermokinetic equation was established for the active recovery and decline phase of metabolism as follows: dP/dt =k _mP (1-SP). The rate constantsk ₁ andk ₂ of two phases of active recovery and decline phase have been also calculated. The metabolism activity of mitochondrial inhibited by a high concentration of trace element selenium has been studied. The metabolic heat released, time of each phase, and rate constants can be significantly influenced by excess of selenite added. These results suggested that a high concentration of selenium can damage the structure and function of mitochondria, and thus influence their metabolism.     

Nitric Oxide Activates or Inhibits Skeletal Muscle Ryanodine Receptors Depending on Its Concentration, Membrane Potential and Ligand Binding

We show that rabbit skeletal RyR channels in lipid bilayers can be activated or inhibited by NO, in a manner that depends on donor concentration, membrane potential and the presence of channel agonists. 10 μm S-nitroso-N-acetyl-penicillamine (SNAP) increased RyR activity at −40 mV within 15 sec of addition to the cis chamber, with a 2-fold increase in frequency of channel opening (F _o ). 10 μm SNAP did not alter activity at +40 mV and did not further activate RyRs previously activated by 2 mm cis ATP at +40 or −40 mV. In contrast to the increase in F _o with 10 μm SNAP, 1 mm SNAP caused a 2-fold reduction in F _o but a 1.5-fold increase in mean open time (T _o ) at −40 mV in the absence of ATP. 1 mm SNAP or 0.5 mm sodium nitroprusside (SNP) induced ∼3-fold reductions in F _o and T _o at +40 or −40 mV when channels were activated by 2 mm cis ATP or in channels activated by 6.5 μm peptide A at −40 mV (peptide A corresponds to part of the II–III loop of the skeletal dihydropyridine receptor). Both SNAP-induced activation and SNAP/SNP-induced inhibition were reversed by 2 mm dithiothreitol. The results suggest that S-Nitrosylation or oxidation of at least three classes of protein thiols by NO each produced characteristic changes in RyR activity. We propose that, in vivo, initial release of NO activates RyRs, but stronger release increases [NO] and inhibits RyR activity and contraction. Received: 27 August 1999/Revised: 25 October 1999     

Mg-Dependent, Zn-ATPase: Enzymatic Characteristics, Ion Specificities and Tissue Distribution

Mucosal crude microsomes, prepared from proximal rat small intestine, exhibited significant Mg-dependent, Zn-ATPase activity; V _max = 23 μmoles Pi/mg protein/hr, K _m = 160 nm, and Hill Coefficient, n= 1.5. Partial purification (∼10-fold) was achieved by detergent extraction, and centrifugation through 250 mm sucrose: V _max = 268 units, K _m = 1 nm, and n= 6. In partially purified preparations, the assay was linear with time to 60 min, and with protein concentration to 1 μg/300 μl. Activities at pH 8 and 8.5 were higher than at pH 7.2. The ATP K _m was 0.7 mm, with an optimal ATP/Mg ratio of ∼2. Ca elicited ATPase activity but did not augment the Zn-dependent activity. In partially purified preparations, the homologous salts of Co, Cd, Cu, and Mn exhibited no detectable activity. Vanadate inhibition studies yielded two component kinetics with a K _i of 12 μm for the first component, and 96 μm for the second component, in partially purified preparations. Tissue distribution analyses revealed gradients of activity. In the proximal half of the small intestine, Mg/Zn activity increased progressively from crypt to villus tip. In long axis studies, this activity decreased progressively from proximal to distal small bowel. Received: 12 September 2000/Revised 6 January 2001     

The metal ion requirements of Arabidopsis thaliana Glx2-2 for catalytic activity

In an effort to better understand the structure, metal content, the nature of the metal centers, and enzyme activity of Arabidopsis thaliana Glx2-2, the enzyme was overexpressed, purified, and characterized using metal analyses, kinetics, and UV–vis, EPR, and ¹H NMR spectroscopies. Glx2-2-containing fractions that were purple, yellow, or colorless were separated during purification, and the differently colored fractions were found to contain different amounts of Fe and Zn(II). Spectroscopic analyses of the discrete fractions provided evidence for Fe(II), Fe(III), Fe(III)–Zn(II), and antiferromagnetically coupled Fe(II)–Fe(III) centers distributed among the discrete Glx2-2-containing fractions. The individual steady-state kinetic constants varied among the fractionated species, depending on the number and type of metal ion present. Intriguingly, however, the catalytic efficiency constant, k _cat/K _m, was invariant among the fractions. The value of k _cat/K _m governs the catalytic rate at low, physiological substrate concentrations. We suggest that the independence of k _cat/K _m on the precise makeup of the active-site metal center is evolutionarily related to the lack of selectivity for either Fe versus Zn(II) or Fe(II) versus Fe(III), in one or more metal binding sites.     

Modulation of the Permeability Transition Pore by Inhibition of the Mitochondrial KATP Channel in Liver vs. Brain Mitochondria

Inhibition of the mitochondrial K_ATP (mitoK_ATP) channel abrogates the beneficial effects of preconditioning induced by a brief episode of sublethal ischemia. We studied the effect of 5-hydroxydecanoate, a well-known inhibitor of the mitoK_ATP channel, on swelling of isolated liver and brain mitochondria. Volume changes were determined by measurement of light absorbance at 540 nm. Mitochondrial swelling induced by adding Ca²⁺ ions correlated with opening of the permeability transition pore as shown by modulation by 1 μM cyclosporin A. In brain mitochondria, 5-hydroxydecanoate did not significantly affect Ca²⁺-induced swelling. In contrast, 50 or 500 μM 5-hydroxydecanoate increased swelling of liver mitochondria by 9.7 ± 5.1% (n = 6, P = 0.057) and 29.4 ± 1.4% (n = 5, P < 0.0001), respectively. The effect of 5-hydroxydecanoate was blocked by cyclosporin A and was dependent on the presence of potassium in the medium. In medium containing 200 μM ATP to inhibit the mitoK_ATP channel, 5–hydroxydecanoate did not further increase Ca²⁺-induced swelling. We conclude that inhibition of the mitoK_ATP channel exerts its detrimental effect by facilitation of permeability transition pore opening.     

A mathematical model for describing light-response curves in <Emphasis Type="Italic">Nicotiana tabacum</Emphasis> L.

A modified exponential model was used to describe light-response curves of Nicotiana tabacum L. The accuracies of an exponential model, a nonrectangular hyperbola model, a rectangular hyperbola model, a modified rectangular hyperbola model and the modified exponential model were evaluated by Mean square error (MSE) and Mean absolute error (MAE). The tests MSE and MAE of the modified exponential model were the lowest among the five models. The light saturation point (LSP) obtained by the exponential model, the nonrectangular hyperbola model and the rectangular hyperbola model were much lower than the measured values, and the maximum net photosynthetic rates (P _max) calculated from these models, were greater than the measured values. P _max at LSP of 1,077 μmol m⁻² s⁻¹ calculated by the modified exponential model was 12.34 μmol(CO₂) m⁻² s⁻¹, which was more accurate than the values obtained from the modified rectangular hyperbola model. The results show that the modified exponential model is superior to other models for describing light-response curves.     

Ceramide induces release of mitochondrial proapoptotic proteins in caspase-dependent and -independent manner in HT-29 cells

In this study, the release of mitochondrial proapoptotic intermembrane space proteins induced by ex-ogenous C2-ceramide in human colon carcinoma (HT-29) cell line was investigated. HT-29 cells were treated with 12.5, 25 and 50 μmol/L C2-ceramide in vitro. Flow cytometer was used to detect the mito-chondrial membrane potential (ΔΨm). Subcellular fractions were extracted by Mitochondrial/Cytosol Fractionation Kit after C2-ceramide treatment for 24 h. SDS-PAGE was used to determine the level of cytochrome c (Cyt c), high temperature requirement A2 (HtrA2) and second mitochondrial-derived ac-tivator of caspases (Smac) released from mitochondria, the expression of X-linked inhibitor of apop-tosis protein (XIAP) and caspase-3 for 24 h. The results showed that ΔΨm began to decrease from 6 h after 25 and 50 μmol/L C2-ceramide treatment (P<0.05) and cyclosporin A (CsA) could inhibit the col-lapse of ΔΨm through regulating mitochondrial membrane permeability transition pore. There was no effect of C2-ceramide on the expression of Cyt c, HtrA2 and Smac in the total levels. 12.5, 25 and 50 μmol/L C2-ceramide could induce Cyt c, HtrA2 and Smac to release from mitochondria to cytosol and down-regulate the expression of XIAP (P<0.05). Also there was expression of cleaved caspase-3 with C2-ceramide treatment. After the treatment with caspase inhibitor, C2-ceramide still induced the release of Cyt c and HtrA2, but Smac did not. Therefore, C2-ceramide could induce apoptosis of HT-29 cells through the mitochondria pathway. The release of Cyt c, HtrA2 and Smac from mitochondria did not occur via the same mechanism, the release of Cyt c and HtrA2 was caspase-independent and the re-lease of Smac was caspase-dependent.     

Photosynthetic acclimation to different light levels in the brown marine macroalga, <Emphasis Type="Italic">Hizikia fusiformis</Emphasis> (Sargassaceae,Phaeophyta)

Hizikia fusiformis thalli experience dynamic incident light conditions during the period of growth. The present study was designed to examine how changing photon irradiance affects the photosynthesis both in the short and long terms by culturing H. fusiformis under three different light levels: 35 μmol photons m^-2 s^-1 (low light, LL), 85 μmol photons m^-2 s^-1 (intermediate light, IL), and 165 μmol photons m^-2 s^-1 (high light, HL). A similar relative growth rate was observed between IL- and HL-grown algae, but the growth rate was significantly reduced in LL-grown algae. The photosynthetic rates (P _n) measured at their respective growth light levels were found to be lowest in the thalli grown at LL and highest at HL. However, LL-grown algae exhibited much higher P _n in comparison with IL- and the HL-grown thalli at the same measuring photosynthetic photon flux density, indicating the photosynthetic acclimation to low growth light in H. fusiformis. The photosynthesis–light curves showed that LL-grown algae had a highest light-saturating maximum P _n (P _max) in comparison with IL- or HL-grown algae when the photosynthetic rates were expressed on the biomass basis. However, P _max was highest in HL-grown algae compared to IL- or LL-grown algae when the rates were normalized to chlorophyll a. The photosynthesis–inorganic carbon (Ci) response curves were also significantly affected by the growth light conditions. The highest value of apparent photosynthetic conductance occurred in LL-grown algae while the lowest value in HL-grown algae. Additionally, the activity of external carbonic anhydrase (CA) tended to increase while the total CA activity inclined to decrease in H. fusiformis thalli when the growth light level altered from 35 to 165 μmol photons per square meter per second. The external CA inhibitors showed a higher inhibition in HL-grown algae compared with LL-grown algae. It was proposed that photosynthetic acclimation to low light condition in H. fusiformis was achieved through an increase in the number of reaction centers and increased capacities of electron transport and of Ci transport within cells. The ability of photosynthetic acclimation to low light confers H. fusiformis thalli to overcome the environmental low light condition as a result of the attenuation of seawater or self-shading through enhancing its photosynthetic performance and carbon assimilation necessary for growth.     

Preliminary investigation of Okhotsk Sea ice algae; taxonomic composition and photosynthetic activity

Okhotsk Sea pack ice from Shiretoko in northern Hokkaido, sampled in March 2007, contained microalgal communities dominated by the centric diatoms Thalassiosira nordenskioeldi and T. punctigera. Domination by this genus is very unusual in sea ice. Communities from nearby fast ice at Saroma-ko lagoon were dominated by Detonula conferavea and Odontella aurita. Average microalgal biomass of the Okhotsk Sea pack ice (surface and bottom) was 1.59 ± 1.09 μg chla l⁻¹ and for fast ice (bottom only) at nearby Saroma-ko lagoon, 16.5 ± 3.2 μg l⁻¹ (=31.1 ± 5.0 mg chla m⁻²). Maximum quantum yield of the Shiretoko pack ice algal communities was 0.618 ± 0.056 with species-specific data ranging between 0.211 and 0.653. These community values are amongst the highest recorded for sea ice algae. Rapid light curves (RLC) on individual cells indicated maximum relative electron transfer rates (relETR) between 20.8 and 60.6, photosynthetic efficiency values (α) between 0.31 and 0.93 and onset of saturation values (E _k) between 33 and 91 μmol photons m⁻² s⁻¹. These data imply that the pack ice algal community at Shiretoko was healthy and actively photosynthesising. Maximum quantum yield of the Saroma-ko fast ice community was 0.401 ± 0.086, with values for different species between 0.361 and 0.560. RLC data from individual Saroma-ko fast ice algal cells indicated relETR between 55.3 and 60.6, α values between 0.609 and 0.816 and E _k values between 74 and 91 μmol photons m⁻² s⁻¹ which are consistent with measurements in previous years.     

A surfactant tolerant laccase of <Emphasis Type="Italic">Meripilus giganteus</Emphasis>

A laccase (Lcc1) from the white-rot fungus Meripilus giganteus was purified with superior yields of 34% and 90% by conventional chromatography or by foam separation, respectively. Size exclusion chromatography (SEC) and sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) yielded a molecular mass of 55 kDa. The enzyme possessed an isoelectric point of 3.1 and was able to oxidize the common laccase substrate 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) at a pH of 2.0, whereas the enzyme was still able to oxidize ABTS and 2,6-dimethoxyphenol (DMP) at pH 6.0. Lcc1 exhibited low K _m values of 8 μM (ABTS) and 80 μM (DMP) and remarkable catalytic efficiency towards the non-phenolic substrate ABTS of 37,437 k _cat/k _m (s⁻¹ mM⁻¹). The laccase showed a high stability towards high concentrations of various metal ions, EDTA and surfactants indicating a considerable biotechnological potential. Furthermore, Lcc1 exhibited an increased activity as well as a striking boost of stability in the presence of surfactants. Degenerated primers were deduced from peptide fragments. The complete coding sequence of lcc1 was determined to 1,551 bp and confirmed via amplification of the 2,214 bp genomic sequence which included 12 introns. The deduced 516 amino acid (aa) sequence of the lcc1 gene shared 82% identity and 90% similarity with a laccase from Rigidoporus microporus. The sequence data may aid theoretical studies and enzyme engineering efforts to create laccases with an improved stability towards metal ions and bipolar compounds.     

Critical Levels of Selenium in Different Crops Grown in an Alkaline Silty Loam Soil Treated with Selenite-Se

Critical levels of selenium in raya (Brassica juncea Czern L.), maize (Zea mays L.), wheat (Triticum aestivum L.) and rice (Oryza sativa L.) were worked out by growing these crops in an alkaline silty loam soil treated with different levels of selenite-Se ranging from 1 to 25 μg g⁻¹ soil. Significant decrease in dry matter yield was observed above a level of 5 μg Se g⁻¹ soil in raya and maize; 4 μg Se g⁻¹ soil in wheat and 10 μg Se g⁻¹ soil in rice shoots. The critical level of Se in plants above which significant decrease in yield would occur was found to be 104.8 μg g⁻¹ in raya, 76.9 μg g⁻¹ in maize, 41.5 μg g⁻¹ in rice and 18.9 μg g⁻¹ in wheat shoots. Significant coefficients of correlation were observed between Se content above the critical level and dry matter yield of raya as well as rice (r = −0.99, P ≤ 0.01), wheat (r = −0.97, P ≤ 0.01) and maize ((r = −0.96, P ≤ 0.01). A synergistic relationship was observed between S and Se content of raya (r = 0.96, P ≤ 0.01), wheat (r = 0.89, P ≤ 0.01), rice (r = 0.85, P ≤ 0.01) and maize (r = 0.84, P ≤ 0.01). Raya, maize and rice absorbed Se in levels toxic for animal consumption (i.e. > 5 mg Se kg⁻¹) when the soil was treated with more than 1.5 μg Se g⁻¹. In case of wheat, application of Se more than 3 μg g⁻¹ soil resulted in production of toxic plants.