Monothiol glutaredoxin cDNA from Taiwanofungus camphorata: a novel CGFS-type glutaredoxin possessing glutathione reductase activity

Glutaredoxins (Grxs) play important roles in the redox system via reduced glutathione as a reductant. A TcmonoGrx cDNA (1039 bp, EU158772) encoding a putative monothiol Grx was cloned from Taiwanofungus camphorata (formerly named Antrodia camphorata). The deduced amino acid sequence is conserved among the reported monothiol Grxs. Two 3-D homology structures of the TcmonoGrx based on known structures of human Grx3 (pdb: 2DIY_A) and Mus musculus Grx3 (pdb: 1WIK_A) have been created. To characterize the TcmonoGrx protein, the coding region was subcloned into an expression vector pET-20b(+) and transformed into E. coli C41(DE3). The recombinant His6-tagged TcmonoGrx was overexpressed and purified by Ni(2+)-nitrilotriacetic acid Sepharose. The purified enzyme showed a predominant band on 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The enzyme exhibited glutathione reductase (GR) activity via dithionitrobenzoate (DTNB) assay. The Michaelis constant (K(M)) values for GSSG and NADPH were 0.064 and 0.041 mM, respectively. The enzyme's half-life of deactivation at 60 °C was 10.5 min, and its thermal inactivation rate constant (k(d)) was 5.37 × 10(-2) min(-1). The enzyme was active under a broad pH range from 6 to 8. The enzyme retained 50% activity after trypsin digestion at 37 °C for 40 min. Both mutants C(40)→S(40) and C(165)→S(165) lost 40-50% GR activity, whereas the mutant S(168)→C(168) showed a 20% increase in its GR activity.     

Correlation of antioxidant capacities to oxygen radical scavenging enzyme activities in blackberry

The activities of the oxygen radical scavenging enzymes [glutathione-peroxidase (GSH-POD), superoxide dismutase (SOD), and guaiacol peroxidase (G-POD)], hydrogen peroxide scavenging enzymes in the ascorbate-glutathione cycle [ascorbate peroxidase (AsA-POD), monodehydroascorbate reductase (MDAR), dehydroascorbate reductase (DHAR), and glutathione reductase (GR)], the nonenzyme components [ascorbate (AsA), dehydroascorbate (DHAsA), glutathione (GSH), and oxidized glutathione (GSSG)], and their antioxidant capacity [oxygen radical absorbance capacity (ORAC)] were measured in the juice of six different thornless blackberry (Rubus sp.) cultivars. The 'Hull Thornless' cultivar contained the highest levels, whereas 'Black Satin' consistently had the lowest activities for all the enzymes tested in this study. ORAC values were also the highest in 'Hull Thornless' and lowest in 'Black Satin'. The highest levels of AsA and DHAsA were in the juice of 'Hull Thornless' blackberries with 1. 09 and 0.15 micromol/g fresh wt, respectively. 'Hull Thornless' also had the highest ratio of AsA/DHAsA among the six blackberry cultivars studied. The 'Smoothstem' cultivar contained the lowest amounts of AsA and DHAsA. 'Hull Thornless' had the highest GSH content with 78.7 nmol/g fresh wt, while 'Chester Thornless' contained the largest amount of GSSG. The highest GSH/GSSG ratio was 4.90 which was seen in the 'Hull Thornless' cultivar. The correlation coefficient between ORAC values and AsA/DHAsA ratios was as high as 0.972. A correlation (r = 0.901) was also detected between ORAC values and GSH content. The antioxidant activity in blackberry juice was positively correlated to the activities of most antioxidant enzymes (r = 0.902 with SOD; r = 0.858 with GSH-POD; r = 0.896 with ASA-POD; and r = 0.862 with GR).     

Tomato (Lycopersicon esculentum) pectin methylesterase and polygalacturonase behaviors regarding heat- and pressure-induced inactivation.

The combined high pressure/thermal (HP/T) inactivation of tomato pectin methyl esterase (PME) and polygalacturonase (PG) was investigated as a possible alternative to thermal processing classically used for enzyme inactivation. The temperature and pressure ranges tested were from 60 degrees C to 105 degrees C, and from 0.1 to 800 MPa, respectively. PME, a heat-labile enzyme at ambient pressure, is dramatically stabilized against thermal denaturation at pressures above atmospheric and up to 500-600 MPa. PG, however, is very resistant to thermal denaturation at 0.1 MPa, but quickly and easily inactivated by combinations of moderate temperatures and pressures. Selective inactivation of either PME or PG was achieved by choosing proper combinations of P and T. The inactivation kinetics of these enzymes was measured and described mathematically over the investigated portion of the P/T plane. Whereas medium composition and salinity had little influence on the inactivation rates, PME was found less sensitive to both heat and pressure when pH was raised above its physiological value. PG, on the other hand, became more labile at higher pH values. The results are discussed in terms of isoenzymes and other physicochemical features of PME and PG.     

外源谷胱甘肽喷施对缺硫胁迫下小白菜谷胱甘肽代谢的影响

[目的]硫是植物生长发育过程中所必需的营养元素,缺硫会阻碍植物生长发育,研究外源谷胱甘肽(GSH)对缺硫胁迫下小白菜GSH代谢的影响,为利用外源GSH减轻缺硫胁迫对植物造成的伤害提供理论依据.[方法]以小白菜为试验材料,采用营养液栽培方法,设正常供硫并喷施蒸馏水对照(CK)、缺硫喷施蒸馏水(H2O)、缺硫喷施25 mg...     

公猪舍夏季温度和流场数值CFD模拟及验证

为研究夏季全漏缝地板公猪舍湿帘风机蒸发降温效果及舍内环境分布规律,该文利用计算流体力学CFD(computational fluid dynamics)对北京养猪育种中心SPF(Specific Pathogen Free Swine)公猪舍进行模拟研究并通过实测数据进行验证。研究中将漏缝地板作为多孔介质简化,基于标准k-?湍流模型对空载及装猪猪舍内的风速场和温度场进行模拟,通过模拟值与实测值的对比验证模型的合理性。结果表明采用该模型模拟空载时猪舍,风速场模拟值与实测值误差较小,相对误差范围在0.25%~30.8%。模拟温度与实测温度最大绝对误差为0.48 K,平均绝对误差为0.11 K,平均相对误差为0.5%。模拟装猪时的猪舍,温度分布结构与装猪前相似,但整体温度略有上升。该研究可对当前常用的含漏缝地板猪舍建模研究提供参考,并为畜禽舍内改造和建筑实践提供理论依据。     

Interpreting pH and nitrate concentration in solutions drained from acid soils. Possible artefacts of lysimeters

Abstract. Solutions collected from lysimeters of acid soils can show pH values close to or even above neutral. Laboratory experiments on an acid soil from Burundi were planned to test if denitrification or CO₂ degassing might explain such a paradox. In the first experiment, soil profiles were reconstituted in columns and leached with 55 μ m Ca(NO₃)₂ solutions at 30 °C and 4 °C. Two drainage regimes were applied: intermittent suction or no suction at the bottom of the columns. In the second experiment, pH values were measured in solutions drained from different horizons at 30 °C, before and after equilibration with ambient air. Sterilized soil was also tested in the same way. Results from experiment 1 showed that despite the accumulation of water in the bottom of soil profiles when no suction was applied, aeration still existed so that reduction reactions, namely denitrification, are not expected to affect greatly the percolate composition. Indeed nitrate concentration was similar in both drainage regimes and was close to the input value. The pH values in percolates were close to 7 at 30 °C and they dropped to about 5.5 when the columns were at 4 °C. In experiment 2, equilibration of percolates with ambient air resulted in pH increase which was greater for the top horizon (C-rich) but negligible when the soil was first sterilized. These convergent results illustrate the very important effect of CO₂ degassing on pH of drained solutions when microbial activity is stimulated at high temperatures, in C-rich soil. This is of prime importance when interpreting results from lysimeter experiments. By chance, this study also showed that large quantities of nitrate can be produced in soil at low temperatures.     

The Cd‐tolerant rice mutant cadH‐5 is a high Cd accumulator and shows enhanced antioxidant activity

To investigate the mechanism of cadmium (Cd) detoxification in rice (Oryza sativa L.), a Cd‐tolerant mutant cadH‐5, obtained by an Agrobacterium tumefaciens‐based gene‐delivery system, was used for a Cd‐tolerance and accumulation study. After 15 d of exposure to 0.75 mM CdCl₂, significant phenotypic differences were observed between the wild type (WT) and cadH‐5. When exposed to 0.5 mM CdCl₂, higher Cd levels were accumulated in cadH‐5 root cell wall, root cytosol, and membranes than those in WT. However, Cd concentrations in root tissues varied in both WT and cadH5. No significant difference of hydrogen peroxide (H₂O₂) concentrations was observed between WT and cadH‐5, while contents of cell‐wall polysaccharides and phytochelatins (PCs) in the mutant were higher compared to WT. The ratios of reduced glutathione to oxidized glutathione (GSH : GSSG) and ascorbate to dehydroascorbate (ASC : DHA) were lower in WT than in cadH‐5, while the NADPH : NADP⁺ ratio was different to the ratios of GSH : GSSG and ASC : DHA; the ascorbate peroxidase (APX, EC 1.11.1.11), glutathione peroxidase (GR, EC 1.6.4.2), dehydroascorbate reductase (DHAR, EC 1.8.5.1), and monodehydroascorbate reductase (MDHAR, EC 1.6.5.4) activities were lower in WT compared to cadH‐5. Our results indicate that under long‐term Cd stress, cadH‐5 plants can accumulate more Cd with more PC. Also, the redox status of ASC‐GSH cycle was more inhibited in WT than in cadH‐5 plants, rendering WT less able to scavenge reactive oxygen species (ROS). The cadH‐5 mutant maintains relatively high ASC, GSH, and NADPH concentrations, ratios of ASC : DHA, GSH : GSSG, and NADPH : NADP⁺, as well as antioxidative enzymatic activities and PC concentrations. Thus, it is tolerant of relatively high Cd accumulation.     

Degradation of malathion and phenthoate by glutathione reductase in wheat germ

Residual malathion in wheat was estimated at a lower value when analysis was performed by extraction with acetone after addition of water to swell the wheat, according to the Japanese Bulletin Method. The supernatant of the wheat homogenate showed degradation not only of malathion but also of phenthoate. Malathion and phenthoate were not degraded by the boiled supernatant of the wheat homogenate. It was presumed for this reason that glutathione reductase (GR; EC 1.6. 4.2) in the wheat degraded malathion. The following results were obtained: (1) GR originating in wheat could degrade malathion and phenthoate. (2) The degradation of malathion by the GR was inhibited by excessive GSSG. (3) There was a high correlation between GR activity and malathion degradation activity of the supernatant of wheat homogenates. It is likely that GR acted on the specific structure of malathion and phenthoate, the S=P-S bond, and the blanch structure bonding with the sulfur atom. Following the above, extraction with acetone after addition of water (the Japanese Bulletin Method) should be replaced by extraction with pure organic solvent and without addition of water for swelling.     

Functional properties of purified vicilins from cowpea (Vigna unguiculata) and pea (Pisum sativum) and cowpea protein isolate

The major storage globulins (vicilins) of cowpea (Vigna unguiculata) and pea (Pisum sativum) seeds were purified by ammonium sulfate precipitation, and a semipurified cowpea protein isolate (CPI) was prepared by isoelectric precipitation. Some of the functional properties of these proteins, including solubility, foaming, and emulsifying capacities, were investigated and compared. The solubility of purified cowpea vicilin was reduced at pH 5.0, increasing markedly below and above this value. Pea vicilin exhibited poor solubility between pH 5.0 and pH 6.0, and CPI was little soluble in the pH range from 4.0 to 6.0. At neutral pH, the emulsifying activity indexes (EAI) of purified pea vicilin and CPI were 194 and 291 m(2)/g, respectively, which compare quite favorably to EAIs of 110 and 133 m(2)/g for casein and albumin, respectively. Remarkably, purified cowpea vicilin exhibited an EAI of 490 m(2)/g, indicating a very high emulsifying activity. Purified cowpea and pea vicilins exhibited lower foaming capacities and foam stablity indexes (FSI) than CPI. FSI values of 80 and 260 min were obtained for purified pea and cowpea vicilin, respectively, whereas a FSI value of 380 min was obtained for CPI. These results are discussed in terms of the possible utilization of purified vicilins or protein isolates from pea and cowpea in the food processing industry.     

Glyphosate-resistant and -susceptible soybean (Glycine max) and canola (Brassica napus) dose response and metabolism relationships with glyphosate

Experiments were conducted to determine (1) dose response of glyphosate-resistant (GR) and -susceptible (non-GR) soybean [Glycine max (L.) Merr.] and canola (Brassica napus L.) to glyphosate, (2) if differential metabolism of glyphosate to aminomethyl phosphonic acid (AMPA) is the underlying mechanism for differential resistance to glyphosate among GR soybean varieties, and (3) the extent of metabolism of glyphosate to AMPA in GR canola and to correlate metabolism to injury from AMPA. GR50 (glyphosate dose required to cause a 50% reduction in plant dry weight) values for GR (Asgrow 4603RR) and non-GR (HBKC 5025) soybean were 22.8 kg ae ha-1 and 0.47 kg ha-1, respectively, with GR soybean exhibiting a 49-fold level of resistance to glyphosate as compared to non-GR soybean. Differential reduction in chlorophyll by glyphosate was observed between GR soybean varieties, but there were no differences in shoot fresh weight reduction. No significant differences were found between GR varieties in metabolism of glyphosate to AMPA, and in shikimate levels. These results indicate that GR soybean varieties were able to outgrow the initial injury from glyphosate, which was previously caused at least in part by AMPA. GR50 values for GR (Hyola 514RR) and non-GR (Hyola 440) canola were 14.1 and 0.30 kg ha-1, respectively, with GR canola exhibiting a 47-fold level of resistance to glyphosate when compared to non-GR canola. Glyphosate did not cause reduction in chlorophyll content and shoot fresh weight in GR canola, unlike GR soybean. Less glyphosate (per unit leaf weight) was recovered in glyphosate-treated GR canola as compared to glyphosate-treated GR soybean. External application of AMPA caused similar injury in both GR and non-GR canola. The presence of a bacterial glyphosate oxidoreductase gene in GR canola contributes to breakdown of glyphosate to AMPA. However, the AMPA from glyphosate breakdown could have been metabolized to nonphytotoxic metabolites before causing injury to GR canola. Injury in GR and non-GR canola from exogenous application of AMPA was similar.     

棉花幼苗AsA-GSH循环对低温胁迫的响应机制研究

为探究棉花幼苗营养器官抗坏血酸-谷胱甘肽(AsA-GSH)循环系统对低温胁迫的响应机制,以新陆早57号为试验材料,研究4℃低温胁迫不同时间(0、1和2 d)对根、茎和叶中抗氧化物质含量和酶活性的影响。结果表明,与对照(0 d)相比,随着低温胁迫时间延长,棉花幼苗根中还原型抗坏血酸(AsA)含量及单脱氢抗坏血酸还原酶(MDHAR)和谷胱甘肽还原酶(GR)活性呈上升趋势,氧化型抗坏血酸(DHA)含量先减少后增加,氧化型谷胱甘肽(GSSG)含量和脱氢抗坏血酸还原酶(DHAR)活性先升高后降低,谷胱甘肽氧化还原状态(GSH/GSSG比率)和抗坏血酸过氧化物酶(APX)活性总体呈下降趋势,抗坏血酸氧化还原状态(AsA/DHA比率)、还原型谷胱甘肽(GSH)含量和谷胱甘肽过氧化物酶(GPX)活性无显著性差异;茎和叶中AsA和GSH含量、AsA/DHA比率及APX、MDHAR和GR活性总体呈上升趋势,且茎中DHAR活性升高,DHA和GSSG含量先减少后增加,GSH/GSSG比率和GPX活性无显著性差异,而叶中DHA含量和GSH/GSSG比率先不变后增加,DHAR活性先升高后降低,GSSG含量和GPX活性无显著性差异。综合分析显示,棉花幼苗通过调节根、茎和叶中抗氧化物质含量和酶活性来清除活性氧,从而适应低温胁迫,减轻低温伤害。此外,低温对根中AsA-GSH循环影响最大。本研究结果为揭示低温胁迫对棉花幼苗AsA-GSH循环代谢机制提供了理论依据。     

Glyphosate effects on photosynthesis,nutrient accumulation,and nodulation in glyphosate‐resistant soybean

Previous greenhouse studies have demonstrated that photosynthesis in some cultivars of first‐ (GR1) and second‐generation (GR2) glyphosate‐resistant soybean was reduced by glyphosate. The reduction in photosynthesis that resulted from glyphosate might affect nutrient uptake and lead to lower plant biomass production and ultimately reduced grain yield. Therefore, a field study was conducted to determine if glyphosate‐induced damage to soybean (Glycine max L. Merr. cv. Asgrow AG3539) plants observed under controlled greenhouse conditions might occur in the field environment. The present study evaluated photosynthetic rate, nutrient accumulation, nodulation, and biomass production of GR2 soybean receiving different rates of glyphosate (0, 800, 1200, 2400 g a.e. ha^–1) applied at V2, V4, and V6 growth stages. In general, plant damage observed in the field study was similar to that in previous greenhouse studies. Increasing glyphosate rates and applications at later growth stages decreased nutrient accumulation, nodulation, leaf area, and shoot biomass production. Thus, to reduce potential undesirable effects of glyphosate on plant growth, application of the lowest glyphosate rate for weed‐control efficacy at early growth stages (V2 to V4) is suggested as an advantageous practice within current weed control in GR soybean for optimal crop productivity.     

Purification and partial characterization of three turnip (Brassicanapus L. var. esculenta D.C.) peroxidases

Three turnip peroxidases (fractions C1, C2, and C3) were partially purified and characterized, to permit study of their feasibility for use in clinical and enzyme immunoassays. These fractions represented 20% of the initial activity, and fractions C1 and C2 were purified to homogeneity. The optimum pH was between 5.0 and 5.5, while optimum temperature ranged from 40 to 55 degrees C. The ABTS K(m) values for the two acidic fractions (C2 and C3) were 0.70 and 0.42 mM, respectively; about 5 times lower than that reported for the acidic commercial horseradish peroxidase (HRP). Fraction C3 had 4 times higher K(m) value than commercial cationic HRP. The molecular weights determined by SDS-PAGE ranged from 39.2 to 42.5 kDa. Activation energies for inactivation were 113 (C1), 130 (C2), and 172 kJ/mol (C3) which are higher or comparable to other peroxidase isoenzymes reported. Fractions C1 and C3 represent an alternative source of peroxidase because of their higher purification yield and specific activity, when compared to fraction C2.     

Reduced-Oxidized Glutathione Status as a Potential Index of Oxidative Stress in Mature Cereal Grain

The purpose of this study was to examine the reduced and oxidized glutathione status of selected cereal grains as a potential index of balance between oxidative stress and antioxidant systems, and the contribution of reduced glutathione to the total antioxidant status in cereal grain extracts. Wheat cultivars Almari and Henika, barley cultivars Gregor and Mobek, rye cultivar Dañkowskie Złote, oat cultivar Sławko, and buckwheat cultivar Kora were used. Total antioxidant status (TAS) was measured by the ABTS (2,2'-azinobis(3-ethyl-benzothiazoline-6-sulphonate)) method. Contents of total phenolic compounds were also determined. Reduced (GSH) and oxidized glutathione (GSSG) (γ-glutamyl-cysteinyl-glycine) were assayed using the spectrofluorimetric method, and results were confirmed by the enzyme recycling method. Correlation coefficient for the GSH/GSSG ratio was r = 0.79. Correlation between TAS and the total phenolic compound content was r = 0.81. Correlation between GSH/GSSG ratio and TAS values was r= 0.46, depending on the extraction system used. The GSH/GSSG ratio may indicate a hierarchy among different cultivars and variance of cereal grains against damage caused by reactive oxygen species. For the main water-soluble antioxidants, our data indicate a potential hierarchy of resistance in investigated cereals against oxidative stress (buckwheat > wheat > barley ≈ rye > oat). This hierarchy was confirmed by the ability of investigated cereal extracts to scavenge superoxide anion radicals in vitro. The reduced-oxidized glutathione status in different cereal grains can be applied as a potential index of balance between oxidative stress and antioxidant systems.     

Simultaneous determination of oxidized and reduced glutathione in eel's (Monopterus albus) plasma by transient pseudoisotachophoresis coupled with capillary zone electrophoresis

Both the reduced form of glutathione (GSH) and the oxidized form of glutathione (GSSG) in eel's ( Monopterus albus) plasma were for the first time determined by transient pseudoisotachophoresis coupled with capillary zone electrophoresis. The method of transient pseudoisotachophoresis coupled with capillary zone electrophoresis has been thoroughly optimized and adequately evaluated for the simultaneous determination of GSH and GSSG in eel's plasma. The detection limits (S/N = 3) of the method developed were 0.2 and 0.05 micromol/L for GSH and GSSG, respectively. The linearity of the calibration curves was valid in the range of 0-10 micromol/L GSH and 0-0.70 micromol/L GSSG. The method was simple, fast, and reproducible. It was found that the respective concentrations of GSH and GSSG were in the range of 9.1-14.5 and 0.31-0.58 micromol/L in the adult eel's plasma, and 10.8-17.9 and 0.49 - 0.68 micromol/L in the juvenile eel's plasma of the three populations determined. Each blood sample was a composite of five eels. For each of the three populations, the concentrations of GSH and GSSG in the adult eel's plasma were lower than those in the juvenile eel's plasma, and the concentrations of GSH and GSSG in the plasma of population 1 (deep yellow finless eels) were higher than those in populations 2 (light yellow finless eels) and 3 (green finless eels) for either the adult or the juvenile eels.     

The elucidation of soil pattern in the Wyre Forest of the West Midlands, England. II. Spatial distribution

The spatial distribution of soil in the Wyre Forest of England was analysed in two phases. In the first the soil was examined at sites chosen using a five-stage nested design with spacings increasing geometrically from 6 m to 600 m. Some 80% of the variance was contributed by components for the spacings between 6 m and 60 m. Measurements were then made on transects at 5 m intervals and semi-variograms estimated to 70 m. Except for those of pH the semi-variograms of the soil properties had the same general transitive form and a common effective range of about 40 m. This short range meant that very intensive sampling, approximately one point per 400 m², is needed to map the soil variation. A survey was made of a small portion of the forest on a 20m × 25 m grid to test the inference. Maps of clay and sand content were made successfully by kriging from the data. The mutual correlations between soil properties and the common range of their semi-variograms enabled a general purpose and spatially coherent soil classification to be created from the data. Its mapping confirmed the intricacy of the soil pattern in the Forest.     

Kinetics and hydrolysis of fenamiphos, fipronil, and trifluralin in aqueous buffer solutions.

Hydrolyses of fenamiphos, fipronil, and trifluralin were studied in aqueous buffer solutions of pH 4.1, 7.1, and 9.1 at different temperatures, 5, 22 +/- 1, 32 +/- 1, and 50 +/- 1 degrees C. Fenamiphos, fipronil, and trifluralin were found to be more stable in acidic and neutral buffer solutions at temperatures of 5 and 22 +/- 1, and dissipation is rapid at 50 +/- 1 degrees C. In basic buffer and at higher temperature, degradation of fenamiphos was found to be very rapid when compared with fipronil and trifluralin. The rate constants calculated at 32 degrees C for fenamiphos were 2349.4 x 10(-)(8) (pH 4.1), 225.2 x 10(-)(8) (pH 7.1), and 30476.0 x 10(-)(8) (pH 9.1); for fipronil 1750.0 x 10(-)(8) (pH 4.1), 3103.0 x 10(-)(8) (pH 7.1), and 3883.0 x 10(-)(8) (pH 9.1); and for trifluralin 2331.0 x 10(-)(8) (pH 4.1), 2360.0 x 10(-)(8) (pH 7.1), and 3188.0 x 10(-)(8) (pH 9.1). On the basis of rate constant values, these pesticides appeared to be more susceptible to hydrolysis than synthetic organophosphorus compounds such as chlorpyriphos, diazinon, malathion, and ronnel. DT(50) values calculated at 32 degrees C were 228 (pH 4.1), 5310.24 (pH 7.1), and 37.68 (pH 9.1) h for fenamiphos; 608.6 (pH 4.1), 373.9 (pH 7.1), and 270.2 (pH 9.1) h for fipronil; and 502.1 (pH 4.1), 496.8 (pH 7.1), and 355.7 (pH 9.1) h for trifluralin.     

CALCIUM DEFICIENCY OF WHEAT GROWN IN ACIDIC SANDY SOIL FROM SOUTHWESTERN AUSTRALIA

Calcium (Ca) deficiency has been associated with acidic very sandy soils that are widespread in Southwestern Australia (WA). There are no documented reports of Ca deficiency of cereals grown in the field. However, there is a possibility that Ca deficiency could be observed on the acidic very sandy soils of WA, particular where fertilizers low in Ca concentration are used.

For previously unfertilized sandy soil collected from under indigenous remnant vegetation in WA, there was no effect on yield of dried shoots of wheat (Triticum aestivum L.) due to addition of fertilizer Ca (0 up to 480 mg Ca/pot) to either the topsoil (top 10 cm of soil, pH 4.6) (1:5 soil 0.01 M CaCl₂, w/v) or subsoil (10–20 cm soil, pH 5.1). However, application of fertilizer Ca increased grain yields, by about 12% for the topsoil and up to about tenfold for the subsoil. For the subsoil, no grain was produced on the nil Ca treatment; about 0.16 g of grain was produced per plant at the second level of applied Ca (60 mg Ca/pot), and about 1.6 g of grain produced per plant at the highest amount of Ca (480 mg Ca/pot), a ten-fold increase in grain production. For the topsoil, grain yield for the highest level of Ca (480 mg Ca/pot) applied was about 1.8 times the amount of grain produced for the nil Ca treatment. The Ca concentration (% Ca) in dried shoots and grain increased with the addition of Ca to both the surface and subsoil. The Ca concentration in the dried tops declined with age of the wheat for both soils. For example, % Ca declined from 0.3% at GS25 to 0.1% at maturity. The highest Ca concentration in dried shoots at harvest always produced the largest amount of grain. There was no grain yield plateau reached in our study so critical concentrations of Ca within the plant tops for grain yield could not be determined.     

Denaturation and aggregation of myosin from two bovine muscle types

The thermal behaviors of myosin from bovine vastus intermedius (VI, predominantly red muscle) and semimembranosus (SM, predominantly white muscle) at pH 6.05 (ultimate pH of VI muscle) and 5.50 (ultimate pH of SM muscle) were compared. Differential scanning microcalorimetry and turbidity measurements were used to monitor changes in myosin during heating from 25 to 80 degrees C at 1 degrees C/min. VI and SM myosin heavy chain isoforms were identified on gradient SDS-PAGE. Endotherms of VI myosin at pH 6.05 had three transition temperatures (T(m)) of 45, 53, and 57 degrees C, whereas at pH 5.50 two transitions were observed at 42 and 59 degrees C. SM myosin had two T(m) values of 46 and 58 degrees C at pH 6.05 and T(m) values of 43 and 62 degrees C at pH 5.5. SM myosin at its ultimate pH was less heat stable than VI myosin at its ultimate pH; however, when SM and VI myosin were compared at the same pH, VI myosin was less stable.     

Bleaching herbicide norflurazon inhibits phytoene desaturase by competition with the cofactors.

Cofactor requirement was determined for the heterologous expressed phytoene desaturases from the cyanobacterium Synechococcus and the higher plant Gentiana lutea. The cyanobacterial enzyme is dependent on either NAD(P) or plastoquinone, whereas only quinones such as plastoquinone can function as a cofactor for the phytoene desaturase from G. lutea. Enzyme kinetic studies were carried out to determine a possible competition between the cofactors and the bleaching herbicide norflurazon. For the Synechococcus enzyme, competition between norflurazon and NADP, as well as plastoquinone, could be demonstrated. The K(m) values for these cofactors were 6.6 mM and 0.23 microM, respectively. Inhibition of the phytoene desaturase from G. lutea by norflurazon was also competitive with respect to plastoquinone. The K(m) values of both enzymes for plastoquinone were very close.