(H)NCAHA and (H)CANNH experiments for the determination of vicinal coupling constants related to the ϕ-torsion angle

Summary A set of three-dimensional triple-resonance experiments is described which provide , , and coupling constants. The pulse sequences generate E.COSY-like multiplet patterns and comprise a magnetization transfer from the amide proton to the α-proton or vice versa via the directly bound heteronuclei. For residues with the ¹H^α spin resonating close to the H₂O signal, a modified HNCA experiment can be employed to measure the vicinal ¹H^N,¹H^α couplings. Ambiguities associated with the conversion of values into ϕ-angle constraints for protein structure determination can be resolved with the knowledge of the heteronuclear ³J-couplings. In favourable cases, stereospecific assignments of glycine α-protons can be obtained by employing the experiments described here in combination with NOE data. The methods are applied to flavodoxin from Desulfovibrio vulgaris.     

Energy requirements of Adélie penguin (Pygoscelis adeliae) chicks

Summary The energy requirements of Adélie penguin (Pygoscelis adeliae) chicks were analysed with respect to body mass (W, 0.145–3.35 kg, n=36) and various forms of activity (lying, standing, minor activity, locomotion, walking on a treadmill). Direct respirometry was used to measure O₂ consumption ( ) and CO₂ production. Heart rate (HR, bpm) was recorded from the ECG obtained by both externally attached electrodes and implantable HR-transmitters. The parameters measured were not affected by hand-rearing of the chicks or by implanting transmitters. HR measured in the laboratory and in the field were comparable. Oxygen uptake ranged from in lying chicks to at maximal activity, RQ=0.76. Metabolic rate in small wild chicks (0.14–0.38 kg) was not affected by time of day, nor was their feeding frequency in the colony (Dec 20–21). Regressions of HR on were highly significant (p< 0.0001) in transmitter implanted chicks (n=4), and two relationships are proposed for the pooled data, one for minor activities ( ), and one for walking ( ). Oxygen consumption, mass of the chick (2–3 kg), and duration of walking (T, s) were related as , whereas mass-specific O₂ consumption was related to walking speed (S, m·s^-1) as .Abbreviations bpm beats per minute - D distance walked (m) - ECG electrocardiogram - HR heart rate (bpm) - ns number of steps - RQ respiratory quotient - S walking speed (m·s^-1) - T time walked (s) - W body mass (kg)     

A quantitative appraisal of the ambivalent metal ion binding properties of cytidine in aqueous solution and an estimation of the<Emphasis Type="Italic"> anti</Emphasis>–<Emphasis Type="Italic">syn</Emphasis> energy barrier of cytidine derivatives

The recently defined versus straight-line plots for L = pyridine-type (PyN) and ortho-aminopyridine-type (oPyN) ligands now allow the evaluation in a quantitative manner of the stability of the 1:1 complexes formed between cytidine (Cyd) and Ca²⁺, Mg²⁺, Mn²⁺, Co²⁺, Ni²⁺, Cu²⁺, Zn²⁺ or Cd²⁺ (M²⁺); the corresponding stability constants, , including the acidity constant, , for the deprotonation of the (N3)H⁺ site had been determined previously under exactly the same conditions as the mentioned plots. Since the stabilities of the M(PyN)²⁺ and M(oPyN)²⁺ complexes of Ca²⁺ and Mg²⁺ are practically identical, it is concluded that complex formation occurs in an outer-sphere manner, and this is in accord with the fact that in the pK_a range 3–7 metal ion binding is independent of or . Ca(Cyd)²⁺ and Mg(Cyd)²⁺ are more stable than the corresponding (outer-sphere) M(PyN)²⁺ complexes and this means that the C2 carbonyl group of Cyd must participate, next to N3 which is most likely outer-sphere, in metal ion binding, leading thus to chelates; these have formation degrees of about 50% and 35%, respectively. Co(Cyd)²⁺ and Ni(Cyd)²⁺ show no increased stability based on the hence, the (C2)O group does not participate in metal ion binding, but the inner-sphere coordination to N3 is strongly inhibited by the (C4)NH₂ group. In the M(Cyd)²⁺ complexes of Mn²⁺, Cu²⁺, Zn²⁺ and Cd²⁺, this inhibiting effect on M²⁺ binding at N3 is partially compensated by participation of the (C2)O group in complex formation and the corresponding chelates have formation degrees between about 30% (Zn²⁺) and 83% (Cu²⁺). The different structures of the mentioned chelates are discussed in relation to available crystal structure analyses. (1) There is evidence (crystal structure studies: Cu²⁺, Zn²⁺, Cd²⁺) that four-membered rings form, i.e. there is a strong M²⁺ bond to N3 and a weak one to (C2)O. (2) By hydrogen bond formation to (C2)O of a metal ion-bound water molecule, six-membered rings, so-called semichelates, may form. (3) For Ca²⁺ and Mg²⁺, and possibly Mn²⁺, and their Cyd complexes, six-membered chelates are also likely with (C2)O being inner-sphere (crystal structure) and N3 outer-sphere. (4) Finally, for these metal ions also complexes with a sole outer-sphere interaction may occur. All these types of chelates are expected to be in equilibrium with each other in solution, but, depending on the metal ion, either the one or the other form will dominate. Clearly, the cytidine residue is an ambivalent binding site which adjusts well to the requirements of the metal ion to be bound and this observation is of relevance for single-stranded nucleic acids and their interactions with metal ions. In addition, the anti–syn energy barrier has been estimated as being in the order of 6–7.5 kJ/mol for cytidine derivatives in aqueous solution at 25 °C.Abbreviations ADP^3– adenosine 5-diphosphate - AMP^2– adenosine 5-monophosphate - ATP^4– adenosine 5-triphosphate - CDP^3– cytidine 5-diphosphate - cl closed - CMP^2– cytidine 5-monophosphate - 3-CMP^2– cytidine 3-monophosphate - CTP^4– cytidine 5-triphosphate - Cyd cytidine - DNA deoxyribonucleic acid - I ionic strength - K_a acidity constant - K_I intramolecular equilibrium constant - L general ligand - M²⁺ general divalent metal ion - NTP^4– nucleoside 5-triphosphate - op open - oPyN ortho-aminopyridine-type ligand - PyN pyridine-type ligand - t-RNA transfer ribonucleic acid - Tu tubercidin (7-deazaadenosine)In honor of Professor Liang-Nian Ji on the occasion of his 70th birthday in friendship and with best wishes.     

A Combined Patch-Clamp and Electrorotation Study of the Voltage- and Frequency-Dependent Membrane Capacitance Caused by Structurally Dissimilar Lipophilic Anions

Interactions of structurally dissimilar anionic compounds with the plasma membrane of HEK293 cells were analyzed by patch clamp and electrorotation. The combined approach provides complementary information on the lipophilicity, preferential affinity of the anions to the inner/outer membrane leaflet, adsorption depth and transmembrane mobility. The anionic species studied here included the well-known lipophilic anions dipicrylamine (DPA⁻), tetraphenylborate (TPB⁻) and [W₂(CO)₁₀(S₂CH)]⁻, the putative lipophilic anion and three new heterocyclic W(CO)₅ derivatives. All tested anions partitioned strongly into the cell membrane, as indicated by the capacitance increase in patch-clamped cells. The capacitance increment exhibited a bell-shaped dependence on membrane voltage. The midpoint potentials of the maximum capacitance increment were negative, indicating the exclusion of lipophilic anions from the outer membrane leaflet. The adsorption depth of the large organic anions DPA⁻, TPB⁻ and increased and that of W(CO)₅ derivatives decreased with increasing concentration of mobile charges. In agreement with the patch-clamp data, electrorotation of cells treated with DPA⁻ and W(CO)₅ derivatives revealed a large dispersion of membrane capacitance in the kilohertz to megahertz range due to the translocation of mobile charges. In contrast, in the presence of TPB⁻ and no mobile charges could be detected by electrorotation, despite their strong membrane adsorption. Our data suggest that the presence of oxygen atoms in the outer molecular shell is an important factor for the fast translocation ability of lipophilic anions.     

Energetics of the growth of Methanobacterium thermoautotrophicum and Methanococcus thermolithotrophicus on ammonium chloride and dinitrogen

Methanobacterium thermoautotrophicum was grown in continuous culture in a fermenter gassed with H₂ and CO₂ as sole carbon and energy sources, and in a medium which contained either NH₄Cl or gaseous N₂ as nitrogen source. Growth was possible with N₂. Steady states were obtained at various gas flow rates with NH₄Cl and with and the maintenance coefficient varied with the gas input and with the nitrogen source. Growth of Methanococcus thermolithotrophicus in continuous culture in a fermenter gassed with H₂, CO₂ as nitrogen, carbon and energy sources was also examined.Abbreviations molecular growth yield (g dry weight of cells per mol of CH₄ evolved) - growth rate (h^-1) - D dilution rate (h^-1) - rate (h_-1); relation of Neijssel and Tempest and of Stouthamer and Bettenhaussen - energy     

Physiology and kinetics of autotrophic denitrification by Thiobacillus denitrificans

Summary A strain of Thiobacillus denitrificans was isolated after enrichment under anaerobic conditions by the continuous culture technique using thiosulfate as energy source and nitrate as electron acceptor and nitrogen source. The isolate was an active denitrifyer, the optimal conditions being 30°C and pH 7.5–8.0. Denitrification was inhibited by sulfate (the reaction product) above 5 g SO ₄ ⁼ /l, whereas high concentrations of the substrates nitrate and thiosulfate were less harmful; nitrite affected denitrification above 0.2 g NO ₂ ^– /l. During the time course of denitrification in a batch culture growth and substrate consumption slowed down already after only half the substrate was utilized due to product inhibition. The following parameters were determined in continuous culture under nitrate limitation: _max=0.11 h^–1, K _S=0.2 mg NO ₃ ^– /l, maximum denitrification rate=0.78 g NO ₃ ^– /g cells·h, g cells/g NO ₃ ^– , g cells/g S₂O ₃ ⁼ . Nitrite did not accumulate during steady state denitrification; the denitrification gas was almost pure N₂. The concentrations of N₂O and NO were below 1 ppm.     

Modification of the data-processing method for the turbidimetric bioassay of nisin

The data processing method of the turbidimetric bioassay of nisin was modified to facilitate its industrial application. The influence of the initial indicator concentration was minimized by a redefined specific dose of the bacteriocin as the quotient between the titer of the added bacteriocin and the initial population density of the indicator in the suspension. It was found that d _c = 0.125 μg ml⁻¹ was the critical dose of nisin that can cause a complete inhibition of the indicator, Pediococcus acidilactici UL5, with an initial OD of 0.135. To eliminate the interference of the cell debris, an equation, , exploiting d _c, was formulated to obtain the intrinsic survival proportion. The use of the specific dose of the bacteriocin and the intrinsic survival proportion as parameters of the dose/response curve greatly enhanced its repeatability and feasibility. A dual-dosage approach was developed to further simplify the conventional standard dose/response curve method.     

Sampling bias in estimating Design II variance components with S1 families

Summary The use of several S₁ individuals to represent an S₀ individual permits the use of a Design II mating scheme for plants with only one pistillate flower per plant. Estimates of additive (V _A ) and dominance (V _D ) variance from this mating scheme will be biased upwards, when a small number (10) of individuals of each S₁ line are used. This bias can be computed, and the additive and dominance estimates can be corrected. Of particular interest is the observation that the additive genetic variance contributes to bias in estimates of V _D . When S₀ plants are non inbred and their selfedprogeny (S₁ lines) are used to represent them in developing families for use in the Design II, where m₁ is the number of individuals used to represent an S₁ line in developing half sib-families and m₂ is the number of individuals used to represent the S₁ line in making up full sib-families. For example, in a 3×3 Design II, with about 10 individuals used to represent each S₁ line in each cross, m₂ = 10 and m₁ = 30. When m₁ = m₂ = 1, and Joint contribution from Department of Agronomy, University of Nebraska 68583, and the S. S. Cameron Laboratory, Werribee, Victoria 3030, Australia. Published as paper No. 7395, Journal Series     

Hydrogen-isotope composition of leaf water in C3 and C4 plants: its relationship to the hydrogen-isotope composition of dry matter

The natural abundance hydrogen-isotope composition of leaf water ( ) and leaf organic matter ( _D ^org ) was measured in leaves of C₃ and C₄ dicotyledons and monocotyledons. The value of leaf water showed a marked diurnal variation, greatest enrichment being observed about midday. However, this variation was greater in the more slowly transpiring C₄ plants than in C₃ plants under comparable environmental conditions. A model based on analogies with a constant feed pan of evaporating water was developed and the difference between C₃ and C₄ plants expressed in terms of either differences in kinetic enrichment or different leaf morphology. Microclimatic and morphological features of the leaves which may be associated with this factor are discussed. There was no daily excursion in the _D ^org value in leaves of either C₃ or C₄ plants. When _D ^org values were referenced to the mean values during the period of active photosynthesis, the discrimination against deuterium during photosynthetic metabolism (D) was greater in C₃ plants (-117 to -121) than in C₄ plants (-86 to -109).These results show that the different water use strategies of C₃ and C₄ plants are responsible for the measured difference in deuterium-isotope composition of leaf water. However, it is unlikely that these physical processes account fully for the differences in hydrogen-isotope composition of the products of C₃ and C₄ photosynthetic metabolism.Symbols Hydrogen-isotope composition of leaf water - _D ^org hydrogen-isotope composition of leaf organic matter     

Survival Chances of Mutants Starting With One Individual

A simple theoretical model of a Darwinian system (a periodic system with a multiplication phase and a selection phase) of entities (initial form of polymer strand, primary mutant and satellite mutants) is given. First case: one mutant is considered. One individual of the mutant appears in the multiplication phase of the first generation. The probabilities to find N individuals of the mutant after the multiplication phase M of the n-th generation (with probability δ of an error in the replication, where all possible errors are fatal errors) and after the following selection phase S (with probability β that one individual survives) are given iteratively. The evolutionary tree is evaluated. Averages from the distributions and the probability of extinction are obtained. Second case: two mutants are considered (primary mutant and new form). One individual of the primary mutant appears in the multiplication phase of the first generation. The probabilities to find N _p individuals of the primary mutant and N _m individuals of the new form after the multiplication phase M of the n-th generation (probability ɛ of an error in the replication of the primary mutant giving the new form) and after the following selection phase S (probabilities β _p and β _m that one individual each of the primary mutant and of the new form survives) are given iteratively. Again the evolutionary tree is evaluated. Averages from the distributions are obtained. The online version of the original article can be found at .     

Kinetic study on oxygenation of Lingula unguis hemerythrin using the stopped-flow 02-jump method

O₂-jump experiments with an improved stopped-flow apparatus have been used to study oxygenation and deoxygenation processes in Lingula unguis hemerythrin. With an O₂ electrode set in the observation cell, O₂ concentration conld be obtained directly. The reliability of this method has been compared with other conventional methods.O₂-jump (up and down) experiments were carried out with L. unguis hemerythrin at pH 6.8 (non-cooperative pH) and at pH 7.6 (cooperative pH). At pH 6.8, both O₂-jump (up) and O₂-jump (down) experiments showed single exponential processes which were consistent with the following scheme: . The value of k _on was estimated to be (4.4 ± 0.5) × 10⁵ M^–1 s ^–1, and k _off was (15 ± 5) s^–1. These values are consistent with those obtained by the temperature-jump method (Zimmer et al. 1986). At pH 7.6, O₂-jump (up) experiments showed two relaxation processes, whereas O₂-jump (down) experiments showed a single exponential process. The faster process in the O₂-jump (up) experiments could be attributed to the same process as that seen in the temperature-jump experiments (Zimmer et al. 1986). The slower process in the O₂-jump (up) experiments corresponds to the process obtained in the O₂-jump (down) experiments. The results are discussed in terms of a state with intermediate affinity in O₂-binding and with the possible existence of a slow step in O₂-binding.Abbreviations Hr hemerythrin - Hb hemoglobin - CM carboxymethyl - T-jump temperature-jump Offprint requests to: H. Kihara     

Thermoregulation,gas exchange,and ventilation in Adelie penguins (Pygoscelis adeliae)

Summary Adelie penguins (Pygoscelis adeliae) experience a wide range of ambient temperatures (T _a) in their natural habitat. We examined body temperature (T _b), oxygen consumption ( ), carbon dioxide production ( ), evaporative water loss ( ), and ventilation atT _a from –20 to 30 °C. Body temperature did not change significantly between –20 and 20°C (meanT _b=39.3°C).T _b increased slightly to 40.1 °C atT _a=30°C. Both and were constant and minimal atT _a between –10 and 20°C, with only minor increases at –20 and 30°C. The minimal of adult penguins (mean mass 4.007 kg) was 0.0112 ml/[g·min], equivalent to a metabolic heat production (MHP) of 14.9 Watt. The respiratory exchange ratio was approximately 0.7 at allT _a. Values of were low at lowT _a, but increased to 0.21 g/min at 30°C, equivalent to 0.3% of body mass/h. Dry conductance increased 3.5-fold between –20 and 30°C. Evaporative heat loss (EHL) comprised about 5% of MHP at lowT _a, rising to 47% of MHP atT _a=30°C. The means of ventilation parameters (tidal volume [VT], respiration frequency [f], minute volume [I], and oxygen extraction [ ]) were fairly stable between –20 and 10°C (VT did not change significantly over the entireT _a range). However, there was considerable inter- and intra-individual variation in ventilation patterns. AtT _a=20–30°C,f increased 7-fold over the minimal value of 7.6 breaths/min, and I showed a similar change. fell from 28–35% at lowT _a to 6% atT _a=30°C.Abbreviations C thermal conductance - EHL evaporative heat loss - oxygen extraction - f respiratory frequency - MHP metabolic heat production - evaporative water loss - LCT lower critical temperature - RE respiratory exchange ratio - T _a ambient temperature - T _b body temperature - rate of oxygen consumption - rate of carbon dioxide production - I inspiratory minute volume - VT tidal volume     

Effect of oxygen on the growth (yield) of Chlorella vulgaris

The growth yield of Chlorella vulgaris, Y _kJ defined as g cells harvested per kJ of light energy absorbed by the cells, was assessed in a turbidostat culture by varying CO₂ and O₂ partial pressures ( and ). The value of Y _kJ ranged from 3.1×10^-3 to 5.0×10^-3 g cells/kJ under light-limited conditions [ = 1.02.4%, = 065%; total pressure of gas (composed of CO₂, O₂ and N₂)=1 atm]. In the light-limited environment, the algal specific growth rate deteriorated appreciably with the increase of . The deterioration accounts for the above range of Y _kJ observed. The growth inhibition due to oxygen that was defined by subtracting from 1.0 the ratio of at given values of to that at = 0% extended from 0.07–0.30 (7–30%). However, glycolate could not be detected in the turbidostat culture. Isotopic experiments on the specific rate of ¹⁴CO₂ uptake also revealed that the inhibition due to oxygen was from 22–38% when was varied from 0 to nearly 100%. These effects of oxygen were discussed, referring to the activity of ribulose-1,5-bisphosphate carboxylase that is inhibited competitively by oxygen.Non-Standard Abbreviations INH isonicotinic acid hydrazide - PPO 2,5-diphenyloxazole - DCMU 3-(-3,4-dichlorophenyl)-1,1-dimetylurea - CA carbonic anhydrase - RuP₂ ribulose-1,5-bisphosphate     

Nitrate reductase in needles,roots and trunk wood of spruce trees [Picea abies (L.) Karst.]

Summary Nitrate reductase (EC 1.6.6.2) activity (NRA), as measured by an in vivo assay, is present in needle leaves and mycorrhizal fine root tips of adult Norway spruce [Picea abies (L.) Karst.] in at least equal amounts on a fresh weight basis, in both adult and 5-year-old trees. NRA could also be demonstrated in trunk wood of deroted trees after fertilization with 5 mM , exhibiting a longitudinal profile in the trunk. Inducibility in needles can more efficiently be achieved by NO₂ (100 g·m^-3) than by 5 mM nitrate, which is effective only in root-amputated trees. A remarkably high level of needle-NRA in unfertilized trees, which are characterized by a very low level of nitrate in the xylem sap, suggests that NRA in spruce needles may in part be constitutive. Organic-N is a major nitrogen source for the needles even in root-amputated trees, indicating pronounced exchange processes between ray parenchyma and trunk xylem, which in turn are modified by the nitrogen source fed to the trunk stump. Intact trees exhibit a very similar amino acid composition of the xylem sap, regardless of whether or has been fed. The amino acid pattern of the needles is not thrown out of balance by flooding with and , which occurs in fertilized derooted trees. This indicates a distinct potential for homoeostasis of nitrogen entrance-metabolism (i.e. NRA and glutamine synthetase activity) in the needles. In the ectomycorrhiza/fine root-system (EMC), marked differences in NRA were observed depending on root-tip diameter and along the longitudinal profile of the fine roots. EMC-nitrate reductase is strongly enhanced by . Needle-NRA exhibits a circannual rhythm. An early summer maximum is followed by a December minimum. This activity pattern matches well the transitory increase of soluble nitrogen in spring and the total protein maximum in winter. In an indirect way assimilatory NRA may well contribute to nitrogen overfertilization (by consumption of NO_X) as one possible cause of the contemporary decline of spruce populations.     

Determination of heteronuclear three-bond J-coupling constants in peptides by a simple heteronuclear relayed E.COSY experiment

Summary A simple heteronuclear relayed E.COSY pulse sequence with a minimum number of pulses is proposed for the quantitative determination of heteronuclear three-bond J-coupling constants in uniformly ¹³C-enriched polypeptide samples. Numerous heteronuclear three-bond coupling constants, including , , , and , can be determined for each residue from a single heteronuclear relayed E.COSY spectrum. Couplings relevant for stereospecific assignments as well as for the determination of dihedral angles in the amino acid backbone and in side chains are obtained. The method is demonstrated on the uniformly ¹³C-enriched decapeptide antamanide (-Val¹-Pro²-Pro³-Ala⁴-Phe⁵-Phe⁶-Pro⁷-Pro⁸-Phe⁹-Phe¹⁰-).     

Suitability of the shaking flask for oxygen supply to microbiological cultures

Due of its simplicity the shaking flask is used in serial studies, e.g. in the screening for secondary metabolites or in the optimization of fermentation processes. Experimental investigations in these small bioreactors are often the first step in developing a large-scale fermentation process.Movement of the flask should produce sufficient mixing, supply of oxygen, and removal of carbon dioxide. In the case of fluids with low or moderate viscosity, gas transport is the most important aspect. This publication summarizes data necessary to calculate the gas transport. These data are derived from the consideration of the gas diffusions through the cotton plug as well as from the substance transport between the gas and liquid phases. As a result suitable fermentation conditions can be selected. Finally, the performance limits of the shaking flask are illustrated using the example of the oxygen supply in a Streptomyces tendae fermentation.List of Symbols A _s Cross section of plug - A Surface area of liquid in flask - a A/V _F specific phase interface area - c Concentration - c ^* Saturation concentration - c Plug diffusion term - D Widest diameter of flask - Diffusion coefficients in multicomponent gas mix tures - Diffusion coefficients in binary gas mixtures - Diffusion coefficient of oxygen in the liquid - d Diameter of neck of flask - e Eccentricity - G Volume-based mass flow - G _m Maximum volume-based mass flow - g Acceleration due to gravity - h Height coordinate - ¯H Mean height of plug - Hy p _i/c ^*, Henry constant - K Consistency index - k D _xy/D _xz, Ratio of diffusion coefficients in binary gas mixtures - k _M Monod constant - k _L a Mass transport coefficient: gas/liquid - M Molecular weight - m Flow exponent - n Speed of shaking - p Pressure - p _i Partial pressure of gas component i - q Area-based flow of volume - R , respiration ratio - Sc , Schmidt number - T Absolute temperature - V Flask volume - V _F Volume of liquid in flask - w Velocity of the Stefan flow - x, y, z Ratios of the partial pressures of the gases O₂, CO₂, N₂ - Rate of shear - Dynamic viscosity of the liquid - Kinematic viscosity of the liquid - Density of the liquid - _x, Density of O₂ gas - Surface tension Indices 0 State in gas volume of shaking flask - 1 State in outside air - G Gas volume - x, y, z O₂, CO₂, N₂     

QTL mapping of resistance to Sclerotinia midstalk rot in RIL of sunflower population NDBLOSsel × CM625

Midstalk rot caused by Sclerotinia sclerotiorum is an important disease of sunflower in its main areas of cultivation. The objectives of this study were to (1) verify quantitative trait loci (QTL) for midstalk-rot resistance found in F₃ families of the NDBLOS_sel × CM625 population in recombinant inbred lines (RIL) derived from the same cross; (2) re-estimate their position and genetic effects; (3) draw inferences about the predictive quality of QTL for midstalk-rot resistance identified in the F₃ families as compared to those in the RIL. Phenotypic data for three resistance (leaf lesion, stem lesion, and speed of fungal growth) and two morphological traits (leaf length and leaf length with petiole) were obtained from 317 RIL following artificial infection in field experiments across two environments. For genotyping the 248 RIL, we selected 41 simple sequence repeat (SSR) markers based on their association with QTL for Sclerotinia midstalk-rot resistance in an earlier study. The resistance traits showed intermediate to high heritabilities and were significantly correlated with each other Genotypic correlations between F₃ families and the RIL were highly significant and ranged between 0.50 for leaf length and 0.64 for stem lesion. For stem lesion, two genomic regions on linkage group (LG) 8 and LG16 explaining 26.5% of the genotypic variance for Sclerotinia midstalk-rot resistance were consistent across generations. For this trait, the genotypic correlation between the observed performance and its prediction based on QTL positions and effects in F₃ families was surprisingly high The genetic effects and predictive quality of these two QTL are promising for application in marker-assisted selection to Sclerotinia midstalk-rot resistance.     

Foam behavior of biological media

Summary The influence of the alcohol concentration on the foaminess, , of-BSA-solutions is considered. This effect is calculated by means of the function (C_BSA . f), where f=1 for pure protein solutions and f>1 for alcohol solutions. f is calculated by f = 2^TT_eff. Here, where T_T is the turbidity temperature change due to solvent structure effects and T_D, the temperature correction due to alcohol-protein interaction. The constants necessary to calculate T_T and T_D are tabulated. The agreement between the calculated and measured foaminess , as a function of the n-propanol concentration is satisfactory and for methanol or ethanol excellent.     

A method for the destruction and analysis of biogenic silicon in two Antarctic diatom species:<Emphasis Type="Italic"> Thalassiosira</Emphasis> sp. and<Emphasis Type="Italic"> Chaetoceros brevis</Emphasis>

Diatoms in the Southern Ocean are limited by iron and light, and therefore produce little biomass. Sufficient biomass for analysis under these conditions requires large sample volumes, and diatom samples are therefore often pre-concentrated on a filter. A method for the digestion of diatom cells on polycarbonate filters, that is also suitable for trace metal analysis, is described here. Additional analysis by inductively coupled plasma-optical emission spectroscopy (ICP-OES) is used for the determination of biogenic silicon. Although several procedures were tested, the method of Hauptkorn et al., which uses tetramethylammonium hydroxide for the destruction of silicon is adapted here [Hauptkorn et al. (2001) Fres J Anal Chem 370:246–250]. Additional nitric acid is added to destroy the polycarbonate filters. The described method results in clear digests and a good correlation between cell numbers and silicon content. Using this procedure, the cellular silicon content for Chaetoceros brevis was determined as 86 ± 4 fmol cell⁻¹. For Thalassiosira sp. a sensitivity effect was observed, and silicon content was determined as . The obtained cellular silicon contents are in good agreement with values presented in the literature.