Pollenfertility and seed formation in theOrnithogalum umbellatum/angustifolium complex (Liliaceae/Scilloideae)

The pollen fertility and seed formation of six species of theOrnithogalum umbellatum/angustifolium complex and of seven related species were studied. Four types of pollen grains could be recognized. The pollen fertility varied greatly in this complex and is not related to the ploidy level. The seed formation ofO. umbellatum showed an adaptation to a subcontinental-Mediterranean climate, that ofO. angustifolium to an Atlantic climate. In both cases raindrops seem to be important for pollination, in view of the absence of insect pollinators. After open pollination 113 seedlings were obtained in four species. Their chromosome numbers were determined. Nearly all the cultivated seedlings were aneuploid, which points to a positive selection of euploids in nature, because aneuploid individuals are rare in the wild.Biosystematic Studies on theOrnithogalum umbellatum/angustifolium Complex III.—Previous parts of this series are Part I: Taxonomy. Proceeding Kon. Ned. Acad. Wet. series C,85 (4), 563–574 (1982) andvan Raamsdonk (1984).     

绿豆幼苗上胚轴伸长生长与细胞渗透势的相关性

在水分供应正常的情况下,切除1/4～1/2的绿豆子叶对上胚轴生长没有明显的抑制作用,但是经0.2M甘露醇处理的幼苗却随着切除的子叶量的增加,上胚轴的生长呈直线剧减,全部切除子叶的幼苗生长几乎完全停止。子叶在水分胁迫状况下对生长的这种作用与它在上胚轴生长区渗透调节中的作用密切相关。幼苗上胚轴的生长与细胞的有效渗透势之间的相关系数为-0.76,有较高的相关性,但是生长与细胞的总渗透势无相关性(相关系数为-0.29)。细胞的渗透势与细胞液中的糖、游离氨基酸和钾离子浓度之间有很高的相关性。表明水分胁迫下生长所需的低渗透势的维持依赖于来自子叶的渗透物质。     

Effect of nonanoic acid and other short chain fatty acids on exchange properties in embryonic axes of Cicer arietinum during germination

Nonanoic acid, which inhibits germination in several seeds, enhanced ion efflux from embryonic axes of Cicer arietinum L., especially at temperatures above 25°C. Other short chain fatty acids had little effect on germination and ion leakage. Nonanoic acid also decreased uptake of ⁸⁶Rb⁺ and ²²Na⁺ and increased efflux of both isotopes from the embryonic axes into the incubation solution. Fusicoccin, which stimulates early germination in C. arietinum , counteracted the effects of nonanoic acid at both 25 and 30°C. These results suggest that nonanoic acid affects the integrity of plasmalemma and other membrane systems. Nonanoic acid thus inhibits cell elongation during early germination by disturbing ion exchange and inhibiting water uptake.     

Sodium-alanine cotransport in oocytes ofXenopus laevis: Correlation of alanine and sodium fluxes with potential and current changes

Summary The sodium-dependentl-alanine transport across the plasma membrane of oocytes ofXenopus laevis was studied by means of [¹⁴C]-l-alanine,²²Na⁺ and electrophysiological measurements. At fixed sodium concentrations, the dependence of alanine transport on alanine concentration follows Michaelis-Menten kinetics; at fixed alanine concentrations, the transport varies with sodium concentration with a Hill coefficient of 2. In the presence of sodium the uptake of alanine is accompanied by a depolarization of the membrane. Under voltage-clamp conditions this depolarization can be compensated by an inward-directed current. Assuming that this current is carried by sodium we arrive at a 21 stoichiometry for the sodium-alanine cotransport. The assumption was confirmed by direct measurements of both sodium and alanine fluxes at saturating concentrations of the two substrates, which also yielded a stoichiometry close to 21. The sodium-l-alanine cotransport is neither inhibited by furosemide (0.5 mmol/liter) nor by N-methyl amino isobutyric acid (5 mmol/liter). A 20-fold excess ofd-alanine overl-alanine caused about 60% inhibition.     

Effect of calcium ion on cytoplasmic streaming during turgor regulation in a brackish water charophyte Lamprothamnium

Abstract The brackish water charophyte Lamprothamnium succinctum regulates its turgor pressure against changes in the external osmotic pressure. Upon hypotonic treatment, the rate of cytoplasmic streaming in the internodal cells fell to almost zero, and then recovered to the original value within 20 min. The decrease could be inhibited by lowering the external Ca²⁺ concentration in the hypotonic medium. Also, cytoplasmic streaming in tonoplast-free cells of L. succintum was sensitive to Ca²⁺ like freshwater charophyte. Thus, the concentration of free Ca²⁺ in the cytoplasm seems to increase transiently upon hypotonic treatment.     

One- and two-dimensional NMR spectral analysis of the consequences of single amino acid replacements in proteins

The traditional approach of using homologous sequences to elucidate the role of specific amino acid residues in protein structure and function becomes more meaningful as the number of differences is minimized, with the limit being alteration of a single residue. For small proteins in solution, NMR spectroscopy offers a means of obtaining detailed information about each residue and its response to a given change in the protein sequence. Extraction of this information has been aided by recent progress in spectrometer technology (higher magnetic fields, more sensitive signal detection, more sophisticated computers) and experimental strategies (new NMR pulse sequences including multiple-quantum and two-dimensional NMR methods). The set of avian ovomucoid third domains, which consists of the third domain proper plus a short leader (connecting peptide) and has a maximum of 56 amino acid residues, offers an attractive system for developing experimental methods for investigating sequence-structure and structure-function relationships in proteins. Our NMR results provide examples of sequence effects on pKa' values, average conformation, and internal motion of amino acid side chains.     

Localized coupling in oxidative phosphorylation by mitochondria from Jerusalem artichoke (Helianthus tuberosus)

Delocalized chemiosmotic coupling of oxidative phosphorylation requires that a single-value correlation exists between the extent of and the kinetic parameters of respiration and ATP synthesis. This expectation was tested experimentally in nigericin-treated plant mitochondria in single combined experiments, in which simultaneously respiration (in State 3 and in State 4) was measured polarographically, FΔψ (which under these conditions was equivalent to ) was evaluated potentiometrically from the uptake of tetraphenylphosphonium⁺ and the rate of phosphorylation was estimated from the transient depolarization of mitochondria during State 4-State 3-State 4 transitions. The steady-state rates of the different biochemical reactions were progressively inhibited by specific inhibitors active with different modalities on various steps of the energy-transducing process: succinate respiration was inhibited competitively with malonate or noncompetitively with antimycin A, or by limiting the rate of transport into the mitochondria of the respiratory substrate with phenylsuccinate; was dissipated by uncoupling with increasing concentrations of valinomycin; ADP phosphorylation was limited with oligomycin. The results indicate generally that when the rate of respiratory electron flow is decreased, a parallel inhibition of the rate of phosphorylation is also observed, while very limited effects can be detected on the extent of . This behavior is in marked contrast to the effect of uncoupling where the decreased rate of ATP synthesis is clearly due to energy limitation. Extending previous observations in bacterial photosynthesis and in respiration by animal mitochondria and submitochondrial particles the results indicate, therefore, that respiration tightly controls the rate of ATP synthesis, with a mechanism largely independent of . These data cannot be reconciled with a delocalized chemiosmotic coupling model.     

Sulfate influx across the rabbit ileal brush border membrane: Sodium and proton dependence,and substrate specificities

Summary In intact ileal mucosa, uptake of SO₄ across the brush border membrane requires the presence of Na and is saturable, withK1/2=1.3mm at 140mm Na (P.L. Smith, S.A. Orellana & M. Field, 1981.J. Membrane Biol. 63:199–206). The present study examines the substrate specificities and transport stoichiometry of the Na-dependent SO₄ uptake process. The effects of variations in medium anion and cation composition on lumen-to-epithelium influx of SO₄ (J _me ^SO4 ) were determined under short-circuit conditions.J _me ^SO4 is inhibited by thiosulfate, but not by phosphate, methylsulfate, vanadate or taurocholate. Cl is weakly inhibitory. Uptake of SO₄ is poorly supported by Li, and is unaffected by K, indicating a specific dependence on Na. At low SO₄ concentration (0.22mm),J _me ^SO4 is a hyperbolic function of medium Na concentration; the corresponding Hill plot is linear with a slope of 1.0, suggesting a transport stoichiometry of 1 Na: 1 SO₄. At high SO₄ concentration (6.7mm), the Na-dependent SO₄ velocity curve is sigmoidal and yields a Hill plot which is again linear but has a slope of 1.56, suggesting transport of more than 1 Na per SO₄. SO₄ uptake in presence of Na exhibits a dependence on medium pH. At 0.22mm SO₄ and 140mm Na,J _me ^SO4 was doubled by lowering pH from 7.4 to 6.8. However, at 6.7mm SO₄ and 140mm Na, changing pH had no effect onJ _me ^SO4 over the range 6.8 to 8.5. The pH dependence ofJ _me ^SO4 at 6.7mm SO₄ was restored when medium Na was lowered to 3mm, suggesting that pH sensitivity is a function of the concentration of preformed NaSO ₄ ^– ion pair. The results suggest that SO₄ influx across the ileal brush border occurs by electroneutral Na⁺/NaSO ₄ ^– or Na⁺/H⁺/SO ₄ ^2– cotransport, the former being favored by high concentrations of Na and SO₄.     

Structural and developmental differences between three types of Na channels in dorsal root ganglion cells of newborn rats

Summary The changes in Na current during development were studied in the dorsal root ganglion (DRG) cells using the whole-cell patch-clamp technique. Cells obtained from rats 1–3 and 5–8 days after birth were cultured and their Na currents were compared. On top of the two types of Na currents reported in these cells (fast-FA current and slow-S current) a new fast current was found (FN). The main characteristics of the three currents are: (i) The voltages of activation are –37, –36, and –23 mV for the FN, FA and S currents, respectively. (ii) The activation and inactivation kinetics of FN and FA currents are about five times faster than those of the S current. (iii) The voltages at which inactivation reaches 50% are –139, –75 and –23 mV for the FN, FA and S currents, respectively.The kinetics and voltage-dependent parameters of the three currents and their density do not change during the first eight days after birth. However, their relative frequency in the cells changes. In the 1–3 day-old rats the precent of cells with S, FA, and mixed S+FN currents is 22, 18, and 60% of the cells, respectively. In the 5–8 day-old, the percent of cells with S, FA, and FN+S is 10, 66 and 22%. The relative increase in the frequency of cells with FA current during development can contribute to the ease of action potential generation compared with cells with FN currents, which are almost completely inactivated under physiological conditions. The predominance of FA cells also results in a significant decrease in the relative frequency of cells with the high-threshold, slow current.Antibodies directed against a part of the S₄ region of internal repeat I of the sodium channel (C ₁ ⁺ , amino acids 210–223, eel channel numbering) were found to shift the voltage dependence of FA current inactivation (but not of FN or S currents) to more negative potentials. The effect was found only when the antibodies were applied externally. The results suggest that FN, FA and S types of Na currents are generated by channels, which are different in the topography of the C ₁ ⁺ region in the membrane.