Assessment of the Respiratory Costs of Adaptation in Plant Species That Differ in Their Responses to Insufficient and Excessive Mineral Nutrition

The effects of insufficient and excessive mineral nutrition on the relative growth rate (RGR); the rate of total dark respiration (R); gross photosynthesis (Pg); and the contents of endogenous IAA, ABA, and cytokinins (CK) were studied in 20–60-day-old seedlings of plant species that differed in their responses to these factors: beet root (Amaranthus retroflexusL.), orchard-grass (Dactylis glomerataL.), and common wheat (Triticum aestivumL.). The changes in the level of mineral nutrition lowered the RGR and Pgvalues and the IAA/ABA and CK/ABA ratios and increased the Rand R/Pgvalues and ABA content, especially in A. retroflexusand T. aestivum. The authors propose a new way to assess the respiratory cost of adaptation to stress based on the relation between the RGR and the R/Pgvalues. The adaptation component of Ris shown to relate to the ABA content.     

Foam behavior of biological media

Summary The surface tension and foaminess of (a) unlimited, (b) substrate limited, and (c) oxygen transfer limited growth media of Hansenula polymorpha were measured using methanol, ethanol or glucose as a substrate.The time dependence of can be described by the Avrami-Überreiter relationship: log (2.3 log V)=n log t+log b, where V = (_O – _eq/(_t – _eq, and _O, _t and _eq are at t_M=0, t_M=t and t_M (equilibrium value).The constants n and b are functions of the fermentation time t_F as long as the growth is unlimited but they are constant in the state of limited growth. With glucose substrate, the foaminess can be presented as a definite function of the time, t_DG, which is necessary to attain _eq. With alcohol as a substrate no definite (t_DG) function was found.Symbols b constant in Eq. (1) - n constant in Eq. (1) - S substrate concentration - T temperature - t_M time h (measured from the beginning of the determination of the surface tension ) - t_F cultivation time h (measured from the time of inoculation) - t_DG time (min) necessary to attain the equilibrium surface tension ) - X dry biomass concentration (gl^–1) - V (_O – _eq)/(_t – _eq) - V_S equilibrium volume of the foam (cm³) - V_G volumetric gas flow rate during the estimation of (cm³ s^–1) - vvm volumetric gas flow rate with regard to the volume of the medium (min^–1) - w_SG superficial gas velocity (cm s^–1) - _m maximum specific growth rate (h^–1) - V_S/V_G foaminess (s) - surface tension, mMm^–1 (milli Newton m^–1) - _O at t_M=0 - _eq equilibrium surface tension ( at t_M) - _t at t_M=t - HP probes from Hansenula polymorpha cultivation - NLG non limited growth - OTLG oxygen transfer limited growth - SLG substrate limited growth     

Average phase difference theory and 1∶1 phase entrainment in interlimb coordination

The dynamics of coupled biological oscillators can be modeled by averaging the effects of coupling over each oscillatory cycle so that the coupling depends on the phase difference between the two oscillators and not on their specific states. Average phase difference theory claims that mode locking phenomena can be predicted by the average effects of the coupling influences. As a starting point for both empirical and theoretical investigations, Rand et al. (1988) have proposed d/dt= — K sin ), with phase-locked solutions =arcsin( /K), where is the difference between the uncoupled frequencies and K is the coupling strength. Phase-locking was evaluated in three experiments using an interlimb coordination paradigm in which a person oscillates hand-held pendulums. was controlled through length differences in the left and right pendulums. The coupled frequency _c was varied by a metronome, and scaled to the eigenfrequency _v of the coupled system K was assumed to vary inversely with _c. The results indicate that: (1) and K contribute multiplicatively to (2) =0 or = regardless of K when =0; (3) 0 or regardless of when K is large (relative to ); (4) results (1) to (3) hold identically for both in phase and antiphase coordination. The results also indicate that the relevant frequency is _c/_v rather than _c. Discussion high-lighted the significance of confirming =arcsin(/K) for more general treatments of phase-locking, such as circle map dynamics, and for the 11 phase-entrainment which characterizes biological movement systems.     

Studies on the chemical composition of isolated soil solution and the cation absorption by plants

Summary The effects were studied of varying amounts and forms of N fertilizers on the chemical composition of soil solution and on the uptake of N, K, Na, Ca and Mg by barley.In isolated soil solution the concentrations as well as the ratios of individual cations depended on form and amount of added nitrogen. The concentration of cations increased proportionally with applications of nitrate+sulphate. Cation-activity ratios in isolated soil solution were governed by the ratios of corresponding exchangeable cations. During the entire growth period the absorption by the plants of individual as well as of cations was independent of their concentrations in soil solution. So long as NH₄ ions were present in the soil solution, plants with NH₄ addition contained less cations than those with NO₃ addition. Thereafter the amounts of cations in the plant were closely correlated with N absorption, but not affected by form of applied N. The ratios K/(Ca+Mg) and Ca/Mg in the plant were closely correlated with corresponding activity ratios but independent of concentration ratios of these ions in isolated soil solution. It is postulated that uptake of cations is governed by N-metabolic processes in the plant, whereas ratios of absorbed cations are influenced by cation-activity ratios in the soil solution.     

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.     

Cell recycling studies for α-amylase production by Bacillus amyloliquefaciens

Production of -amylase by a strain of Bacillus amyloliquefaciens was investigated in a cell recycle bioreactor incorporating a membrane filtration module for cell separation. Experimental fermentation studies with the B. amyloliquefaciens strain WA-4 clearly showed that incorporating cell recycling increased -amylase yield and volumetric productivity as compared to conventional continuous fermentation. The effect of operating conditions on -amylase production was difficult to demonstrate experimentally due to the problems of keeping the permeate and bleed rates constant over an extended period of time. Computer simulations were therefore undertaken to support the experimental data, as well as to elucidate the dynamics of -amylase production in the cell recycle bioreactor as compared to conventional chemostat and batch fermentations. Taken together, the simulations and experiments clearly showed that low bleed rate (high recycling ratio) various a high level of -amylase activity. The simulated fermentations revealed that this was especially pronounced at high recycling ratios. Volumetric productivity was maximum at a dilution rate of around 0.4 h^–1 and a high recycling ratio. The latter had to exceed 0.75 before volumetric productivity was significantly greater than with conventional chemostat fermentation.List of Symbols a proportionality constant relating the specific growth rate to the logarithm of G (h) - a ₁ reaction order with respect to starch concentration - a ₂ reaction order with respect to glucose concentration - B bleed rate (h^–1) - C starch concentration (g/l) - C ₀ starch concentration in the feed (g/l) - D dilution rate (h^–1) - D _E volumetric productivity (KNU/(mlh)) - e intracellular -amylase concentration (g/g cell mass) - E extracellular -amylase concentration (KNU/ml) - F volumetric flow rate (l/h) - G average number of genome equivalents of DNA per cell - k _l intracellular equilibrium constant - k ₂ intracellular equilibrium constant - k _s Monod saturation constant (g/l) - k ₃ excretion rate constant (h^–1) - k _d first order decay constant (h^–1) - k _gl rate constant for glucose production - k _st rate constant for starch hydrolysis - k _t1 proportionality constant for -amylase production (gmRNA/g substrate) - k ₁ translation constant (g/(g mRNAh)) - KNU kilo Novo unit - m maintenance coefficient (g substrate/(g cell massh)) - n number of binding sites for the co-repressor on the cytoplasmic repressor - Q repression function K₁/K₂Q1.0 - R ratio of recycling - R _s rate of glucose production (g/lh) - r _c rate of starch hydrolysis (g/(lh)) - R _eX retention by the filter of the compounds X: starch or -amylase - r intracellular -amylase mRNA concentration (g/g cell mass) - r _C volumetric productivity of starch (g/lh) - r _E volumetric productivity of intracellular -amylase (KNU/(g cell massh)) - r _r volumetric productivity of intracellular mRNA (g/(g cell massh)) - r _e volumetric productivity of extracellular -amylase (KNU/(mlh)) - r _s volumetric productivity of glucose (g/(lh)) - r _X volumetric productivity of cell mass (g/(lh)) - S ₀ free reducing sugar concentration in the feed (g/l) - S extracellular concentration of reducing sugar (g/1) - t time (h) - V volume (l) - X cell mass concentration (g/l) - Y yield coefficient (g cell mass/g substrate) - Y _E/S yield coefficient (KNU -amylase/g substrate) - Y _E total amount of -amylase produced (KNU) - substrate uptake (g substrate/(g cell massh)) - specific growth rate of cell mass (h^–1) - _d specific death rate of cells (h^–1) - _m maximum specific growth rate of cell mass (h^–1) This study was supported by Bioprocess Engineering Programme of the Nordic Industrial Foundation and the Center for Process Biotechnology, the Technical University of Denmark.     

Photosynthetic glow peaks and their relationship with the free energy changes

This paper is concerned with relating thermoluminescence to the total free-energy change, G, involved in detrapping a particular electron-hole pair as a photosynthetic sample is warmed from an initial low temperature. It extends a mathematical discussion of four possible mechanisms introduced in an earlier paper [DeVault, Govindjee and Arnold, Proc Nat'l Acad Sci USA 80: 983–987 (1983)]; here, particular attention is paid to the dependence of the absolute temperature of the maximum of a glow-peak, T _m , on the total free-energy change, G. The conclusion from the cases studied is that T _m =G/(k _B W) where G is evaluated at T _m , W is a complicated function of temperature and of thermodynamic parameters in the steps of the mechanism, and k _B is the Boltzmann constant. If the rate limiting step in the mechanism of detrapping is not preceded by any step in which G is appreciably negative, W is likely to have a value of about 33 and T _m is approximately proportional to G. Otherwise W can become much smaller and more strongly dependent on temperature and T _m is no longer proportional to G. These conclusions are of significance in lending theoretical support to the practice of inferring redox midpoint potential changes from shifts in T _m .     

Enzymic synthesis of the trisaccharide core region of the carbohydrate chain ofN-glycoprotein

Transmannosylation from mannotriose (Man1-4Man1-4Man) to the 4-position at the nonreducing end N-acetylglucosaminyl residue ofN,N-diacetylchitobiose was regioselectively induced through the use of -d-mannanase fromAspergillus niger. The enzyme formed the trisaccharide Man1-4GlcNAc1-4GlcNAc (3.7% of the enzyme-catalysed net decrease ofN,N-diacetylchitobiose) from mannotriose as a donor andN,N-diacetylchitobiose as an acceptor. Mannobiose (Man1-4Man) was also shown to be useful as a donor substrate for the desired trisaccharide synthesis.Abbreviations Man d-mannose - (M _n) (n=1–5) -linkedn-mer of mannose - GlcNAc₂ 2-acetamido-2-deoxy--d-glucopyranosyl-(1–4)-2-acetamido-2-deoxy-d-glucose     

A new mouse T-cell receptorα chain variable region family

Sequence analysis of the rearranged T-cell receptor a chain gene segments from an influenza reactive T-cell clone T2.5-5 and a hemin chloride reactive T-cell hybrid SJL-HE-1.1 have revealed a previously undescribedV gene family. We have designated this familyV 15. Southern hybridization analysis has indicated that this family most probably contains only two members, and that these are conserved in each of six mouse strains representing three previously describedV haplotypes:V ^a ,V ^b , andV ^c .     

Distribution of greenhouse gases, nitrite, and δ13C of dissolved inorganic carbon in Lake Biwa: Implications for hypolimnetic metabolism

The vertical distribution of dissolved greenhouse gases (CH₄,CO₂ and N₂O), NO ,and ¹³C of CO₂ in Lake Biwa during a stagnantperiod was precisely determined. CO₂ as well as NO was accumulated in the hypolimnion, whereas NO and CH₄concen\-trations were generally higher in theepilimnion than in the hypolimnion. In August, NO andCH₄ were ephemerally accumulated at the thermocline. Theconcentration of CH₄ always exceeded equilibrium with respectto air/water exchange. N₂O was rather uniformly distributed inboth time and space, and remained near equilibrium with respect toair/water exchange. All of these observations are similar to otherstratified, oligotrophic lakes, in which the hypolimnia were welloxygenated. The ¹³C of CO₂ became morenegative with increasing depth, and showed a strong negativecorrelation with apparent oxygen utilization. From the data, the ¹³Cvalue of organic matter decomposed into CO₂ inthe hypolimnion was calculated by isotope mass-balance, and found tobe in a similar range to ¹³C of phytoplankton and benthic algaeand distinctively higher than ¹³C of both terrestrial andsedimentary organic matters. This suggests that autochthonous organicmatter was the major source of CO₂.     

High irradiance response promotion of a subsequent light induction response in Sinapis alba L.

Relative quantum responsivity curves for inhibition of hypocotyl elongation in Sinapis alba L. seedlings previously grown in white light confirm that a marked end of day inhibition response can be induced by a monochromatic light treatment (30 min) at the end of the light period. In dark grown seedlings, however, no growth inhibition can be induced by a 30 min monochromatic light treatment. A prerequisite for an induction response appears to be a pretreatment with continuous light. Far red light is most effective with blue and red light showing a lesser effectiveness. The light pretreatment also shows a marked fluence rate dependency with respect to its ability to allow an induction response to manifest itself. The pretreatment required shows all the characteristics of a classical HIR response. The appearance of the effect in plants treated with the herbicide SAN 9789 seems to exclude chlorophyll as being the photoreceptor.Abbreviations SAN 9789 4-chloro-5-(methylamino)-2-(, , -trifluoro-m-tolyl)-3(2H)-pyridazinone - RG9 light long wavelength far red light (Schott RG9 colour glass) - FR far red light - WL white light - BL blue light - RL red light - D darkness - P_tot total phytochrome - P_fr far red absorbing form of phytochrome - HSR high irradiance response     

Constant-time NOESY: An aid in the analysis of protein NMR spectra

Summary A constant-time version of the homonuclear NOESY experiment (CT-NOESY) is described. The experiment yields simplified protein spectra, in which cross peaks arising from protons with zero or small couplings are differentiated from other cross peaks, thus partially overcoming the problem of signal overlap. In addition, the CT-NOESY spectrum provides information on the magnitude of³J_NH- and³J coupling' constants, and is thus useful to determine torsion angle constraints and to perform stereospecific assignments of CHH protons in the case of³J constants.     

Ultrastructure of spermatogenesis and sperm ofMulticotyle purvisi (Plathelminthes,Aspidogastrea)

Summary The ultrastructure of spermatogenesis is described for the first time in an aspidogastrean. The zone of differentiation which is usually formed during digenean spermiogenesis was not observed inMulticotyle purvisi. Instead, spermatid components are assembled within the common cytoplasmic mass before the outgrowth of spermatids. Microtubules, mitochondrion, nucleus and axonemes including their basal body regions, migrate from the cytoplasm into the spermatid which is pinched off at the level of the arching membrane. An unusual, complex structure of the basal body region is described. Intercentriolar bodies and striated rootlets are left behind and quickly disappear from the residual cytoplasm. Despite these atypical aspects, spermiogenesis results in the formation of mature sperm with the classical structure common to Digenea and Monogenea Polyopisthocotylea with the addition of some extra, non-cortical microtubules and a dense rod along part of the length of the sperm.Abbreviations used in the figures A cell type A, primary spermatogonium - AM arching membrane - AX axoneme - AZ attachment zone - B cell type B — spermatogonium - BB basal body region - C cell type C — spermatogonium - CEL central element - CI cisternae - CY cytophore - D cell type D — primary spermatocyte - DO doublet of microtubules - DR dark rod - E cell type E — multinucleate condensed cytophore - ER endoplasmic reticulum - G glycogen - GO Golgi body - I intercentriolar body - LB lamellate body - M mitochondrion - ME remnants of arching membranes - MT microtubules - N nucleus - R rootlet - S spermatid     

Osmotropotaxis inDrosophila melanogaster

Summary Drosophila melanogaster is able to perform osmotropotaxis under open-loop conditions. With an optimal stimulus the average turning tendency to the side of higher concentration corresponds to a circular track with radiusr=0.8 cm. The response amplitude does not decrease within 1 or 2 h. Unilaterally antennectomized flies in an homogeneous odor field show a permanent turning tendency towards their intact side.The smallest concentration ratio to elicit osmotropotaxis in normal flies ranges between 610 and 910 at high and between 210 and 510 at an about 50 times lower odor intensity. No negative tropotaxis (i.e. turning to the side of lower concentration) is observed, even with strong repellents.     

The α2 cDNA sequence of human haptoglobin carries a bacterial promoter functionalin vivo

Various constructions of human haptoglobin (Hp) cDNA coding either for the complete ^2FS precursor protein or only for the subunit have been placed under the control of the P_R promoter in the bacterial expression vector pCQV2 (Queen, 1983). In addition to the expected 45,000 dalton polypeptide synthesized after induction of the P_R promoter, the complete ^2FS constructions constitutively express a smaller polypeptide of 30,000 dalton corresponding to a truncated Hp protein. Computer analysis of the HpcDNA revealed the presence of two strong potential bacterial promoters (²P_F and ²P_S) located in the duplicated ^2FS sequence. Both Hp promoter signals are followed by potential mRNA start sites and ribosome binding sites at a compatible distance from initiation codons. In addition, the Hp² cDNA sequence, when fused upstream to the cDNA coding for ¹-antitrypsin, constitutively promotesin vivo the efficient expression of an hybrid protein specifically recognized by antibodies raised against ¹-antitrypsin or haptoglobin.     

Organization of α-chain genes among Hb G-Philadelphia heterozygotes in association with Hb S, β-thalassemia,and α-thalassemia-2

The percentages of the -chain variant Hb G-Philadelphia (Hb G) or ₂ 68 AsnLys₂ were evaluated in 84 adult and 18 newborn heterozygotes. These included members of three families who were studied in more detail by nucleic acid hybridization techniques. The adult heterozygotes fell in two categories, one with a higher proportion of Hb G [46.5±1.0% (SD), N=21] and another with lower values (33.9±3.4%, N=63). Among the newborn heterozygotes, two babies fell in the category with the higher proportion of Hb G while 16 babies gave values between 25 and 34%. Studies of -chain gene organization on the parents of one neonate with a Hb G level of 27% at birth and 37% at 8 months excluded the presence of chromosomes with triplicated -chain genes which could lead to the ^0G/ genotype. Rather, these studies on five Hb G heterozygotes from three families confirmed the linkage between Hb G and a specific type of -thalassemia-2 associated with the presence of a 16-kbp Bgl II fragment which most probably carries the ^G locus since it has been found in 19 Hb G heterozygotes studied to date. The presence of an -thal-2 heterozygosity and three -chain genes (^0G/) was confirmed among Hb G heterozygotes with lower proportions of this variant. It is likely that the even lower values found in some newborn could arise through defective assembly of ^G- dimers. The presence of an -thal-2 homozygosity and two active -chain genes, one on each chromosome (^0G/⁰), was confirmed among heterozygotes with the higher proportion of Hb G. One of each of these categories was present in each of the three families investigated. This type of variability in the number of active -chain genes due to a heterozygosity or a homozygosity for -thalassemia-2 explains the trimodality of Hb S percentages among heterozygotes and the atypical hematological or biosynthetic features among patients with -thalassemia and sickle-cell syndromes.This research was supported by USPHS Research Grants HLB-05168 and HLB-15158 and by designated research funds of the Veterans Administration. This is Contribution No. 0693 of the Department of Cell and Molecular Biology, Medical College of Georgia, Augusta.     

Soluble Expression and Affinity Purification of Functional Domain of Human Acetylcholine Receptor <Emphasis Type="Italic">α</Emphasis>-subunit by the Modulation of Maltose Binding Protein

An open reading frame of the -subunit 1-205 residues (₂₀₅) of human acetylcholine receptor (AchR) was amplified by PCR with pUC-AChR₂₀₅ as the template and inserted into vector pMAL-c2X. The constructed pMAR₂₀₅ was transferred into E. coli BL21 which were then grown in LB medium. The amount of soluble MBP-AChR₂₀₅ protein reached about 25% of total soluble proteins from the cell lysate. Using amylose-affinity chromatography, about 35 mg MBP-AChR₂₀₅ could be obtained from 1 l culture. Western blot analysis and ELISA showed that immunoreactivities of both MBP-AChR₂₀₅ and AChR₂₀₅ were similar to that of AChR -subunit from Torpedo.Revisions received 23 September 2004     

Belastung des Kiebitzes (Vanellus vanellus) mit Dieldrin und anderen chlorierten Kohlenwasserstoffen

Zusammenfassung 45 Eier und 8 Kükenlebern des Kiebitzes aus den Jahren 1984 und 1986 wurden auf -, -, -HCH, HCB, DDT, Dieldrin, Endrin und PCB untersucht. Hohe Konzentrationen an Dieldrin wurden in den Eiern gefunden, während die Lebern von Kiebitzküken geringer belastet waren. Die geringen Konzentrationen der übrigen Pestizide und die mäßige an PCB entsprachen der niedrigen Trophieebene des Kiebitzes. Die Frage, ob die Belastung mit Dieldrin einen Einfluß auf den Rückgang von Kiebitzpopulationen in einigen Gebieten haben könnte, wird kurz diskutiert.

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The burden of Dieldrin and other chlorinated hydrocarbons on the Lapwing (Vanellus vanellus)

Summary -, -, -HCH, HCB DDT, Dieldrin, Endrin, and PCB were determined in 45 eggs and 8 livers of young Lapwings in 1984 and 1986. High concentrations of Dieldrin were found in eggs while those in livers of young Lapwings were lower. The low residues of the other pesticides and the moderate ones of PCB are in accordance with the low trophic level of the Lapwing. The probable influence of the Dieldrin burden on the decline of Lapwing populations in several regions is discussed briefly.

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Gefördert mit Hilfe von Forschungsmitteln des Landes Niedersachsen     

Delivery of foreign genes to intact barley cells by high-velocity microprojectiles

Summary Foreign DNA was introduced through the cell walls of intact suspension culture cells of barley (Hordeum vulgare L.) by utilizing the particle acceleration approach. DNA-coated microscopic tungsten particles were accelerated to velocities that permitted their penetration of intact cells. Chimaeric constructs of -glucuronidase and neomycin phosphotransferase II under the control of the dual Agrobacterium T_R 12 promoter or the cauliflower mosaic virus 35S promoter served as reporter genes. Three days after particle delivery, high-level expression of both reporter genes was observed. That plasmid size could be critical for stabilizing DNA in the course of particle delivery will be discussed.     

Conformational dynamics of the anticodon loop in yeast tRNAPhe as sensed by the fluorescence of wybutine

Conformational and dynamic properties of the anticodon loop of yeast tRNA^Phe were investigated by analyzing the time resolved fluorescence of wybutine serving as a local structural probe adjacent to the anticodon GmAA on its 3 side. The influence of Mg²⁺, important for stabilizing the tertiary structure of tRNA, and of the complementary anticodon s²UUC of E. coli tRNA ₂ ^Glu were investigated.Fluorescence lifetimes and anisotropies were measured with ps time resolution using time correlated single photon counting and a mode locked synchronously pumped and frequency doubled dye laser as excitation source. From the analysis of lifetimes () and rotational relaxation times ( _R ) we conclude that wybutine occurs in various structural states: (i) one stacked conformation where the base has no free mobility and the only rotational motion reflects the mobility of the whole tRNA molecule (=6 ns, _R =19 ns), (ii) an unstacked conformation where the base can freely rotate (=100 ps, _R = 370 ps) and (iii) an intermediary state (=2 ns, _R = 1.6 ns).Under biological conditions, i. e. in the presence of Mg²⁺ and neutral salts, wybutine is found in a stacked and immobile state which is consistent with the crystallographic picture. In the presence of the complementary codon however, as exemplified by the E. coli-tRNA ₂ ^Glu anticodon, our analysis indicates that the codon-anticodon complex exists in an equilibrium of structural states with different rotational mobility of wybutine. The conformation with wybutine freely mobile is the predominant one and suggests that this conformation of the codon-anticodon structure differs from the canonical 3–5 stack.