Membrane docking of the C2 domain from protein kinase Cα as seen by polarized ATR-IR. The role of PIP₂

We have used attenuated total internal reflection infrared spectroscopy (ATR-IR) spectroscopy to study the association of the C2 domain from protein kinase Cα (PKCα) with different phospholipid membranes, so as to characterise the mode of membrane docking and its modulation by the second-messenger lipid PIP?. In parallel, we have also examined the membrane interaction of the C2 domain from cytosolic phospholipase A?. PIP? did not induce significant changes in secondary structure of the membrane-bound PKCα-C2 domain, nor did binding of the PKCα-C2 domain change the dichroic ratios of the lipid chains, whereas the C2 domain from phospholipase A? did perturb the lipid chain orientation. Measurements of the dichroic ratios for the amide I and amide II protein bands were combined so as to distinguish the tilt of the β-sheets from that of the β-strands within the sheet. When associated with POPC/POPS membranes, the β-sandwich of the PKCα-C2 domain is inclined at an angle α=35° to the membrane normal, i.e., is oriented more nearly perpendicular than parallel to the membrane. In the process of membrane docking, the tilt angle increases to α=44° in the presence of PIP?, indicating that the β-sandwich comes closer to the membrane surface, so confirming the importance of this lipid in determining docking of the C2 domain and consequent activation of PKCα.     

Transport of ribosomal RNA into the oöcytes of the milkweed bug, Oncopeltus fasciatus

The accumulation of detectable amounts of newly synthesized ribosomes has been shown to be prevented by ligaturing the nutritive cords between the oöcytes and the trophic syncytium of the Oncopeltus ovary. Unligatured control oöcytes on the opposite side of the same bug accumulated normal quantities of newly synthesized ribosomes under the same conditions. It is concluded that the oöcytes are inactive in the synthesis of ribosomes, this function being entirely the province of the trophic syncytium. Evidence is presented which supports the contention that the oöcytes are synthesizing transfer RNA as well as a high molecular weight, polydisperse, fraction which may be the maternal messenger RNA of the egg.     

The ‘true’ growth efficiency of phytoplankton as influenced by light attenuation and insolation: implications of the photosynthesis-irradiance relationship

The true growth efficiency (c) relates the light energy absorbed by phytoplankton to the production of biomass corrected for constant energy requirement of maintenance. Continuous culture studies have shown that, at constant incident irradiance, the value of c for both prokaryotic and eukaryotic species is constant. Culture data for the relevant conditions of incident light may be used for directly estimating the growth rate from daily insolation of optically deep, fully mixed lakes, when the light absorption by the phytoplankton can be established. In order to examine the influence of vertical light attenuation and daily insolation on c, computations were made on a basis of a photosynthesis-irradiance curve of light-limited Oscillatoria limnetica. For steady state growth, the true growth efficiency is linearly related to the areal quantum efficiency of photosynthesis ( _a ). The computations showed that _a remains constant at fluctuating vertical light attenuation, no matter whether the concentration of tripton or phytoplankton changes. The effect of insolation is great: _a is 0.108 mol O₂/E at very low light, but only 0.014 mol O₂/E at 400 W m^–2 subsurface downward irradiance. The results imply that a c-value obtained from cultures for summer averaged insolation must be corrected: between cloudy and clear days the value may vary by a factor of 2. The true growth efficiency for cultures will decrease by about 10% when the same daily irradiation is dosed sinusoidally instead of constantly.     

Heteroplasmy of the chloroplast genome of Medicago sativa L. cv ‘Regen S’' confirmed by sequence analysis

The heteroplasmy of chloroplast DNA (cpDNA) observed in Medicago sativa L., which involves the presence (type B) or absence (type A) of an Xba I restriction site, was examined using closed fragments covering the variable XbaI site from type-A and type-B cpDNA. The 6.2-kb PstI fragment of DNA from type-A cpDNA (–XbaI) and from type-B cpDNA (+XbaI) was cloned into pUC19 plasmids. EcoRI fragments bearing the variable XbaI site from the type-A and type-B 6.2-kb PstI fragments were subcloned into pUC19. DNA sequences of both types of the 696-bp EcoRI fragments were determined and computer-assisted analysis of the sequence data carried out. Type-A cpDNA was found to differ from type-B cpDNA by 1 base, a G to T conversion, which results in a non-recognition site for XbaI in the type-A cpDNA. The sequence difference was in a non-coding region. Cloning and sequencing of the fragments verified the individual identity of the type-A and type-B cpDNA.     

The genius of E. E. metchnikoff— Discoveries over the centuries

On the 15 of May we celebrated the 150th anniversary of the outstanding Russian biologist Elias E. Metchnikoff (1845–1916)—Nobel Prize winner (1908), full and honorary member of many scientific academies of the world. His main works were applied to the zoology of invertebtates, evolution, embryology, immunology, microbiology, infectious pathology, gerontology, etc. Elias Metchnikoff published essays on anthropology, theory of orthobiosis, role of social and social-hygienic factors in solving the problems of old age and life elongation. On 30 May-2 June 1995 an International Symposium dedicated to Metchnikoff's 150th anniversary was held in Moscow. This is a text of the lecture given by us at the opening ceremony.     

Micropropagation of Calluna vulgaris cv. ‘H.E. Beale’

Axillary shoot producing cultures were obtained from microcuttings and shoot tips of Calluna vulgaris cv. H.E. Beale. For cultures derived from microcuttings the highest multiplication rate of 38 shoots (5 mm or longer) was obtained on a reduced salt medium with the addition of 0.5 mgl^-1 2-isopentenyladenine (2iP) during an 8 week subculture. For shoot tip derived cultures 0.2 mgl^-1 6-benzyladenine (BA) was the best cytokinin and led to a multiplication rate of 26 for a 6 week subculture. The addition of 1 g/l casein hydrolysate to a multiplication medium enhanced shoot proliferation in presence of 0.5 mgl^-1 BA.Despite various auxin treatments shoots formed no roots in vitro but rooted readily if transferred to a peat substrate ex vitro. A high rooting percentage (80%) was also obtained with shoots taken from the end of a multiplication phase and rooted directly. An additional subculture on low auxin containing media before transfer to peat substrate is recommended because the shoot condition can be improved in this way. A high number of rooted plantlets was produced, so the methods described will allow mass propagation.     

Conformation of the C-terminal pentapeptide—the ‘message sequence’—of tachykinins as determined by consensus dynamic

The tachykinins are a family of gastrointestinal peptides comprising eight members: substance P, neurokinin A, neurokinin B, eledoisin, physalemin, uperolein, kassinin and phyllomedusin. Consensus dynamics was carried out on an ensemble of seven tachykinins to determine the binding conformation of the common C-terminal fragment: Phe-X-Gly-Leu-Met-NH ₂ ,the ’message sequence’ of tachykinins. Three binding modes for the C-terminal pentapeptide were determined. The first binding conformation is folded due to an intramolecular H-bond between the NH of the variable residue (X) and CO of Met. Other features include γ-bends at both the variable amino acid (X) and at Gly. The global minimum of the simulation has this conformation for the C-terminal pentapeptide. The other two binding modes have slightly higher energies. The second is chiefly characterized by a β-turn around the segment X-Gly-Leu-Met, with additional β-bends at the variable amino acid (X) and Met. The final binding conformation is composed of β-bends around the variable amino acid (X) and Leu, and a ’pseudo’ γ-bend at the terminal Met. This paper was presented at the MBU Silver Jubilee Symposium on Structural Biology and 24th Annual Meeting of the Indian Biophysical Society held at Indian Institute of Science, Bangalore, Dec. 9–12, 1996.     

‘The vain speculation disillusioned by the sense’: the Italian painter Agostino Scilla (1629–1700) called ‘The Discoloured’, and the correct interpretation of fossils as ‘lithified organisms’ that once lived in the sea

The search for the true nature and origin of fossils gave rise, starting about the beginning of the sixteenth century and throughout the seventeenth century, to a heated debate between the supporters of the inorganic nature of these objects and proponents of an interpretation of fossils as ex-vivi. Among the latter, a key figure is the Italian painter Agostino Scilla (1629–1700), who represents one of the first naturalists who was able to accumulate substantial evidence to support the interpretation of fossils as once-living marine organisms. The world view of Scilla and his real contribution to paleontology are here analysed and contextualised, taking into account the state of knowledge characterising the time of his activities. Scilla was not only a pioneer in the field of paleontology, but he prepared the groundwork for the achievement of milestone geological concepts such as actualism, taphonomy and recognition of internal and external moulds (in the instance of non-representation of original hard parts); Scilla deserves further recognition as one of the first to attempt sedimentological interpretation of rocky bodies observed directly in the field.     

The Fate of the Method of ‘Paradigms’ in Paleobiology

An earlier article described the mid-twentieth century origins of the method of “paradigms” in paleobiology, as a way of making testable hypotheses about the functional morphology of extinct organisms. The present article describes the use of “paradigms” through the 1970s and, briefly, to the end of the century. After I had proposed the paradigm method to help interpret the ecological history of brachiopods, my students developed it in relation to that and other invertebrate phyla, notably in Euan Clarkson’s analysis of vision in trilobites. David Raup’s computer-aided “theoretical morphology” was then combined with my functional or adaptive emphasis, in Adolf Seilacher’s tripartite “constructional morphology.” Stephen Jay Gould, who had strongly endorsed the method, later switched to criticizing the “adaptationist program” he claimed it embodied. Although the explicit use of paradigms in paleobiology had declined by the end of the century, the method was tacitly subsumed into functional morphology as “biomechanics.”     

Morpho-Physiological and Antioxidant Traits of Marigold cv. ‘Sparse Petal’ and ‘Compact Petal’ as Influenced by Irrigation Intervals

The effect of irrigation intervals was studied on physiological, morphological, and antioxidant traits of two marigold (Calendula officinalis L.) cultivars in Karaj, Iran, in a split-plot experiment based on a randomized complete block design with three replications. The experimental treatments included irrigation at three levels of I1 (irrigation interval of 3 days), I2 (irrigation interval of 5 days), and I3 (irrigation interval of 7 days) as the main plot and cultivar at two levels of V1 (cv. ‘sparse petal’) and V2 (cv. ‘compact petal’) as the sub-plot. The results based on the comparison of the means showed that the increase in irrigation interval from 3 to 7 days decreased the leaf area index, crop growth rate, relative growth rate, and net assimilation rate by 73.53, 85.76, 93.47, and 94.81%, respectively. It also decreased the flower yield, plant height, flower number, and leaf number by 71.92, 41.84, 99.31, and 58.67%, respectively. The interaction between irrigation and cultivar revealed that I3V1 had the highest total phenol content (3013.59 g gallic acid 100 g^?1 tissue) and antioxidant capacity (60.8%). It can be inferred that the treatment of I1 and cv. ‘compact petal’ give the best results for flower yield and physiological and morphological traits, and the treatment of I3 and both cultivars provide the best results for antioxidant traits in the climatic conditions of the Karaj region.

     

Reconsideration of the term ‘vitrification’ as used in micropropagation

The term vitrification is currently used to describe two types of processes related to tissue-cultured plant material. The first is used to describe organs and tissues having an abnormal morphological appearance and physiological function. The second is used to describe the transition from liquid to solid state, i.e. the formation of ice during low temperature storage of in vitro cultured cells, tissues and organs. Use of the same term to define two greatly different processes in the same research area can only lead to confusion, especially for key words. Thus it is appropriate to reconsider the usage of vitrification in the first sense mentioned above. It is recommended that the term vitrification should no longer be used to indicate plant material with an abnormal morphological appearance and physiological function, and should be substituted by the term hyperhydricity.This paper was written as a follow up of the discussions held during the workshop on Vitrification at the IAPTC-Congress in Amsterdam (1990). Different eminent research workers in the field of vitrification were invited to formulate their preference for a new terminology, and they came to a consensus.     

The ‘fluid mosaic model’ and the ‘lattice model’ as two nonequilibrium states of the same membrane and the significance for biochemical control

Summary Biochemical control involves steep and hysteretic response. But the law of mass action does not allow for cooperativity. Therefore resort is classically made to concerted conformational change of protomers. This explanation of steepness and hysteresis by cooperativity is supported by regular surface patterns often observed by electron microscopy. But at other times the lattice appearance which gave rise to the lattice model is not observed. By contrast, a random appearance is observed and the fluid mosaic model of the membrane is assumed. So we are faced with the choice between the fluid mosaic model and the lattice model. Recently the fluid mosaic model is favoured but unlike the lattice model it does not explain the steep hyteretic response.It is suggested that the lattice model and the fluid mosaic model are in fact expression of two states of the very same membrane. The random state corresponds to a resting state. The lattice state corresponds to an active or inhibited state. Thus the transition from random distribution to hexagonal distribution provides simultaneously for triggering and hysteretic cycle with respect to both chemical production and transport across the membrane. This is a universal mechanism for rapid responsiveness and cyclic activity which is largely independent of the chemical mechanism assumed. It is based on the law of mass action supplemented by lateral diffusion. Conformational change and cooperativity are not invoked at all.     

The genetic control of tolerance to aluminum toxicity in the ‘Essex’ by ‘Forrest’ recombinant inbred line population

Aluminum (Al) toxicity to plant roots is a major problem of acidic soils. The main chemical reaction involved is Al hydrolysis. Application of lime or nitrate fertilizers to raise soil pH reduces Al toxicity but not as economically as a plant genotypes with natural tolerance against this stress. Ammonium fertilization of crops and assimilation of ammonium (even that derived from dinitrogen) are particularly acidifying of the root zone. The aims of the present study were to find genotypes of soybean tolerant to aluminum stress and identify QTL underlying that trait. Used were recombinant inbred lines (RILs) derived from the cross of ‘Essex’ by ‘Forrest’. RILs were grown in a greenhouse for 3 weeks and then transferred to hydroponics in a growth chamber. Root lengths (RL) were measured before and 72 h after Al treatment. RL before and after Al treatment were measured and used to calculate root tolerance index (RTI) and relative mean growth (RMG). RILs 1, 85, 40 and 83 had significant (P < 0.005) tolerance to Al stress judged by RL after Al, RTI and RMG. Eleven minor but significant marker–trait associations (P < 0.05) were detected using one-way ANOVA but only two major loci were significant in composite interval maps (LOD >3.0). The QTL on linkage group F (chromosome 13) was in the interval Satt160–Satt252 with a peak at 24 cM (peak LOD was 3.3). The QTL underlay 31% of trait variation and the Essex allele provided an additional 1.61 cm of root growth over 72 h in the presence of Al. The QTL on linkage group C2 (probably chromosome 4) was in the interval from Satt202 to Satt371 with a peak at 3.2 cM (peak LOD was 14.7). The QTL underlay 34% of trait variation or 1.81 cm of growth over 72 h in the presence of Al. Both loci encompassed genes implicated in citrate metabolism, a method of aluminum detoxification known to vary among soybean cultivars. Two major loci and at least nine minor loci were inferred to underlie tolerance to Al. RILs and markers may be used to select alleles that increase tolerance to soybean against Al stress.