Aerenchyma and lenticel formation in pine seedlings: A possible avoidance mechanism to anaerobic growth conditions

Seedlings of pond pine ( Pinus serotina Michx.), sand pine [ P. clausa (Engelm.) Sarg.], and loblolly pine ( P. taeda L., wet-site and drought-hardy seed sources) were grown in hydroponic solution culture using a non-circulating, continuously flowing design under anaerobic or aerobic conditions to determine whether flooding tolerance was correlated with enhanced internal root aeration. Transport of atmospheric O₂ from the shoot to the root of anaerobically grown loblolly and pond pine seedlings was demonstrated via rhizosphere oxidation, using both reduced indigo-carmine solution and a polarographic, ensheathing Pt-electrode. Stem and root collar lenticels were the major sites of atmospheric O₂ entry for submerged roots in these seedlings. No O₂ leakage was detected from roots of aerobically grown pine seedlings. Longitudinal and radial pathways for gaseous diffusion via intercellular air spaces in the pericycle and between ray parenchyma cells, respectively, were demonstrated histo-logically in anaerobically grown loblolly and pond pines. Rhizosphere oxidation, and lenticel and aerenchyma development in roots of flood-intolerant sand pine seedlings grown in anaerobic solutions were minimal. Only 15 days of anaerobic growth conditions were necessary to increase internal root porosities of loblolly and pond pine seedlings – although not to the extent found in seedlings treated for 30 or 75 days. Histological results indicated that root tissue in the secondary stage of growth was capable of forming intercellular air spaces, demonstrating a degree of internal plasticity – at least in the more flood-tolerant loblolly and pond pine seedlings.     

Photosynthetic characteristics of the submerged macrophyte Ceratophyllum demersum

The photosynthetic and growth characteristics of Ceratophyllum demersum L. were investigated under laboratory conditions which simulated those encountered in the plants' normal environment. The carbon fixation rate of C. demersum measured with ¹⁴C at light and carbon saturation at pH 8.0 was 4.48 mg C (g ash-free dry weight)⁻¹ h⁻¹. It was lower at pH 6.5 than at pH 8.0. The light use efficiencies in quiescent plants and actively growing plants were 6.3 and 8.7 × 10⁻⁹ kg CO₂ J⁻¹, respectively, with corresponding maximum photosynthetic rates of 2.67 and 4.36 mg C (g ash-free dry weight)⁻¹ h⁻¹. Photorespiration in actively growing plants consumed 24% of the carbon fixed. Incubation with DCMU demonstrated that about one-third was refixed. The optimum temperature for carbon fixation was 25°C. The C₃-photosynthetic pathway was the main operational route as indicated by the early photosynthetic products (largely C₃-acids) and the absence of Krantz anatomy and the chlorophyll a:b ratio (2.7). The maximum relative growth rates ranged from 0.025 to 0.041 g ash-free dry weight (g ash-free dry weight)⁻¹ day⁻¹ in the field (Lake Vechten, 1 to 3 m depth classes).     

A method to quantitate Coomassie blue-stained proteins in cylindrical polyacrylamide gels

A method for the quantitation of Coomassie blue-stained proteins in cylindrical polyacrylamide gels is described. It involves an elution of the dye with an 80% methanol solution in a sealed Pyrex tube at 100 degrees C for 3 h and a measurement of its concentration at 585 nm. Using a 6.5% polyacrylamide gel and bovine serum albumin as a protein standard, the curve of absorbance of the dye solution as a function of the amount of protein was observed to be linear up to 30-40 micrograms of protein and as little as 0.8-1.0 micrograms of protein could be measured. The validity of the method was indicated by the values obtained for the relative proportions of the human erythrocyte membrane proteins. Using this method, the color yields of several proteins varying widely with respect to their size, amino acid composition, and carbohydrate content were determined in a 6.5% polyacrylamide gel. The results showed that they were generally the same except for proteins having a high carbohydrate content which were significantly lower.     

Genetic reconstruction and functional analysis of the repeating lipoyl domains in the pyruvate dehydrogenase multienzyme complex of Escherichia coli

The dihydrolipoamide acetyltransferase component (E2p) of the pyruvate dehydrogenase complex of Escherichia coli contains three highly homologous sequences of about 100 residues that are tandemly repeated to form the N-terminal half of the polypeptide chain. All three sequences include a lysine residue that is a site for lipoylation and they appear to form independently folded functional domains. These lipoyl domains are in turn linked to a much larger (about 300 residues) subunit-binding domain of the E2p chain that aggregates to form the octahedral inner core of the complex and also contains the acetyltransferase active site. In order to investigate whether individual lipoyl domains play different parts in the enzymic mechanism, selective deletions were made in vitro in the dihydrolipoamide acetyltransferase gene (aceF) so as to excise one or two of the repeating sequences. This was facilitated by the high degree of homology in these sequences, which allowed the creation of hybrid lipoyl domains that closely resemble the originals. Pyruvate dehydrogenase complexes incorporating these genetically reconstructed E2p components were purified and their structures were confirmed. It was found that the overall catalytic activity, the system of active site coupling, and the ability to complement pyruvate dehydrogenase complex mutants, were not significantly affected by the loss of one or even two lipoyl domains per E2p chain. No special role can be attached thus far to individual lipoyl domains. On the other hand, certain genetic deletions affecting the acetyltransferase domain caused inactivation of the complex, highlighting particularly sensitive areas of that part of the E2p chain.     

Localization of a base-paired interaction between small nuclear RNAs U4 and U6 in intact U4/U6 ribonucleoprotein particles by psoralen cross-linking

The small nuclear RNAs U4 and U6 display extensive sequence complementarity and co-exist in a single ribonucleoprotein particle. We have investigated intermolecular base-pairing between both RNAs by psoralen cross-linking, with emphasis on the native U4/U6 ribonucleoprotein complex. A mixture of small nuclear ribonucleoproteins U1 to U6 from HeLa cells, purified under non-denaturing conditions by immune affinity chromatography with antibodies specific for the trimethylguanosine cap of the small nuclear RNAs was treated with aminomethyltrioxsalen. A psoralen cross-linked U4/U6 RNA complex could be detected in denaturing polyacrylamide gels. Following digestion of the cross-linked U4/U6 RNA complex with ribonuclease T1, two-dimensional diagonal electrophoresis in denaturing polyacrylamide gels was used to isolate cross-linked fragments. These fragments were analysed by chemical sequencing methods and their positions identified within RNAs U4 and U6. Two overlapping fragments of U4 RNA, spanning positions 52 to 65, were cross-linked to one fragment of U6 RNA (positions 51 to 59). These fragments show complementarity over a contiguous stretch of eight nucleotides. From these results, we conclude that in the native U4/U6 ribonucleoprotein particle, both RNAs are base-paired via these complementary regions. The small nuclear RNAs U4 and U6 became cross-linked in the deproteinized U4/U6 RNA complex also, provided that small nuclear ribonucleoproteins were phenolized at 0 degree C. When the phenolization was performed at 65 degrees C, no cross-linking could be detected upon reincubation of the dissociated RNAs at lower temperature. These results indicate that proteins are not required to stabilize the mutual interactions between both RNAs, once they exist. They further suggest, however, that proteins may well be needed for exposing the complementary RNA regions for proper intermolecular base-pairing in the course of the assembly of the U4/U6 RNP complex from isolated RNAs. Our results are discussed also in terms of the different secondary structures that the small nuclear RNAs U4 and U6 may adopt in the U4/U6 ribonucleoprotein particle as opposed to the isolated RNAs.     

The characteristics of light in floral induction in vitro of Cichorium intybus. The possible role of phytochrome

The action of light in the initiation of floral buds in vitro has been studied using monochromatic light qualities on root explants of a long day plant, Cichorium intybus L. cv. Witloof. Red light (660 nm, 0.30 W m^-2) promotes flowering, while far-red (730 nm, 0.31 W m^-2) and irradiation with combined red + far-red (0.20 + 0.41 W m^-2) have no effect. In short day conditions floral response can be obtained in two ways: 1) by interrupting the dark period with 5 brief irradiations of red light (0.45 W m^-2, 12 min) at regular intervals, although these are counteracted by far-red irradiations of equal intensity and duration; 2) by interrupting the long night with 5 h red light applied during the second third of the night, while at the beginning or at the end it is ineffective. Red light efficiency appears to depend on the photosynthetic activity of the tissues, so that flowering increases with increasing intensity of white light and is suppressed if no white light is supplied. The reproductive development is determined by the coordination of proper irradiation conditions with sufficient sensitivity of the perceiving meristematic cells. The period of highest sensitivity to environmental light conditions in the life cycle of a Cichorium root explant occurs between the 8th and the 16th day after the start of the culture. The data strongly suggest that phytochrome is involved in flower induction of Cichorium in vitro.     

Relative effectiveness and interaction of ultraviolet-B, red and blue light in anthocyanin synthesis of apple fruit

The effect of light on anthocyanin production in apple ( Malus pumila Mill. cv. Jonathan) skin disks was investigated, with prolonged irradiation from different light sources. High fluence rates of white light provided from a xenon lamp were unable to produce large amounts of anthocyanin, and anthocyanin production became saturated at about 30 W m⁻². When UV-B light, provided by a fluorescent lamp which had an emission peak at 312 nm, was combined with the white light, anthocyanin production was synergistically stimulated and increased up to the highest fluence rates of white light tested (44 W m⁻²). This UV-B light was more effective than red and blue light provided from fluorescent lamps, but anthocyanin production became saturated at about 1.7 W m⁻². However, simultaneous irradiation with red and UV-B light had a synergistic effect. UV-B light was also effective in increasing anthocyanin production in whole fruit. Therefore this synergism seemed to have an important role in the development of the desirable red skin color under field light conditions. The results of aminoethoxyvinylglycine treatment suggested that ethylene was not involved in the stimulative effect of UV-B light.     

Chloroplast movement in Mougeotia induced by blue light pulses

The profile-to-face chloroplast movement in the green alga Mougeotia has been induced by strong blue and near-ultraviolet light pulses (6 J m^-2). Simultaneously, strong red or far-red light (10 W m^-2) was applied perpendicularly to the inducing beam. The response was measured photometrically. Against the far-red background the reciprocity law was found to hold for pulse durations varying two orders of magnitude. The action spectrum exhibited a maximum near 450 nm and a distinct increase in near-ultraviolet. The time-course and the spectral dependence of pulse responses of chloroplasts in Mougeotia were similar to those recorded for other plants which are sensitive only to blue. This points to an alternative sensor system active in the short-wavelength region in addition to the phytochrome system.Abbreviations FR far-red light - Pr red absorbing form of phytochrome - Pfr far-red absorbing form of phytochrome - R red light This paper is dedicated to the memory of Professor Jan Zurzycki     

Studies on the mechanism of light-dependent germination of akinetes of blue-green algae

This paper deals with the role of light in the germination of akinetes of Anabaena azollae. The two maxima action spectra are situated at 385 and 615 nm and the stimulation of the germination process by photosynthate was confirmed. The photoreceptor absorbing at 385 nm was identified as a flavin and that at 615 nm as a phytochrome. A model is suggested for the mode of action of light in the germination of akinetes of blue-green algae.C. Tsui     

Effects of suspensoids (turbidity) on penetration of solar radiation in aquatic ecosystems

In mainland Australia and in southern Africa, the aridity of the climate and sparse vegetative cover increase the susceptibility of the soils to erosion, and as a consequence surface waters are usually turbid. The inanimate suspensoids in such waters, the tripton fraction of the limnologist, are responsible for virtually all the light scattering, and also, by virtue of the yellow-brown humic materials adsorbed on their surface, for a substantial part of the light absorption. Spectral absorption data for suspensoids in terms of theirin situ absorption coefficient values, and the contribution of suspensoids to absorption of photosynthetically available radiation (PAR) are given for certain Australian water bodies.To understand the effect of suspensoids on attenuation of the solar flux with depth, the scattering coefficient must also be known, and this can be determined from the nephelometric turbidity or from up- and down-welling irradiance measurements. The effect of particle size on scattering efficiency is discussed.An equation expressing the vertical attenuation coefficient for downward irradiance as a function of absorption coefficient, scattering coefficient and solar altitude is presented, and is used to explore the effects of absorption due to dissolved colour and suspensoids, and the effects of scattering by suspensoids, on the penetration of PAR.Suspensoids, by increasing the rate of attenuation of the solar flux with depth, can greatly diminish the euphotic depth of a water body, with a consequent decrease in the ratio of the euphotic to the mixed depth: thus turbidity can reduce productivity of a water body substantially below that which might be expected on the basis of nutrient availability. Shallow turbid waters of low intrinsic colour can, however, be highly productive. By diminishing the depth of the layer within which solar energy is dissipated as heat, suspensoids can greatly modify the hydrodynamic behaviour of water bodies, and this also has far-reaching ecological consequences.Suspensoids drastically impair the visual clarity of water, a fact of major significance for the aquatic fauna, as well of aesthetic significance for humanity. The reciprocal of the Secchi depth is more correctly thought of as a guide to the vertical contrast attenuation coefficient rather than to the vertical attenuation coefficient for irradiance. The reflectivity of a water body, being at any wavelength proportional to the backscattering coefficient divided by the absorption coefficient, is highly dependent on the concentration, and optical character, of the suspensoids present. This has implications not only for the appearance (colour, muddiness) of the water to an observer, but also for the remote sensing of water composition by air- or satellite-borne radiometric sensors.