Long-term effects of metals in sewage sludge on soils,microorganisms and plants

Summary This paper reviews the evidence for impacts of metals on the growth of selected plants and on the effects of metals on soil microbial activity and soil fertility in the long-term. Less is known about adverse long-term effects of metals on soil microorganisms than on crop yields and metal uptake. This is not surprising, since the effects of metals added to soils in sewage sludge are difficult to assess, and few long-term experiments exist. Controlled field experiments with sewage sludges exist in the UK, Sweden, Germany and the USA and the data presented here are from these long-term field experiments only. Microbial activity and populations of cyanobacteria,Rhizobium leguminosarum bv.trifolii, mycorrhizae and the total microbial biomass have been adversely affected by metal concentrations which, in some cases, are below the European Community's maximum allowable concentration limits for metals in sludge-treated soils. For example, N₂-fixation by free living heterotrophic bacteria was found to be inhibited at soil metal concentrations of (mg kg^–1): 127 Zn, 37 Cu, 21 Ni, 3.4 Cd, 52 Cr and 71 Pb. N₂-fixation by free-living cyanobacteria was reduced by 50% at metal concentrations of (mg kg^–1): 114 Zn, 33 Cu, 17 Ni, 2.9 Cd, 80 Cr and 40 Pb.Rhizobium leguminosarum bv.trifolii numbers decreased by several orders of magnitude at soil metal concentrations of (mg kg^–1): 130–200 Zn, 27–48 Cu, 11–15 Ni, and 0.8–1.0 Cd. Soil texture and pH were found to influence the concentrations at which toxicity occurred to both microorganisms and plants. Higher pH, and increased contents of clay and organic carbon reduced metal toxicity considerably. The evidence suggests that adverse effects on soil microbial parameters were generally found at surpringly modest concentrations of metals in soils. It is concluded that prevention of adverse effects on soil microbial processes and ultimately soil fertility, should be a factor which influences soil protection legislation.     

Genetic dissection of carotenoid synthesis in arabidopsis defines plastoquinone as an essential component of phytoene desaturation.

Carotenoids are C40 tetraterpenoids synthesized by nuclear-encoded multienzyme complexes located in the plastids of higher plants. To understand further the components and mechanisms involved in carotenoid synthesis, we screened Arabidopsis for mutations that disrupt this pathway and cause accumulation of biosynthetic intermediates. Here, we report the identification and characterization of two nonallelic albino mutations, pds1 and pds2 (for phytoene desaturation), that are disrupted in phytoene desaturation and as a result accumulate phytoene, the first C40 compound of the pathway. Surprisingly, neither mutation maps to the locus encoding the phytoene desaturase enzyme, indicating that the products of at least three loci are required for phytoene desaturation in higher plants. Because phytoene desaturase catalyzes an oxidation reaction, it has been suggested that components of an electron transport chain may be involved in this reaction. Analysis of pds1 and pds2 shows that both mutants are plastoquinone and tocopherol deficient, in addition to their inability to desaturate phytoene. Separate steps of the plastoquinone/tocopherol biosynthetic pathway are affected by these two mutations. The pds1 mutation affects the enzyme 4-hydroxyphenylpyruvate dioxygenase because it can be rescued by growth on the product but not the substrate of this enzyme, homogentisic acid and 4-hydroxyphenylpyruvate, respectively. The pds2 mutation most likely affects the prenyl/phytyl transferase enzyme of this pathway. Because tocopherol-deficient mutants in the green alga Scenedesmus obliquus can synthesize carotenoids, our findings demonstrate conclusively that plastoquinone is an essential component in carotenoid synthesis. We propose a model for carotenoid synthesis in photosynthetic tissue whereby plastoquinone acts as an intermediate electron carrier between carotenoid desaturases and the photosynthetic electron transport chain.     

Photosynthetic activity and population dynamics of Amoebobacter purpureus in a meromictic saline lake

Abstract A dense population of the purple sulfur bacterium Amoebobacter purpureus in the chemocline of meromictic Mahoney Lake (British Columbia, Canada) underwent consistent changes in biomass over a two year study period. The integrated amount of bacteriochlorophyll reached maxima in August and declined markedly during early fall. Bacteriochlorophyll was only weakly correlated with the light intensity and water temperature in the chemocline. In the summer, bacterial photosynthesis was limited by sulfide availability. During this period the intracellular sulfur concentration of A. purpureus cells decreased. A minimum concentration was measured at the top of the bacterial layer in August, when specific photosynthetic rates of A. purpureus indicated that only 14% of the cells were photosynthetically active. With the exception of a time period between August and September, the specific growth rates calculated from CO₂ fixation rates of A. purpureus were similar to growth rates calculated from actual biomass changes in the bacterial layer. Between August and September 86% of the A. purpureus biomass disappeared from the chemocline and were deposited on the littoral sediment of Mahoney Lake or degraded within the mixolimnion. This rise of cells to the lake surface was not mediated by an increase in the specific gas vesicle content which remained constant between April and November. The upwelling phenomenon was related to the low sulfur content of A. purpureus cells and a low resistance of surface water layers against vertical mixing by wind.     

Vascular Development and Sap Flow in Apple Pedicels

Xylem and phloem tissues of the pedicel of apple fruit increasein cross-sectional area throughout development. The increasein phloem is similar in the two cultivars examined (Cox's OrangePippin and Royal Gala) and reflects a steadily increasing phloemsap flow to the fruit. The increase in xylem tissue is due toa proliferation of non-conducting, structural, components sinceclose examination reveals no increase in the number of vesselelements from just after flowering onwards. The greater number,and the larger diameter, of the vessels in Cox's explains theinitially higher xylem conductance found in this cultivar. In vitro measurements of xylem exudation reveal a decline duringthe growing season in the xylem conductance of both cultivarsand an increasing proportion of fruit (particularly in Cox's)in which the xylem comes to be totally non-conducting. Thisobservation is in line with previously reported measurementsof xylem sap flow in vivo. The straightforward techniques used in this study offer a feasiblealternative to more arduous methods of assessing xylem and phloemsap flows and their balance during growth.Copyright 1994, 1999Academic Press Apple, xylem, phloem, vascular development, sap flow, Malus domestica Borkh     

Some rumen ciliates have endosymbiotic methanogens

Abstract Most of the small ciliate protozoa, including Dasytricha ruminantium and Entodinium spp. living in the rumen of sheep, were found to have intracellular bacteria. These bacteria were not present in digestive vacuoles. They showed characteristic coenzyme F₄₂₀ autofluorescence and they were detected with a rhodamine-labelled Archaea-specific oligonucleotide probe. The measured volume percent of autofluorescing bacteria (1%) was close to the total volume of intracellular bacteria estimated from TEM stereology. Thus it is likely that all of the bacteria living in the cytoplasm of these ciliates were endosymbiotic methanogens, using H₂ evolved by the host ciliate to form methane. Intracellular methanogens appear to be much more numerous than those attached to the external cell surface of ciliates.     

Reduction in omega-3 fatty acids by UV-B irradiation in microalgae

UV-B irradiation reduced the levels of omega-3 fatty acid, eicosapentaenoic acid (EPA, 20:53) and docosahexaenoic acid (DHA, 22:63), in microalgae; the degree of reduction varied with species.Chaetoceros calcitrans andSkeletonema costatum were high UV-B tolerant species, followed byPhaeodactylum tricornutum, Chroomonas salina, Pavlova lutheri, andThalassiosira pseudonana.Isochrysis galbana (T.ISO) andProrocentrum micans were UV- B sensitive. Cells in logarithmic phase were most sensitive to UV- B irradiation. Nitrate-, phosphate-, or sulphate-starved cells were more UV-B sensitive than non-starved cells grown in a complete basal medium. A relatively short exposure to high UV-B was more damaging than a longer exposure to lower irradiance. Visible light intensity levels had a profound impact on the sensitivity of microalgal cultures to UV-B, with high levels decreasing UV-B dependent damage. Addition of polyamines (putrescine, spermidine or spermine) or an amino acid (cysteine) to the culture medium minimized the reduction of omega-3 fatty acid content in microalgae caused by UV-B irradiation.Author for correspondence     

Effects of virus infection on demographic traits of an agamospermous population of Eupatorium chinense (Asteraceae)

There are few studies of the interaction between wild plants and viruses. In this paper, the incidence of a geminivirus (tobacco leaf curl virus, TLCV) infection, and its effects on mortality, growth and reproduction of its host-plant, Eupatorium chinense, are reported. A total of 221 plants of an agamospermous population of E. chinense were chosen and their demographic behaviour followed over 2 years (1991–1992). The proportion of infected plants differed between years, with fewer plants infected in 1991 than in 1992. Under low virus incidence (35.3% in 1991), infection was significantly associated with taller plants (>80 cm). However, when the incidence of infected plants increased by almost two times (69.1%) in 1992, this tendency disappeared and small plants were also infected. Virus infection had significant effects on mortality of agamospermous plants. Almost half of the initial number of marked plants (n=221) died after 1 year of observations. Of those dead plants (n=105), 86 plants (82%) were infected in 1991, indicating that virus infection was an important, but not the sole cause of mortality. In 1992, 116 plants were alive, and of these, 40% were infected in 1991, indicating that some infected plants survived 1 year. Agamospermous plants were classified in three groups according to the extent of virus infection (plants infected in 2 years, infected in 1 year and uninfected plants) to detect the effect of virus infection on growth of plants of E. chinense. Infected plants had significantly lower growth rates than healthy plants. Infected plants also produced significantly fewer seeds than uninfected plants. Virus infection, however, had no significant effect on the probability of reproduction in plants of E. chinense, suggesting that infected plants may reproduce but with a lower seed output. In this study, we showed that virus infection may have a strong effect on demographic traits and, as a consequence, on fitness components of plants of E. chinense. These effects were higher than those sometimes observed in other plant-herbivore or plant-pathogen interactions.     

Laminin A, B1, and B2 Chain Gene Expression in Transected and Regenerating Nerves: Regulation by Axonal Signals

Abstract: Laminin A, B1, and B2 chain mRNA levels in degenerating and regenerating mouse sciatic nerves were examined using northern blot analysis. In normal intact nerves, B1 and B2 mRNA steady-state levels were high, but when the nerves were crushed, the steady-state levels of B1 and B2 mRNA per milligram wet tissue weight of the distal segments of the nerves increased five- to eightfold over that of control levels as the total RNA and β-actin mRNA levels increased, suggesting that these increases were the consequence of Schwann cell proliferation after axotomy. When the steady-state levels of B1 and B2 mRNA were normalized as the ratio to total RNA or β-actin mRNA levels, however, they drastically decreased to about 20% of the normal nerve levels in the nerve segments distal to both the crush and transaction sites 1 day after injury. In the crushed nerves, B1 and B2 mRNA levels gradually increased as the regenerating nerves arrived at the distal segments and reestablished normal axon–Schwann cell contact, and then returned to normal levels on the 21 st day. In the transected nerves, where Schwann cells continued to be disconnected from axons, both B1 and B2 mRNA levels remained low. Cultured Schwann cells expressed detectable levels of B1 and B2 chain mRNA which significantly increased when the cells were cocultured with sensory neurons. However, mRNA for A chain was not detectable in the normal, axotomized nerves or in cultured Schwann cells. These data indicate that Schwann cells express laminin B1 and B2 chain mRNA that are up-regulated by axonal or neuronal contact, but they do not express A chain mRNA.     

Electrophoretic karyotypes of the elm tree pathogen Ophiostoma ulmi (sensu lato)

Pulsed field gel electrophoresis using OFAGE, TAFE, and CHEF systems has been used to more fully characterize karyotypic variation within the two closely related fungal species of Ophiostoma ulmi sensu lato. Twelve wild-type and laboratory strains, representing the less agressive species O. ulmi and both of the biotypes of the more aggressive species O. novo-ulmi were studied and their karyotypes determined. Depending on the strain, a minimum of four to a minimum of eight chromosomal DNA bands were present that fall into three distinct size classes, with one exception. Strain CESSI6K (O. novo-ulmi, North American aggressive subgroup) contains a unique chromosomal DNA band which comigrated near a Saccharomyces cerevisiae chromosome of 0.95 Mb. This unique band was the smallest O. ulmi s. l. chromosomal DNA observed. Seven of the twelve strains shared a common chromosomal DNA banding pattern, whereas each of the other five had a unique karyotype. There was no correlation between chromosome profile and species, as some O. novo-ulmi and O. ulmi strains shared common electrophoretic karyotypes.     

Phospholipase A2 inhibitors from marine algae

Twelve out of twenty-nine compounds isolated from benthic marine algae from the phyla Chlorophyta, Phaeophyta and Rhodophyta have been found to be potent inhibitors of bee venom derived phospholipase A₂ (PLA₂) (> 50%) in the M range. The compounds investigated were from: Bryopsis pennata, Rhipocephalus phoenix, Caulerpa prolifera, C. racemosa, C. bikinensis, Cymopolia barbata, Laurencia cf. palisada, Laurencia sp., Ochtodes crockeri, Liagora farinosa, Sphaerococcus coronipifolius, Phacelocarpus labillardieri, Dictyota sp., B furcaria galapagensis, Stypopodium zonale, Dictyopteris undulata, Stoechospermum marginatum, Dictyopteris divaricata, Dilophus fasciola and Dilophus sp. This is the first report of bee venom PLA₂ inhibition in vitro by pure compounds isolated from marine algae.