Comparative biochemical and immunological studies of the glycine betaine synthesis pathway in diverse families of dicotyledons

Members of the Chenopodiaceae can accumulate high levels (>100 mol·(g DW)^-1) of glycine betaine (betaine) in leaves when salinized. Chenopodiaceae synthesize betaine by a two-step oxidation of choline (cholinebetaine aldehyde betaine), with the second step catalyzed by betaine aldehyde dehydrogenase (BADH, EC 1.2.1.8). High betaine levels have also been reported in leaves of species from several distantly-related families of dicotyledons, raising the question of whether the same betaine-synthesis pathway is used in all cases.Fast atom bombardment mass spectrometry showed that betaine levels of >100 mol·(g DW)^-1 are present in Lycium ferocissimum Miers (Solanaceae), Helianthus annuus L. (Asteraceae), Convolvulus arvensis L. (Convolvulaceae), and Amaranthus caudatus L. (Amaranthaceae), that salinization promotes betaine accumulation in these plants, and that they can convert supplied choline to betaine aldehyde and betaine. Nicotiana tabacum L. and Lycopersicon lycopersicum (L.) Karst. ex Farw. (Solanaceae), Lactuca sativa L. (Asteraceae) and Ipomoea purpurea L. (Convolvulaceae) also contained betaine, but at a low level (0.1–0.5 mol·(g DW)^-1. Betaine aldehyde dehydrogenase activity assays, immunotitration and immunoblotting demonstrated that the betaine-accumulating species have a BADH enzyme recognized by antibodies raised against BADH from Spinacia oleracea L. (Chenopodiaceae), and that the M_r of the BADH monomer is in all cases close to 63 000. These data indicate that the cholinebetaine aldehydebetaine pathway may have evolved by vertical descent from an early angiosperm ancestor, and might be widespread (albeit not always strongly expressed) among flowering plants. Consistent with these suggestions, Magnolia x soulangiana was found to have a low level of betaine, and to express a protein of M_r 63 000 which cross-reacted with antibodies to BADH from Spinacia oleracea.Abbreviations BADH Betaine aldehyde dehydrogenase - DCIMS desorption chemical ionization mass spectrometry - FABMS fast atom bombardment mass spectrometry - M_r relative molecular mass - PAGE polyacrylamide gel electrophoresis - SDS sodium dodecyl sulfate - TLC thin-layer chromatography     

Osmoregulation in Dunaliella tertiolecta: effects of salt stress,and the external pH on the internal pH

The intracellular pH of the halotolerant green algae Dunaliella tertiolecta, was determined by the distribution of 5,5-dimethyl-2(¹⁴C)-oxalolidine-2,5-dione (DMO) between the cell and the surrounding medium. 5,5-dimethyl-2(¹⁴C)oxalolidine-2,4-dione was not metabolized by the algal cells. The intracellular pH of Dunaliella tertiolecta was 6.8 in the dark and 7.4 in the light. During a salt stress, after two hours, the intracellular pH was increased by 0.2 pH units in both light and dark. The salt stressed cells maintained a constant pH of about 7.5 over the pH range of 6.5 to 8.5. Because of the relatively low permeability coefficient of the plasma membrane for DMO, this technique does not permit rapid pH determinations during the induction period after a salt stress. The magnitude of the salt induced pH changes measured 2 h after the salt stress implies a minor importance of this alkalization in this time range, but does not exclude a larger importance of pH changes for osmoregulation during the induction period.Abbreviations Chl chlorophyll - DMO 5,5-dimethyl-2(¹⁴C)oxalolidine-2,4-dione - PCV packed cell volume - SDS sodium dodecyl sulfate     

Effect of acetaminophen on hepatic content and biliary efflux of glutathione disulfide in mice

The increased expiration of ethane and pentane by mice treated with hepatotoxic doses of acetaminophen suggests the possibility of oxidant mechanisms associated with the necrosis. However, studies in rats are not consistent with oxidant stress mechanisms causing the damage, because acetaminophen given to rats does not increase GSSG efflux, a sensitive index of intrahepatic oxidant stress. To compare the extent of oxidant stress generated by acetaminophen in mice versus rats, hepatic content and biliary efflux of GSSG and GSH in mice have been examined. Bile was collected from anesthetized male ICR mice before and after intraperitoneal administration of acetaminophen (325 mg/kg, 2.15 mmol/kg), t-butyl hydroperoxide (TBHP) (1.5 mmol/kg), diethyl maleate (400 mg/kg, 2.33 mmol/kg, in corn oil) or saline (control) and GSH and GSSG were measured by the enzymatic recycling method of Tietze. An increase in biliary GSSG efflux was produced by t-butyl hydroperoxide, but not by the other agents. Biliary GSH/GSSG ratios decreased in acetaminophen-treated animals, presumably reflecting the marked depletion of hepatic GSH, since a similar decrease was observed with non-hepatotoxic doses of diethyl maleate. The failure of acetaminophen to increase the hepatic content or biliary efflux of GSSG in ICR mice is not consistent with the view that oxidant stress mechanisms cause the damage, despite the increases in alkanes expired after acetaminophen administration in this specific animal model.     

Host plant resistance in Brachiaria grasses to the spittlebug Zulia colombiana

Twelve forage grass accessions including 11 accessions of Brachiaria Griseb, were evaluated in a glasshouse for host plant resistance to nymphs and tolerance to feeding damage caused by adults of Zulia colombiana (Lallemand) (Homoptera: Cercopidae). Resistance to nymphs was evaluated with a technique that provided uniform environmental conditions and abundant feeding sites. B. brizantha Stapf (cv. Marandú) was the most resistant of the accessions tested based on nymphal mortality, duration of nymphal stadia, and weight of adult females. Andropogon gayanus Kunth, resistant to spittlebug attack in the field, was susceptible under the conditions of this study. While growth habit and rooting characteristics may contribute to field resistance, other resistance factors are present within the genus Brachiaria, particularly in the case of B. brizantha cv. Marandú. The number of insect-days causing severe damage in the most tolerant species (B. dictyoneura Stapf and B. humidicola Schweick) was approximately six times greater than that necessary to cause the same level of damage to the most susceptible species (B. ruziziensis Germain & Evrard and B. decumbens Stapf). No difference was found in regrowth capacity between infested and noninfested plants within accessions. There was a significant positive correlation between number of insect-days causing severe damage (tolerance) and regrowth of infested plants.

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Résumé Brachiaria est une graminée fourragère prometteuse pour les sols tropicaux acides, saturés d'aluminium. Z. colombiana est un Cercopidae très répandu, limitant l'utilisation de Brachiaria en Amérique Latine. La résistance (antibiose et tolérance) à Z. colombiana, de Brachiaria d'origines diverses a été examinée. B. brizantha cv. Marandù s'est révélé le plus résistant d'après la forte mortalité larvaire, la prolongation du développement larvaire, et le poids réduit des femelles adultes de Z. colombiana. Andropogon gayanus, résistant dans la nature, s'est révélé sensible. Ces résultats suggèrent que cette résistance de A. gayanus dans la nature pourrait être due à la structure du végétal et à son mode de croissance. Dans le cas de B. brizantha cv. Marandù, des facteurs supplémentaires de résistance, mis en évidence à partir de différents modes de croissance, ont été éliminés, de façon à identifier les mécanismes de l'antibiose présents chez Brachiaria. Une grande gamme de résistance aux attaques alimentaires a été observée chez Brachiaria. Les plus résistants ont besoin de 6 fois plus de jours d'attaque par Z. colombiana pour provoquer les dégâts observés sur individus sensibles.

     

Is the ‘microbial loop’ an early warning indicator of anthropogenic stress?

Various components of the Microbial loop such as bacteria, heterotrophic nanoflagellates and autotrophic picoplankton were analyzed, for the first time across the Great Lakes, during a cruise in the summer of 1988. In addition, the size fractionated primary productivity using carbon-14 techniques was also determined. The statistical analysis indicated that bacteria, autotrophic picoplankton and ultraplankton/picoplankton productivity were significantly higher in Lakes Ontario and Erie than Lakes Huron and Michigan. The autotrophic picoplankton and ultraplankton/picoplankton productivity was higher in Lake Erie compared to Lake Ontario.The autotrophic picoplankton showed sensitivity to nutrients and contaminants in various types of environments. A dramatic decrease of autotrophic picoplankton in eutrophic-contaminated areas, such as Ashbridges Bay, Hamilton Harbour and western Lake Erie was observed. Conversely, in Saginaw Bay, another eutrophic environment, the autotrophic picoplankton were significantly higher than in Lake Huron. The sensitivity of autotrophic picoplankton to nutrients/contaminants might have implications to trophic interactions. Our results suggest that structural and functional characteristics of the microbial loop may be operating differently in stressed versus unstressed ecosystems. The possibility of using autotrophic picoplankton as an early warning indicator of environmental perturbation is proposed.     

Acid shock proteins of Escherichia coli

Synthesis of total cellular proteins of Escherichia coli was studied after transfer of cultures from pH 6.9 to pH 4.3. Proteins induced by such an external pH shift down were identified by mono- and bi-dimensional electrophoresis. 30 to 45 min after an acid shift, a group of at least sixteen polypeptides was markedly induced. Four of these polypeptides corresponded to the well known heat shock proteins GroEL, DnaK, HtpG and HtpM. Their pH induction was RpoH-dependent. Three other pH-induced proteins were previously identified as stress proteins induced either by osmolarity or aerobiosis or low temperature (proteins 32 (defined in this paper), C70.0 and C62.7). Seven other proteins were specifically induced after an acid shift and were called acid shock proteins (ASP). The induction of one of these proteins was RpoH-dependent, whereas that of others was RpoH-independent.     

Effect of seaweed concentrate on the growth and mineral nutrition of nutrient-stressed lettuce

The effects of the seaweed concentrate Kelpak on the growth and mineral nutrition of lettuce plants grown under conditions of varying nutrient supply were investigated. Kelpak significantly increased the yield and the concentration and amounts of Ca, K and Mg in the leaves of lettuce receiving an adequate supply of nutrients, but had little effect on nutrient stressed plants. Results are discussed in relation to the physiological mechanisms that have been proposed to explain the beneficial effects of seaweed concentrate on plants.author for correspondence     

Tolerance ofFestuca rubra L. to zinc in relation to mycorrhizal infection

Summary Plant yield of mycorrhizal and non-mycorrhizalFestuca rubra L. was linearly decreased with increasing zinc concentrations in nutrient solution. In all cases, non-mycorrhizal plant growth was significantly greater than that of mycorrhizal plants. Zinc and phosphorus concentrations of root and shoot of mycorrhizal plants were greater in all zinc treatments while mycorrhizal plants showed equal or lower tolerance indices to zinc than non-mycorrhizal plants. Yield depressions of mycorrhizal plants may be the result of enhanced zinc and phosphorus concentrations combined with the cost for growth and maintenance of the mycorrhizal fungi.     

Bacterial interactions with silver

Summary This review examines interactions between bacteria and the biologically non-essential metal, silver. Aspects of silver toxicity, tolerance and accumulation (possible binding and uptake as opposed to energy-dependent transport) in bacteria are discussed. In addition, plasmid biology is examined briefly since little information is available on the exact mechanism(s) of plasmid-endoced silver resistance in bacteria.     

The influence of CO2 enrichment, phosphorus deficiency and water stress on the growth, conductance and water use of Pinus radiata D. Don

Abstract. Seedlings of Pinus radiata D. Don were grown in growth chambers for 22 weeks with two levels of phosphorus, under either well-watered or water-stressed conditions at CO₂ concentrations of either 330 or 660mm³ dm^?3. Plant growth, water use efficiency and conductance were measured and the relationship between these and needle photosynthetic capacity, water use efficiency and conductance was determined by gas exchange at week 22. Phosphorus deficiency decreased growth and foliar surface area at both CO₂concentrations; however, it only reduced the maximum photosynthetic rates of the needles at 660 mm³ CO₂ dm^?3 (plants grown and measured at the same CO₂ concentration). Water stress reduced growth and foliar surface area at both CO₂ concentrations. Increases in needle photosynthetic rates appeared to be partly responsible for the increased growth at high CO₂ where phosphorus was adequate. This effect was amplified by accompanying increases in needle production. Phosphorus deficiency inhibited these responses because it severely impaired needle photosynthetic function. The relative increase in growth in response to high CO₂ was higher in the periodically water-stressed plants. This was not due to the maintenance of cell volume during drought. Plant water use efficiency was increased by CO₂ enrichment due to an increase in dry weight rather than a decrease in shoot conductance and, therefore, transpirational water loss. Changes in needle conductance and water use efficiency in response to high CO₂ were generally in the same direction as those at the whole plant level. If the atmospheric CO₂ level reaches the predicted concentration of 660 mm³ dm^?3 by the end of next Century, then the growth of P. radiata will only be increased in areas where phosphorus nutrition is adequate. Growth will be increased in drought-affected regions but total water use is unlikely to be reduced.