DESMID-BACTERIAL ASSOCIATIONS IN SPHAGNUM-DOMINATED WISCONSIN PEATLANDS1

In a survey employing epifluorescence microscopy with the DNA fluorochrome DAPI, associations between bacteria and filamentous desmids were found to be commonplace in acidic, Sphagnum-dominated Wisconsin peat-lands. Bacteria were associated with all genera of filamentous desmids encountered including Desmidium, Hyalotheca, Onychonema, Spondylosium, and Teilingia. Although only associations involving filamentous desmids having mucilaginous sheaths are illustrated here, bacteria were also noted on taxa lacking sheaths as well as some unicellular forms. Bacteria on Desmidium majus Lagerheim, D. grevillii (Kütz.) De Bary, and Hyalotheca dissiliens (Smith) Bréb. ex Ralfs tended to be concentrated in small pockets in the sheath material located near the isthmus and in the region between adjacent cells in the filament, whereas those associated with Spondylosium pulchrum (Bail.) Archer were more evenly distributed throughout the sheath. Most bacteria were rodshaped. Those associated with S. pulchrum, D. grevillii, and D. majus ranged from 1.1 to 11.2 μm in length. Bacteria within the sheaths of H. dissiliens and D. grevillii were Gram-negative. A second morphologically distinct population of bacteria was found at the sheath margin in D. majus and D. grevillii. Field collections containing filamentous desmids were examined with scanning electron microscopy and bacteria associated with Desmidium majus were investigated with transmission electron microscopy.     

Potential canopy influences on the isotopic composition of nitrogen and sulphur in atmospheric deposition

Isotopic studies of nitrogen and sulphur inputs to plant/soil systems commonly rely on limited published data for the ¹⁵N/¹⁴N and ³⁴S/³²S ratios of nitrate, ammonium and sulphate in rainfall. For systems with well-developed plant canopies, however, inputs of these ions from dry deposition or particulates may be more important than rainfall. The manner in which isotopic fractionation between ions and gases may lead to dry deposition and particulates having ¹⁵N/¹⁴N or ³⁴S/³²S ratios different from those of rainfall is considered. Data for rainfall and throughfall in coniferous plantations are then discussed, and suggest that: (1) in line with expectations, nitrate washed from the canopy has ¹⁵N/¹⁴N ratios higher than those in rainfall; (2) the ¹⁵N/¹⁴N ratios of ammonium washed from the canopy are variable, with high ratios being found for canopies of higher pH in conditions of elevated ambient ammonia gas concentrations; and (3) in accord with expectations and previous work, ³⁴S/³²S ratios of sulphate washed from the canopy are not substantially different from those in rainfall. The study suggests that if atmospheric inputs are relevant to isotopic studies of the sources of nitrogen for canopied systems, then confident interpretation will require analysis of these inputs. Received: 3 March 1996 / Accepted: 28 September 1996     

Cloning and characterization of chloroplast ribosomal protein-encoding genes, rpl16 and rps3, of the marine macro-algae, Gracilaria tenuistipitata

In Gracilaria tenuistipitata, a highly differentiated multicellular member of the marine red algae, Rhodophyta, chloroplast (cp) DNA can be separated as a satellite band from the nuclear DNA in a CsCl gradient. Using a heterologous probe from Chlamydomonas, the ribosomal protein-encoding gene, rpl16, was located on a 4.5-kb EcoRI fragment of cp DNA. The fragment was cloned and a 1365-bp region around rpl16 was sequenced. The gene order around rpl16, 5′ rpl22-rps3-rpl16, is identical to that detected in the chloroplast DNA of liverwort, tobacco and maize. Both the nucleotide sequence and the amino-acid sequence of rpl16 are more conserved than that of rps3. The rpl16 gene contains no intron, a feature which shows more similarity to the unicellular green algae, Chlamydomonas, than the other land plants. Sequences that may form a stable stem-loop structure were detected within the coding sequence of rpl16.     

Immune response of U.D.N.-infected salmon to Saprslegnia

Sera from salmon Sulnio salar L. suspected of U.D.N.-infection were tested (by double gel diffusion) against antigens prepared from diseased salmon tissues and Suprolegniu . Precipitating antibodies to the fungus were found in 93 % of the salmon, but only 66% of these fish were colonized by fungus. The antibodies were detected in parr and adult salmon.     

The response of sweet corn to HF and SO2 under field conditions

“Marcorss” sweet corn plants grown in field plots were exposed continuously in open-top chambers for 32 days to ambient air, charcoal-filtered air or charcoal-filtered air containing HF (ca. 0.5 μgF m⁻³), SO₂ (ca. 235 μg m⁻³), or the two pollutants combined. Elliptical chlorotic leasions appeared after 23 days on leaves of plants exposed to SO₂/HF, and shortly thereafter on plants exposed to all other treatments. At harvest, the number of plants with lesions was significantly greater in chambers supplied with SO₂/HF than in chambers with SO₂, HF, or filtered air.

The different treatments had no effect on fresh or dry weights of leaves, husks, or tassels, height of plants, or number of kernels per ear. Exposure to SO₂/HF reduced the fresh and dry weights of stalks. There were fewer mature ears in the SO₂/HF and unfiltered air treatments than in the others. The reduction in yield from SO₂/HF was about the same as that ascribed to ambient photochemical oxidants in the unfiltered air treatment.

HF combined with SO₂ had no effect on accumulation of S as compared with SO₂ alone, but there was a striking reduction in accumulation of foliar F in plants exposed to SO₂/HF as compared with HF alone.     

The effects of culture media on Saprolegnia antigens

In a previous paper (Hodkinson &Hunter, 1970a) we reported that salmon, suffering from a disease called ulcerative dermal necrosis, appeared to have precipitating antibodies to the mycelium of a fungus associated with their diseased skin lesions. This was confirmed when antigens were prepared from pure cultures of the fungus, a species ofSaprolegnia (Hodkinson &Hunter, 1970b). In this paper we describe the effects of culture media on the antigenicity ofSaprolegnia mycelium, as detected by double gel diffusion with salmon sera.     

Periodic shedding and replacement of venomous caudal spines,with special reference to South American freshwater stingrays,Potamotrygon spp.

Synopsis The number of venomous caudal spines and their length and position relative to one another were determined in seven species of South American freshwater rays (Potamotrygonidae) and eight marine or euryhaline species of four families from the Caribbean Coast of South and Central America. Most species have two visible spines at certain stages in the shedding-replacement cycle and only one visible spine at other stages (following shedding). If we include the embryological beginnings of the spines before they erupt and become visible, the spine counts of most rays are actually 2 rather than 1 or 2. Since most species apparently follow this pattern, spine counts are of little use in distinguishing between species except in the relatively few that may have only one, or no spines. Eight captive Potamotrygon specimens maintained in simulated tropical temperature conditions over 12 months showed periodic shedding and replacement of spines. The molts were biannual for a given ray but annual for a given spine. They alternated between two spine loci and their cycles were approximately six months out of phase with each other. Recent studies on Dasyatis sabina by others report only one molt per year, with replacement spines forming always posterior to the primary spine rather than alternating between posterior and anterior. Supernumerary spines (counts of more than two, up to five) are also discussed, as are counts of one and zero.     

Polynucleotide Phosphorylase Plays an Important Role in the Generation of Spontaneous Mutations in Escherichia coli

Polynucleotide phosphorylase (PNP) plays a central role in RNA degradation, generating a pool of ribonucleoside diphosphates (rNDPs) that can be converted to deoxyribonucleoside diphosphates (dNDPs) by ribonucleotide reductase. We report here that spontaneous mutations resulting from replication errors, which are normally repaired by the mismatch repair (MMR) system, are sharply reduced in a PNP-deficient Escherichia coli strain. This is true for base substitution mutations that occur in the rpoB gene leading to Rif^r and the gyrB gene leading to Nal^r and for base substitution and frameshift mutations that occur in the lacZ gene. These results suggest that the increase in the rNDP pools generated by polynucleotide phosphorylase (PNP) degradation of RNA is responsible for the spontaneous mutations observed in an MMR-deficient background. The PNP-derived pool also appears responsible for the observed mutations in the mutT mutator background and those that occur after treatment with 5-bromodeoxyuridine, as these mutations are also drastically reduced in a PNP-deficient strain. However, mutation frequencies are not reduced in a mutY mutator background or after treatment with 2-aminopurine. These results highlight the central role in mutagenesis played by the rNDP pools (and the subsequent dNTP pools) derived from RNA degradation.     

Zinc enhances adiponectin oligomerization to octadecamers but decreases the rate of disulfide bond formation

Adiponectin, a hormone secreted from adipocytes, has been shown to protect against development of insulin resistance, ischemia–reperfusion injury, and inflammation. Adiponectin assembles into multiple oligomeric isoforms: trimers, hexamers and several higher molecular weight (HMW) species. Of these, the HMW species are selectively decreased during the onset of type 2 diabetes. Despite the critical role of HMW adiponectin in insulin responsiveness, its assembly process is poorly understood. In this report, we investigated the role of divalent cations in adiponectin assembly. Purified adiponectin 18mers, the largest HMW species, did not collapse to smaller oligomers after treatment with high concentrations of EDTA. However, treatment with EDTA or another chelator DTPA inhibited the oligomerization of 18mers from trimers in vitro. Zn²⁺ specifically increased the formation of 18mers when compared with Cu²⁺, Mg²⁺, and Ca²⁺. Distribution of adiponectin oligomers secreted from zinc chelator TPEN-treated rat adipocytes skewed toward increased proportions of hexamers and trimers. While we observed presence of zinc in adiponectin purified from calf serum, the role of zinc in disulfide bonding between oligomers was examined because the process is critical for 18mer assembly. Surprisingly, Zn²⁺ inhibited disulfide bond formation early in the oligomerization process. We hypothesize that initial decreases in disulfide formation rates could allow adiponectin subunits to associate before becoming locked in fully oxidized conformations incapable of further oligomerization. These data demonstrate that zinc stimulates oligomerization of HMW adiponectin and possibly other disulfide-dependent protein assembly processes.