Freeze-Thawing of Aquaspirillum magnetotacticum Cells Selectively Releases Periplasmic Proteins

Cells of the gram-negative bacterium Aquaspirillum magnetotacticum, when suspended in buffer and freeze-thawed, produced pinkish orange supernatant fluid. The fluid contained ≤2.0% of total extractable outer membrane component 2-keto-3-deoxyoctonate or of the cytoplasmic membrane marker succinic dehydrogenase. Electrophoretic banding patterns and difference spectra of proteins and hemoproteins released by freeze-thawing cells were distinct from those of membrane-associated substances and similar to those of periplasmic substances obtained by applying conventional fractionation methods to this organism.     

UV irradiation inhibits initiation of DNA replication from oriC in Escherichia coli

Summary Irradiation of Escherichia coli with UV light causes a transient inhibition of DNA replication. This effect is generally thought to be accounted for by blockage of the elongation of DNA replication by UV-induced lesions in the DNA (a cis effect). However, by introducing an unirradiated E. coli origin (oriC)-dependent replicon into UV-irradiated cells, we have been able to show that the environment of a UV-irradiated cell inhibits initiation of replication from oriC on a dimer-free replicon. We therefore conclude that UV-irradiation of E. coli leads to a trans-acting inhibition of initiation of replication. The inhibition is transient and does not appear to be an SOS function.     

Identification of cDNA clones for tomato (Lycopersicon esculentum Mill.) mRNAs that accumulate during fruit ripening and leaf senescence in response to ethylene

Changes in gene expression during foliar senescence and fruit ripening in tomato (Lycopersicon esculentum Mill.) were examined using in-vitro translation of isolated RNA and hybridization against cDNA clones.During the period of chlorophyll loss in leaves, changes occurred in mRNA in-vitro translation products, with some being reduced in prevalence, whilst others increased. Some of the translation products which changed in abundance had similar molecular weights to those known to increase during tomato fruit ripening. By testing RNA from senescing leaves against a tomato fruit ripening-related cDNA library, seven cDNA clones were identified for mRNAs whose prevalence increased during both ripening and leaf senescence. Using dot hybridization, the pattern of expression of the mRNAs corresponding to the seven clones was examined. Maximal expression of the majority of the mRNAs coincided with the time of greatest ethylene production, in both leaves and fruit. Treatment of mature green leaves or unripe fruit with the ethylene antagonist silver thiosulphate prevented the onset of senescence or ripening, and the expression of five of the seven ripening- and senescence-related genes.The results indicate that senescence and ripening in tomato involve the expression of related genes, and that ethylene may be an important factor in controlling their expression.Abbreviations cDNA copy-DNA - MW molecular weight - PAGE polyacrylamide gel electrophoresis - SDS sodium dodecyl sulphate     

The analysis of hypervariable DNA profiles: problems associated with the objective determination of the probability of a match

Summary A computerised system has been used to store DNA profiles from 3 hypervariable loci. This initial survey illustrates that band matching is only possible after analysis of the errors associated with electrophoretic systems. A number of databases have been constructed with the three probes investigated and two methods of frequency determination, binning and sliding window fitting, have been compared.     

AN AMPLIFICATION OF THE PHYLOGENETIC SPECIES CONCEPT

Abstract— The goal of a phylogenetic species concept is to reveal the smallest units that are analysable by cladistic methods and interpretable as the result of phylogenctic history. We define species as the smallest aggregation of populations (sexual) or lineagcs (asexual) diagnosable by a unique combination of character states in comparable individuals (semaphoronts). A character state is an inherited attribute distributed among all comparable individuals (semaphoronts) of the same historical population, clade, or terminal lineage. This definition of species is character-based and pattern oriented. Evolutionary explanations of phylogenetic species are consistent with contemporary explanations of processes of speciation, but require only the assumption of nested hierarchical pattern. We discuss the compatibility of the phylogenetic species concept with various biological needs for species and justify its use at the exclusion of alternative species concepts.     

Basement-membrane components associated with the extracellular matrix of the lymph node

Summary Lymph nodes contain an extensive array of extracellular matrix fibers frequently referred to as reticular fibers because of their reticular pattern and positive reaction with silver stains. These fibers are known to contain primarily type-III collagen. In the present study, frozen and plastic-embedded sections of mouse and human lymph nodes were subjected to immunostaining with a panel of monospecific antibodies directed against type-IV collagen, type-III collagen, laminin, entactin, and heparan sulfate proteoglycan. Immunofluorescent staining revealed that, in addition to being uniformly stained with antibodies to type-III collagen, these fibers also stained positively with antibodies to type-IV collagen and to other basement-membrane-specific components. Furthermore, the basement-membrane-specific antibodies stained the outer surface of individual fibers. These same type-III collagen-rich fibers were distinct from blood vascular basement membranes since they did not react with antibodies to factor VIII-related antigen, an endothelial-cell-specific marker. The role of these basement-membrane-specific components associated with the reticular fibers of lymphoid tissue is unknown. However, it is possible that the ligands promote attachment of reticular fibroblasts as well as macrophages and lymphocytes to the extracellular matrix fibers.     

The importance of Paracallisoma coecus and myctophid fishes to nesting fork-tailed and Leach's storm-petrels in the Queen Charlotte Islands, British Columbia

The diet of fork-tailed (Oceanodroma furcata) and Leach's storm-petrel(O.leucorhoa) was studied at Petrel Island, a small island northof Hippa Island in the Queen Charlotte Islands, British Columbiain 1983. The diet of the two storm-petrel species consistedmostly of the amphipod Paracallisoma coecus and fish. Most fisheswhich could be identified were myctophids, and may have beenacquired by storm-petrels at night. Fishes increased while P.coecusdeclined in importance as a food over the nestling season ofthe storm-petrels. The amount of oil also declined in the Leach'sstorm-petrel diet over the season. Since P.coecus contains muchoil, the seasonal decline of both P.coecus and oil was probablyassociated. Other less important foods were jellyfishes, copepods,isopods, euphausiids, shrimp, squid and octopus. Jellyfishes(Velella velella) only occurred in the diet of Leach's storm-petrels,which may relate to that storm-petrel feeding in open oceanwhere V. velella occurs most frequently. The most numerous copepodwas Neocalanus cristatus, which was only observed in the storm-petreldiet early on during the study period. An abyssal copepod, Bathycalanusbradyi, and the isopod Cirolana californiensis (tentativelyidentified, the latter only previously known from sea canyonsoff California) were surprising food items of storm-petrels.Most euphausiid prey consisted of Thysanoessa spinifera andEuphausia pacifica.     

Ac transposition in transgenic tomato plants

Summary As an initial step towards developing a transposon mutagenesis system in tomato, the maize transposable element Ac was transformed into tomato plants via Agrobacterium tumefaciens. Southern analysis of leaf tissue indicated that in nine out of eleven transgenic plants, Ac excised from the T-DNA and reintegrated into new chromosomal locations. The comparison of Ac banding pattern in different leaves of the same primary transformant provided evidnece for transposition during later stages of transgenic plant development. There was no evidence of Ds mobilization in tomato transformants.     

Salicyloylaspartate as an endogenous component in the leaves of Phaseolus vulgaris

An amide conjugate of o-methoxybenzoic acid and aspartic acid has been isolated from bean leaves. After extraction and methylation of plant material, this compound was isolated as two isomeric monoethyl monomethyl esters. The ethylation of the aspartyl carboxyl groups was shown to be a likely result of an extraction procedure utilising acidified ethanol, the methylation of the aromatic hydroxy of the methoxy group to be due to the derivatisation procedure. Studies with pentafluorobenzylation confirmed that the endogenous compound is o-hydroxybenzoylaspartate.     

Simultaneous degradation of the herbicides 2,4-dichlorophenoxyacetic acid and 2-(2-methyl-4-chlorophenoxy)propionic acid by mixed bacterial cultures

The simultaneous degradation of 2,4-dichlorophenoxyacetic acid (2,4-D) and 2-(2-methyl-4-chlorophenoxy)propionic acid (mecoprop) was achieved by two mixed cultures in the absence of any additional carbon or energy substrates. Mecoprop was not completely degraded by either of the two cultures, nor did addition of 2,4-D affect the degradation of mecoprop. The cultures completely degraded 2,4-D, and the degradation was uninfluenced by the addition of mecoprop. Nearly complete dechlorination of the mixture of two herbicides was achieved by both cultures, on the basis of the total amount of the two herbicides degraded. During the course of the reaction, however, the expected values of chloride were not met. Cell growth continued after the degradation of the parent substrates ceased. Although the mecoprop degradation did not continue to completion, spectral and growth data indicated that the metabolites which had accumulated during the reaction were degraded upon further incubation.