Further studies on seed transmission in the ecology of some aphid-transmitted viruses

A small proportion (1–4%) of the seeds of Stellaria media extracted from fallow soil from three widely separated areas contained cucumber mosaic virus (CMV). S. media seeds buried for 21 months produced 5 % infected seedlings. S. media plants from Britain, N. America and Australia were least severely affected by the CMV strain obtained from their country of origin and showed more severe reactions when infected with two alien strains. Several weed species were experimentally infected with lettuce mosaic, turnip mosaic and cauliflower mosaic viruses but, although virus was detected in the seeds of some species, it was not transmitted to any of their seedlings.     

Chromosomal introgression in house mice from the hybrid zone between M. m. domesticus and M. m. musculus in Denmark

The recent discovery of Robertsonian (Rb) translocations in Danish mice from the hybrid zone between Mus musculus musculus and M. m. domesticus stimulated the chromosomal analysis of populations along a north-south transect through this zone. G-Banding identified the Rb fusions as Rb(3.8), Rb(2.5) and Rb(6.9). The cytogenetic results show that there is a gradual decrease in the number of fusions as one proceeds north, the translocations abruptly ending in populations from the centre of the hybrid zone determined by seven diagnostic allozymic markers. These results indicate that Rb fusions are present only in domesticus or predominantly domesticus-genotype mice and that they do not introgress into M. m. musculus . To test if genie incompatibilities between the musculus genetic background and Rb fusions were involved in the systematic elimination of the latter, predominantly musculus mice from the hybrid zone were crossed with Rb domesticus mice carrying Rb(3.8). The karyotypic analysis of the progeny showed no distortion of the transmission ratio of this fusion.
The chromosomal and allozymic analysis of these mice further indicates that (i) recombination is not suppressed between metacentrics and their acrocentric homologues and (ii) specific domesticus chromosomal segments are tolerated in the musculus genomes whereas the Rb centromeres are not.     

APPLICATIONS OF TIME-RESOLVED FLUOROIMMUNOASSAY TO DETECT MAGNETIC BEAD CAPTURED ESCHERICHIA COLI O157:H7

A time-resolved fluorescence technique was developed to detect Escherichia coli O157:H7 in ground beef burger. After a 4.5 h enrichment period, streptavidin coated magnetic beads conjugated with biotin-labeled anti E. coli O157:H7 were used to capture the bacteria. The bacteria were, at the same time, also labeled by a nonfluorescent, europium (Eu)-tagged anti-E. coli O157:H7 antibody. The sandwiched bacterial complexes were then concentrated using a magnetic particle concentrator and washed to remove other solution components. Upon addition of an enhancement buffer, the Eu-labels were then released from the antibodies and chelated to nitrilo-triacetic acid (NTA) and trioctylphosphine oxide (TOPO) to form highly fluorescent Eu-(2-NTA)₃(TOPO)_2–3 micellar complexes. Delayed fluorescence associated with these complexes was measured and its intensity was used to estimate the original bacterial concentration spiked in hamburger. This approach was applied to detect E. coli O157:H7 spiked in hamburgers. The results indicated this method is able to detect 1 CFU/g of the bacteria after a brief enrichment for four and half hours at 37C. Specificity studies indicated that the approach exhibited no or limited cross reactivity to Salmonella typhimurium, E. coli K-12 or Shigella dysenteriae spiked in hamburgers. Thus, the developed approach may be used as a rapid screening procedure for E. coli O157 bacteria in foods.     

Summer drought alters plant-mediated competition between foliar- and root-feeding insects

Summer droughts are predicted to increase in severity and frequency in the United Kingdom, due to climate change. Few studies have addressed the impacts of drought on interactions between species, and the majority have focussed on increases in CO₂ concentration and changes in temperature. Here, the effect of experimental summer drought on the strength of the plant‐mediated interaction between leaf‐mining Stephensia brunnichella larvae and root‐chewing Agriotes larvae was investigated. Agriotes larvae reduced the abundance and performance of S. brunnichella feeding on a mutual host plant, Clinopodium vulgare, as well as the rate of parasitism of the leaf‐miner. The interaction did not, however, occur on plants subjected to a severe drought treatment, which were reduced in size. Changes to summer rainfall, due to climate change, may therefore reduce the occurrence of plant‐mediated interactions between insect herbivores.     

A Comparison of Experimental and Computational Methods for Mapping the Interactions Present in the Transition State for Folding of FKBP12

The folding pathway of FKBP12, a 107 residue / protein, has been characterised in detail using a combination of experimental and computational techniques. FKBP12 follows a two-state model of folding in which only the denatured and native states are significantly populated; no intermediate states are detected. The refolding rate constant in water is 4 s^-1 at 25 °C. Two different experimental strategies were employed for studying the transition state for folding. In the first case, a non-mutagenic approach was used and the unfolding and refolding of the wild-type protein measured as a function of experimental conditions such as temperature, denaturant, ligand and trifluoroethanol (TFE) concentration. These data suggest a compact transition state relative to the unfolded state with some 70% of the surface area buried. The ligand-binding site, whichis mainly formed by two long loops, is largely unstructured in the transition state. TFE experiments suggest that the -helix may be formed in the transition state. The second experimental approach involved using protein engineering techniques with -value analysis. Residue-specific information on the structure and energetics of the transition state can be obtained by this method. 34 mutations were made at sites throughout the protein to probe the extent of secondary and tertiary structure in the transition state. In contrast to some other proteins of this size, no element of structure is fully formed in the transition state, instead, the transition state is similar to that found for smaller, single-domain proteins, such as chymotrypsin inhibitor 2 and the SH3 domainfrom -spectrin. For FKBP12, the central three strands of the -sheet (2, 4 and 5), comprise the most structured region of the transition state. In particular Val 101, which is one of the most highly buried residues and located in the middle of the central -strand,makes approximately 60% of its native interactions. The outer -strands, and the ends of the central -strands are formed to a lesser degree. The short -helix is largely unstructured in the transition state as are the loops. The data are consistent with a nucleation-condensation model of folding, the nucleus of which is formed by side chains within -strands 2, 4 and 5 and the C-terminus of the -helix. These residues are distant in the primary sequence, demonstrating the importance of tertiary interactions in the transition state. High-temperature molecular dynamic simulations on the unfoldingpathway of FKBP12 are in good agreement with the experimental results.     

Influence of GSTM1 and NAT2 genotypes on the relationship between personal exposure to PAH and biomarkers of internal dose

This study examined the interaction of glutathione S-transferase (GSTM1) and N acetyltransferase (NAT2) genotypes and personal exposure to carcinogenic polycyclic aromatic hydrocarbons (PAH) with biomarkers of exposure in a cohort of 51 non-smoking women from Bohemia, CZ. The biomarkers included urinary PAH metabolities and white blood cell DNA adducts. Personal PAH exposure was significantly correlated with urinary PAH metabolites for all individuals (r 0.36, p 0.01, n 46). After stratifying by genetic polymorphism the correlation between personal PAH exposure and urinary PAH metabolites increased for individuals with NAT2 slow acetylators (r 0.58, p 0.001, n 29) and the combination of GSTM1 null and NAT2 slow acetylators (r 0.60, p 0.01, n 16). DNA adduct levels were not significantly correlated with personal PAH exposure (r 0.16, p 0.32, n 51), unless restricted to individuals with the GSTM1 gene (r 0.59, p 0.005, n 21). Personal exposure data were essential for elucidating the possible effect of genotypes on the relationship between PAH exposure and these two classes of internal biomarkers. \[This abstract does not necessarily reflect EPA policy.]     

Thermal inactivation of cherry leaf roll virus in tissue cultures of Nicotiana rustica raised from seeds and meristem-tips

Nicotiana rustica tissue cultures derived from seeds or embryos infected with cherry leaf roll virus (CLRV), remained infected after culture at 22 ^oC. No infectivity was found in cultures held at 32 ^oC for 5 days but it was readily detected after such cultures were transferred to 25 ^oC for 8 days. Virus was permanently eradicated from most plants after 20 days incubation at 32 ^oC and from all plants after 7 days incubation at 40 ^oC. Partially purified preparations of CLRV lost infectivity after 9–12 days at 22^oC, 5 days at 32^oC and 3 days at 40^oC.     

Electrical Characteristics as Discriminant Criteria for Three Forms of Skeleton Weed (Chondrilla juncea L.)

Using impedance parameters obtained from an electrophysiologicaltechnique it was possible to discriminate between the threeAustralian forms of skeleton weed with high statistical significance,thus providing new criteria within the field of quantitativetaxonomy.     

Phase Change in Hedera helix L: II. THE POSSIBLE BOLE OF ROOTS AS A SOURCE OF SHOOT GIBBERELLIN-LIKE SUBSTANCES

When synthesis was estimated by the agar diffusion techniqueboth basal and lateral adventitious roots of Hedera helix L.in the juvenile growth phase were shown to synthesize gibberellin-likesubstances. Seedlings and cuttings from juvenile ivy could begrown in water culture for several weeks. When roots were excisedfrom these plants shoot growth was reduced in comparison withthat of intact plants. The stems, apices, and leaves of derootedseedlings and cuttings contained lower levels of extractablegibberellin-like substances than comparable organs of intactplants. The major zone of gibberellin-like activity in intactplants co-chromatographed with gibberellins A₁ and A₃. Whenthese gibberellins were applied to plants grown in culture theywere found to promote growth of intact but not derooted plants.These findings are discussed in relation to the role of rootfactors and particularly gibberellins in phase change.     

Zearalenone production in barley

A plot of barley cv. Golden Promise was sprayed with an isolate of Fusarium culmorum known to produce zearalenone, an oestrogenic toxin. The grain was harvested and stored under conditions that were known to induce toxin production in sterilized grain inoculated with the same strain. Toxin was not found in the harvested grain but appeared 20 wk after storage. A plot of unsprayed barley was harvested at the same time as the sprayed plot. F. culmorum was isolated from the grain at the time of harvesting but there was no toxin detectable. This appeared at the same time as toxin in the sprayed plot. Higher concentrations of zearalenone were produced in the control suggesting that the F. culmorum isolated was either a different strain from that used to inoculate the sprayed plot or that the microbial flora of the grain affected toxin production.