Use of real-time PCR to examine the relationship between disease severity in pea and <Emphasis Type="Italic">Aphanomyces euteiches</Emphasis> DNA content in roots

Aphanomyces euteiches causes severe root rot of peas. Resistance is limited in commercial pea cultivars. Real-time fluorescent PCR assay specific for A. euteiches was used to study the relationship between disease severity and pathogen DNA content in infected peas. Five pea genotypes ranging in levels of resistance were inoculated with five isolates of A. euteiches. Plants were visually rated for disease development and the amount of pathogen DNA in roots was determined using the PCR assay. The susceptible genotypes Genie, DSP and Bolero tended to have significantly more disease and more pathogen DNA than the resistant genotypes 90-2079 and PI 180693. PI 180693 consistently had less disease, while 90-2079 had the lowest amount of pathogen DNA. The Spearman correlation between pathogen DNA quantity and disease development was positive and significant (P < 0.05) for three isolates, but was not significant for two other isolates. This suggests that the real-time PCR assay may have limited application as a selection tool for resistance in pea to A. euteiches. Its utility as a selection tool would be dependent on the correlation between disease development and pathogen DNA content for a given pathogen isolate. The accuracy and specificity of the real-time PCR assay suggests considerable application for the assay in the study of mechanisms of disease resistance and the study of microbial population dynamics in plants.     

Capsid protein sequence gene analysis of <Emphasis Type="Italic">Apple mosaic virus</Emphasis> infecting pears

Apple mosaic virus (ApMV, genus Ilarvirus) was detected in pears, a previously non-reported virus host. No symptoms were visible on the hosts leaves. Seventeen out of 22 randomly selected pear trees in Italy (Lombardy) and in three regions in the Czech Republic were ApMV-infected. All nine newly sequenced ApMV isolates from pears had a 15-nucleotide insertion in the capsid protein gene in identical position of that of apple isolates compared with isolates from hop and prunes. The insertion is the most prominent (but not essential) modification of the capsid protein gene, which results in a phylogenetic separation of ApMV isolates into three clusters. Sequence analysis data of an additional 15 isolates revealed a sequence correlation with kernelled fruit trees (apple and pear).     

The use of Verticillium dahliae and Diplodia scrobiculata to induce resistance in Pinus halepensis against Diplodia pinea infection

The pathogenic fungi Verticillium alboatrum and Diplodia scrobiculata were assayed for biological control of Diplodia pinea on Aleppo pine (Pinus halepensis) in Catalonia (north-eastern Spain). Young shoots were pre-treated with inoculations of either V. dahliae or D. scrobiculata, by placing colonized agar plugs on wounds made by removing a single needle fascicle. An inoculation with D. pinea was performed 15 days later. Two months after the shoot inoculations, the canker length on the stems was measured and the percentage of shoot dieback calculated. Verticillium dahliae and D. scrobiculata were found to significantly reduce the canker length of D. pinea (P < 0.05) when compared with positive controls. Diplodia pinea was slightly more sensitive to V. dahliae than to D. scrobiculata, but no significant differences (P > 0.05) were observed in the mean canker length between the two treatments. Trees pre-inoculated with V. dahliae resulted in 31.12% shoot dieback, while those pre-inoculated with D. scrobiculata resulted in 32.18% shoot dieback, compared with positive controls (42.85%).     

Variable response of open-pollinated seedling progeny of avocado to Phytophthora root rot

Phytophthora root rot, caused byPhytophthora cinnamomi Rands, is the most important disease of avocado (Persea americana Miller). In an attempt to identify root rot-resistant rootstocks that could ultimately be used under conditions in southern Florida, we screened open-pollinated progeny of avocado from the National Germplasm Repository in Miami. From 1996 to 1998, a total of 2,355 seedlings from 51 accessions were examined in potting mix artificially infested withP. cinnamomi. Most seedlings developed severe root rot, but tolerance was observed in some families (i.e., progeny of certain accessions). Although the most susceptible families developed mean disease ratings of up to 97% root necrosis, mean ratings for the most tolerant families were less than 60%. There was also a strong relationship between the racial background of the female parent and the tolerance of seedlings. Seedlings of the West Indian race and hybrids between it and the Guatemalan race were significantly more tolerant than those from other parents (P< 0.05). Individuals in several families developed < 50% root necrosis, the arbitrary standard of tolerance in this study. Twelve families accounted for 82% (188 of 229) of the tolerant seedlings, and only two of these did not have a West Indian or Guatemalan × West Indian pedigree. Broad-sense heritability for PRR tolerance was 0.45. This is the first report on the inheritance of PRR tolerance in avocado and on the influence of genotype and racial pedigree under controlled conditions.     

Environmental persistence of 2,3,7,8-tetrachlorodibenzo-<Emphasis Type="Italic">p</Emphasis>-dioxin in soil around hardstand 7 at eglin air force base,Florida

Background and Goal  A number of global events have generated intense scientific scrutiny and public concern of polychlorinated dibenzo-p-dioxins (dioxin). DIoxins have been associated with a range of adverse health effects. 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is recognized as among the most dangerous of the dioxin compounds, and was a contaminant found In one of the herbicides used for vegetation control during the Vietnam conflict: Agent Orange. As a result of purging spray systems and leaking drums of Agent Orange concentrate, TCDD contamination occurred in the soils surrounding Hardstand 7 at Eglin Air Force Base, Florida. This research uses a multimedia model to estimate the TCDD concentrations in surface soil around Hardstand 7 for a 70-year time Interval beginning with observed surface soil concentrations 1984. Methods  Hardstand 7 is a nearly 40-m circular concrete and asphalt aircraft parking area. The hardstand was used as a staging area for spraying equipment used to disseminate herbicide In a test area called C-52A within EAFB. Concentrated herbicide was also stored in 208-L barrels for use with the equipment, and later, for disposal. In 1984, a field investigation characterized the extent of TCDD contamination in surface soils around Hardstand 7 using a radial sampling protocol. The 1984 observed concentrations and locations was used in a multimedia model, CalTOX, as an initial source term concentrations and locations to estimate expected concentrations during the subsequent 70-years. Results  The results indicate that more than 94% of the TCDD observed in surface soils 1984 will remain after a 70-year period. Access restrictions and remediation activities at the site eliminate bar verification of the CalTOX estimates. Conclusions. TCDD is highly persistent in the soil medium and natural attenuation may not produce a significant decrease In soil concentrations. Recommendation  Active remediation actions may be required to prevent exposure to TCDD contamination surface soils. Verifying CalTOX concentration estimates an Important step that should be performed, however, the, model provides an easy to use tool to estimate TCDD surface soil contamination at herbicide storage or dispersion staging sites.     

A new plant-based bioassay for aquatic sediments

Background, Goal and Scope  To date, standardised bioassays for the assessment of the ecotoxicological potential in sediments and dredged material use test organisms like bacteria, algae and crustaceae. This paper presents the development and application of a novel sediment contact test (whole sediment) withMyriophyllum aquaticum, a representative of rooted aquatic macrophytes. The aim of the present study is to demonstrate the value of a sediment contact test with rooted macrophytes as a supplement to existing test batteries in order to improve the assessment of sediment toxicity. Methods  The newly developed sediment contact test withMyriophylhim aquaticum was applied to natural whole sediments. For performing the test, whorls ofMyriophyllum aquaticum were directly planted in the native sediment and incubated in the light at 24°C (cf. section results and discussion). The end points of the test were the number of the shoots and the fresh weight of the whole plants. The duckweed growth inhibition test withLemna minor according to ISO/DIS 20079 was performed in pore waters from sediment samples. The results of the sediment contact test withMyriophyllum aquaticum were compared with each other and with those of the aquatic duckweed test. Results and Discussion  A test protocol for the new plant-based sediment contact test using the aquatic plantMyriophyllum aquaticum as an indicator was developed. The best control sediment proved to be the OECD sediment (OECD 207). A test period of 10 days appeared to be sufficient for the test. The increase of biomass and the derived growth rate were found to be the most suitable evaluation parameters. The growth behaviour ofMyriophyllum aquaticum differed depending on the origin of sediments. Therefore, plant-affecting contamination, that is bound in sediments, was indicated. Conclusions  The novel sediment contact test withMyriophyllum aquaticum can indicate phytotoxic effects in sediments. Therefore, it allows a better assessment of the overall-toxicity in whole sediments. Recommendations and Outlook  The sediment contact test withMyriophyllum aquaticum is a valuable tool for the evaluation of the ecotoxicological risk potential of waters and sediments. It should become a complement to a standardised test battery generally used for the assessment of sediment toxicity.     

Hematopoiesis as a competitive exclusion process: Estimation of a stem cell selective advantage

Female Safari cats are offspring of domestic and Geoffroy parents, and are balanced heterozygotes with equal numbers of Blood cells containing domestic and Geoffroy-type glucose 6-phosphate dehydrogenase (G6PD),an X-chromosome derived enzyme. In previous studies, however, we observed increasing percentages of Blood cells with Geoffroy G6PD in cats aged 4–12 years. Similarly, ratios of parental X-chromosome phenotypes greater than three to one are common in women over age 60, while women under 40 typically exhibit one to one ratios. Using a two-compartment hidden-Markov model of the distribution of phenotype in samples taken during the second stage of hematopoiesis (Blood cell production), we estimate, through simulation and mathematical calculation, the Geoffroy selective advantages necessary to reproduce the data obtained from female Safari cats. It is shown that small differences in the kinetics of hematopoietic stem cells (HSC), less than might be detected by in vitro assays, can explain the “clonal dominance” observed with aging in cats, and by extension, human females. Conceptually, hematopoiesis in females can be viewed as a competitive exclusion process in which two populations of HSC (defined by parental X-chromosome phenotype) compete for environmental resources. As in many ecological examples, dominance may occur only after long periods of time.     

Recovery of cover-crop-N in the soil-plant system in the Guinea savannah zone of Ghana

In order to understand the efficiency of residue-N use and to estimate the minimum input required to obtain a reasonable level of crop response, it is important to quantify the fate of the applied organic-N. The recovery of N from ¹⁵N-labelled Crotalaria juncea was followed in the soil and the succeeding maize crop. Apparent N recovery (ANR) by maize from unlabelled Crotalaria juncea, Crotalaria retusa, Calopogonium mucunoides, Mucuna pruriens and mineral fertilizer at three locations were also evaluated. The maize crop recovered 4.7% and 7.3% of the ¹⁵N-labelled C. juncea-N at 42 days after sowing (DAS) and at final harvest, respectively. The corresponding ¹⁵N recovery from the soil was 92.4% and 58.5%. The highest mean ANR of 57.4% was with mineral fertilizer, whereas the mean ANR of 14.3% from C. retusa was the lowest. A large pool substitution and added-N interaction effect was observed when comparing N recovery from the labelled and unlabelled C. juncea. The amount of residue-N accounted for by the isotope dilution method at 42 DAS was 97.1% and at final harvest 65.8%. The large residue-N recovery in the soil organic-N pool explains the residual effect usually observed with organic residue application.     

The contribution of<Emphasis Type="Italic"> Phragmites australis</Emphasis> litter to methane (CH<Subscript>4</Subscript>) emission in planted and non-planted fen microcosms

The contribution of Phragmites australis (Cav.) Trin. ex Steud. (common reed) litter as an origin of CH ₄-C was studied in a microcosm experiment. ¹⁴C-labelled, dried and ground P. australis root and shoot litter was buried in waterlogged, planted or non-planted fen microcosms. The evolution of ¹⁴CO ₂ and ¹⁴CH ₄ from the pots was monitored during the 35-day experiment. The ¹⁴C activity in the shoots and roots of the plants, soil, and inundation water was also analysed at the end of the experiment. Up to 40% of the released CH ₄-C originated from the added litter, whereas the rest originated from old soil organic matter. The comparison of planted and non-planted pots suggested that the contribution of recent plant-derived C (i.e. root exudates) to CH ₄ emission was negligible. The proportion of litter-derived CH ₄-C was significantly higher in the planted pots, suggesting that the presence of plants enhanced the formation of CH ₄ from litter. The major part of the initial ¹⁴C activity was recovered from the soil. About 3% was recovered from the inundation water, about 10% was emitted as CO ₂, and only <0.01% as CH ₄. However, these results demonstrated that plant litter and old soil organic matter are the major sources of CH ₄-C in fens during the early growth stage of P. australis.