Isolation,characterization, and properties of factor F1 from mitochondrial preparations of housefly (Musca domestica L.) thorax

A water-soluble Mg²⁺-dependent ATPase (coupling factor F₁) was isolated from the mitochondria of housefly thorax. It comprised about 14% of the proteins from a crude preparation. The F₁ preparation was nearly homogeneous as assessed by gel electrophoresis, isoelectric focusing, and electron microscopy. It was composed of five subunits with the following apparent molecular weights: α, 68,000; β, 61,000; γ, 38,000; δ, 27,000; and ?, 17,500. The isoelectric pH (pI) of this protein was 7.3. F₁ had a pH optimum of 8.2 and a temperature optimum between 37 and 45°C. The enzyme was fairly stable at 25°C. Nearly complete loss of activity was noticed at 0°C, while at 0 or 25°C, glycerol (20%) partially stabilized the enzyme activity against such inactivation. The K_m value of the enzyme with respect to ATP was 0.4 mM. The activity was stimulated by low concentrations of 2,4-dinitrophenol. The enzyme was inhibited by azide, p-hydroxymercuribenzoate, and guanidine hydrochloride. Oligomycin and the pesticides pyrethrin, cyhexatin, and DDT have no effect on the enzyme activity. However, all of these chemicals inhibited intact Mg²⁺- ATPase. The results are discussed in the light of differential responses of soluble and intact ATPase to these pesticides.     

Effectiveness of the resistance inducer prohexadione‐Ca against fireblight in shoots of apple trees inoculated with Erwinia amylovora1

Due to the lack of effective, non‐phytotoxic and publicly acceptable materials for controlling fireblight in pome fruit trees, novel strategies against Erwinia amylovora are being sought. Resistance‐inducing compounds, such as prohexadione‐Ca, represent promising alternatives. Prohexadione‐Ca is the active substance of the bioregulator Regalis, currently being introduced in several European countries and overseas. Prohexadione‐Ca reduces shoot elongation due to inhibition of gibberellin biosynthesis. Furthermore, it leads to significant changes in the spectrum of flavonoids and their phenolic precursors in pome fruits, which causes reduced susceptibility to fireblight and other pathogens. In 2002 and 2003, container‐grown apple trees of the cultivars ‘Idared’ and ‘Freedom’ were treated with different dosages of prohexadione‐Ca two weeks before inoculation with E. amylovora. The effect of prohexadione‐Ca against shoot blight was determined by measuring the lengths of necrotic lesions and symptoms on vascular bundles caused by the pathogen. Treatments with prohexadione‐Ca turned out to be much superior to the ones with streptomycin, kasugamycin and a bacterial antagonist, which were used for comparison. Acibenzolar‐S‐methyl (Bion), another resistance‐inducing compound, was included in some of the experiments and gave intermediate results. The simultaneous control of excessive shoot growth and shoot infections by fireblight is seen as a major advantage of using prohexadione‐Ca in pome fruit trees.     

PCR-based detection of Xanthomonas campestris pathovars in Brassica seed

Xanthomonas campestris is a seedborne bacterium that causes black rot of crucifers. Substantial crop losses may result from the rapid spread of the bacteria under favourable conditions, especially those occurring during seedling production. A PCR-based method has been developed for the rapid and sensitive detection of the pathovars of X. campestris that affect crucifers. Primers were designed to specifically amplify a 619 bp fragment of the hrpF gene from X. campestris . Amplification products were not detected from other Xanthomonas species, or from other pathogenic or epiphytic bacteria occurring on these plants. To avoid false-negative results arising from the presence of amplification inhibitors in plant extracts, primers targeting a 360 bp section of the internal transcribed spacer (ITS) region from Brassica spp. were included in a multiplex PCR. The assay readily detected X. campestris pv. campestris infections in diseased plants and from bacterial colonies isolated on growth media, and was more sensitive and specific than traditional plating methods and a commercially available ELISA. A seed-washing protocol was optimized to allow the detection of a single artificially infected seed among 10 000 healthy seeds using the multiplex PCR.     

Note: Comparison of three laboratory methods to evaluate the pathogenicity and virulence of tenPseudomonas syringae pv.syringae strains on apple, pear, cherry and peach trees

The pathogenicity and virulence of ten GreekPseudomonas syringae pv.syringae strains from different hosts (citrus, pear, apple, peach and cherry) were evaluated using three different laboratory methods, which produced results in good agreement. All ten strains were virulent on apple, pear, cherry and peach trees. The extent of tissue colonized varied considerably among strains and cultivars. On excised shoots and twigs of apple and pear, strains BPI 176, BPI 203, PI 2 and PI 14 were the most virulent and strains BPI 689, BPI 992, BPI 4, BPI 20, PI 18 and PI 19 were the least virulent. On excised shoots and twigs of peach and cherry, strains BPI 176, BPI 203, PI 2, PI 14, PI 18 and PI 19 were the most virulent and strains BPI 4 and BPI 20 were the least virulent. Moderate virulence was evinced by strains BPI 689 and BPI 992. These pathogenicity assays are proposed as rapid and reproducible screening systems to evaluate the susceptibility of apple, pear, cherry and peach cultivars to this bacterial pathogen.     

Metabolism, uptake and translocation of 14C-paraquat in resistant and susceptible biotypes of Crassocephalum crepidioides (Benth.) S. Moore

The metabolism, uptake and translocation of paraquat in resistant (R) and susceptible (S) biotypes of Crassocephalum crepidioides (Benth.) S. Moore (redflower ragleaf) at the 10-leaf stage was investigated. A study on the properties of leaf surface was carried out to examine the relationship between leaf surface characters and paraquat absorption. The extractable paraquat was not metabolized by the leaf tissue of either the resistant or susceptible biotypes. Differential metabolism, therefore, does not appear to play a role in the mechanism of resistance. Both biotypes did not show any significant difference in the amount of cuticle and trichome densities. Furthermore, both biotypes are identical in the structure of stomata, trichomes and epicuticular wax. The results of the leaf surface studies are in agreement with the findings of the uptake study. Both biotypes demonstrated no significant difference in absorption between the resistant and susceptible biotypes. However, 10% of the absorbed ¹⁴C-paraquat into the S biotype was translocated basipetally, but not in the R biotype. The results of this study suggest that in C. crepidioides , differential translocation may contribute to the mechanism of resistance at the 10-leaf stage.     

Risk Analysis for Latent Infection of Prune by Monilinia fructicola in California

ABSTRACT The quantitative relationships between incidence of latent infection (ILI) of prune by Monilinia fructicola and wetness duration (WD) for different bloom and fruit developmental stages and different inoculum concentrations were obtained. Three levels of ILI were considered as criteria for low, moderate, and high risks of latent infection, respectively. Seasonal patterns of WD leading to different risk levels of latent infection were obtained for low (IP(L)) and high (IP(H)) inoculum potential conditions in orchards. Longer WD was needed at a resistant than at a susceptible fruit developmental stage to induce similar levels of latent infection. The curves of WD leading to different levels of ILI over the growing season (risky WD curves) were used in risk analysis for latent infection. Multi-year historical WD data from 10 prune-growing locations in California were compared with risky WD curves. The percentage of days (P) with WD leading to a certain risk level of latent infection was calculated for each month from historical weather data. Under the IP(L) condition, the P distributions for low risk of latent infection were higher in March and April than in May and were the lowest in June for most locations. Under the IP(H) condition, the number of days that WD leading to low risk of latent infection in May increased compared with those under the IP(L) condition. The risk analysis approach was evaluated by using separate experimental data as incidence of fruit brown rot obtained from different prune orchards over years. Consistency between predicted overall risk levels of latent infection and observed incidence of fruit brown rot was obtained. The results demonstrated the usefulness of the risk analysis in decision support system for disease management.     

Evaluation of PCR,IEF and ELISA techniques for the detection and identification of potato cyst nematodes from field soil samples in England and Wales

Effective management of potato cyst nematodes (PCNs) requires simple, rapid and accurate identification and quantification of field populations. Soil samples from a survey of 484 fields in potato rotations in England and Wales were used to compare the identification and quantification of PCNs using IEF, PCR, ELISA and bait plant tests. The cyst counts and bait plant test revealed that 64.3% of field samples contained PCNs. Bait plant tests increased the detection rate of PCNs in field samples by 4–6.4%. This means that some infestations are cryptic and would not normally be detected by standard counts. IEF, PCR and ELISA methods distinguished between Globodera rostochiensis and G pallida and were able to register mixed populations; however they were not in full agreement. All methods suggested that G pallida is the dominant species in the field samples tested. The PCR results indicated that 66% of field samples contained pure G pallida, 8% contained pure G rostochiensis and 26% contained mixtures of the two species. Estimates of the relative process times taken per sample in the PCR, IEF and ELISA techniques are given. © 2001 Society of Chemical Industry     

Interference between triazine-resistant Brassica napus and Panicum miliaceum

Atrazine carryover often limits growers to production of atrazine-tolerant crops the year following application, and allows the increase of triazine-tolerant weed species such as Panicum miliaceum L. (wild proso millet). Tiriazine-resistant Brassica napus L. cv. ‘Triton’ (oilseed rape) was tested to characterize the nature of interspecific interference with P. miliaceum. In a greenhouse study, atrazine at 2.2 kg ha^?1 depressed oilseed rape fruit (siliqua) number and fruit dry weight, and delayed flowering, but did not significantly affect height or weight of shoots, Oilseed rape fruit weight was reduced at 200 P. miliaceum plants m^?2. fruit number and shoot weight were inhibited at 400 weeds m^?2. and height was reduced and flowering delayed at 600 weeds m^?2. Number and weight of fruits were reduced by one-third after 8 weeks of interference as compared to oilseed rape grown with the weed for 4 weeks. Oilseed rape height was reduced by 29% and shoot weight by 55% by 600 weeds m^?2 and 2–2 kg ha^?1 atrazine, while fruit number and weight were reduced by 72%. Oilseed rape shoot weight was reduced by 74% by 600 weeds m^?2 for 12 weeks of interference, while fruit number and weight were reduced by 85% and 82%. respectively. In a field study, fluazifop reduced early season P. miliaceum cover by 72%, but did not increase oilseed rape cover. Mid-season P. miliaceum shoot weight was decreased by 97% by fluazifop and oilseed rape shoot weight was increased by 34%. P. miliaceum control increased oilseed rape biomass by 38% at 89 days, but biomass of oilseed rape sown at 11.2 kg ha^?1 with 2.2 kg atrazine ha^?1 was not decreased by P. miliaceum interference at 89 days.     

The occurrence of carbendazim resistance in Rhynchosporium secalis on winter barley in England and Wales in 1992 and 1993

In 1992, samples of Rhynchosporium secalis from 19 winter barley crops in England and Wales were examined for carbendazim resistance. Of the 120 isolates obtained, 14·2% were resistant to carbendazim. A larger survey was carried out in England and Wales in 1993 when samples from 74 crops were examined. On this occasion 16·6% of the 639 isolates obtained were resistant to carbendazim, and resistant isolates were detected in 46% of crops.