Inhibition of insect cytochrome P-450 by some metyrapone analogues and compounds containing a cyclopropylamine moiety and their evaluation as inhibitors of juvenile hormone biosynthesis

Two metyrapone analogues, 2-(l-imidazolyl)-2-methyl-l-phenyl-l-pro-panone (A-phenyl-B-imidazolyl-metyrapone; III) and 2-methyl-l-phenyl-2-{1,2,4-triazol-l-yl)-l -propanone (A-phenyl-B-triazolyl-meiyrapone; IV) as well as two cyclopropylamine derivatives. N-cyclopropyl-4-icrt-butylbenzylamine (V) and N-cyclopropyl-4-(3,7-dimethyl-7-methoxy-octyloxy)benzamide (cyclopropylamine acylated with a JH analogue acid of known structure; VI) were synthesized and evaluated in biological assays for JH biosynthesis on cockroach, Diploptera punctata corpora allata and egg growth in adult cockroach as well as for mixed function oxidase activities, i.e. epoxidation of aldrin to dieldrin and O-demethylation of 7-methoxy-4-methylcoumarin to 7-hydroxy-4-methylcoumarin on microsomes from housefly, Musca domestica, abdomen and from cockroach midgut. Compound VI was a good in-vitro inhibitor of JH biosynthesis, but it had significantly lower activities in the assays for inhibition of microsomal cytochrome P-450. Compound IV and metyrapone had moderate activity as inhibitors of oocyte growth. Compounds III, IV and V were more potent inhibitors of housefly aldrin epoxidation than metyrapone and they inhibited the enzyme activity by almost 100% at 02mM, while in cockroach midgut microsome assay metyrapone was more potent than these three compounds.     

The growth of nematode 'tolerant' and 'intolerant' soyabeans as affected by phosphorus, Glomus intraradices and light

The effects of the vesicular-arbuscular mycorrhizal (VAM) fungus, Glomus intraradices , and the soyabean cyst nematode (SCN), Heterodera glycines , were studied singly and in combination on two soyabean cultivars, cv. Bragg (nematode intolerant) and cv. Wright (moderately nematode tolerant) grown in the greenhouse in soils with low (35 μg/g) and high (70 μg/g) phosphorus (P). Cultivar Wright grew better than cv. Bragg, showing a greater response to P and VAM, and was damaged less by SCN. These differences were not apparent, or were reversed, when the same cultivars were grown in growth chambers where lighting was sub-optimal. Cultivar Wright had a larger shoot:root ratio than cv. Bragg. This finding and the observed growth responses indicate that cv. Wright has a more efficient root system than cv. Bragg. This, we suggest, is the basis of both the greater VAM response observed in. and the greater nematode tolerance ascribed to, cv. Wright compared to cv. Bragg.     

The relation of mansonones to resistance against dutch elm disease and their accumulation,as induced by several agents

No evidence was found that mansonones E and F, which accumulated in elms after infection withCeratocystis ulmi, were responsible for resistance against Dutch elm disease inUlmus hollandica cl. 390. Furthermore, production of mansonones inU. americana was only about one-fifteenth of that inU. hollandica cl. Belgica. Yet both are susceptible to Dutch elm disease.Induction of these compounds was not specific, for accumulation occurred not only after introduction of crude toxin ofC. ulmi, but also after inoculation withVerticillium albo-atrum and after introduction of H₂SO₄ or ethanol.The dark discolouration of the wood after toxin treatment was restricted to the lower part of the stem. No mansonones could be detected in the non-discoloured higher parts of the stem or from the necrotic or wilted leaves.Samenvatting Er werd geen aanwijzing gevonden dat mansononen E en F, die na infectie metC. ulmi gevormd worden, verantwoordelijk zijn voor de resistentie tegen de iepziekte inU. hollandica cl. 390 (Tabel 1). U. americana bevat ongeveer 15 maal zo weinig van deze stoffen alsU. hollandica cl. Belgica, terwijl beide vatbaar zijn.De inductie van de synthese van deze stoffen bleek niet specifiek te zijn, omdat bij chemische beschadiging met H₂SO₄ of ethanol ook mansononen gevormd werden.De donkere verkleuring van het hout bleek na toxine toediening beperkt tot het onderste gedeelte van de stam. Geen mansononen werden gevonden in hoger gelegen, niet verkleurde stamgedeelten of in de necrotische en verwelkte bladeren.     

Evaluation of models to predict take-all incidence in winter wheat as a function of cropping practices,soil, and climate

The incidence and severity of take-all, caused by Gaeumannomyces graminis var. tritici (Ggt), in susceptible crops depend on climate, soil characteristics and cropping practices. Take-all can be controlled by modifying crop rotation, crop management and fungicide treatment. When available, fungicides are used as a seed treatment and are partially effective. There is currently no reliable method for helping farmers to optimise their choice of cropping system to improve take-all control. In this study, we defined 16 models, based on various mathematical functions and input variables, for predicting disease incidence in a wheat crop as a function of soil characteristics, climate, crop rotation and crop management. The parameters of these models were estimated from field experiments carried out at six sites in the north of France over a ten-year period. The root mean squared error of prediction (RMSEP) values of the models were estimated by cross validation and compared. RMSEP was in the range 16.34–65.93% and was higher for the models based on multiplicative functions. The lowest RMSEP value was obtained for a dynamic model simulating disease incidence during the crop cycle and which included among input variables the percentage of diseased plants determined at GS30.     

Evaluation of methyl iodide as a soil fumigant in container and small field plot studies

Methyl iodide was evaluated as a soil fumigant as a potential replacement for the widely used soil fumigant methyl bromide. In container trials, methyl iodide was significantly more effective than methyl bromide against the plant parasitic nematodes Meloidogyne incognita, Heterodera schachtii and Tylenchulus semipenetrans and the plant pathogenic fungus Rhizoctonia solani. In small field plots, soil populations of root-knot nematodes were no longer detected after methyl iodide fumigation at an application rate of 112 kg ha^-1. However, after growing a susceptible lima bean host for two months, substantial root-knot galling occurred, while Rhizobium nodulation was absent. At 168 kg ha^-1 of methyl iodide, root-knot galling was reduced to less than 1%, and no Pythium propagules were recovered on selective detection media. These efficacy data support the conclusion that methyl iodide is a likely candidate for replacing methyl bromide as a soil fumigant. © 1998 SCI.     

Evaluation of Trichoderma spp. as a Biocontrol Agent Against Soilborne Fungi and Plant-parasitic Nematodes in Israel

The optimal conditions required to market Trichoderma as a biocontrol agent against soilborne fungi and nematodes are discussed. These include a proper formulation, an efficient delivery system, and alternative methods for Trichoderma's application.The implementation of Trichoderma in integrated pest management (IPM) can be achieved using a soil treatment which combines reduced amounts of biocides/fungicides and the Trichoderma preparation. Biocontrol activity can be increased by combining two (or more) types of biocontrol agents. Moreover, the construction of a genetically modified Trichoderma can lead to the improvement of certain traits which are absent or not highly expressed in the native microorganism isolated from its natural habitat.Different Trichoderma harzianum and T. lignorum isolates were tested for their nematicidal activity against the root-knot nematode Meloidogyne javanica. In short-term experiments, improved growth of nematode-infected plants and decreases in the root-galling index and the number of eggs per gram of root were achieved when nematode-infested soils were pre- exposed to the T. harzianum preparations. A long-term experiment resulted in improved growth and higher yield of nematode-infected plants, but no significant change in the galling index, either by pre-exposure of the fungus to the soil or by enrichment in the root-ball.As biocontrol is an integral part of the IPM philosophy, judicious use of Trichoderma against soilborne pathogens, when demonstrated to be consistently effective, practical and economic, can serve as a model for the introduction and implementation of other biocontrol means into IPM.     

Communication to the Editor Degradation of Bromoxynil,Ioxynil, Dichlobenil and their Mixtures by Agrobacterium radiobacter 8/4

Degradation of three benzonitrile herbicides, bromoxynil (3,5-dibromo-4-hydroxybenzonitrile), ioxynil (3,5-diiodo-4-hydroxybenzonitrile), dichlobenil (2,6-dichlorobenzonitrile), and their mixtures by the soil micro-organism Agrobacterium radiobacter 8/4 was studied in batch cultures. Bromoxynil was found to be most rapidly degraded, while dichlobenil had the lowest toxicity to our strain. All transformations of studied benzonitriles were performed by the nitrile hydratase which has been shown to act on a broad range of substituted aromatic nitriles. © 1997 SCI.     

陕甘宁接壤区发展潜力评价和生态经济发展模式——以平凉地区为例

选取平凉地区为陕甘宁接壤区典型地区 ,对于其发展潜力从区位、资源、地缘人文、地缘生态等方面做出评价 ,也分析了发展的限制因子。认为平凉地区具有发展成为次一级市场中心的可能。给出了适合本区的生态经济的发展模式 ,并且提出相应的产业对策。     

Weed populations in winter wheat as affected by crop sequence, intensity of tillage and time of herbicide application in a cool and humid climate

Summary There is a lack of information on the combined effects of preceding crop, reduced tillage (especially no-tillage) and the time of herbicide application on the development of weed populations and the efficiency of weed control in winter wheat in humid temperate climates. An experiment was conducted with a crop rotation (winter wheat – oilseed rape – winter wheat – maize) on a sandy loam and a loamy silt soil in the Swiss midlands to investigate the impact of different preceding crops and pre- and post-emergence control of weeds in conventional tillage (CT; mouldboard plough), minimum tillage (MT; chisel plough) and no-tillage (NT; no soil disturbance systems). When winter wheat was grown after maize and winter wheat was grown after oilseed rape, the ranking order of weed density in treatments without herbicide application was NT < MT < CT and CT < MT < NT respectively. Analysis of variance and canonical discriminant analysis showed that Epilobium spp., Sonchus arvensis , Myosotis arvensis and volunteer crops were more abundant in NT than in MT and CT. The efficiency of post-emergence weed control was generally better than that of pre-emergence weed control, regardless of tillage intensity.     

Competition between sugarbeet and Sinapis arvensis and Chenopodium album, as affected by timing of nitrogen fertilization

The effects of timing of N fertilization (early, standard or late) on competition between two sugarbeet cultivars (Ritmo, semi-prostrate; and Rizor, erect) and two weeds ( Sinapis arvensis and Chenopodium album ) were studied over 2 years at Viterbo. In both years, time of N fertilization did not influence biomass, yield and yield quality of the weed-free crop, but early N fertilization gave higher crop biomass reduction in the presence of S. arvensis and lower crop biomass reduction in the presence of C. album . Root and sucrose yield responded to competition in the same way as biomass. However, percentage reductions were higher, as both weeds affected harvest index. The two cultivars showed the same response to competition. At the weed densities studied, crop competitive ability was favoured by late N fertilization in the presence of S. arvensis and by early N fertilization in the presence of C. album . Crop competition only reduced seed production by C. album but lowered seed germination of both weeds.     

Diversity and detection of Korean Erwinia pyrifoliae strains as determined by plasmid profiling, phylogenetic analysis and PCR

Twenty-five strains of Erwinia pyrifoliae were investigated for their plasmid profiles and genetic relatedness. Four types of plasmid profile were observed for the first time, suggesting intraspecific plasmid profile diversity in E. pyrifoliae . Moreover, BOX-PCR and phylogenetic analysis based on the 16S-23S intergenic transcribed spacer (ITS) region showed genetic variations among E. pyrifoliae strains, although all strains were clustered in one group and separated from E. amylovora . On the other hand, ERIC-PCR and phylogenetic analysis based on partial groEL gene sequences revealed close genetic relatedness among the strains. Amplification with EpSPF and EpSPR primers of a fragment of approximately 0·65 kb from the genomic DNA of all E. pyrifoliae strains, but not from E. amylovora strains, suggested that this primer set is useful for identification of this pathogen.     

Molecular analysis and virus transmission tests place Olpidium virulentus, a vector of Mirafiori lettuce big-vein virus and tobacco stunt virus, as a distinct species rather than a strain of Olpidium brassicae

The rDNA-ITS sequences of ten single-sporangium isolates of Olpidium virulentus (a noncrucifer strain of Olpidium brassicae), which transmits Mirafiori lettuce big-vein virus (MLBVV) and tobacco stunt virus (TStV), were compared with those of six single-sporangium isolates of O. brassicae. The sequence similarity within isolates of O. virulentus or O. brassicae was almost identical (98.5%–100.0%), but was low between the two species (79.7%–81.8%). In a phylogenetic analysis of the rDNA-ITS region, O. virulentus and O. brassicae fell into two distinct clusters, indicating that O. virulentus, a vector of MLBVV and TStV, is a distinct species rather than a strain of O. brassicae.     

Characterization of a potyvirus from eggplant (Solanum melongena) as a strain of potato virus Y by N-terminal serology and sequence relationships

A potyvirus (eggplant mottle virus, EMoV) causing mosaic mottling in eggplant ( Solanum melongena ) was characterized on the basis of biological, serological and partial nucleotide sequence properties. EMoV infected Chenopodium amaranticolor and members of the Solanaceae. Polyclonal antiserum against EMoV showed antigenic relationship with henbane mosaic potyvirus (HMV) and potato Y potyvirus (PVY). Virus-specific antibodies directed to the N-terminal region of EMoV cross-reacted only with PVY. Determination and comparison of nucleotide sequence of the coat protein (CP) and the 3'-untranslated region (UTR) of EMoV with other potyviruses showed that the level of homology was highest with PVY isolates. Comparative sequence analyses of the CP amino acid and 3'-UTR sequences with distinct PVY isolates placed EMoV within the PVY^O subgroup.     

Influence of soil type and pH on the colonisation of sugar beet seedlings by antagonistic Pseudomonas and Bacillus strains,and on their control of Pythium damping-off

In five different soils originating from Scotland (Craibstone and Cruden Bay), Germany (Magdeburg and Uelzen) and Greece (Tymbaki), Pseudomonas fluorescens B5 reached higher population sizes (4.7–5.7logCFU/plant) on 12-day-old sugar beet seedlings than Bacillus subtilis MBI 600 (4.1–4.8logCFU/plant). Total population size per plant was not affected by soil type. In all five soils, the antagonists reached highest population densities in the hypocotyl and the upper 2cm root section (P. fluorescensB5: 5.2–6.8log₁₀CFU/g plant fresh weight, Bacillus subtilisMBI 600: 5.2–6.1log₁₀CFU/g plant fresh weight) and declined to 0–3log₁₀CFU below 4cm root depth. Colonisation by P. fluorescens B5 down the root was slightly increased in the soils from Craibstone, Magdeburg, and Uelzen compared to the sandy clay loam from Tymbaki. In lux-marked P. fluorescens B5, population density was positively correlated with light emission in all soils; the light emission indicated physiological activity of the strains. However, P. fluorescens B5 reduced Pythium damping-off (measurement after 14 days plant growth) only in three of the five soils (Craibstone, Cruden Bay and Magdeburg). Co-inoculation of B. subtilis MBI 600 increased downward colonisation of the root by P. fluorescens B5, but not the total population ofP. fluorescens B5 per plant. Bacillus subtilisMBI 600 did not reduce Pythium damping-off in any of the soils nor did it influence the efficiency of co-inoculated P. fluorescens B5; its population consisted mainly of physiologically inactive spores. In Craibstone soil, pH did not affect population density, distribution along the root or biocontrol activity against P. ultimum of P. fluorescens B5 or B. subtilis MBI 600.     

Relevance of the deposit structure for the uptake and bio-efficacy of diquat,as monitored by the spatially resolved chlorophyll fluorescence

Aim of our study was to exploit the relation between deposit structure at the microscale and the uptake and biological efficacy of herbicides. For this purpose, we analysed the relevance of the deposit structure of diquat dibromide, as affected by surfactants, on the spatially resolved chlorophyll fluorescence (ChlF) and the desiccation of the leaves. The present study is a sequential work to our studies with the systemic compound glyphosate. On that basis, we hypothesized here that larger deposits of diquat are negatively related to the bio-efficacy of the compound. By using selected ethoxylated rapeseed oil adjuvants (RSO 5, RSO 10, RSO 30, RSO 60) we influenced the deposit properties of diquat dibromide droplet residue on the leaves of easy-to-wet Viola arvensis and the difficult-to-wet Chenopodium album species. With the spatially-resolved pulse amplitude modulated (PAM) ChlF technique we demonstrated the effect of diquat on the physiology of the tissue. As shown, the RSO surfactants did not affect the area of diquat residue on the easy-to-wet leaves of V. arvensis; this trend is similar to those observed for ChlF and the herbicide desiccation potential. In contrary, on C. album, decreased deposit area of diquat droplet was associated with increased effect on ChlF parameters and increased desiccation potential of the herbicide, thus explaining its higher foliar uptake.     

Root and Collar Rot of Milkwort Caused by Cylindrocladium pauciramosum, a New Record for Europe

A new disease of milkwort (Polygala myrtifolia) was observed on several commercial nurseries in southern Italy. Diseased plants showed wilting, stunting, chlorosis or loss of foliage, and rotting of the basal stem as well as the crown and roots. A Cylindrocladium species was consistently found associated with crown, basal stem, and root lesions. The etiology of this disease was proved on milkwort, by fulfilling Koch's postulates. Two hundred Cylindrocladium isolates were collected from the most important Sicilian and Calabrian ornamental production areas from different host plants. Isolates were identified as Cylindrocladium pauciramosum (teleomorph Calonectria pauciramosa) on the basis of their obpyriform to broadly ellipsoidal terminal vesicles, conidiophore branching pattern, conidium morphology, as well as mating type studies with tester strains of C. pauciramosum for selected isolates. This is the first record of this pathogen from Europe and it is the first report of C. pauciramosum on milkwort.     

Evaluation of a rapid diagnostic field test kit for identification of Phytophthora species, including P. ramorum and P. kernoviae at the point of inspection

Plant health regulations to prevent the introduction and spread of Phytophthora ramorum and P. kernoviae require rapid, cost effective diagnostic methods for screening large numbers of plant samples at the time of inspection. Current on-site techniques require expensive equipment, considerable expertise and are not suited for plant health inspectors. Therefore, an extensive evaluation of a commercially available lateral flow device (LFD) for Phytophthora species was performed involving four separate trials and 634 samples. The assay proved simple to use, provided results in a few minutes and on every occasion a control line reacted positively confirming the validity of the test. LFD results were compared with those from testing a parallel sample, using laboratory methods (isolation and real-time PCR). The diagnostic sensitivity of the LFD (87·6%) compared favourably with the standard laboratory methods although the diagnostic specificity was not as stringent (82·9%). There were a small number ( n  = 28) of false negatives, but for statutory purposes where all positive samples must be identified to species level by laboratory testing, overall efficiency was 95·6% as compared with visual assessment of symptoms of between 20-30% for P. ramorum and P. kernoviae . This work demonstrates the value of the LFD for diagnosing Phytophthora species at the time of inspection and as a useful primary screen for selecting samples for laboratory testing to determine the species identification.     

Detection of the German grapevine yellows (Vergilbungskrankheit) MLO in grapevine,alternative hosts and a vector by a specific PCR procedure

A polymerase chain reaction procedure was developed which enables specific amplification of a ribosomal sequence from the mycoplasmalike organism (MLO) associated with German grapevine yellows (Vergilbungskrankheit, VK) and stolbur-related diseases of solanaceous plants. Successful amplification from all samples prepared from various cultivars collected in different viticultural areas indicates that the causal agent is a relatively homogeneous organism. Amplification was also achieved with template DNA prepared from naturally infected weeds in vineyards such asConvolvolus arvensis andSolanum nigrum, and from the planthopperHyalesthes obsoletus that was collected in the vineyards. Feeding of insects of this species on grapevine seedlings resulted in the development of typical yellows symptoms by the grapes.H. obsoletus could therefore be identified as a vector of Vergilbungskrankheit.Abbreviations FD Flavescence dorée - GY Grapevine yellows - MLO Mycoplasmalike organism - PCR Polymerase chain reaction - RFLP restriction fragment length polymorphism - VK Vergilbungskrankheit (German grapevine yellows)     

Induction of peroxidase, scopoletin, phenolic compounds and resistance in Hevea brasiliensis by elicitin and a novel protein elicitor purified from Phytophthora palmivora

Elicitin and a new protein 75 kDa elicitor were purified from the culture filtrate of Phytophthora palmivora, a pathogen of Hevea brasiliensis (rubber plant). Elicitin was obtained by using a one step of DEAE cellulose chromatography and the new elicitor was obtained by two steps of chromatography: a DEAE cellulose column followed by a hydrophobic column. Both elicitors were stable to heat and a wide range of pH values, but were sensitive to ProteaseK. Both elicitors induced scopoletin, peroxidase isozymes (with substrate o-dianisidine and scopoletin) and total phenolic compounds in cell suspension of H. brasiliensis with similar kinetics. In addition, both elicitors induced peroxidase enzyme (o-dianisidine), total phenolic compounds and enhanced local resistance against P. palmivora on young rubber tree seedlings. However, the increase of peroxidase enzyme and total phenolic compounds in rubber tree seedlings was different from those in cell suspension. Furthermore, during the expression of local resistance the zoospore of P. palmivora induced the peroxidase enzyme (o-dianisidine) more rapidly and with higher level than the control plants. H. brasiliensis is more responsive to the new elicitor than elicitin in triggering defense responses. That is the new elicitor was active at a concentration lower than those required for elicitin, about a 30-fold decrease for activation defense responses in cell suspension. For induction of peroxidase enzyme (o-dianisidine), phenolic compounds and local resistance of rubber plants against P. palmivora, the 75 kDa protein was active at about a 2-fold lower concentration when compared to elicitin.     

Priming of the oxidative burst in rice and wheat cell cultures by ulvan,a polysaccharide from green macroalgae,and enhanced resistance against powdery mildew in wheat and barley plants

The capacity of ulvan, a sulphated heteropolysaccharide, to prime the chitin‐ and chitosan‐elicited oxidative burst in wheat and rice cells was tested. Gas‐liquid chromatography showed that ulvan was composed of rhamnose, xylose, glucuronic acid, glucose and galactose. It contained very low amounts of proteins and ca. 19% sulphate groups. The polymer did not elicit the production of hydrogen peroxide in suspension‐cultured wheat or rice cells. Furthermore, in both cell cultures, the simultaneous addition of ulvan and chitin hexamer or chitosan polymer did not significantly alter the intensity of the oxidative burst caused by the elicitors alone. However, pretreatment of wheat cells with ulvan increased the chitin‐elicited oxidative burst about five‐ to sixfold, and that elicited by chitosan about twofold. In rice cells, the production of H₂O₂ elicited by chitin or chitosan was strongly primed by pretreatment with the same concentration of ulvan, increasing the burst triggered by the elicitors alone by 150 and 80 times, respectively. Pretreatment of whole plants with ulvan significantly reduced the symptom severity of Blumeria graminis infection, by 45% in wheat and by 80% in barley. Thus, the priming activity of ulvan on the oxidative burst correlates with a decrease of disease symptoms in infected plants. This is apparently the first report on priming activity of a natural algal polysaccharide.