Microsomal oxidase activity of three blowfly species and its induction by phenobarbital and β-naphtoflavone

Induction of the microsomal oxidase system by dietary phenobarbital and β-naphthoflavone was examined in three blowflies, Phormia regina (Mg.), Lucilia illustris (Mg.), and Eucalliphora lilica (Walk.). Responses were similar in adults and larvae of all species. Phenobarbital increased cytochrome P-450 levels up to 9-fold and aldrin epoxidase up to 138-fold. Increases in cytochrome P-450 and aldrin epoxidase caused by β-naphthoflavone were minor relative to those produced by phenobarbital. In toxicity experiments with carbaryl and propoxur tolerance was associated with the amount of microsomal oxidase activity. Using piperonyl butoxide to synergize carbaryl and propoxur there was no clear indication for the use of either the synergist ratio or synergist difference as an indicator of microsomal oxidase activity.     

The effect of α-hexachlorocyclohexane on liver growth in intact and adrenalectomized rats

α-Hexachlorocyclohexane was administered as a single oral dose (100 mg/kg body wt) to both intact and adrenalectomized male Sprague-Dawley rats. In the intact rats there was a peak in [³H]thymidine incorporation into DNA at 30 hr and an increase in relative liver weight (liver weight/body wt × 100) which peaked at 60 hr post α-HCH dose. However, in α-HCH-dosed adrenalectomized rats [³H]thymidine incorporation into DNA remained elevated throughout the study period and relative liver weight was not significantly increased. In corticosterone-supplemented α-HCH-dosed Adx rats, [³H]thymidine incorporation into DNA was significantly reduced and RLW was increased comparable to that seen in intact animals. These results suggest that α-HCH-induced liver growth is mediated by corticosterone.     

Studies on BHC isomers and related compounds: VIII. Urinary metabolites produced from γ- and β-BHC in the mouse: Chlorophenol conjugates

Metabolites produced from γ-BHC and β-BHC in the mouse urine were purified and characterized. Most metabolites from the both isomers were not extractable by organic solvents, and were conjugates such as sulfates and glucuronides. After hydrolysis with an appropriate enzyme, the conjugates gave chlorophenols, among which 2,4,6-trichlorophenol existed most abundantly—about 25% in the total urine metabolites. 2,4-Dichlorophenol constituted also a significant portion.     

Efficacy and dose–response relationship in biocontrol of Fusarium disease in maize by Streptomyces spp.

Two isolates of Streptomyces spp. DAUFPE 11470 and DAUFPE 14632 were evaluated to determine the antagonist–pathogen inoculum concentration relationship under greenhouse conditions. Pathogen and antagonist concentration, significantly (P < 0.05) affected development of Fusarium disease in maize with a significant interaction between pathogen and antagonist concentration. Dose–response relationship also differed significantly (P < 0.05) between the two isolates, but both isolates demonstrated effective control of Fusarium disease, regardless of pathogen concentration. The isolate DAUFPE 11470 provided the most effective control. The lowest value for disease incidence occurred at low pathogen (10³ chlamydospore g⁻¹ soil) and high antagonist concentration (10⁶ cfu ml⁻¹) for both isolates. The disease incidence for control plants ranged from 19% to 76%. However, in relation to control the lowest disease reduction occurred at low pathogen (10³ chlamydospore g⁻¹ soil) and high antagonist concentrations (10⁶ cfu ml⁻¹). These reductions were 10.6% and 13% for DAUFPE 14632 and DAUFPE 11470, respectively. The highest disease reductions, in relation to control plants, occurred at high pathogen (10⁶ chlamydospore g⁻¹ soil) and antagonist (10⁶ cfu ml⁻¹) concentrations for both isolates. These values were 55% and 62.2% for DAUFPE 14632 and DAUFPE 11470, respectively. The chlamydospore germination of Fusarium moniliforme was affected by glucose addition, antagonist isolates and type of inoculum. The lowest chlamydospore germination was observed with bacterial suspensions of the isolates, for all glucose additions. The results suggested that both Streptomyces spp. isolates were effective at different doses as biocontrol agents against F. moniliforme. Also, there was evidence for at least two mechanisms of biocontrol and that apparently, both isolates showed the same mechanisms of biocontrol action related to production of bioactive compounds and competition for carbon. Further studies will be developed to improve the level and effectiveness of control by these isolates.     

Characterization of phytoplasmas related to peanut witches’-broom and stolbur groups associated with alfalfa diseases in Iran

Journal of Plant Diseases and Protection - In the central and south eastern provinces of Iran, alfalfa plants showing phytoplasma diseases-like symptoms categorized into three major types, namely...     

Induction of Chitinase and β-1,3-Endoglucanase Proteins by UV Irradiation and Wounding in Grapefruit Peel Tissue

UV irradiation enhanced the resistance of grapefruit against the development of green mold décay caused byPenicillium digitatum, the main postharvest pathogen of citrus fruit, and significantly inhibited the fungus’ growth at the fruit wound sites. Immunoblotting analysis using specific citrus chitinase and β-1,3-endoglucanase antibodies, showed that UV irradiation, wounding of the fruit, or a combination of these two treatments, induced the accumulation of a 25 kD chitinase protein in the fruit’s peel tissue. On the other hand, UV irradiation or wounding of the fruit alone was unable to induce the accumulation of 39 and 43 kD β-1,3-endoglucanase proteins, but the combination of the two treatments increased these protein levels. It is suggested that both chitinase and β-1,3-endoglucanase may play a role in the UV-induced resistance of grapefruit againstP. digitatum. Contribution from the Agricultural Research Organization, The Volcani Center, Bet Dagan, Israel. No. 403/99. http://www.phytoparasitica.org posting June 3, 1999.     

The response of phenolic compounds in grapes of the variety ‘Chardonnay’ (Vitis vinifera L.) to the infection by phytoplasma Bois noir

The study was carried out on phytoplasma susceptible grapevine variety ‘Chardonnay’ (Vitis vinifera L.). The changes in total and individual phenolics, with a focus on hydroxycinnamic acids, flavanols and flavonol contents, were studied in phytoplasma-symptomatic and non-symptomatic berries of Bois noir (BN) infected and uninfected vines. Evident responses to BN infection at veraison have been monitored in a decreased accumulation of caftaric and coutaric acids, p-coumaroyl hexose, procyanidin B1, procyanidin trimer as well as of quercetin-3-O-glucoside, quercetin-3-O-glucuronide and quercetin-3-O-xyloside. At berry softening BN infection statistically increased the content of total phenolics, hydroxycinnamic acids and flavanols, but decreased the flavonol contents, especially at phytoplasma-symptomatic berry skins. Later, at harvest, the BN infection caused an additional significant decrease of coutaric acid and p-coumaroyl pentose contents, moreover of procyanidin B1 and procyanidin dimmers (1, 2, and 3), trimer, kaempferol-3-O-glucoside and of most identified quercetins, except of quercetin-3-O-xyloside. At harvest, non-symptomatic berries from infected plants showed similar dynamics in the total phenolic content compared to berry skins from uninfected plants, but in total flavanols and flavonols content similarity to those symptomatic was observed. The latter decreases grape quality and its antioxidant potential. The Bois noir disease showed specific, local and growth-phase-induced responses regarding the content of phenolics in berry skins, where in particular the differences between phytoplasma-symptomatic and non-symptomatic grapes have to be underlined.     

Recombination-like sequences in the upstream region of the phage-type DNA polymerase in ‘<Emphasis Type="Italic">Candidatus</Emphasis> Liberibacter asiaticus’

We determined and compared nucleotide sequences in the upstream region of the bacteriophage-type DNA polymerase in 15 Asian isolates of the citrus-greening bacterium ‘Candidatus Liberibacter asiaticus’ using the genomic information of American isolate Psy62. Some isolates had nucleotide changes and a large 156-bp deletion, which were concentrated in the putative bacteriophage genes ORF3 and ORF4 and corresponding intergenic region. Eight partial recombination-like patterns were observed in this variable region. Most were confirmed to have occurred between the original types 1 and 2, but a fragment, only several dozen nucleotides long and possibly derived from a third type, was also identified.     

Effect of α tocopherol and selenium on antioxidant status, lipid peroxidation and hepatopathy induced by malathion in chicks

The objective of the present study was to investigate the role of α tocopherol and selenium on malathion induced hepatic damage, and antioxidant defense in chicks. The chicks were divided into three groups. First group received malathion 10 mg/kg BW, orally for 60 days, the second group received the same dose of malathion but was supplemented with α tocopherol and selenium for 60 days and the third group served as the control. A compromised antioxidant capacity as evidenced by increased levels of erythrocytic lipid peroxidation and decreased concentration of vitamin E and decreased activity of glutathione peroxidase was observed in chicks following the administration of malathion. An improved antioxidant status was observed in chicks of second group with α tocopherol and selenium supplementation including higher concentration of vitamin E, increased activity of glutathione peroxidase and lower levels of lipid peroxidation. Histopathological studies of liver in the chicks which received malathion exhibited, moderate to severe degenerative and necrotic changes in the hepatocytes. The correlation of decreased antioxidant status of chicks with degenerative changes in liver suggests that lipid peroxidation may be one of the important mechanism in the chronic toxicity of malathion. The results indicate that α tocopherol and selenium were effective in partially alleviating degenerative changes induced by malathion in the liver of chicks by attenuating processes leading to lipid peroxidation.     

Characterization of α-amylase and its activity from mustard aphid Lipaphis erysimi (Kalt.) (Hemiptera) and its inhibition by Rorippa indica defensin

Abstract

Lipaphis erysimi Kaltenbach is one of the economically important pests of Indian mustard Brassica juncea. Plant defense against these pests include impairing digestive processes in the insect gut. Thus, for an effective pest management, it is imperative to understand the digestive enzymes present in the pest. The present study involves the characterization of mustard aphid α-amylase and the efficacy of a potent insecticidal agent, RiD (Rorippa indica defensin) against it. The total protein of L. erysimi was analysed to determine the α-amylase activity present in it. The relative α-amylase activity in the total protein was assayed by the dinitrosalicylic acid (DNS) procedure, using 1% soluble starch as substrate and was observed to be 2.32?±?0.025?U/mL of the total protein. The optimum temperature for α-amylase activity was found to be 35?°C and optimum pH was recorded as 6.5. Presence of RiD showed a decrease in the α-amylase activity in a non-competitive way. Moreover, cryo-sections of the aphid midgut also revealed bound RiD in the confocal analysis.     

Detection of ‘Candidatus Liberibacter asiaticus’ in citrus by concurrent tissue print-based qPCR and immunoassay

‘Candidatus Liberibacter asiaticus’ (CLas) is associated with the most destructive disease of citrus, huanglongbing (HLB). The most widely used methods for detection of CLas are PCR-based and require purification of DNA from plant samples. Elution of DNA from tissue prints made on nitrocellulose membranes followed by qPCR (TPE-qPCR) was compared to DNA extraction of plant tissue followed by PCR (X-PCR) by testing the same tissue samples. The former estimated a higher CLas population in tissue prints than the latter (t-test; P = 0.009). All extracts prepared for TPE-qPCR throughout the experiment were also tested by conventional PCR and 80.8% were identified as positive. A similar set of stem and petiole tissue samples was tested by TPE-qPCR and immunoassay. Although the detection rate by TPE-qPCR was higher than by immunoassay, about 6% of tissue prints were positive by immunoassay but not by TPE-qPCR. Thus, a higher detection rate would be achieved by combining TPE-qPCR with immunoassay. Significant differences were observed in the performance of nitrocellulose membranes from different manufacturers in these assays. Immunotissue prints showed that the spatial distribution pattern of CLas infection varied widely from one sample to another, but the patterns were highly correlated among serial sections from the same sample, suggesting that CLas preferentially colonizes adjacent phloem cells in a vertical rather than horizontal direction.     

λ-cyhalothrin and cypermethrin induced in vivo alterations in the activity of acetylcholinesterase in a freshwater fish, Channa punctatus (Bloch)

In the present study, the in vivo effects of λ-cyhalothrin and cypermethrin on the activity of acetylcholinesterase (AChE, EC 3.1.1.7) were evaluated for 96 h in brain, muscle and gills of Channa punctatus. Both compounds exhibited tissue specific as well as dose dependent decrease in the activity of AChE. The treated fish showed a significant decrease in the activity of AChE in brain and a lesser inhibition in muscle and gills in response to the increasing concentrations of λ-cyhalothrin as well as cypermethrin. Our results indicated that the brain was the main target organ for both insecticides, followed by muscle and gills, as determined by AChE inhibition study. However, these organs showed variations in the degree of AChE inhibition for separate treatments of both insecticides. The λ-cyhalothrin was a more potent AChE inhibitor as compared to cypermethrin. These findings indicated that apart from the established mechanism of delayed closure of sodium ion channels, these pyrethroids inhibit the activity of AChE in C. punctatus which could further aggravate their neurotoxic effects.     

Effect of the disodium salt of cyclopentene β,β′-triketone on the development of infections induced by TMV in the leaves of hypersensitive and susceptible tobacco plants

Journal of Plant Diseases and Protection - We show here that the disodium salt of 2-acetyl-5-chloro-4- hydroxycarbonylmethylthiocyclopent-4-ene-1,3-dione (salt 2) inhibits tobacco mosaic virus...     

Spatial pattern analysis of citrus canker-infected plantings in são paulo, Brazil, and augmentation of infection elicited by the asian leafminer

ABSTRACT Eradication of Asiatic citrus canker (ACC) has become increasingly difficult over the last decade, following the introduction of the Asian leafminer into Brazil and Florida, which has led to changes in the eradication protocols. The present study, undertaken in Brazil, was aimed at characterizing the spatial patterns of ACC in commercial citrus plantings to gain better understanding of the dynamics of the disease subsequent to introduction of the leafminer. The spatial patterns of ACC were mapped in 326 commercial citrus plantings and statistically assessed at various spatial dimensions. The presence of "within-group" aggregation in each plot was examined via beta-binomial analysis for groups of trees parsed into three-by-three-tree quadrats. The relative intensity of aggregation was expressed as a binomial index of dispersion (D) and heterogeneity among plots expressed as the intracluster correlation coefficient, rho. The population of data sets was found to fall into three D categories, D < 1.3, 1.3 3.5. These categories then were related to other spatial characteristics. The binary form of Taylor's power law was used to assess the overdispersion of disease across plots and was highly significant. When the overall population of plots was parsed into D categories, the Taylor's R (2) improved in all cases. Although these methods assessed aggregation well, they do not give information on the number of foci or aggregations within each plot. Therefore, the number of foci per 1,000 trees was quantified and found to relate directly to the D categories. The lowest D category could be explained by a linear relationship of number of foci versus disease incidence, whereas the higher two categories were most easily explained by a generalized beta function for the same relationship. Spatial autocorrelation then was used to examine the spatial relationships "among groups" composed of three-by-three-tree quadrats and determine common distances between these groups of ACC-infected trees. Aggregation was found in >84% of cases at this spatial level and there was a direct relationship between increasing D category and increasing core cluster size, and aggregation at the among-group spatial hierarchy was generally stronger for the within-row than for the across-row orientation. Clusters of disease were estimated to average between 18 and 33 tree spaces apart, and the presence of multiple foci of infection was commonplace. The effectiveness of the eradication protocol of removing all "exposed" trees within 30 m surrounding each "ACC-infected tree" was examined, and the distance of subsequent infected trees beyond this 30-m zone from the original focal infected tree was measured for each plot. A frequency distribution was compiled over all plots to describe the distance that would have been needed to circumscribe all of these outliers as a theoretical alternative protocol to the 30-m eradication protocol. The frequency distribution was well described by a monomolecular model (R(2) = 0.98) and used to determine that 90, 95, and 99% of all newly infected trees occurred within 296, 396, and 623 m of prior-infected trees in commercial citrus plantings, respectively. These distances are very similar to previously reported distances determined for ACC in residential settings in Florida.     

Assessment of the high-dose concept and level of control provided by MON 87701 × MON 89788 soybean against Anticarsia gemmatalis and Pseudoplusia includens (Lepidoptera: Noctuidae) in Brazil

BACKGROUND: Genetically modified MON 87701 × MON 89788 soybean (Glycine max), which expresses the Cry1Ac and EPSP‐synthase proteins, has been registered for commercial use in Brazil. To develop an Insect Resistance Management (IRM) program for this event, laboratory and field studies were conducted to assess the high‐dose concept and level of control it provides against Anticarsia gemmatalis and Pseudoplusia includens. RESULTS: The purified Cry1Ac protein was more active against A. gemmatalis [LC₅₀ (FL 95%) = 0.23 (0.15–0.34) µg Cry1Ac mL^?1 diet] than P. includens [LC₅₀ (FL 95%) = 3.72 (2.65–4.86) µg Cry1Ac mL^?1 diet]. In bioassays with freeze‐dried MON 87701 × MON 89788 soybean tissue diluted 25 times in an artificial diet, there was 100% mortality of A. gemmatalis and up to 95.79% mortality for P. includens. In leaf‐disc bioassays and under conditions of high artificial infestation in the greenhouse and natural infestation in the field, MON 87701 × MON 89788 soybean showed a high level of efficacy against both target pests. CONCLUSIONS: The MON 87701 × MON 89788 soybean provides a high level of control against A. gemmatalis and P. includes, but a high‐dose event only to A. gemmatalis. Copyright © 2012 Society of Chemical Industry     

Rapid and quantitative detection of citrus huanglongbing bacterium ‘Candidatus Liberibacter asiaticus’ by real-time fluorescent loop-mediated isothermal amplification assay in China

Huanglongbing (HLB) is a devastating citrus disease that is associated with bacteria of the species ‘Candidatus Liberibacter asiaticus (Las)’ in China. All currently popular cultivars are susceptible to HLB, and there are no effective approaches to control the disease once the plants have been infected. Therefore, rapid and easy diagnostic tools are needed for maintaining clean nursery stock and inoculum removal strategies to control HLB. A real-time fluorescence loop-mediated isothermal amplification (RealAmp) targeting16S rDNA was developed for the rapid and quantitative detection of Las in China. The detection sensitivity of the RealAmp assay was approximately 1 pg/μl template DNA (equal to 2.4 × 10⁵ target DNA copies/μl) and no cross-reaction was observed with other pathogens or virus-free seedlings. Besides the quantitative detection using SYTO-9 fluorescent dye, an improved closed-tube visual inspection technique using SYBR Green I staining was developed for rapid screening of samples and minimize the risk of cross-contamination. The RealAmp assay is an alternative quantitative method, which would be used as a routine detection service for nursery materials in China.     

Biological control of the millet head miner Heliocheilus albipunctella in the Sahelian region by augmentative releases of the parasitoid wasp Habrobracon hebetor: effectiveness and farmers’ perceptions

On-farm augmentative releases of the parasitoid Habrobracon hebetor (Say) for controlling the millet head miner (MHM) Heliocheilus albipunctella (de Joannis) were tested in Burkina Faso, Mali and Niger from 2007 to 2009. In addition, a survey of farmers’ perceptions of insect pests, with particular focus on MHM, and the biological control program (BCP) was carried out. There was a significant increase of MHM parasitization rate after the releases, with up to 97% mortality. The survey on farmers’ perceptions revealed a fair knowledge of the MHM and the ability of farmers to describe the pest and the damage it caused. Farmers claimed that the biocontrol agent H. hebetor is effective and perceived a significant gain in grain yield due to this control strategy. Implications of these findings for a large extension of the MHM biocontrol program are discussed.     

Involvement of the β-cinnamomin elicitin in infection and colonisation of cork oak roots by <Emphasis Type="Italic">Phytophthora cinnamomi</Emphasis>

The virulence of two wild type (PA45 and PA37) and two genetically modified (13C: hygromycin resistant; FATSS: hygromycin resistant and β-cin knock-down) Phytophthora cinnamomi strains towards cork oak (Quercus suber) was assessed via a quantitative evaluation of disease symptoms arising from a soil infestation assay, and by a histological analysis of root colonization. Comparison of virulence, as expressed by symptom severity, resulted in the following ranking: highly virulent (wild type strains), medium virulence (strain 13C) and weakly virulent (FATSS). Both transgenic strains were compromised in their virulence, as expressed by symptom severity, but strain 13C was much less affected than FATSS. Microscopic observation showed that the FATSS strain was unable to effectively invade the root, while 13C and the two wild type strains were all able to rapidly colonize the whole root, including the vascular tissue. These results strengthen the notion that elicitins are associated, either directly or indirectly, with the infection process of Phytophthora.