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Duduk B.,Serbian Institute of Pesticides and Environmental Protection
Methods in molecular biology (Clifton, N.J.) | Year: 2013

Current phytoplasma detection and identification methods are primarily based on nested polymerase chain reaction followed by restriction fragment length polymorphism analysis and gel electrophoresis. These methods can potentially detect and differentiate all phytoplasmas including those previously not described. The present protocol describes the application of this method for identification of phytoplasmas at 16S rRNA (16Sr) group and 16Sr subgroup levels. Source


Marcic D.,Serbian Institute of Pesticides and Environmental Protection
Journal of Pest Science | Year: 2012

This review focuses on biological profiles of contemporary acaricides, acaricide resistance, and other up-to-date issues related to acaricide use in management of plant-feeding mites. Over the last two decades a considerable number of synthetic acaricides emerged on the global market, most of which exert their effects acting on respiration targets. Among them, the most important are inhibitors of mitochondrial electron transport at complex I (METI-acaricides). Discovery of tetronic acid derivatives (spirodiclofen and spiromesifen) introduced a completely new mode of action: lipid synthesis inhibition. Acaricide resistance in spider mites has become a global phenomenon. The resistance is predominantly caused by a less sensitive target site (target site resistance) and enhanced detoxification (metabolic resistance). The major emphasis in current research on acaricide resistance mechanisms deals with elucidation of their molecular basis. Point mutations resulting in structural changes of target site and leading to its reduced sensitivity, have recently been associated with resistance in Tetranychus urticae Koch and other spider mites. The only sustainable, long-term perspective for acaricide use is their implementation in multitactic integrated pest management programs, in which acaricides are applied highly rationally and in interaction with other control tactics. Considering that the key recommendation for effective acaricide resistance management is reduction of the selection for resistance by alternations, sequences, rotations, and mixtures of compounds with different modes of action, the main challenge that acaricide use is facing is the need for new active substances with novel target sites. Besides implementation of advanced technologies for screening and design of new synthetic compounds, wider use of microbial and plant products with acaricidal activity could also contribute increased biochemical diversity of acaricides. © 2012 Springer-Verlag. Source


Siewert C.,Humboldt University of Berlin | Luge T.,Max Planck Institute for Molecular Genetics | Duduk B.,Serbian Institute of Pesticides and Environmental Protection | Seemuller E.,Julius Kuhn Institute | And 3 more authors.
PLoS ONE | Year: 2014

'Candidatus Phytoplasma mali' is a phytopathogenic bacterium of the family Acholeplasmataceae assigned to the class Mollicutes. This causative agent of the apple proliferation colonizes in Malus domestica the sieve tubes of the plant phloem resulting in a range of symptoms such as witches'- broom formation, reduced vigor and affecting size and quality of the crop. The disease is responsible for strong economical losses in Europe. Although the genome sequence of the pathogen is available, there is only limited information on expression of selected genes and metabolic key features that have not been examined on the transcriptomic or proteomic level so far. This situation is similar to many other phytoplasmas. In the work presented here, RNA-Seq and mass spectrometry shotgun techniques were applied on tissue samples from Nicotiana occidentalis infected by 'Ca. P. mali' strain AT providing insights into transcriptome and proteome of the pathogen. Data analysis highlights expression of 208 genes including 14 proteins located in the terminal inverted repeats of the linear chromosome. Beside a high portion of house keeping genes, the recently discussed chaperone GroES/GroEL is expressed. Furthermore, gene expression involved in formation of a type IVB and of the Sec-dependent secretion system was identified as well as the highly expressed putative pathogenicity-related SAP11-like effector protein. Metabolism of phytoplasmas depends on the uptake of spermidine/putescine, amino acids, co-factors, carbohydrates and in particular malate/citrate. The expression of these transporters was confirmed and the analysis of the carbohydrate cycle supports the suggested alternative energy-providing pathway for phytoplasmas releasing acetate and providing ATP. The phylogenetic analyses of malate dehydrogenase and acetate kinase in phytoplasmas show a closer relatedness to the Firmicutes in comparison to Mycoplasma species indicating an early divergence of the Acholeplasmataceae from the Mollicutes. © 2014 Siewert et al. Source


Marcic D.,Serbian Institute of Pesticides and Environmental Protection | Medo I.,Serbian Institute of Pesticides and Environmental Protection
Systematic and Applied Acarology | Year: 2015

The effects of the biopesticide azadirachtin-A (commercial product NeemAzal T/S; Trifolio-M GmbH, Germany) on behavior, survival, reproduction and population growth of the two-spotted spider mite (Tetranychus urticae Koch) were investigated in laboratory bioassays. The biopesticide was applied to bean leaf discs or primary bean leaves positioned on moistened cotton wads in Petri dishes by using a Potter spray tower. In a choice test, T. urticae females preferred the untreated halves of leaves over the halves treated with 50, 25, 12.5, 6.25, 3.12, and 1.56 mg/L azadirachtin-A and their oviposition in those treatments was significantly greater on untreated halves in the first 24 h, as well as its sum over 72 h (the only exception was oviposition after treatment with 6.25 mg/L). A run-off effect, ranging 4-32%, was also observed in the choice test. Viability and reproduction of survivors, as well as population growth, were affected after treatments of T. urticae females with 70, 35, and 17.5 mg/L of azadirachtin-A during their preovipositional period. All biopesticide treatments significantly reduced both gross fecundity and net fecundity (summed over seven days), the reduction ranging from 20-30% and 39-53%, respectively, compared with control. Female longevity was significantly reduced as well: treated females lived 1.4-2 days shorter than control females. All concentrations significantly lowered the instantaneous rate of increase, 11-18% against control. In females that hatched from eggs treated with 15, 7.5, and 3.75 mg/L azadirachtin-A and completed juvenile development exposed to its residues, gross fecundity was reduced 14-32%, but a significant reduction was found only after treatment with the highest concentration. On the other hand, all concentrations significantly reduced net fecundity (26-43%). Treated females lived 0.8-1.2 days shorter than control females and the reduction was significant after all treatments. The instantaneous rate of increase was significantly lowered by 15 and 7.5 mg/L concentrations, i.e. by 12% and 9%, respectively, compared with control. Sublethal effects of azadirachtin-A (NeemAzal-T/S) and its impact on the management of T. urticae populations are discussed. © Systematic & Applied Acarology Society. Source


Tomasevic A.,Serbian Institute of Pesticides and Environmental Protection | Mijin D.,University of Belgrade | Kiss E.,University of Novi Sad
Separation Science and Technology | Year: 2010

This article describes the photolytic and photocatalytic removal of the insecticide methomyl at low concentration from different types of water, upon UV, visible or natural solar light radiation, in the presence of TiO2 and ZnO, as well as using Fe-ZSM-5 zeolite and AlFe-pillared montmorillonite (photo-Fenton process). The rate of photodecomposition of methomyl was measured using UV spectrometry and HPLC, while its mineralization was investigated by ion chromatography (IC) and total organic carbon (TOC) analysis. The photochemical removal of methomyl is a natural and applicable model for the purification of water. © Taylor & Francis Group, LLC. Source

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