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Genomic selection is a technology that allows for the determination of the genetic value of varieties of agricultural plants and animal breeds, based on information about genotypes and phenotypes. The measured breeding value (BV) for varieties and breeds in relation to the target trait allows breeding stages to be thoroughly planned and the parent forms suitable for crossing to be chosen. In this work, the BLUP method was used to assess the breeding value of 149 Russian varieties and introgression lines (4 measurements for each variety or line, 596 phenotypic points) of spring wheat according to the content of seven chemical elements in the grain - K, Ca, Mg, Mn, Fe, Zn, Cu. The quality of the evaluation of breeding values was assessed using cross-validation, when the sample was randomly divided into five parts, one of which was chosen as a test population. The following average values of the Pearson correlation were obtained for predicting the concentration of trace elements: K - 0.67, Ca - 0.61, Mg - 0.4, Mn - 0.5, Fe - 0.38, Zn - 0.46, Cu - 0.48. Out of the 35 models studied, the p-value was below the nominal significant threshold (p-value < 0.05) for 28 models. For 11 models, the p-value was significant after correction for multiple testing (p-value < 0.001). For Ca and K, four out of five models and for Mn two out of five models had a p-value below the threshold adjusted for multiple testing. For 30 varieties that showed the best varietal values for Ca, K and Mn, the average breeding value was 296.43, 785.11 and 4.87 mg/kg higher, respectively, than the average breeding value of the population. The results obtained show the relevance of the application of genomic selection models even in such limited-size samples. The models for K, Ca and Mn are suitable for assessing the breeding value of Russian wheat varieties based on these characteristics.
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An increase in genetic diversity of bread wheat caused by spring x winter forms leads to an alteration of genetic control of maturity time. Maturity time (MAT) is one of major yield components in wheat, which has two components: the heading date (HD) and grain-filling period (GFP). Using the Illumina Infinium 25k platform we analyzed the genetic control of the HD, GFP and MAT in the F2 and F2:3 populations from a cross between late-ripening spring/winter line 124-1 and spring wheat cultivar Novosibirskaya 31, possessing the same allelic composition of the VRN1 and PPD-D1 genes. The phenotypic evaluation of the populations studied was performed during three years. A total of 17 QTLs were mapped, out of which 4 QTLs for MAT or its components were confirmed over two years. Two common MAT and HD QTLs were identified on the 4A chromosome, and two loci controlling GFP and MAT were found on 6B chromosome. An environmentally stable HD QTL QHd.icg-7B.1 was associated with the FT-B1 gene having a non-synonymous polymorphism [G/C] in its coding region. A novel ÐD QTL was identified on 7D chromosome. QTL dissection allowed to propose putative genes for QMat.icg4-A and QMat.icg6-B, namely the SPL family gene (TraesCS4A02G359500) and the TCP transcription factor (TraesCS6B02G462100), respectively. The results of this study provide information for further investigation into wheat development.
Assuntos
Fenótipo , Locos de Características Quantitativas , Triticum , Triticum/genética , Triticum/crescimento & desenvolvimento , Locos de Características Quantitativas/genética , Mapeamento Cromossômico , Grão Comestível/genética , Fatores de Tempo , Estações do AnoRESUMO
The F3h gene encodes the flavonoid synthesis enzyme flavanone 3-hydroxylase. Unlike most plant genomes, the bread wheat (Triticum aestivum L.) B genome has two, rather than just one F3h copy. The paralogous F3h-B2 sequence was isolated by PCR and shown to be transcribed, but its predicted polypeptide differed from the typical F3H sequence at a number of the conserved residues associated with its putative substrate-binding sites. The F3h-B2 promoter region was highly divergent from that of F3h-B1, and the transcriptional profiles of the two genes were distinct. Among a panel of 95 Triticeae accessions, representing 24 species, an F3h-2 copy was only detected within those carrying a B, S, G, or an R genome. Analysis of the coding sequence divergence suggested that a small segmental duplication occurred early in the evolution of the Triticeae tribe. The duplicated F3h copy appears to have acquired a novel function in the progenitor of the closely related B, G, and S genomes, as well as in the R genome. In other Triticeae genomes, the F3h-2 paralog may have been lost following pseudogenization.
Assuntos
Genes Duplicados , Genes de Plantas , Oxigenases de Função Mista/genética , Poaceae/genética , Sequência de Aminoácidos , Sítios de Ligação , Evolução Biológica , Sequência Conservada , Duplicação Gênica , Genoma de Planta , Oxigenases de Função Mista/metabolismo , Dados de Sequência Molecular , Triticum/genéticaRESUMO
The genetic diversity of common wheat hybrid lines Triticum aestivum/Triticum durum and Triticum aestivum/Triticum dicoccum (2n = 42, F(6-7)) using chromosome-specific microsatellite (SSR) markers and C-staining of chromosomes was studied. Cluster analysis of data obtained by 42 SSR markers indicated that the hybrid lines can be broken into three groups according to their origin. There were two cases of complete genetic similarity between lines 183(2)-2/184(1)-6 and-208-3/213-1, which were obtained using common wheat as the parental plants. In cross combinations, when the stabilization of the nuclear genome of hexaploid lines occurred against a background of the cytoplasmic genome of tetraploid wheats, there was a high level of divergence between sister lines, in some cases exceeding 50%. The evaluation of the degree of susceptibility of the lines to powdery mildew, leaf and stem rust, and septoria leafblotch was performed under different environmental conditions. It was shown that resistance to powdery mildew and leaf rust significantly depended on the region where assays were conducted. An evaluation of the field data showed that he lines 195-3, 196-1, and 221-1 with T. durum genetic material displayed complex resistance to fungal pathogens in Western Siberia and the Republic of Belarus. For lines 195-3 and 196-1, one shows a possible contribution of chromosomes 4B and 5B in the formation of complex resistance to diseases. Hybrid lines with complex resistance can be used to expand the genetic diversity of modern common wheat cultivars for genes of immunity.
Assuntos
Resistência à Doença/genética , Meio Ambiente , Genoma de Planta , Triticum/genética , Fungos/patogenicidade , Repetições de Microssatélites , Ploidias , Triticum/imunologia , Triticum/microbiologiaRESUMO
Wild and domesticated emmer (ÐÐÐÐ, 2n = 28) are of significant interest for expanding the genetic diversity of common wheat as sources of a high protein and microelement grain content, resistance to many biotic and abiotic factors. Particular interest in these species is also determined by their close relationship with Triticum aestivum L., which facilitates interspecific hybridization. The objective of this work was to analyze the nature of alien introgressions in hybrid lines from crossing common wheat varieties with T. dicoccoides and T. dicoccum, and to assess the effect of their genome fragments on the cytological stability of introgression lines. A C-banding technique and genotyping with SNP and SSR markers were used to determine localization and length of introgression fragments. Assessment of cytological stability was carried out on the basis of chromosome behavior in microsporogenesis. A molecular cytogenetic analysis of introgression wheat lines indicated that the inclusion of the genetic material of wild and domesticated emmer was carried out mainly in the form of whole arms or large fragments in the chromosomes of the B genome and less extended inserts in the A genome. At the same time, the highest frequency of introgressions of the emmer genome was observed in chromosomes 1A, 1B, 2B, and 3B. The analysis of the final stage of meiosis showed a high level of cytological stability in the vast majority of introgression wheat lines (meiotic index was 83.0-99.0 %), which ensures the formation of functional gametes in an amount sufficient for successful reproduction. These lines are of interest for the selection of promising material with agronomically valuable traits and their subsequent inclusion in the breeding process.
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The relationship between a variety's genotype, environmental conditions and phytopathogenic load are the key factors contributing to high yields that should be taken into account in selecting donors for resistance and high manifestation of valuable traits. The study of leaf rust resistance in 49 common wheat varieties was carried out in the field against the natural pathogen background and under laboratory conditions using single-pustule isolates with virulence to Lr9 and Lr24. It has been shown that the varieties carrying alien genes Lr6Agi2 (Tulaikovskaya 10) and Lr6Agi1 (Voevoda) were resistant to leaf rust infection both in the field and in the laboratory. Varieties KWS Buran, KWS Akvilon, KW 240-3-13, and Etyud producing crop yields from 417 to 514 g/m2 comparable to the best standard variety Sibirskaya 17 can be reasonably used as Lr24 resistance gene donors under West Siberian conditions. Omskaya 44 variety showing crop yield of 440g/m2 can be used as a donor for Lr19 and partially effective Lr26. Varieties Tuleevskaya and Altayskaya 110 with Lr9 in their genomes are recommended for the development of resistance gene-pyramided genotypes. The highest protein and gluten contents were observed in the CS2A/2M sample, while KWS Buran, Altayskaya 110, Volgouralskaya, and KWS Akvilon showed the lowest values. Varieties CS2A/2M, Tulaikovskaya 10, Pavon, and Tuleevskaya were ranked the highest in micro- (Cu, Mn, Zn, Fe) and macronutrient (Ca, Mg, K) contents among the common wheat samples from the collection, while the lowest values for most elements were observed in KWS Buran, Novosibirskaya 15, and Volgouralskaya. Winter varieties demonstrating leaf rust resistance against the infectious background typically carry adult plant resistance genes (Lr34, Lr12, and Lr13), particularly combined with the juvenile Lr26 gene. The presence of Lr41 in a winter type line (KS 93 U 62) allowed it to maintain resistance against a leaf rust pathogen clone kLr24, despite the presence of Lr24 in the genotype. Varieties Doka and Cheshskaya 17 may act as donors of resistance genes Lr26 + Lr34 and Lr9 + Lr12 + Lr13 + Lr34, as well as sources of dwarfing without losses in winter hardiness and yield under West Siberian conditions.
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The yield and grain quality of spring and winter wheat signif icantly depends on varieties' resistance to lodging, the genetic basis of this trait being quantitative and controlled by a large number of loci. Therefore, the study of the genetic architecture of the trait becomes necessary for the creation and improvement of modern wheat varieties. Here we present the results of localization of the genomic regions associated with resistance to lodging, plant height, and upper internode diameter in Russian bread wheat varieties. Phenotypic screening of 97 spring varieties and breeding lines was carried out in the f ield conditions of the West Siberian region during 2017-2019. It was found that 54 % of the varieties could be characterized as medium and highly resistant to lodging. At the same time, it was noted that the trait varied over the years. Twelve varieties showed a low level of resistance in all years of evaluation. Plant height-based grouping of the varieties showed that 19 samples belonged to semi-dwarfs (60-84 cm), and the rest were included in the group of standard-height plants (85-100 cm). Quantitative trait loci (QTL) mapping was performed by means of genome-wide association study (GWAS) using 9285 SNP markers. For lodging resistance, plant height, and upper internode diameter, 26 signif icant associations (-log p > 3) were found in chromosomes 1B, 2A, 3A, 3D, 4A, 5A, 5B, 5D, 6A, and 7B. The results obtained suggest that the regions of 700-711 and 597-618 Mb in chromosomes 3A and 6A, respectively, may contain clusters of genes that affect lodging resistance and plant height. No chromosome regions colocalized with the QTLs as sociated with lodging resistance or upper internode diameter were found. The present GWAS results may be important for the development of approaches for creating lodging-resistant varieties through marker-assisted and genomic selection.
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Lodging is one of the main factors in reducing the yield and grain quality of winter and spring wheat varieties. The resistance of wheat cultivars to lodging largely depends on environmental factors, biological and morphological features of the stem and root systems. Selection of the varieties for resistance to lodging is relevant in many countries of the world and has a number of achievements. Plant height is one of the most important morphological characters associated with lodging resistance. Breeding of the varieties carrying the dwarfing genes (Rht) is the main direction to reduce the risk of lodging. The Rht-B1b, Rht-D1b, Rht8 and Rht11 genes are widely used throughout the world due to their significant influence on agronomically valuable traits, including lodging. It turned out to be important to study the anatomical and morphological features and chemical composition of stem tissues, which complement the assessment of resistance to lodging and allow the varietal material to be more fully characterized. The thickness of stem internodes and their anatomical structure play an important role in the stem strength. The diameter of the stem, its thickness and weight, a large number of vascular bundles and a wide ring of mechanical tissues correlate with resistance to lodging. The content of lignin, silicon and cellulose are important structural components and provide the stem strength of wheat plants. Molecular genetic analysis and mapping of genes and quantitative trait loci are of great importance in identifying the genetic basis of the relationship between the anatomical and morphophysiological characters of the stem and root system and lodging. Genetic factors reflecting correlations between the lodging and the thickness of the stem wall, the number of vascular bundles and other characters were mapped to chromosomes 1A, 1B, 2A, 2D, 3A, 4B, 4D, 5A, 5D, 6D and 7D. It has been found that loci with high phenotypic effects on lodging tolerance are colocalized with loci responsible for plant height, stem diameter and stem strength. To increase resistance to lodging, it is necessary to develop a set of agrotechnical methods that reduce the influence of soil and climatic factors and create wheat varieties tolerant to lodging.
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A total of 40 introgressive lines of common wheat (2n = 42) Triticum aestivum L x T. timopheevii Zhuk., resistant to brown rust and partly to powdery mildew, were examined. Based on cytological analysis of meiosis in pollen mother cells (PMC), hybrid lines were subdivided into two groups characterized by either stable or unstable meiosis. In cytologically stable lines, chromosome configuration at the MI stage of meiosis was mostly bivalent (21II) with small proportion of defect cells (almost 10%), which at most contained two univalents (20II + + 21). Cytologically unstable group was comprised of the lines, containing high proportions of cells with abnormal chromosome pairing in meiotic PMC, as well as the cells with multivalents, and the lines containing aneuploid plants. Localization of the T. timopheevii fragments performed with the use of SSR markers showed that the lines with unstable meiosis were characterized by higher numbers of introgressions compared to stable lines. The influence of certain chromosomes of T. timopheevii on chromosome pairing stability was also demonstrated. In cytologically unstable lines, the increased frequency of 2A substitutions along with the high frequency of introgression of T. timopheevii genetic material into chromosome 7A was observed. Multivalents were scored in all cases of introgression in chromosome 7A. It was suggested that the reason for the genome instability in hybrid forms lied in insufficient compensating ability of certain T. timopheevii chromosomes and/or their parts, involved into recombination processes.
Assuntos
Quimera/genética , Pareamento Cromossômico/genética , Cromossomos de Plantas/genética , Instabilidade Genômica , Meiose/genética , Triticum/genética , Aneuploidia , Doenças das Plantas/genética , Doenças das Plantas/microbiologiaRESUMO
Introgressive lines resulting from crossing common wheat Triticum aestivum with the tetraploid T. timopheevii are characterized by effective resistance to leaf rust caused by Puccinia triticina Eriks. Molecular analysis using 350 specific short sequence repeat (SSR) markers was used to locate the T. timopheevii genome to chromosomes 1A, 2A, 2B, 5A, 5B, and 6B. A population of F2 offspring of crossing hybrid line 842-2 with common wheat cultivar Skala was obtained for mapping the loci controlling leaf rust resistance. Analysis of association of phenotypic and genotypic data by means of simple interval mapping (SIM) and composite interval mapping (CIM) has shown that the resistance of adult plants is determined by two loci in chromosomes 5B and 2A. The major locus QLr.icg-5B transferred from T. timopheevii chromosome 5G mapped to the interval of microsatellite loci Xgwm408-Xgwm1257 controls 72% of the phenotypic diversity of the trait. The other, minor locus QLr.icg-2A located to chromosome 2A at a distance of 10 cM from Xgwm312 accounts for 7% of the trait expression. Microsatellite markers located near these loci may be used for controlling the transfer of commercially valuable loci when new lines and cultivars are created.
Assuntos
Quimera/genética , Genoma de Planta/genética , Doenças das Plantas/genética , Poliploidia , Locos de Características Quantitativas/genética , Triticum/genética , Quimera/microbiologia , Mapeamento Cromossômico/métodos , Cromossomos de Plantas/genética , Doenças das Plantas/microbiologia , Triticum/microbiologiaRESUMO
Hexaploid triticale (x Triticosecale Wittmack) lines were examined using molecular markers and the hybridization in situ technique. Triticale lines were generated based on wheat varieties differing by the Vrn gene systems and the earing times. Molecular analysis was performed using Xgwm and Xrms microsatellite markers with the known chromosomal localization in the common wheat Triticum aestivum, and rye Secale cereale genomes. Comparative molecular analysis of triticale lines and their parental forms showed that all lines contained A and B genomes of common wheat and also rye homeologous chromosomes. In the three lines the presence of D genome markers, mapped to the chromosomes 2D and 7D, was demonstrated. This was probably the consequence of the translocations of homeologous chromosomes from wheat genomes, which took part during the process of triticale formation. The data obtained by use of genomic in situ hybridization supported the data of molecular genetic analysis. In none of the lines wheat--rye translocations or recombinations were observed. These findings suggest that the change of the period between the seedling appearance and earing time in triticale lines compared to the initial wheat lines, resulted from the inhibitory effect of rye genome on wheat vernalization genes.
Assuntos
Cromossomos de Plantas/genética , Genes de Plantas/genética , Poliploidia , Secale/genética , Triticum/genética , Produtos Agrícolas/genética , Recombinação Genética , Especificidade da EspécieRESUMO
The interaction of a spin labeled compound carrying an alkylating group 4-(3-iodo-2-oxopropylidene)-2,2,3,5,5-pentamethylimidozolydene-1-oxyl (RJ) and capable of binding covalently to mixed function oxidases (MFO) was studied. Measurements of the difference spectrum of cytochrome P-450 demonstrated that RJ induces spectral changes characteristic of type I substrates (lambda max = 403 nm; lambda min = 418 nm). The spectral binding constant (Ks) was 66 microM as determined from the difference spectrum. RJ inhibited the microsomal oxidation of substrates of cytochrome P-450 (aniline, aminopyrine and benzo [a]pyrene). This inhibition was shown not to be associated with the conversion of cytochrome P-450 to cytochrome P-420, or with the suppression of the activities of NADPh-cytochrome c reductase and NADPH-cytochrome P-450 reductase. Thus, evidence was obtained for the possible interaction of RJ with cytochrome P-450. RJ injected to rats (5 mg/100 g body wt, i.p.), inhibited the hydroxylation of benzo[a]pyrene, a type I substrate, (21%) and aniline, a type II substrate, (40%) in the microsomes from their livers. The presence of a paramagnetic center in RJ made it possible to study its interaction with microsomes. The electron paramagnetic resonance (EPR) spectrum of RJ was recorded in the rat liver microsomal fraction after in vivo administration of RJ. In rats treated with RJ (5 mg/100 g), hexobarbital sleeping time was prolonged 1.5-fold. Alkylating analogs of substrates of cytochrome P-450 are suggested as agents for structural studies of the active center of cytochrome P-450 and the development of efficient inhibitors of reactions catalyzed by this enzyme.
Assuntos
Sistema Enzimático do Citocromo P-450/metabolismo , Oxigenases de Função Mista/metabolismo , Oxirredutases/metabolismo , Aminopirina/metabolismo , Compostos de Anilina/metabolismo , Animais , Benzo(a)pireno , Benzopirenos/metabolismo , Técnicas In Vitro , Cinética , Masculino , Microssomos Hepáticos/enzimologia , NADPH-Ferri-Hemoproteína Redutase/metabolismo , Oxirredução , Ratos , Ratos EndogâmicosRESUMO
Homologous 1-naphtoxyalcanthiols of the type 1-C10H7O(CH2)nSH (n = 2-7) are used for structural studies of the microsomal cytochrome P450 active centre. It was found that the strongest complex of thiol with P450 is formed for n = 3. Microsomal oxidation of P450 substrates aminopyrine and benz(a) pyrene is inhibited by the 1-naphtoxyalcanthiols studied. A non-monotonous dependence of pI50 on n was found, the compound with a chain length n = 3 appeared to be the most effective inhibitor. The interaction of this thiol (n = 3) with both the heme group of P450 and the hydrophobic substrate zone is supposed and the distance between these points was estimated. It is possible to employ this approach for structural studies on the active centers of different isoforms of P450.
Assuntos
Sistema Enzimático do Citocromo P-450/análise , Membranas Intracelulares/enzimologia , Microssomos Hepáticos/enzimologia , Naftalenos , Compostos de Sulfidrila , Animais , Sítios de Ligação , Isoenzimas/análise , Cinética , Masculino , Oxirredução , Ratos , Ratos Endogâmicos , Especificidade por SubstratoRESUMO
Twenty-four Triticum eastivum x T. timopheevii hybrid lines developed on the basis of five varieties of common wheat and resistant to leaf rust were analyzed by the use of microsatellite markers specific for hexaploid common wheat T. aestivum. Investigation of intervarietal polymorphism of the markers showed that the number of alleles per locus ranged from 1 to 4, depending on the marker (2.5 on average). In T. timopheevii, amplification fragments are produced by 80, 55, and 30% of primers specific to the A, B, and D common wheat genomes, respectively. Microsatellite analysis revealed two major areas of introgression of the T. timopheevii genome: chromosomes of homoeological groups 2 and 5. Translocations were detected in the 2A and 2B chromosomes simultaneously in 11 lines of 24. The length of the translocated fragment in the 2B chromosome was virtually identical in all hybrid lines and did not depend on the parental wheat variety. In 15 lines developed on the basis of the Saratovskaya 29, Irtyshanka, and Tselinnaya 20, changes occurred in the telomeric region of the long arm of the 5A chromosome. Analysis with markers specific to the D genome suggested that introgressions of the T. timopheevii genome occurred in chromosomes of the D genome. However, the location of these markers on T. timopheevii chromosomes is unknown. Our data suggest that the genes for leaf-rust resistance transferred from T. timopheevii to T. aestivum are located chromosomes of homoeological group 2.
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Cruzamentos Genéticos , Poliploidia , Triticum/genética , Cromossomos , Fungos/patogenicidade , Predisposição Genética para Doença , Repetições de Microssatélites , Doenças das Plantas/genética , Especificidade da Espécie , Triticum/fisiologiaRESUMO
The search for STS (sequence-tagged site) and RAPD (random amplified polymorphic DNA) markers tightly linked to some genes of homeologous group 5 chromosomes of common wheat Triticum aestivum L., more specifically, awns inhibitor genes (B1), vernalization response gene (Vrn1), and homeologous chromosome pairing gene (Ph1), was conducted. To estimate the linkage of the gene with the marker, wheat lines marked with recessive alleles b1 and vrn1 were used. RELP (restriction fragment length polymorphism) and SSR (simple sequence repeat) analyses of isogenic wheat lines were conducted to characterize the chromosomal region transferred to the isogenic line from the donor parent. In RAPD analysis of isogenic wheat lines marked with recessive alleles b1 and vrn1, 95 arbitrary primers were used. To develop STS markers, analysis of the primary structure of RELP markers Xpsr426 and Xcdo504, tightly linked to the Vrn1 gene, and the Xpsr1201 marker, located at the Ph1 locus, was carried out. Two markers that are tightly linked to the Vrn1 gene (5AL)--RAPD marker Xr405 and STS marker Xsts426--were obtained in this work. In addition, there is every reason to believe that Xsts426 can be used as a PCR marker of genes Vrn2 (5BL) and Vrn3 (5DL), while Xsts1201, of the gene Ph1 (5BL).
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Cromossomos , Triticum/genética , Sequência de Bases , DNA de Plantas , Ligação Genética , Marcadores Genéticos , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , Sitios de Sequências RotuladasRESUMO
The cytochrome P-450 content, activity of microsomal monooxygenases, nonspecific esterases and glutathione S-transferases were studied at different stages of development of the Colorado beetle, cotton bollworm, cabbage butterfly, wax moth from the laboratory and natural populations. The data obtained demonstrate significant species, sexual and age differences in the activity of enzyme systems of insecticide detoxication. The toxic efficiency of insecticides at certain developmental stages depends on the level of activity of the enzyme systems involved in their metabolism. These data are discussed with respect to the problem of insects' sensitivity to insecticides at different developmental stages.
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Insetos/crescimento & desenvolvimento , Metamorfose Biológica , Microssomos/enzimologia , Preparações Farmacêuticas/metabolismo , Animais , Biotransformação , Insetos/metabolismo , Inseticidas/metabolismo , Inseticidas/farmacologiaRESUMO
The activity of three enzymatic systems of xenobiotic metabolism (cytochrome P-450-dependent monooxygenases, non-specific esterases and glutathione S-transferases) was studied in sensitive (S) and resistant to tetrametrin (Rtetr.), permetrin (Rperm.), mecarbenyl (Rmec.) and chlorophos (Rchlor.) strains of the housefly M. domestica L. In Rtetr. and Rmec., the activity of microsomal monooxygenases was increased 2.7- and 2.3-fold, respectively, as compared to S. The position of maxima of CO-difference spectra of cytochrome P-450 in all resistant strains (with the exception of Rchlor.) were shifted towards the short-wave region by 1-2 nm. The activity of glutathione S-transferase in Rtetr. was increased as compared to S. Analysis of the total esterase activity and electrophoresis in starch gel revealed quantitative and qualitative differences between the strains under study. In all resistant strains, except for Rmec., additional bands corresponding to the esterase activity were observed. The experimental results are discussed in terms of resistance of insects to insecticides.
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Moscas Domésticas/enzimologia , Inseticidas/farmacologia , Animais , Sistema Enzimático do Citocromo P-450/metabolismo , Resistência a Medicamentos , Eletroforese em Gel de Amido , Esterases/metabolismo , Glutationa Transferase/metabolismo , Inativação Metabólica , Inseticidas/metabolismo , Microssomos/enzimologiaRESUMO
The activity of three enzymatic systems of xenobiotic metabolism (cytochrome P-450-dependent monooxygenases, non-specific esterases and glutathione S-transferases) was studied at different stages of development of the Colorado potato beetle (Leptinotarsa decemlineata). Significant sex and ontogenetic differences in the content of cytochrome P-450, position of maxima of CO-difference spectra of the cytochrome P-450 reduced form and substrate specificity of cytochrome P-450 were revealed. The activity of non-specific esterases was shown to increase depending on the age of larvae. The insecticide 1-naphtholenol methylcarbamate which is metabolized by the system of cytochrome P-450-dependent monooxygenases is more toxic in the larvae than at the imago stage, which is correlated with the activity of this system at different stages of ontogenesis. The microsomal monooxygenase inhibitor, piperonylbutoxide, increases the insecticide toxicity in the Colorado beetle imago more than 2-fold. The experimental results testify to different contribution of the detoxication systems to the sensitivity of the Colorado beetle to insecticides at various metamorphosis stages.
Assuntos
Besouros/enzimologia , Inseticidas/farmacologia , Animais , Sistema Enzimático do Citocromo P-450/metabolismo , Resistência a Medicamentos , Esterases/metabolismo , Glutationa Transferase/metabolismo , Inativação Metabólica , Inseticidas/metabolismo , Larva/enzimologiaRESUMO
The activity of enzymes providing for permetrin detoxication in the imago resistant natural population of the Colorado potato beetle Leptinotarsa decemlineata was studied with the view of elucidating the biochemical mechanisms of resistance to the insecticide. It was demonstrated that the activity of the main enzymes of insect detoxication, i.e., microsomal monooxygenases, nonspecific esterases and glutathione-S-transferases in the permetrin-resistant population of L-decemlineata is enhanced as compared with the permetrin-sensitive population. It was demonstrated that the inhibitors of microsomal monooxygenases, piperonyl butoxide, and of nonspecific esterases, butifos, significantly increase the sensitivity of the resistant population to permetrin. The experimental results suggest that the activity of microsomal monooxygenases and nonspecific esterases is the main factor which determines the resistance of the Colorado potato beetle to permetrin.
Assuntos
Besouros/enzimologia , Resistência a Inseticidas , Inseticidas/farmacocinética , Piretrinas/farmacocinética , Animais , Inativação Metabólica , Inseticidas/farmacologia , Permetrina , Piretrinas/farmacologiaRESUMO
It was shown that the alkylating analog of cytochrome P-450 substrate - 4-bromomethyl-2,2,5,5-tetramethyl-3-imidazoline-3-oxide-1-oxyl effectively inhibits in vitro the activities of monooxygenases, glutathione-S-transferases and esterases from the abdomen of the house fly Musca domestica L. The non-alkylating analog (RH) appeared to be a very weak inhibitor of these enzymes. It was demonstrated that the inhibitory action of the alkylating analog consists in its covalent binding to the enzyme protein. Topical application of the cytochrome P-450 alkylating analog on insects led to a decrease of the enzyme activity by approximately 20%.