DNA-GENOTYPING OF SPRING BARLEY TO LOOSE SMUT RESISTANCE BY PCR METHOD

2015 ◽  
Vol 10 (3) ◽  
pp. 98-101
Author(s):  
Блохин ◽  
Vasiliy Blokhin ◽  
Вильданова ◽  
Gulusa Vildanova ◽  
Нижегородцева ◽  
...  
Keyword(s):  
Resistance Gene ◽  
Resistance Genes ◽  
Spring Barley ◽  
Loose Smut ◽  
Gene Encoding ◽  
Accounting Data ◽  
Environmentally Safe ◽  
Pcr Method ◽  
Dna Genotyping ◽  
Research Institute

The most economical and environmentally safe way to control the disease is to use resistant barley varieties of plants. The use of molecular markers, associated with disease resistance genes, predetermine great possibilities, being used as a tool for the selection of breeders. The study aim is to identify varietal specimen, carrying genes, which are encoding resistance to loose smut of barley. A method of DNA-genotyping by PCR markers was used in the study. During the process of the research 83 varietal specimen have been tested, which were selected in the nursery Tatar Research Institute of Agriculture. The following two SCAR-markers, resistant to loose smut: Un8-700R and Un8-700 were used to identify the U8 gene. The studies found, that 36.2% have the gene, encoding the susceptibility to loose smut, in the genome alleles. 5.3% of the total number of varietal specimen was characterized by the presence of the resistance gene Un8 in the genome allele. The other specimen was observed no resistance gene to loose smut. The comparison with accounting data of loose smut spread, provided by the Barley breeding Department, showed no infestation of barley lines with resistance gene of loose smut in natural infectious background. As a result of molecular and genetic evaluation, the five specimen of nursery of Tatar Research Institute of Agriculture were identified with resistance genes in the genome alleles, which had been recommended, as a valuable genotypes for selection according to the loose smut resistance of barley.

scholarly journals Screening of Antimicrobial Resistance Genes and Epidemiological Features in Hospital and Community-Associated Carbapenem Resistant Pseudomonas aeruginosa Infections

2020 ◽  
Author(s):  
ayse erturk ◽  
Ayşegül Çopur ÇİÇEK ◽  
Nebahat EJDER ◽  
Uğur KOSTAKOĞLU ◽  
İlknur Esen YILDIZ ◽  
...  
Keyword(s):  
Resistance Genes ◽  
Multidrug Resistant ◽  
Gene Encoding ◽  
Pfge Typing ◽  
Carbapenem Resistant ◽  
Antimicrobial Resistance Genes ◽  
Genes Encoding ◽  
Pcr Method ◽  
Vitek 2 ◽  
Hospital Acquired

Abstract Background: Researching carbapenem-resistant isolates and the use of antibiotics and following infection control policies enable the identification of carbapenemase-producing bacteria and prevent their spread.Methods: P. aeruginosa isolates were recovered from Medicine Faculty of Recep Tayyip Erdoğan University between April 2015 and October 2016 and identified by conventional methods and the automated Vitek 2 Compact (BioMerieux, France) system. Antimicrobial susceptibility experiments were performed in accordance with CLSI criteria and the automated Vitek 2 Compact system. The PCR method was investigated for the presence of β-lactamase resistance genes. PFGE typing was performed to show clonal relation among samples.Results: Seventy P. aeruginosa strains were isolated from seventy patients. The median age of 70 cases was found 66 with minimum 17 and maximum 92 years old. 67.1% of the patients had contact with the health service in the last 90 days and 75.7% of the patients had received antimicrobial therapy in the previous 90 days. The most common comorbidity was cardiovascular diseases. Twenty-four (34.3%) strains were carbapenem resistant, 2 strains were multidrug-resistant except colistin, and none of the samples had colistin resistance. The gene encoding β-lactamase or metallo-β-lactamase was found in a total of 36 strains. The bla VEB gene was identified in only 1 strain alone, but in combination with other resistance genes in a total of 17 strains. While the bla PER gene was detected in 5 samples alone, it was found in 13 samples in combination with other genes. Among the genes encoding metallo-β-lactamase, the most bla NDM positive was detected (n=22), followed by 14 positive samples of bla KPC . bla IMP and bla VIM were detected in 5 and 1 samples, respectively. Also, the association of bla VEB - bla PER and bla VEB - bla KPC - bla NDM was found to be very high. Much more resistance genes and associations were detected in hospital-acquired samples than community-acquired samples, both proportionally and in terms of co-occurrence. Most of the community-associated strains were collected in the F2 clade, while most of the hospital-associated strains were collected in the G1 clade. However, no difference was found between the community and hospital-associated strains according to PFGE results. Simultaneously, other microorganisms were also isolated from patients from which these 6 P. aeruginosa strains were isolated. Of these patients, 5 patients died, except the number 70.Conclusions: The median length of stay (days) was found to be significantly higher in the group with HAI than in the group with CAI. Compared to sample 28 and 37, which carried 5 β-lactamase coding genes, the death of these 5 patients with fewer or no resistance genes showed that the coexistence of other factors - especially other microorganisms in addition to resistance genes, was important.

scholarly journals A Novel Tryptophanyl-tRNA Synthetase Gene Confers High-Level Resistance to Indolmycin

2009 ◽  
Vol 53 (9) ◽  
pp. 3972-3980 ◽  
Author(s):  
James J. Vecchione ◽  
Jason K. Sello
Keyword(s):  
Resistance Gene ◽  
Resistance Genes ◽  
Point Mutations ◽  
Trna Synthetase ◽  
Sensitive Strain ◽  
Antibacterial Drug ◽  
Gene Encoding ◽  
Trna Synthetases ◽  
High Level ◽  
Level Resistance

ABSTRACT Indolmycin, a potential antibacterial drug, competitively inhibits bacterial tryptophanyl-tRNA synthetases. An effort to identify indolmycin resistance genes led to the discovery of a gene encoding an indolmycin-resistant isoform of tryptophanyl-tRNA synthetase. Overexpression of this gene in an indolmycin-sensitive strain increased the indolmycin MIC 60-fold. Its transcription and distribution in various bacterial genera were assessed. The level of resistance conferred by this gene was compared to that of a known indolmycin resistance gene and to those of genes with resistance-conferring point mutations.

scholarly journals Genetic Characterization of Barley Net Blotch Resistance Genes

Plant Disease ◽  
2011 ◽  
Vol 95 (1) ◽  
pp. 19-23 ◽  
Author(s):  
Patrick D. O'Boyle ◽  
Wynse S. Brooks ◽  
Brian J. Steffenson ◽  
Erik L. Stromberg ◽  
Carl A. Griffey
Keyword(s):  
Resistance Gene ◽  
Resistance Genes ◽  
Wide Spectrum ◽  
Spring Barley ◽  
Dominant Gene ◽  
Test Results ◽  
Net Blotch ◽  
Barley Net Blotch ◽  
Dominant Genes

Net blotch, caused by Pyrenophora teres f. teres, is one of the most devastating diseases of barley (Hordeum vulgare). Efficient utilization of available resistance sources is dependent upon successful characterization of genes conditioning resistance in diverse sources. Five net-blotch-resistant parents and one susceptible parent were intercrossed to identify novel resistance genes and postulate gene number and mode of inheritance. Seedling response to isolate ND89-19 was evaluated in a greenhouse test. Results indicate that the resistant spring barley lines CIho 2291 and CIho 5098 and the winter barley cv. Nomini each have single dominant genes for resistance. Resistance in CIho 5098 is governed by the same dominant gene conferring resistance in Nomini. Resistance in CIho 2291 is controlled by one dominant gene which, putatively, is the same gene conferring resistance in ND B112 but differs from the resistance genes carried by the other parents in this study. The resistance gene in Nomini or CIho 5098 could be pyramided with the resistance gene in CIho 2291 or ND B112 to enhance the durability of resistance against a wide spectrum of P. teres isolates.

scholarly journals Metagenomics Analysis Reveals the Microbial Communities, Antimicrobial Resistance Gene Diversity and Potential Pathogen Transmission Risk of Two Different Landfills in China

Diversity ◽  
2021 ◽  
Vol 13 (6) ◽  
pp. 230
Author(s):  
Shan Wan ◽  
Min Xia ◽  
Jie Tao ◽  
Yanjun Pang ◽  
Fugen Yu ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Microbial Communities ◽  
Resistance Gene ◽  
Resistance Genes ◽  
Gene Diversity ◽  
Antibiotic Resistance Gene ◽  
Antibiotic Resistance Genes ◽  
Pathogen Transmission ◽  
Transmission Risk ◽  
Antimicrobial Resistance Gene

In this study, we used a metagenomic approach to analyze microbial communities, antibiotic resistance gene diversity, and human pathogenic bacterium composition in two typical landfills in China. Results showed that the phyla Proteobacteria, Bacteroidetes, and Actinobacteria were predominant in the two landfills, and archaea and fungi were also detected. The genera Methanoculleus, Lysobacter, and Pseudomonas were predominantly present in all samples. sul2, sul1, tetX, and adeF were the four most abundant antibiotic resistance genes. Sixty-nine bacterial pathogens were identified from the two landfills, with Klebsiella pneumoniae, Bordetella pertussis, Pseudomonas aeruginosa, and Bacillus cereus as the major pathogenic microorganisms, indicating the existence of potential environmental risk in landfills. In addition, KEGG pathway analysis indicated the presence of antibiotic resistance genes typically associated with human antibiotic resistance bacterial strains. These results provide insights into the risk of pathogens in landfills, which is important for controlling the potential secondary transmission of pathogens and reducing workers’ health risk during landfill excavation.

Organization, Expression and Evolution of a Disease Resistance Gene Cluster in Soybean

Genetics ◽  
2002 ◽  
Vol 162 (4) ◽  
pp. 1961-1977
Author(s):  
Michelle A Graham ◽  
Laura Fredrick Marek ◽  
Randy C Shoemaker
Keyword(s):  
Disease Resistance ◽  
Resistance Gene ◽  
Resistance Genes ◽  
Developmental Stages ◽  
Gene Evolution ◽  
Pcr Amplification ◽  
Disease Resistance Genes ◽  
Disease Resistance Gene ◽  
R Genes ◽  
Specific Primers

Abstract PCR amplification was previously used to identify a cluster of resistance gene analogues (RGAs) on soybean linkage group J. Resistance to powdery mildew (Rmd-c), Phytophthora stem and root rot (Rps2), and an ineffective nodulation gene (Rj2) map within this cluster. BAC fingerprinting and RGA-specific primers were used to develop a contig of BAC clones spanning this region in cultivar “Williams 82” [rps2, Rmd (adult onset), rj2]. Two cDNAs with homology to the TIR/NBD/LRR family of R-genes have also been mapped to opposite ends of a BAC in the contig Gm_Isb001_091F11 (BAC 91F11). Sequence analyses of BAC 91F11 identified 16 different resistance-like gene (RLG) sequences with homology to the TIR/NBD/LRR family of disease resistance genes. Four of these RLGs represent two potentially novel classes of disease resistance genes: TIR/NBD domains fused inframe to a putative defense-related protein (NtPRp27-like) and TIR domains fused inframe to soybean calmodulin Ca2+-binding domains. RT-PCR analyses using gene-specific primers allowed us to monitor the expression of individual genes in different tissues and developmental stages. Three genes appeared to be constitutively expressed, while three were differentially expressed. Analyses of the R-genes within this BAC suggest that R-gene evolution in soybean is a complex and dynamic process.

scholarly journals Genotype Heterogeneity in Accessions of a Winter Barley Core Collection Assessed on Postulated Specific Powdery Mildew Resistance Genes

Agronomy ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 513
Author(s):  
Antonín Dreiseitl
Keyword(s):  
Resistance Genes ◽  
Core Collection ◽  
Powdery Mildew Resistance ◽  
Gene Bank ◽  
Winter Barley ◽  
The Core ◽  
Genotype Identification ◽  
Environmentally Safe ◽  
Mla Locus ◽  
Resistance Tests

Gene bank accessions are necessary for implementing many research and breeding projects. However, a great number of accessions are contaminated or confused. If such accessions are used, the results obtained from these projects are inaccurate and non-reproducible. There are methods that allow almost perfect genotype identification; nevertheless, they are relatively recent and results cannot be compared with the characteristics of the original accessions. Growing resistant cultivars is an environmentally safe and cheap way of disease management and knowledge of diverse resistance genes and their combinations can be used to identify varieties and verify their authenticity and homogeneity. For this purpose, all 172 accessions of the core collection (CC) of the Czech winter barley (Hordeum vulgare) gene bank, originating from 35 countries, were studied. For resistance tests, 51 reference isolates of Blumeria graminis f. sp. Hordei, collected in all nonpolar continents over a period of 63 years and representing the global virulence/avirulence diversity of the pathogen, were used. Only 25 barley accessions were homogeneous (genetically uniform), whereas 147 accessions were heterogeneous due to presence of different genotypes. In total, 17 resistance genes were found singly or in combinations; 76.3% of accessions with identified resistance genes carried alleles at the Mla locus. To purify the CC, progenies of individual plants must be multiplied and authenticity and homogeneity of the seed should be confirmed with resistance tests, and subsequently can be studied with more advanced methods.

scholarly journals Effect of Bentonite and Barley Straw on the Restoration of the Biological Quality of Agriculture Soil Contaminated with the Herbicide Successor T 550 SE

Agriculture ◽  
2021 ◽  
Vol 11 (1) ◽  
pp. 27
Author(s):  
Jadwiga Wyszkowska ◽  
Monika Tomkiel ◽  
Agata Borowik ◽  
Małgorzata Baćmaga ◽  
Jan Kucharski
Keyword(s):  
Agricultural Soil ◽  
Plant Protection ◽  
Spring Barley ◽  
Biochemical Properties ◽  
Barley Straw ◽  
Negative Effects ◽  
Soil Quality Index ◽  
Oligotrophic Bacteria ◽  
Organotrophic Bacteria ◽  
Environmentally Safe

Environmentally safe ways are sought to prevent the accumulation and to accelerate the degradation of herbicide active substances in agricultural soil. This study aimed to determine the effectiveness of finely-ground barley straw and bentonite in mitigating the effects of agricultural soil contamination with Successor T 550 SE. This herbicide was applied in the following doses: 0, 0.73, and 14.63 mg of the active substance per kg. The bentonite and spring barley straw were used at 10 g/kg. The action of these additives was compared to soil without the addition of straw and bentonite. The application of the experimental herbicide disturbed microbial systems, such as organotrophic bacteria, oligotrophic bacteria and their spores, actinobacteria, and fungi. A positive response to the herbicide dose of 14.63 mg a.s./kg was observed only for spores of oligotrophic bacteria. Further disturbances were observed in the agricultural soil biochemical properties, i.e., in the activity of dehydrogenases, urease, catalase, acid, and alkaline phosphatase, arylsulfatase, and β-glucosidase. A significant decrease in the activity of dehydrogenases, acid phosphatase, and arylsulfatase was observed following the application of 14.63 mg a.s./kg. The yield of maize decreased following the application of the analysed plant protection agent. Based on the soil quality index (BA), the addition of straw was more effective in restoring soil homeostasis than bentonite. Both bentonite and straw can be successfully used to improve agricultural soil biological activity. However, more effective mitigation of the negative effects of the herbicide in soil was observed in objects supplemented with barley straw. This improved the microbiological and biochemical properties of the soil. Barley straw was more effective than bentonite in restoring soil biological balance.

scholarly journals Characterization of a Unique Chromosomal Copper Resistance Gene Cluster from Xanthomonas campestris pv. vesicatoria

2005 ◽  
Vol 71 (12) ◽  
pp. 8284-8291 ◽  
Author(s):  
Huseyin Basim ◽  
Gerald V. Minsavage ◽  
Robert E. Stall ◽  
Jaw-Fen Wang ◽  
Savita Shanker ◽  
...  
Keyword(s):  
Pseudomonas Syringae ◽  
Resistance Genes ◽  
Xanthomonas Campestris ◽  
Sinorhizobium Meliloti ◽  
The United States ◽  
Copper Resistance ◽  
Pcr Analysis ◽  
Pepper Plant ◽  
Gene Encoding ◽  
Copper Resistance Genes

ABSTRACT We characterized the copper resistance genes in strain XvP26 of Xanthomonas campestris pv. vesicatoria, which was originally isolated from a pepper plant in Taiwan. The copper resistance genes were localized to a 7,652-bp region which, based on pulsed-field gel electrophoresis and Southern hybridization, was determined to be located on the chromosome. These genes hybridized only weakly, as determined by Southern analysis, to other copper resistance genes in Xanthomonas and Pseudomonas strains. We identified five open reading frames (ORFs) whose products exhibited high levels of amino acid sequence identity to the products of previously reported copper genes. Mutations in ORF1, ORF3, and ORF4 removed copper resistance, whereas mutations in ORF5 resulted in an intermediate copper resistance phenotype and insertions in ORF2 had no effect on resistance conferred to a copper-sensitive recipient in transconjugant tests. Based on sequence analysis, ORF1 was determined to have high levels of identity with the CopR (66%) and PcoR (63%) genes in Pseudomonas syringae pv. tomato and Escherichia coli, respectively. ORF2 and ORF5 had high levels of identity with the PcoS gene in E. coli and the gene encoding a putative copper-containing oxidoreductase signal peptide protein in Sinorhizobium meliloti, respectively. ORF3 and ORF4 exhibited 23% identity to the gene encoding a cation efflux system membrane protein, CzcC, and 62% identity to the gene encoding a putative copper-containing oxidoreductase protein, respectively. The latter two ORFs were determined to be induced following exposure to low concentrations of copper, while addition of Co, Cd, or Zn resulted in no significant induction. PCR analysis of 51 pepper and 34 tomato copper-resistant X. campestris pv. vesicatoria strains collected from several regions in Taiwan between 1987 and 2000 and nine copper-resistant strains from the United States and South America showed that successful amplification of DNA was obtained only for strain XvP26. The organization of this set of copper resistance genes appears to be uncommon, and the set appears to occur rarely in X. campestris pv. vesicatoria.

scholarly journals Regulation of the Bacillus subtilis bcrC Bacitracin Resistance Gene by Two Extracytoplasmic Function σ Factors

2002 ◽  
Vol 184 (22) ◽  
pp. 6123-6129 ◽  
Author(s):  
Min Cao ◽  
John D. Helmann
Keyword(s):  
Bacillus Subtilis ◽  
Resistance Gene ◽  
Mutant Strain ◽  
Stress Responses ◽  
Double Mutant ◽  
De Novo ◽  
De Novo Synthesis ◽  
Gene Encoding ◽  
Single Promoter ◽  
Higher Sensitivity

ABSTRACT Bacitracin resistance is normally conferred by either of two major mechanisms, the BcrABC transporter, which pumps out bacitracin, or BacA, an undecaprenol kinase that provides C55-isoprenyl phosphate by de novo synthesis. We demonstrate that the Bacillus subtilis bcrC (ywoA) gene, encoding a putative bacitracin transport permease, is an important bacitracin resistance determinant. A bcrC mutant strain had an eightfold-higher sensitivity to bacitracin. Expression of bcrC initiated from a single promoter site that could be recognized by either of two extracytoplasmic function (ECF) σ factors, σX or σM. Bacitracin induced expression of bcrC, and this induction was dependent on σM but not on σX. Under inducing conditions, expression was primarily dependent on σM. As a consequence, a sigM mutant was fourfold more sensitive to bacitracin, while the sigX mutant was only slightly sensitive. A sigX sigM double mutant was similar to a bcrC mutant in sensitivity. These results support the suggestion that one function of B. subtilis ECF σ factors is to coordinate antibiotic stress responses.

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