scholarly journals Molecular Characterization and Identification of Biocontrol Isolates of Trichoderma spp

2000 ◽  
Vol 66 (5) ◽  
pp. 1890-1898 ◽  
Author(s):  
M. R. Hermosa ◽  
I. Grondona ◽  
E. A. Iturriaga ◽  
J. M. Diaz-Minguez ◽  
C. Castro ◽  
...  

ABSTRACT The most common biological control agents (BCAs) of the genusTrichoderma have been reported to be strains ofTrichoderma virens, T. harzianum, and T. viride. Since Trichoderma BCAs use different mechanisms of biocontrol, it is very important to explore the synergistic effects expressed by different genotypes for their practical use in agriculture. Characterization of 16 biocontrol strains, previously identified as “Trichoderma harzianum” Rifai and one biocontrol strain recognized asT. viride, was carried out using several molecular techniques. A certain degree of polymorphism was detected in hybridizations using a probe of mitochondrial DNA. Sequencing of internal transcribed spacers 1 and 2 (ITS1 and ITS2) revealed three different ITS lengths and four different sequence types. Phylogenetic analysis based on ITS1 sequences, including type strains of different species, clustered the 17 biocontrol strains into four groups: T. harzianum-T. inhamatum complex, T. longibrachiatum, T. asperellum, and T. atroviride-T. koningii complex. ITS2 sequences were also useful for locating the biocontrol strains inT. atroviride within the complex T. atroviride-T. koningii. None of the biocontrol strains studied corresponded to biotypes Th2 or Th4 of T. harzianum, which cause mushroom green mold. Correlation between different genotypes and potential biocontrol activity was studied under dual culturing of 17 BCAs in the presence of the phytopathogenic fungi Phoma betae,Rosellinia necatrix, Botrytis cinerea, andFusarium oxysporum f. sp. dianthi in three different media.

2003 ◽  
Vol 81 (3) ◽  
pp. 255-266 ◽  
Author(s):  
Tesfaye Wubet ◽  
Michael Weiß ◽  
Ingrid Kottke ◽  
Franz Oberwinkler

Taxus baccata L. roots collected from two sites in southern Germany were heavily colonized by arbuscular mycorrhizal fungi (AMF). The colonization pattern was of the Paris type. The diversity of the colonizing AMF species was investigated using polymerase chain reaction based molecular techniques. The internal transcribed spacer (ITS) region of the DNA from AMF within the roots was amplified using Glomeromycota-specific primers and then cloned and sequenced. Phylogenetic analysis using a data set of 5.8S rDNA sequences from a wide range of glomeralean taxa as well as data sets of partial ITS2 sequences from glomeralean subgroups indicated root colonization by four sequence types of Glomus and one sequence type of Archaeospora. These sequence types are distinct from any previously published sequences and differed between the two study sites.Key words: arbuscular mycorrhiza, Paris-type AM, molecular diversity, ribosomal internal transcribed spacers, Taxus baccata.


Parasitology ◽  
1999 ◽  
Vol 118 (6) ◽  
pp. 541-551 ◽  
Author(s):  
N. E. COLLINS ◽  
B. A. ALLSOPP

We sequenced the rRNA genes and internal transcribed spacers (ITS) of several Theileria parva isolates in an attempt to distinguish between the causative agents of East coast fever and Corridor disease. The small subunit (SSU) and large subunit (LSU) rRNA genes from a cloned T. p. lawrencei parasite were sequenced; the former was identical to that of T. p. parva Muguga, and there were minor heterogeneities in the latter. The 5·8S gene sequences of 11 T. parva isolates were identical, but major differences were found in the ITS. Six characterization oligonucleotides were designed to hybridize within the variable ITS1 region; 93·5% of T. p. parva isolates examined were detected by probe TPP1 and 81·8% of T. p. lawrencei isolates were detected by TPL2 and/or TPL3a. There was no absolute distinction between T. p. parva and T. p. lawrencei and the former hybridized with fewer of the probes than did the latter. It therefore seems that a relatively homogenous subpopulation of T. parva has been selected in cattle from a more diverse gene pool in buffalo. The ITSs of both T. p. parva and T. p. lawrencei contained different combinations of identifiable sequence segments, resulting in a mosaic of segments in any one isolate, suggesting that the two populations undergo genetic recombination and that their gene pools are not completely separate.


Antibiotics ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 709
Author(s):  
Marta Jorba ◽  
Marina Pedrola ◽  
Ouldouz Ghashghaei ◽  
Rocío Herráez ◽  
Lluis Campos-Vicens ◽  
...  

This work reports a detailed characterization of the antimicrobial profile of two trimethoprim-like molecules (compounds 1a and 1b) identified in previous studies. Both molecules displayed remarkable antimicrobial activity, particularly when combined with sulfamethoxazole. In disk diffusion assays on Petri dishes, compounds 1a and 1b showed synergistic effects with colistin. Specifically, in combinations with low concentrations of colistin, very large increases in the activities of compounds 1a and 1b were determined, as demonstrated by alterations in the kinetics of bacterial growth despite only slight changes in the fractional inhibitory concentration index. The effect of colistin may be to increase the rate of antibiotic entry while reducing efflux pump activity. Compounds 1a and 1b were susceptible to extrusion by efflux pumps, whereas the inhibitor phenylalanine arginyl β-naphthylamide (PAβN) exerted effects similar to those of colistin. The interactions between the target enzyme (dihydrofolate reductase), the coenzyme nicotinamide adenine dinucleotide phosphate (NADPH), and the studied molecules were explored using enzymology tools and computational chemistry. A model based on docking results is reported.


Foods ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 491
Author(s):  
Alejandra Ramirez-Hernandez ◽  
Ana K. Carrascal-Camacho ◽  
Andrea Varón-García ◽  
Mindy M. Brashears ◽  
Marcos X. Sanchez-Plata

The poultry industry in Colombia has implemented several changes and measures in chicken processing to improve sanitary operations and control pathogens’ prevalence. However, there is no official in-plant microbial profile reference data currently available throughout the processing value chains. Hence, this research aimed to study the microbial profiles and the antimicrobial resistance of Salmonella isolates in three plants. In total, 300 samples were collected in seven processing sites. Prevalence of Salmonella spp. and levels of Enterobacteriaceae were assessed. Additionally, whole-genome sequencing was conducted to characterize the isolated strains genotypically. Overall, the prevalence of Salmonella spp. in each establishment was 77%, 58% and 80% for plant A, B, and C. The mean levels of Enterobacteriaceae in the chicken rinsates were 5.03, 5.74, and 6.41 log CFU/mL for plant A, B, and C. Significant reductions were identified in the counts of post-chilling rinsate samples; however, increased levels were found in chicken parts. There were six distinct Salmonella spp. clusters with the predominant sequence types ST32 and ST28. The serotypes Infantis (54%) and Paratyphi B (25%) were the most commonly identified within the processing plants with a high abundance of antimicrobial resistance genes.


Author(s):  
Fatma Ben Abid ◽  
Clement K. M. Tsui ◽  
Yohei Doi ◽  
Anand Deshmukh ◽  
Christi L. McElheny ◽  
...  

AbstractOne hundred forty-nine carbapenem-resistant Enterobacterales from clinical samples obtained between April 2014 and November 2017 were subjected to whole genome sequencing and multi-locus sequence typing. Klebsiella pneumoniae (81, 54.4%) and Escherichia coli (38, 25.5%) were the most common species. Genes encoding metallo-β-lactamases were detected in 68 (45.8%) isolates, and OXA-48-like enzymes in 60 (40.3%). blaNDM-1 (45; 30.2%) and blaOXA-48 (29; 19.5%) were the most frequent. KPC-encoding genes were identified in 5 (3.6%) isolates. Most common sequence types were E. coli ST410 (8; 21.1%) and ST38 (7; 18.4%), and K. pneumoniae ST147 (13; 16%) and ST231 (7; 8.6%).


Pathogens ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 405
Author(s):  
Anna Matysiak ◽  
Michal Kabza ◽  
Justyna A. Karolak ◽  
Marcelina M. Jaworska ◽  
Malgorzata Rydzanicz ◽  
...  

The ocular microbiome composition has only been partially characterized. Here, we used RNA-sequencing (RNA-Seq) data to assess microbial diversity in human corneal tissue. Additionally, conjunctival swab samples were examined to characterize ocular surface microbiota. Short RNA-Seq reads, obtained from a previous transcriptome study of 50 corneal tissues, were mapped to the human reference genome GRCh38 to remove sequences of human origin. The unmapped reads were then used for taxonomic classification by comparing them with known bacterial, archaeal, and viral sequences from public databases. The components of microbial communities were identified and characterized using both conventional microbiology and polymerase chain reaction (PCR) techniques in 36 conjunctival swabs. The majority of ocular samples examined by conventional and molecular techniques showed very similar microbial taxonomic profiles, with most of the microorganisms being classified into Proteobacteria, Firmicutes, and Actinobacteria phyla. Only 50% of conjunctival samples exhibited bacterial growth. The PCR detection provided a broader overview of positive results for conjunctival materials. The RNA-Seq assessment revealed significant variability of the corneal microbial communities, including fastidious bacteria and viruses. The use of the combined techniques allowed for a comprehensive characterization of the eye microbiome’s elements, especially in aspects of microbiota diversity.


2000 ◽  
Vol 6 (3) ◽  
pp. 197-205 ◽  
Author(s):  
T. Jimenez ◽  
M.A. Martinez-Anaya

Water soluble pentosans (WSP) from doughs and breads made with different enzyme preparations are characterized according to extraction yield, sugar composition, xylose/arabinose ratio and molecular weight (MW) distribution. Extraction yield was greater for dough than for bread samples, ranging from 0.94 to 1.64%, but bread extracts had a higher purity. Percent of pentoses in purified WSP was greater in pentosanase supplemented samples (28-55%) than in control and amylase containing samples (23-32%). Major sugars were xylose and arabinose, but glucose and mannose also appeared in the extracts. The xylose/arabinose (Xyl/Ara) ratio was 1.3-1.6 and underwent small changes during processing. Enzyme addition caused an increase in Xyl/Ara ratio, attributable to a debranching of arabinoxylans (AX) with higher degree of Ara substitution by arabinofuranosidase. Addition of pentosanases had a significant effect in increasing WSP with MW over 39 000, whereas those of low MW changed only slightly. MW distribution depended on enzyme source, and whereas some enzymes showed activity during fermentation others increased their activity during baking. No synergistic effects were observed in studied variables due to the combination of amylases with pentosanases. Protein in WSP extracts eluted together with ferulic acid suggesting they were linked, but not associated with a determined carbohydrate fraction.


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