scholarly journals The Aspergillus niger Major Allergen (Asp n 3) DNA-Specific Sequence Is a Reliable Marker to Identify Early Fungal Contamination and Postharvest Damage in Mangifera indica Fruit

2021 ◽  
Vol 12 ◽  
Author(s):  
Jorge Martínez ◽  
Ander Nevado ◽  
Ester Suñén ◽  
Marta Gabriel ◽  
Ainara Vélez-del-Burgo ◽  
...  

The aim of this work was to study the value of the main allergen Asp n 3 of Aspergillus niger as a molecular marker of allergenicity and pathogenicity with the potential to be used in the identification of A. niger as a contaminant and cause of spoilage of Mangifera indica. Real-time polymerase chain reaction (RT-PCR) was used for the amplification of Asp n 3 gene. Two pairs of primers were designed: one for the amplification of the entire sequence and another one for the amplification of the most conserved region of this peroxisomal protein. The presence of A. niger was demonstrated by the early detection of the allergenic protein Asp n 3 coding gene, which could be considered a species-specific marker. The use of primers designed based on the conserved region of the Asp n 3 encoding gene allowed us to identify the presence of the closely related fungal species Aspergillus fumigatus by detecting Asp n 3 homologous protein, which can be cross-reactive. The use of conserved segments of the Asp n 3 gene or its entire sequence allows us to detect phylogenetically closely related species within the Aspergilaceae family or to identify species-specific contaminating fungi.

Genome ◽  
1998 ◽  
Vol 41 (2) ◽  
pp. 148-153 ◽  
Author(s):  
Monique Abadon ◽  
Eric Grenier ◽  
Christian Laumond ◽  
Pierre Abad

An AluI satellite DNA family has been cloned from the entomopathogenic nematode Heterorhabditis indicus. This repeated sequence appears to be an unusually abundant satellite DNA, since it constitutes about 45% of the H. indicus genome. The consensus sequence is 174 nucleotides long and has an A + T content of 56%, with the presence of direct and inverted repeat clusters. DNA sequence data reveal that monomers are quite homogeneous. Such homogeneity suggests that some mechanism is acting to maintain the homogeneity of this satellite DNA, despite its abundance, or that this repeated sequence could have appeared recently in the genome of H. indicus. Hybridization analysis of genomic DNAs from different Heterorhabditis species shows that this satellite DNA sequence is specific to the H. indicus genome. Considering the species specificity and the high copy number of this AluI satellite DNA sequence, it could provide a rapid and powerful tool for identifying H. indicus strains.Key words: AluI repeated DNA, tandem repeats, species-specific sequence, nucleotide sequence analysis.


2020 ◽  
Vol 26 (2) ◽  
Author(s):  
Ritu Kunjam ◽  
V.K. Kanungo ◽  
S.K. Jadhav

Increased urbanization and industrialization in recent time has made a significant impact on air quality of the area. The atmosphere is rich in propagule of different fungal species. The investigation on airborne Aspergillus contribution was conducted in Periphery of Raipur city from February, 2018 to March, 2019 with the help of gravity petriplate containing PDA (Potato Dextrose Agar) medium. In this study, total 11 species of Aspergillus were recorded. The percentage frequency and percentage contribution of different Aspergillus species were different in different seasons. Aspergillus niger was most frequent throughout the year followed by Aspergillus fumigatus, A. flavus, and A. nidulans etc. While Aspergillus clavatus, and A. versicolor, A. aculeatus were the least frequent species. The result indicated the highest percentage contribution of Aspergillus niger (43.29 percent) followed by A. fumigatus (9.02percent), A. flavus (8.42 percent) while A. clavatus (0.21 percent). The objective of the studies was to determine a seasonal variation in concentrations of Aspergillus on the basis of meteorological parameters.


2013 ◽  
Vol 65 (3) ◽  
pp. 955-962 ◽  
Author(s):  
Milica Ljaljevic-Grbic ◽  
M. Stupar ◽  
Jelena Vukojevic ◽  
Ivana Maricic ◽  
Natasa Bungur

Pieces of art stored in museum depots and display rooms are subject to fungal colonization that leads to bio-deterioration processes. Deteriorated wooden sculptures and art photographs temporarily stored in the quarantine room of the Cultural Center of Belgrade were subject to mycological analyses. Twelve fungal species were identified on the wooden substratum and five species were detected on photograph surfaces. Trichoderma viride, Chaetomium globosum and Alternaria sp. were the fungi with proven cellulolytic activity detected on the examined cellulose substrata. Indoor air mycobiota were estimated to 210.09 ? 8.06 CFU m-3, and the conidia of fungus Aspergillus niger were the dominant fungal propagules in the air of the examined room.


Author(s):  
Kassim Dekhil

 Objective: This study was aimed to identify the public pattern of presentation, influencing factors, and sort the fungal species, distribution of sex of patients with otomycosis.Results: The predominant complaints were pruritus and found in 76 patients (88.73%), discomfort and pain found in 62 patients (72.09%), aural fullness in 48 patients (55.81%), tinnitus in 34 patients (39.53%), hearing impairment in 50 cases (58.31%), ear discharge in 22 patients (25.58%), and most of the symptoms seen in 36 patients (68.14%). The results showed a total of eight fungal species belong to six different genera, namely, Aspergillus, Candida, Penicillium, Rhizopus, Alternaria, and Cephalosporium were isolated during this study. Among identified fungi, Aspergillus niger was found to be the most prevalent fungal species with 35.71% followed by Candida albicans (27.55%), Aspergillus flavus (10.20%), Aspergillus fumigatus (8.16), Penicillium digitatum (6.12%) and Cephalosporium species (4.08%), and Rhizopus species (5.1%), while Alternaria alternata had the lowest percentage (6.54%).Conclusion: Otomycosis/mycotic otitis externa is still a common problem and there is a rise in the occurrence of otomycosis in latest years, especially in tropical and subtropical humid climates.


2021 ◽  
Vol 4 ◽  
Author(s):  
Christina Wiebe ◽  
Petra Nowak ◽  
Hendrik Schubert

Assessing the biodiversity of an ecosystem plays a major role in ecosystem management. However, proper determination on species-level is often tricky when morphological features are scarce and especially rare species require huge sampling efforts to be detected in the aquatic realm. As an alternative to conventional methods, environmental samples can be examined via the eDNA method, allowing for large-scale integration as well as taxa resolution independent from expression of morphological characters. However, to apply this technique genetic markers that are specific to a species or at least a genus are required. Such markers until now have been successfully developed only for a few well studied taxonomic groups like, e.g., fishes and amphibians, but are still missing for others, especially plants and algae (e.g. Bista et al. 2017). This project focusses on the development of species-specific markers for the macrophytic green algae Tolypella canadensis (Characeae, Charophyta), a rare alga preferring deep water and known so far mainly from remote places. Tolypella canadensis is a circumpolar species and prefers oligotrophic lakes, where it grows in depths up to 13 m (Langangen 2002; Romanov and Kopyrina 2016). In addition, proper determination of Tolypella-species is a field of a few specialists, further complicating monitoring or even detection of this rare species. The design of the species-specific primers was based on reference nucleotide sequences of the chloroplast genes rbcL, psbC and atpB and of the ribosomal internal transcribed spacer regions ITS1 and ITS2, obtained from GenBank (Perez et al. 2017). To determine the specificity of the newly designed primers, DNA isolates obtained from T. canadensis specimens collected from the Torneträsk (Sweden, 2018) and other charophyte species were prepared in different proportions. The sensitivity of the primers was experimentally assayed by using serial dilutions of T. canadensis DNA. Additionally, a mock test comprised of a sample with the DNA of several charophyte species was conducted and finally, the markers were tested on environmental samples from the Torneträsk. Tolypella canadensis-specific primers of the ITS2 region yielded positive PCR amplifications of one single band when T. canadensis was present in a sample. Cross-amplification was not found during the mock test; other charophyte species did not yield positive amplification. The eDNA samples from the Torneträsk validated the performance of the ITS2 marker. The T. canadensis-specific marker designed in this project was proven to be sensitive and accurate. It could be recommended as a useful tool to detect the presence of T. canadensis DNA, even at low concentration and in complex samples containing other charophyte species.


LWT ◽  
2021 ◽  
Vol 137 ◽  
pp. 110336
Author(s):  
Irini Bosmali ◽  
Georgios Lagiotis ◽  
Evangelia Stavridou ◽  
Nadia Haider ◽  
Maslin Osathanunkul ◽  
...  

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Mochamad Syaifudin ◽  
Michaël Bekaert ◽  
John B. Taggart ◽  
Kerry L. Bartie ◽  
Stefanie Wehner ◽  
...  

Abstract Tilapias (family Cichlidae) are of importance in aquaculture and fisheries. Hybridisation and introgression are common within tilapia genera but are difficult to analyse due to limited numbers of species-specific genetic markers. We tested the potential of double digested restriction-site associated DNA (ddRAD) sequencing for discovering single nucleotide polymorphism (SNP) markers to distinguish between 10 tilapia species. Analysis of ddRAD data revealed 1,371 shared SNPs in the de novo-based analysis and 1,204 SNPs in the reference-based analysis. Phylogenetic trees based on these two analyses were very similar. A total of 57 species-specific SNP markers were found among the samples analysed of the 10 tilapia species. Another set of 62 species-specific SNP markers was identified from a subset of four species which have often been involved in hybridisation in aquaculture: 13 for Oreochromis niloticus, 23 for O. aureus, 12 for O. mossambicus and 14 for O. u. hornorum. A panel of 24 SNPs was selected to distinguish among these four species and validated using 91 individuals. Larger numbers of SNP markers were found that could distinguish between the pairs of species within this subset. This technique offers potential for the investigation of hybridisation and introgression among tilapia species in aquaculture and in wild populations.


2018 ◽  
Vol 16 (4) ◽  
Author(s):  
Leandro Marajó ◽  
Patrik F. Viana ◽  
Milena Ferreira ◽  
Lúcia H. Rapp Py-Daniel ◽  
Eliana Feldberg

ABSTRACT Farlowella is one of the most diverse genera of the Loricariinae, restricted to South America rivers. The taxonomic and phylogenetic relationships among its species are contentious and, while genetic studies would contribute to the understanding of their relationships, the only available datum refer to the karyotype description of only one species. In the present study two Amazonian species, Farlowella cf. amazonum and F. schreitmuelleri, were analyzed using conventional and molecular cytogenetic procedures. Both species had diploid chromosome number 58, but different fundamental numbers (NF) 116 and 112, respectively, indicative of chromosomal rearrangements. C-banding is almost poor, especially in F. cf. amazonum, and occurs predominantly in the centromeric and in some telomeric regions, although genome of F. schreitmuelleri possessed a much larger heterochromatin amount then those of F. cf. amazonum. The chromosomes bearing the NOR sites were likely the same for both species, corresponding to the 1st metacentric pair in F. cf. amazonum and to the 28th acrocentric in F. schreitmuelleri. The location of the 5S rDNA was species-specific marker. This study expanded the available cytogenetic data for Farlowella species and pointed the remarkable karyotype diversity among species/populations, indicating a possible species complex within genus.


2020 ◽  
Vol 6 (4) ◽  
pp. 308
Author(s):  
Joana Carvalho-Pereira ◽  
Filipa Fernandes ◽  
Ricardo Araújo ◽  
Jan Springer ◽  
Juergen Loeffler ◽  
...  

A new and easy polymerase chain reaction (PCR) multiplex strategy, for the identification of the most common fungal species involved in invasive fungal infections (IFI) was developed in this work. Two panels with species-specific markers were designed, the Candida Panel for the identification of Candida species, and the Filamentous Fungi Panel for the identification of Aspergillus species and Rhizopusarrhizus. The method allowed the correct identification of all targeted pathogens using extracted DNA or by colony PCR, showed no cross-reactivity with nontargeted species and allowed identification of different species in mixed infections. Sensitivity reached 10 to 1 pg of DNA and was suitable for clinical samples from sterile sites, with a sensitivity of 89% and specificity of 100%. Overall, the study showed that the new method is suitable for the identification of the ten most important fungal species involved in IFI, not only from positive blood cultures but also from clinical samples from sterile sites. The method provides a unique characteristic, of seeing the peak in the specific region of the panel with the correct fluorescence dye, that aids the ruling out of unspecific amplifications. Furthermore, the panels can be further customized, selecting markers for different species and/or resistance genes.


Plant Disease ◽  
2008 ◽  
Vol 92 (11) ◽  
pp. 1480-1487 ◽  
Author(s):  
Guiping Yan ◽  
Richard W. Smiley ◽  
Patricia A. Okubara ◽  
Andrea Skantar ◽  
Sandra A. Easley ◽  
...  

A species-specific polymerase chain reaction (PCR) method was developed to detect and identify the root-lesion nematodes Pratylenchus neglectus and P. thornei from soil. A primer set was designed from Pratylenchus 28S rRNA gene sequences of the D3 expansion domain. Primer specificity was confirmed with 23 isolates of 15 nematode species and other plant-parasitic and non-plant-parasitic nematodes typically present in the soil communities, and with six fungal species commonly associated with wheat root rot. DNA obtained using a commercially available kit and a method developed in our laboratory gave comparable amplification. PCR conditions were optimized and the two species were differentiated by PCR products of 144 bp for P. neglectus and 288 bp for P. thornei. With this assay, we detected a single juvenile in 1 g of sterile, inoculated soil. Examination of 30 field soil samples revealed that this method was applicable to a range of soils naturally infested with these two pathogens in Oregon. This PCR-based method is rapid, efficient, and reliable, does not require expertise in nematode taxonomy and morphology, and could be used as a rapid diagnostic tool for commercial and research applications for disease forecasting and management.


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