Differentiation of Aspergillus niger by random amplification of polymorphic DNA

Abed, Kawther

doi:10.1007/s10295-008-0378-x

Cited by 7 publications

(3 citation statements)

References 19 publications

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“…This difference may also be attributed to a possible genetic diversity among strains from different regions. The genetic variability and the phylogenetic characterization of Aspergillus section Nigri isolated from vineyards have been assessed by using DNA fingerprints generated by PCR (Abed, 2008;Bau et al, 2006;Chiotta et al, 2011;Esteban et al, 2008Esteban et al, , 2006Martínez-Culebras and Ramón, 2007;Martínez-Culebras et al, 2009;Oliveri et al, 2008;Perrone et al, 2006b;Spadaro et al, 2012;Susca et al, 2013). Different molecular marker techniques such as restriction fragment length polymorphisms (RFLPs), randomly amplified polymorphic DNA (RAPDs), amplified fragment length polymorphisms (AFLPs), minisatellites or variable number tandem repeats (VNTRs) and microsatellites or simple sequence repeats (SSRs) were used in the aforementioned studies.…”

Section: Introductionmentioning

confidence: 99%

Ecophysiological characterization of Aspergillus carbonarius, Aspergillus tubingensis and Aspergillus niger isolated from grapes in Spanish vineyards

García-Cela

Crespo-Sempere

Ramos

et al. 2014

International Journal of Food Microbiology

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The aim of this study was to evaluate the diversity of black aspergilli isolated from berries from different agroclimatic regions of Spain. Growth characterization (in terms of temperature and water activity requirements) of Aspergillus carbonarius, Aspergillus tubingensis and Aspergillus niger was carried out on synthetic grape medium. A. tubingensis and A. niger showed higher maximum temperatures for growth (>45 °C versus 40-42 °C), and lower minimum aw requirements (0.83 aw versus 0.87 aw) than A. carbonarius. No differences in growth boundaries due to their geographical origin were found within A. niger aggregate isolates. Conversely, A. carbonarius isolates from the hotter and drier region grew and produced OTA at lower aw than other isolates. However, little genetic diversity in A. carbonarius was observed for the microsatellites tested and the same sequence of β-tubulin gene was observed; therefore intraspecific variability did not correlate with the geographical origin of the isolates or with their ability to produce OTA. Climatic change prediction points to drier and hotter climatic scenarios where A. tubingensis and A. niger could be even more prevalent over A. carbonarius, since they are better adapted to extreme high temperature and drier conditions.

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Section: Introductionmentioning

confidence: 99%

Ecophysiological characterization of Aspergillus carbonarius, Aspergillus tubingensis and Aspergillus niger isolated from grapes in Spanish vineyards

García-Cela

Crespo-Sempere

Ramos

et al. 2014

International Journal of Food Microbiology

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“…Alternatively, random amplification of polymorphic DNA (RAPD) and restriction fragment length polymorphism PCR (RFLP-PCR) are used, where the internal transcribed spacer regions between 18S, 5.8S and 28S rRNA genes are investigated [ 50 , 72 – 74 ]. Both methods are suitable for a reliable identification of Aspergillus species, but the correct interpretation of the resulting complex DNA fragment patterns is difficult [ 73 ]. Furthermore, these methods cannot be used for analyzing several species within a single multiplex PCR, as it was done in this study.…”

Section: Discussionmentioning

confidence: 99%

Genotyping bacterial and fungal pathogens using sequence variation in the gene for the CCA-adding enzyme

Franz¹,

Betat²,

Mörl³

2016

BMC Microbiol

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BackgroundTo allow an immediate treatment of an infection with suitable antibiotics and bactericides or fungicides, there is an urgent need for fast and precise identification of the causative human pathogens. Methods based on DNA sequence comparison like 16S rRNA analysis have become standard tools for pathogen verification. However, the distinction of closely related organisms remains a challenging task. To overcome such limitations, we identified a new genomic target sequence located in the single copy gene for tRNA nucleotidyltransferase fulfilling the requirements for a ubiquitous, yet highly specific DNA marker. In the present study, we demonstrate that this sequence marker has a higher discriminating potential than commonly used genotyping markers in pro- as well as eukaryotes, underscoring its applicability as an excellent diagnostic tool in infectology.ResultsBased on phylogenetic analyses, a region within the gene for tRNA nucleotidyltransferase (CCA-adding enzyme) was identified as highly heterogeneous. As prominent examples for pro- and eukaryotic pathogens, several Vibrio and Aspergillus species were used for genotyping and identification in a multiplex PCR approach followed by gel electrophoresis and fluorescence-based product detection. Compared to rRNA analysis, the selected gene region of the tRNA nucleotidyltransferase revealed a seven to 30-fold higher distinction potential between closely related Vibrio or Aspergillus species, respectively. The obtained data exhibit a superb genome specificity in the diagnostic analysis. Even in the presence of a 1,000-fold excess of human genomic DNA, no unspecific amplicons were produced.ConclusionsThese results indicate that a relatively short segment of the coding region for tRNA nucleotidyltransferase has a higher discriminatory potential than most established diagnostic DNA markers. Besides identifying microbial pathogens in infections, further possible applications of this new marker are food hygiene controls or metagenome analyses.Electronic supplementary materialThe online version of this article (doi:10.1186/s12866-016-0670-2) contains supplementary material, which is available to authorized users.

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“…The DNA of the 23 isolates used in this study was isolated by the protocol described by Doyle & Doyle (1990). Random Amplified Polymorphic DNA (RAPD) analysis was done with primers A1, A6 and OPA4 according to Abed (2008). The program FreeTree (Hampl et al, 2001) was used to construct the dendrogram using the distances of Jaccard and the UPGMA method.…”

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Additional species of Aspergillus causing bole rot disease in Agave sisalana

Santos

Silva

Corrêa

et al. 2014

Trop. plant pathol.

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The production of sisal in Bahia, Brazil, has been declining due to the occurrence of a disease know as bole rot. Aspergillus niger was regarded as the only causal agent. In this study A. brasiliensis and A. tubingensis, in addition to Aspergillus niger, identified on the basis of morphological and molecular analyses, were shown to cause bole rot of sisal. Their pathogenicity was confirmed but their significance for the epidemiology of the disease in the field remains unclear.

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Differentiation of Aspergillus niger by random amplification of polymorphic DNA

Cited by 7 publications

References 19 publications

Ecophysiological characterization of Aspergillus carbonarius, Aspergillus tubingensis and Aspergillus niger isolated from grapes in Spanish vineyards

Ecophysiological characterization of Aspergillus carbonarius, Aspergillus tubingensis and Aspergillus niger isolated from grapes in Spanish vineyards

Genotyping bacterial and fungal pathogens using sequence variation in the gene for the CCA-adding enzyme

Additional species of Aspergillus causing bole rot disease in Agave sisalana

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