Clinical Microbiology: Open Access

Molecular detection and characterization of Fusarium sporotrichioides based on ITS2 rDNA polymorphism

3^rd Global Microbiologists Annual Meeting

August 15-17, 2016 Portland, Oregon, USA

Afaf I Shehata, Ekram A M Al-Sanae, Ali H Bahkali, Mohammed Abdo Yahya and Amal A Al Hazzani

King Saud University, KSA

Posters & Accepted Abstracts: Clin Microbiol

Abstract:

The genus Fusarium contains a number of soil borne species with worldwide distribution. The presented PCR assays are highly selective and sensitive in detecting the Fusarium genus. In order to identify the 18 Fusarium isolates obtained at the molecular level, PCR analysis using primer specific for the conserved ITS DNA region of Fusarium genus was conducted. The data indicated that, all of the 18 isolates showed a clear band corresponding to the expected molecular size of the ITS region (431 bp). These results confirmed that all the tested samples belong to the genus Fusarium. Also, when all eighteen isolates of Fusarium species were analyzed by PCR for fumonisin producing ability using FUM1 gene based primers, the expected DNA fragment of 183 bp was amplified only in Fusarium verticillioides (3 isolates), Fusarium avenaceum (3 isolates), Fusarium semitectum (1 isolate) and Fusarium culmorum (2 isolates) showed a positive result with FUM1 gene set of primers. No bands were seen in other isolates of Fusarium spp., and the standard (Fusarium graminearum). In case of zearalenone, the PKS4 gene of F. graminearum has been reported to be essential in the production of zearalenone. The result indicated that the expected DNA fragment of 280 bp was amplified only in Fusarium verticillioides (3 isolates), Fusarium avenaceum (3 isolates) and Fusarium culmorum (2 isolates) and Fusarium graminearum. Microsatellite-primed PCR resembles the well-known RAPD technique but is advantageous because of the ability to generate more complex banding patterns and a high degree of reproducibility. The discriminating powers of the three MP-primers [(CTG)5, (M13) and (T3B)] used in this study were nearly the same. Cluster analyses were performed on the genomic fingerprints generated by each of the primers tested. Three dendrograms were generated with the UPGMA method. The patterns resulting from the T3B and (CTG)5 test were more distinct and T3B was the most successful primer because it always led to high polymorphic banding patterns that were suitable for interspecies comparisons. Our results indicated that there was no association between clustering in the MP-PCR dendrogram and the geographic origin and morphological identification of the tested isolates.

Biography :

Email: afafsh@ksu.edu.sa