Multiplex-PCR detection of fumonisin-producing F.verticillioides and F.proliferatum in corn of feedstuff

Cereals contamination of feedstuff such as corn with mycotoxigenic species of Fusarium is considered as a main source of fumonisin mycotoxin groups which cause worldwide serious diseases in human and animal health. According, in this study, a total of 100 Fusarium isolates were recovered from corn samples collected from 16 dairy farms in Razavy Khorasan Province, Iran, during 2011-2012. Fusarium isolates were identified on morphological and using species-specific PCR. Considering that, early detection of Fusarium species could be very important to prevent mycotoxin contaminating food and feed grains, a Multiplex-PCR assay was designed on the base of four sets of specific primers including: ITS-F/R, Vert1/2, FUM1-F/R and FP3-F/FP4-R which has been developed for the discovery of the two common Fusarium species (F.verticillioides and F.proliferatum) that they can potentially produce fumonisin. In this study, PCR analysis comfirmed 78% of morphological identification to species level as F.proliferatum (53.8%) and 29.5% as F.verticillioides. 42 (53.84%) of total (78) F.verticillioides and F.proliferatum isolated were fumonisin producing out of which 24 (57.14%) were F.proliferatum while 18 (42.86 %) were F.verticillioides. Moreover, we have established the method to discriminate between the two populations F.verticillioides and F.proliferatum, simultaneously enable us to discriminate fumonisin-producing strains from non-fumonisin-producing strains in the corn grains of animal feedstuff.

Keywords:

Corn , Multiplex-PCR , F.verticillioides , F.proliferatum , fumonisin