Nested PCR Method for Early Detection of Fumonisin Producing Fusarium verticillioides in Pure Cultures, Cereal Samples and Plant Parts

Abstract

ABSTRACTFumonisin-producing Fusarium verticillioides was detected in cereal samples by semi-nested PCR using one forward, VERTF-1, and two reverse primers, VERTR and VERTF-2. A total of 326 Fusarium species isolated from maize, paddy, sorghum, wheat and pearl-millet samples were subjected to a first round of n-PCR with the species specific VERTF-1 and VERTR set of primers which recorded 59.50% of F. verticillioides. Further, second round of n-PCR scored 53.98% of fumonisin-producing F. verticillioides with fumonisin specific VERTF-1 and VERTF-2 set of primers. Maize samples recorded the highest frequency of fumonisin-producing F. verticillioides 40.98%, followed by paddy 33.33%, and sorghum 12.50%. Sensitivity of nested PCR was conducted by whole grain experiment of the cereals, roots and leaves of the cereals by diluting the DNA 10-100 times, in which 1:50, 1:75, and 1:100 diluted samples recorded positive. This method can be used for the early detection of fumonisin producing F. verticillioides occurring on cereals.