Multiplex PCR for the early detection of fumonisin producing Fusarium verticillioides

In the present study, fumonisin producing Fusarium verticillioides was specifically detected in pure cultures, cereal samples and plant materials by multiplex PCR using one forward VERTF-1 and two reverse primers VERTR and VERTF-2. A total of 326 Fusarium isolates were obtained from maize, sorghum, paddy wheat and pearl millet samples collected from different districts of Karnataka, India. All Fusarium species were subjected to single round of PCR with species specific and fumonisin specific primers which recorded 59.50% of F. verticillioides and 53.98% of fumonisin producing F. verticillioides. Maize samples recorded highest frequency 34.42% of fumonisin producing F. verticillioides followed by paddy 28.57% and sorghum 16.66%. Sensitivity of multiplex PCR experiment was conducted by whole grain experiment of the collected cereals, roots and leaves of the cereal samples by diluting the DNA 10 to 100 times in which 1:50, 1:75 and 1:100 diluted samples recorded positive. The developed multiplex PCR assay provided a powerful tool for the accurate detection, identification and discrimination of potential fumonisin producing F. verticillioides strains among the population. The present study is the first report of developing the multiplex PCR method for early detection of fumonisin producing F. verticillioides from cereal samples, pure cultures and plant parts.