PCR METHOD FOR THE DETECTION OF GENUS FUSARIUM AND FUMONISIN‐PRODUCING ISOLATES FROM FRESHLY HARVESTED SORGHUM GRAINS GROWN IN KARNATAKA, INDIA

Abstract

ABSTRACT Sixty‐four isolates of Fusarium species isolated from 44 sorghum samples collected during 2004–2005 were subjected to polymerase chain reaction (PCR) analysis. PCR detection was performed on all Fusarium species using two different sets of primers, namely, internal transcribed spacer (ITS) and FUM1. The developed protocol is rapid for small‐scale extraction of DNA from fungal mycelia for the detection of Fusarium species using diagnostic PCR. Sixty‐four Fusarium isolates, namely, Fusarium verticillioides (45), Fusarium proliferatum (4), Fusarium anthophilum (4), Fusarium sporotrichioides (3), Fusarium pallidoroseum (6) and Fusarium oxysporum (2), were analyzed by PCR using the ITS and FUM1 set of primers. All Fusarium species scored positive with the ITS set of primers. Among the 64 isolates, 53 have scored positive with the FUM1 set of primers. These 53 isolates represent F. verticillioides (45), F. proliferatum (4) and F. anthophilum (4), respectively. The results of the study revealed that PCR‐based technique could be used to identify a group of potential fumonisin‐producing Fusarium species.PRACTICAL APPLICATIONSPolymerase chain reaction (PCR) method provides the basis for a simple, accurate, rapid and precise detection of potential fumonisin producing Fusarium species, which are of considerable risk to animal and human health. The detection of Fusarium species is therefore crucial for the prevention of toxins entering the food chain. The protocols developed in this study are helpful for a rapid and small‐scale extraction of nucleic acid from fungal mycelia, occurring on contaminated sorghum samples. A large number of samples can be screened in relatively less time using this PCR protocol when compared with the conventional methods.