Molecular Detection of Aflatoxin Producing Fungal Gene, aflQ Gene in Infected Rice Samples Using PCR

Rice (Oryza sativa L.) is the staple food of over half the world's population. According to FAO, 25% of the world's food crops are affected by mycotoxins. Mycotoxin detection is important for quality control, especially while exporting food grains. Mycotoxin producing gene fragments in fungal strains were amplified using PCR and realtime PCR along with Bioanalyzer in an attempt to develop a rapidassay for mycotoxin detection. As many as 120 samples of rice were collected from various storage areas like godowns, wholesale and retail shops and farmer's fields in major rice growing districts in Andhra Pradesh (A.P). Fungal DNAwas extractedusing DNA extraction kits. DNA from fungal contaminated rice samples was extracted by the CTAB method. Forward and reverse primers were designed by identifying homologous regions from original gene sequences (obtained from GENBANK) of the aflQ gene, which is the key gene involved in the production of the mycotoxin, aflatoxin respectively. Sequences from different species of fungi were aligned using megalign software from DNASTAR Lasergene 8.0 version, and homologous regions were identified. Primer 3.0 software was used for designing primers from these homologous regions. Results showed that out of 120 rice samples tested for the detection of the genes producing aflatoxin, aflQ gene was detected in 35 samples.