Ecology, Environment and Conservation Paper

Vol. 19 Issue 04, 2013; Page No.(1241-1246)

DETECTION OF MYCOTOXIN PRODUCING FUNGAL SPECIES FOR MAIZE (ZEA MAYSL.) SEEDS

S. Nagalakshmi , K. Manorama, Arun Prem Kumar and P. Jayamma, M. Sreedhar, Ch. V. Durgaran, V.L.N. Reddy, Sunil Kulkarni, Anurag Chaturvedi, A. Ravicharan

Abstract

A simple, rapid, and efficient method for isolating genomic DNA from germinated seeds of Maize that is free from polysaccharides and polyphenols is reported. DNA was extracted, treated with RNase, measured and tested for completeness using agarose gel electrophoresis. DNA purification from Maize grains yielded abundant, amplifiable DNA with yields typically between 100 and 200 ng DNA/mg. The effectiveness and reliability of the method was tested by assessing quantity and quality of the isolated DNA using Nano drop method. Three gene specific PCR-based markers procedure were used to detect genomic DNA of mycotoxins aflatoxins deoxynivalenol, fumonisins in contaminated Maize seeds. In this method there is no need to use liquid nitrogen for crushing germinated seedlings. In combination with the PCR assay it is a fast and cost-effective alternative to traditional diagnostic methods for the early detection of toxigenic mycotoxins in maize seeds.

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