Asian J Agric & Biol. 2019;7(2):197-203.

A simple, rapid, safe and low-cost method to extract DNA from phytopathogenic fungi

Adnan A. Lahuf1*, Ola H. Jaafar1, Zainab L. Hameed2

1Department of Plant protection, College of Agriculture, University of Kerbala, Kerbala, Iraq

2Department of Field crops, College of Agriculture, University of Kerbala, Kerbala, Iraq

Abstract

The aim of this study was to develop an easy, fast, non-hazardous and inexpensive technique for extraction of genomic DNA from multiple plant fungal pathogens. Samples of pure fungal growth of Fusarium equesti , Neoscytalidium dimidiatum , Fusarium proliferatum and Alternaria alternata isolated from diseased wheat, grapevine, potato and lily plants respectively were ground with sterilized sand and NaOH (2N), followed by a centrifuging process to separate the sand grains and cellular components of fungi from the DNA. Subsequently, the DNA was mixed with Tris  buffer (1 M) pH 8. The ITS region of rDNA was successfully amplified, sequenced and analyzed from the extracted DNA of the four pathogenic fungi. This new approach provides a simple, rapid, safe and low cost way to obtain DNA samples of sufficient quantity and quality for use in molecular assays for the identification of plant fungi.

Keywords: DNA extraction, Fungi, PCR, Sequence, Phylogeny analysis