Efficient DNA extraction method from soil infested with Fusarium sp. for metagenomics study
DOI:
https://doi.org/10.59797/journaloffoodlegumes.v29i1.551Keywords:
Fusarium, Soil samples, Restriction digestion, PCR, NCBI-BLASTAbstract
Simple and rapid identification of pathogenic fungi from infested soil is pre-requisite to study the fungal dynamics in soil. The traditional methods of fungal characterization are more cumbersome and non-specific. Therefore, a rapid and specific method of fungal characterization through genomic DNA is more precise. Hence, in the present study, samples from infested soil were collected randomly for fungal DNA isolation. The DNA concentration ranged between 240 to 410 ng/ìlwith A260/280 values of 1.48 to 1.69. The quality of DNA was confirmed by restriction digestion with Hind III and Hae III enzymes and further confirmed by PCR amplification with ITS specific primers. Sequencing and NCBI-BLAST analysis of 420bp amplicon confirmed presence of Fusarium spp. and sequence was submitted to NCBI database (Accession no.: KP153547 to KP153550). Homology search in NCBI-BLAST, found 94-96% homologues with Fusarium spp. Thus, the protocol used in present study helps in rapid detection of soil borne fungi and also gives information about fungal dynamics and facilitates in proper disease management.
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