Development of a sensitive molecular detection assay for mango malformation disease caused by Fusarium mangiferae.

Abstract

To develop a sensitive and specific molecular assay for detection of mango malformation disease (MMD), which is caused primarily by Fusarium mangiferae. We screened 100 ISSR primers and identified one (UBC888) that directed the stable amplification of a specific gene fragment of 479bp (GenBank accession number KJ526382). Based on the DNA sequence of this fragment, a pair of SCAR primers (W342 and W1772) were designed to amplify another gene fragment of 1376bp (GenBank accession number KJ526383), demonstrating the successful conversion of an ISSR marker to a SCAR marker. An effective and simple detection assay for MMD was established based on this pair of PCR primers, with a high level of specificity and sensitivity to the DNA of F. mangiferae and other species of Fusarium both in vitro and in vivo. It can detect as little as 10pg fungal DNA from the DNA of mango's tissues. Our assay provides a practical method for the early diagnosis so that proper prevention of the mango malformation disease can be developed.