THE DEVELOPMENT OF A RAPID PCR ASSAY FOR DETECTION OF FUSARIUM-MONILIFORME

The fungus Fusarium moniliforme infects a wide range of crops througho ut the world. In maize (Zea mays L.) it causes seedling blight and roo t, stalk, and ear rots. A simple procedure that can be used to detect infection by F. moliliforme from infected plant tissues has been devel oped. A F. moniliforme genomic library was prepared and used to identi fy the recombinant clones containing fungal DNA sequences not hybridiz ing with the DNA of the host plant, maize. Based on the nucleotide seq uence information obtained from the E moniliforme pUCF2 genomic clone, specific oligonucleotides were designed and used as primers for in vi tro DNA amplification by the polymerase chain reaction. An amplificati on product was obtained with R moniliforme DNA preparations whereas no amplified DNA was detected with DNAs from other fungal pathogens, inc luding various Fusarium species, or from the host plant. This PCR anal ysis was successfully employed to identify E moniliforme directly from the mycelia that develop from naturally infected maize seeds, with no need to obtain pure fungal cultures for reliable diagnosis. The proto col can be used for the diagnosis of infected plants and soils in epid emiological studies of Fusarium diseases, for seed health testing, and for evaluation of susceptibility to colonization in commercial maize hybrids.