Species‐specific PCR Assays for the Fungal Pathogens Fusarium moniliforme and Fusarium subglutinans and their Application to Diagnose Maize Ear Rot Disease

Abstract

Fusarium moniliforme Sheldon (syn. F. verticillioides (Sacc.) Nirenberg) and F. subglutinans (Wollenweber & Reinking) Nelson Toussoun & Marasas comb. nov., two anamorphs of the so‐called‘Gibberella fujikuroi species complex', are important maize pathogens. Together with F. proliferatum, F. culmorum, and F. graminearum (teleomorph: Gibberella zeae) they are involved in the stalk rot and ear rot disease of maize. All species produce secondary metabolites (mycotoxins) which are a potential health hazard for humans and animals that consume maize and maize products frequently. In this study the development of polymerase chain reaction (PCR) assays for an easy and sensitive identification of G. fujikuroi anamorphs in maize kernels are described. The primer pairs are based on sequences of randomly amplified polymorphic DNA (RAPD) fragments and are specific for F. moniliforme and F. subglutinans respectively. The PCR assays are independent of the high phenotypic variability of traits which may complicate classification by morphological characters. They detect approximately 100 to 200 fungal genomes in the presence of an excess of maize DNA. For the analysis of infected maize kernels a rapid and easy DNA extraction was used which does not introduce inhibitory substances into the PCR. Hence the assays enable an early identification and detection of the two pathogens in host tissue by plant breeders and plant health inspection services. The assays were successfully applied to identify field isolates from Poland and to detect the pathogens in maize ears of various hybrids in Germany.

New PCR Assays for the Identification of Fusarium verticillioides, Fusarium subglutinans, and Other Species of the Gibberella fujikuroi Complex

“…The results obtained (Figure 4A) indicate the presence of F. subglutinans in the PG 1 sample and the presence of F. verticillioides in all three naturally contaminated seed samples tested, what is in accordance with the literature that reports the presence of F. verticillioides in 100% of the maize seeds in Brazil [26]. Möller et al [13] have also set up a multiplex PCR to detect F. verticillioides and F. subglutinans simultaneously, but there are no data on the specificity of these primers with the genomic DNA of F. circinatum , F. konzum , and F. andiyazi . The present multiplex PCR reaction represents a significant advance for the simultaneous molecular detection of F. verticillioides and F. subglutinans .…”

Early PCR-based detection of Fusarium culmorum, F. graminearum, F. sporotrichioides and F. poae on stem bases of winter wheat throughout Poland

“…Fusarium diseases of wheat are very important factors contributing to economic losses and deterioration in grain quality (McMullen et al 2012). The PCR-based assays can be used for the routine detection and identification of pathogenic fungi from genus Fusarium without morphological determination (Murillo et al 1998;Moeller et al 1999;Mulé et al 2004). The development and use of PCR assays could be also very helpful for early diagnosis and control of Fusarium population on wheat ear and stem base (Ben-Amar et al 2012).…”

Development of PCR assay based on ITS2 rDNA polymorphism for the detection and differentiation ofFusarium sporotrichioides

“…Most of these techniques are based on species specific primers (SCARs) for F. avenaceum [15], F. culmorum, F. graminearum [8,12,18], F. moniliforme, F. subglutinans [7], F. poae [10], F. sambucinum [18], F. sporotrichioides [17], F. torulosum and F. venenatum [18]. Most of these techniques are based on species specific primers (SCARs) for F. avenaceum [15], F. culmorum, F. graminearum [8,12,18], F. moniliforme, F. subglutinans [7], F. poae [10], F. sambucinum [18], F. sporotrichioides [17], F. torulosum and F. venenatum [18].…”