Fate of deoxynivalenol in contaminated wheat grain during preparation of Egyptian 'balila'.

Soaking and boiling whole wheat kernels in water are the key steps in the preparation of an Egyptian dish called 'balila'. The effects of washing, soaking and boiling wheat kernels in tap water or in 0.1 M Na2CO3 solution on the deoxynivalenol (DON) content of the wheat kernels were studied. Boiling contaminated wheat kernels in water reduced the DON content of the grain by 70%. The mechanism of decontamination due to boiling is probably a leaching of DON out of the grain into the boiling medium. A combined treatment of soaking in 0.1 M Na2CO3 solution (pH 11) with subsequent boiling reduced the DON content of the grain by 93%. Data suggest that apart from leaching DON out of the kernels into the boiling medium, a degradation of DON occurred in alkaline medium. Modifying the traditional process of 'balila' preparation by using Na2CO3 solution may be useful to reduce the risk of mycotoxin exposure via 'balila'.

Presymptomatic and quantitative detection of Mycosphaerella graminicola development in wheat using a real-time PCR assay.

Presymptomatic and accurate diagnosis of Mycosphaerella graminicola leaf blotch is desirable for the disease prediction and the timely application of fungicides. To develop a sensitive PCR assay, four specific primer pairs were designed. They were more specific than three known specific primer pairs. Three of them could detect as little as 0.5 pg M. graminicola DNA in a conventional PCR. A real-time PCR assay was applied for monitoring the disease progression in both inoculated and naturally infected wheat plants using the primer pair ST-rRNA F/R. In inoculated plants, M. graminicola DNA could be detected immediately after inoculation and a steady increase was detected before visible symptoms appeared at 8 days. The rapid growth period took place between 6 and 16 days postinoculation. In the field, the disease progression in the top three leaf layers was followed during the epidemic period. The results were significantly correlated to the disease indices (R=0.8986) and also to the number of pycnidia per leaf (R=0.9227). These suggest that the real-time PCR assay is a reliable approach for the presymptomatic and accurate detection of M. graminicola development in the field.

Comparative assessment of genotyping methods for study genetic diversity of Fusarium oxysporum isolates.

In this study, we evaluated three PCR-based methods for the molecular typing of nonpathogenic Fusarium oxysporum isolates: random amplified polymorphic DNA (RAPD), polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) and amplified fragment length polymorphism (AFLP). The analyses were performed using 64 isolates of F. oxysporum collected from cotton-producing areas in Egypt. A number of polymorphic RAPD, PCR-RFLP and AFLP bands were scored in all isolates and the genetic similarity among them was assessed. Clustering analysis separated the isolates into two main groups, with similarities ranging from 87 to 100% for RAPD, 80 to 100% for PCR-RFLP and 88 to 97% for AFLP, respectively. The obtained data suggested that all three types of markers are equally informative, but the three assays differed in the amount of detected polymorphic bands. AFLP fingerprinting was also found to be more differentiating than other techniques for the typing of F. oxysporum populations.