Journal of Agronomy and Field Innovations

Background: Fusarium head blight causes so much destruction that it has been given the name wheat head blight. The destruction is caused by a pest called Fusarium graminearum, which causes immense financial damage. It is estimated that the damages amount to over $2 billion. In addition to monetary losses, this fungus contaminates wheat with deoxynivalenol (DON) toxin that poses a danger to human health. Traditional diagnostic techniques such as morphology studies and culture isolation are slow and laborious, as well as ineffective. 
Objective: This study aims to create and validate the multiplex PCR method that facilitates the process of detecting Fusarium graminearum in infected wheat. 
Methods: The new multiplex PCR method developed in this research enables the detection of three DNA regions that makes identification of F. graminearum easy. The method was validated in 8 different varieties of wheat grown in different regions during two growing seasons. 480 wheat spike samples were tested and compared against the traditional culture method.
Results: The sensitivity of the multiplex-PCR method was found to be 96.2% and specificity to be 99.1%. Detecting the target DNA with this method takes only 6-8 hours, which is much shorter than the 10-14-day period necessary for classical culture methods of isolation. The study strongly correlates with the reference method of diagnosis (Cohen's κ = 0.938, P < 0.001). Fusarium graminearum was identified in 87.5% of the symptomatic samples as well as in 12.8% of the asymptomatic samples, indicating its ability to show pathogen presence at early stages of infection.
Conclusion: Multiplex-PCR is a fast, effective, high-quality method that could be used for the identification of Fusarium graminearum in wheat. High specificity and sensitivity along with the ability to detect silent infections make it widely applicable for monitoring wheat head blight, controlling the disease as well as in integrated pest management strategies in order to ensure sustainable cereal production.