Detection of nivalenol synthesis gene in madder seeds infected by Fusarium species by PCR

Abstract

Introduction: The madder (Rubia tinctorum) is one of the most important crops for medical and industrial applications. This product maybe infected by Fusarium species that produces potentially fatal mycotoxins. The purpose of this current research is to identify toxins produced by madder seeds-associated Fusarium fungi using molecular and biochemical methods. Methods: From major areas of cultivation of madder seeds in Yazd province including, Ardekan and Bafgh, sampling was done. Culture and purification of Fusarium isolates were taken place in specific media. Detection of fungi with the ability to produce trichothecenes mycotoxins such as nivalenol (NIV) through gene-specific primers for Tri13 by the polymerase chain reaction method (PCR) was performed. To confirm the NIV production potential, high performance liquid chromatography (HPLC) was applied. Results: In this study, five Fusarium species including F.solani, F.oxysporum, F. poae, F. semitectum, and F. eqiuesti were identified from madder seeds. The results showed that among Fusarium species isolated from madder seeds, F. poae  and F. equiseti have the ability to produce NIV. The gene involved in NIV synthesis, Tri13, was detected in two species so that all these isolates were identified as NIV producing type. The HPLC performance showed that all studied Fusarium species, have the potential to produce NIV mycotoxin. Conclusion: Based on results, Tri13 gene has a crucial role in trichothecene produce. Thus, the PCR method can be used in various products for faster detection of fungi have the potential mycotoxin production. Given that this is the first report of NIV trichothecene in seeds of madder in Yazd, a proscriptive measure should be taken to control and reduce the fungal and toxin production in these products.

Keywords

• madder
• seeds
• Fusarium
• NIV