Browsing by Author "Ghimire, Sadikshya"

Fusarium proliferatum has recently become a major threat to onion, which is an important food crop for food security and has a significant role in the agricultural sector. This fungus is found causing rots and producing mycotoxin fumonisin that, if ingested, can cause carcinogenic effects in humans and fatal diseases in animals. F. proliferatum has been identified as a pathogen causing rots and wilts in many plants in several countries, and recently some isolates of this pathogen were also found in Finland causing basal rot in onion. Though F. proliferatum has wide adaptability and pathogenicity, there are research gaps on this newly emerging pathogen, which is mostly limited to some specific hosts such as maize. There is a lack of knowledge of its infection mechanisms and mycotoxin production dynamics in onion. This experimental work was conducted at the University of Helsinki, Finland, to study the nature of pathogenicity and toxin gene expression of F. proliferatum in laboratory conditions as a function of time. Spore suspensions of three isolates of F. proliferatum, Fpr047, Fpr049, and Fpr919, were prepared and used as inoculums that were injected into healthy organically grown onions, which were then stored in dark for five weeks. Control bulbs were inoculated with sterile water. Pathogen virulence, based on symptom development, and fungal colonization in the onion tissues were determined at five different time points and the toxin gene expression was determined at three time points. Colonization levels were determined by real-time PCR using primers binding to the intergenic spacer (IGS) region of F. proliferatum. A part of the IGS region of the three isolates was sequenced to study the diversity between the isolates. All the tested isolates were found to be virulent, and they colonized the onions after one week from inoculation. However, the isolate Fpr919 appeared different from the other two in terms of symptom severity. It was more aggressive than the other two, causing disease symptoms earlier and causing more severe rot symptoms in the infected bulbs. It also had nucleotide sequence variations in the IGS region in comparison with the other two isolates, suggesting genetic diversity. No significant differences were observed between the isolates in the fungal colonization levels. Expression of FUM1 gene and a putative virulence gene SIX2-1 was detected by RT-RT-PCR in most of the infected tissue samples. The results obtained signify that F. proliferatum is a pathogen with the potential of producing fumonisin toxin in onion, suggesting the need for further molecular study on this fungus to control the disease and prevent mycotoxin contamination in plant products.