مرکز منطقه ای اطلاع رساني علوم و فناوري - Determination of Fusarium oxysporum Schltdl by Molecular Methods on Flower Bulbs

Abstract :

Objective: In this study, diagnoses of Fusarium oxysporum which is casual agent of the most common disease on flower bulbs Fusarium wilt have been carried out by using PCR-based techniques. Thereby, a valid protocol is prepared for the pathogen to use in practice. Material ve Methods: The material of the research consists of isolates of Fusarium oxysporum which is isolated from flower bulbs, reference isolate Fusarium oxysporum f. sp. tulipae and TEF1 ve FOW1 primer pairs. In the study DNA isolation was carried out with Fusarium isolates, which are isolated from flower bulbs, and reference isolate. The presence of Fusarium oxysporum was determined by Real-Time PCR and sequencing. Results: As a result of the study, TEF1 primer pairs amplified on the 20th cycle and demonstrated the presence of F.oxysporum. On the other hand, FOW1 primer pairs didn t amplified. There is no contaminaton on the negative controls. As a result of the sequence, samples were justified that they are Fusarium. Conclusion: Current diagnostic methods in the determination of the Fusarium wilt have been lasting approximately for a week, and their diagnosis on the basis of species have been difficult. Besides, this period extends because of pathogenicity. With this study, Fusarium oxysporum which is casual agent of the most common disease on flower bulbs Fusarium wilt, was determined in a fast and correct way by conventional and real-time PCR and validity of the method has been pointed out with sequencing. Thereby, a valid protocol is prepared for the pathogen to use in practice. When we consider these reasons, preference of PCR techniques which are more sensitive, more reliable and faster will lead advantages.