Abstract
Fusarium head blight (FHB) caused by several Fusarium species is one of the most serious diseases affecting wheat throughout the world. The in vitro production of the toxins deoxynivalenol, zearalenone fumonisin, T-2, and HT-2 was quantitvely evaluated in 8 different isolates of Fusarium species collected from feed samples. It was possible to detect zearalenone and the other mycotoxins in 100% and 50% of the isolates, respectively. In the present study, loop-mediated isothermal amplification method (LAMP) was designed for diagnosing Fusarium garmanirum infections and testing against feed samples, infested samples and pure cultures. The LAMP amplicon was directly visualized in the reaction tubes by the naked eye following the addition of calcein fluorescence. The LAMP products appeared as DNA marker pattern, with many bands of different sizes from 145 base pairs up to the loading well. Loop-LAMP procedure was used to detect genomic DNA of F. graminearum in fungal pure culture and in contaminated feed samples. In the future, this assay will support plant quarantine programs in Saudi Arabia and Gulf Cooperation Council states, to prevent the introduction of foreign FHB species. Key words: LAMP-PCR, Fusarium head blight, feed samples.
Highlights
Head blight is caused by several Fusarium spp.; distribution and predominance of species significantly vary among climatic conditions, geographical zones, countries, and years (Doohan et al, 2003; Xu, 2003)
The current study describes the loop-mediated isothermal amplification method (LAMP) for the detection of F. garmanirum in infested feed sample
Fumonisins were produced by a number of Fusarium species, notably F. verticillioides and F. graminearum
Summary
Head blight is caused by several Fusarium spp.; distribution and predominance of species significantly vary among climatic conditions, geographical zones, countries, and years (Doohan et al, 2003; Xu, 2003). Morphological identification of Fusarium spp. is difficult due to their similarities. F. avenaceum are very difficult to separate by the morphological characteristics of their spores (Yli Mattila et al, 2004). The novel nucleic acid amplification method loop-mediated isothermal amplification (LAMP) has been reported as a simple rapid diagnostic tool for early detection of microorganisms (Parida et al, 2008). LAMP was developed for rapid detection of pathogenic or allergenic fungal in the environment (Sun et al, 2010). LAMP-PCR using internal labeled probes have been developed for the major FHB pathogens in different plant materials (Niessen and Vogel, 2010; Abd-Elsalam et al, 2011; Niessen et al, 2012).
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