کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
2091490 1081732 2012 8 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Rapid identification of Fusarium graminearum species complex using Rolling Circle Amplification (RCA)
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی بیوتکنولوژی یا زیست‌فناوری
پیش نمایش صفحه اول مقاله
Rapid identification of Fusarium graminearum species complex using Rolling Circle Amplification (RCA)
چکیده انگلیسی

Rolling Circle Amplification (RCA) of DNA is a sensitive and cost effective method for the rapid identification of pathogenic fungi without the need for sequencing. Amplification products can be visualized on 1% agarose gel to verify the specificity of probe-template binding or directly by adding fluorescent dyes. Fusarium Head Blight (FHB) is currently the world's largest threat to the production of cereal crops with the production of a range of mycotoxins as an additional risk. We designed sets of RCA padlock probes based on polymorphisms in the elongation factor 1-α (EF-1α) gene to detect the dominant FHB species, comprising lineages of the Fusarium graminearum species complex (FGSC). The method also enabled the identification of species of the Fusarium oxysporum (FOSC), the Fusarium incarnatum–equiseti (FIESC), and the Fusarium tricinctum (FTSC) species complexes, and used strains from the CBS culture collection as reference. Subsequently probes were applied to characterize isolates from wheat and wild grasses, and inoculated wheat kernels. The RCA assays successfully amplified DNA of the target fungi, both in environmental samples and in the contaminated wheat samples, while no cross reactivity was observed with uncontaminated wheat or related Fusarium species. As RCA does not require expensive instrumentation, the technique has a good potential for local and point of care screening for toxigenic Fusarium species in cereals.


► Rolling Circle Amplification (RCA) of Fusarium graminearum species complex.
► Rolling Circle Amplification (RCA) probes specific for different Fusarium spp.
► Identification of Fusarium in pure culture and contaminated wheat samples.
► Gel electrophoretic and direct visualization of RCA products.
► Sensitive and cost effective identification of Fusarium Head Blight species.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Microbiological Methods - Volume 89, Issue 1, April 2012, Pages 63–70
نویسندگان
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