کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
2092676 1081808 2010 7 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Detection of Thielaviopsis basicola in soil with real-time quantitative PCR assays
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی بیوتکنولوژی یا زیست‌فناوری
پیش نمایش صفحه اول مقاله
Detection of Thielaviopsis basicola in soil with real-time quantitative PCR assays
چکیده انگلیسی

SummaryThielaviopsis basicola is a soil-borne fungus with a wide host range and a cosmopolitan distribution. It causes disease on many agricultural crops and in China it is the causal agent of black root rot on tobacco plant. Early diagnosis and detection of the pathogen in soil are critical to control this disease in field. The objective of this study was to develop sensitive and effective methods suitable for large-scale detection and quantification of T. basicola. Based on the nucleotide sequences of the internal transcribed spacer (ITS) regions of rDNA genes of Thielaviopsis spp, primers and TaqMan probe were designed specifically to amplify DNA from T. basicola and real-time, quantitative PCR (qPCR) assays were developed for rapid, specific and sensitive detection and quantification of T. basicola. It was sensitive with the detection limit of 100 fg μl−1 genomic DNA of T. basicola in qPCR assays. By combining the qPCR assays with the efficient protocol to extract DNA from soil, it was possible to achieve real-time detection of T. basicola in soil in 4–5 h and the detection limit of 3 conidia per reaction in qPCR was recorded. The assays were applied to survey soils from tobacco fields in China for the presence of T. basicola and the analyses of naturally infested soil showed the reliability of the qPCR assays.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Microbiological Research - Volume 165, Issue 5, 20 July 2010, Pages 411–417
نویسندگان
, ,