PJB-2012-222
MOLECULAR DETECTION OF FUSARIUM OXYSPORUM IN THE INFECTED CUCUMBER PLANTS AND SOIL
SHUMEI ZHANG1,2, XIAOYU ZHAO2, YUXIA WANG2, JING LI2, XIULING CHEN1, AOXUE WANG1* AND JINGFU LI1*
Abstract
In this study, a one-step PCR protocol was developed for rapid and accurate detection of the pathogen Fusarium oxysporum in infected cucumber plants and soil. The primers Fc-1 and Fc-2 were designed according to F. oxysporum internal transcribed spacer (ITS) regions of ribosomal DNA (rDNA). A specific 315-bp PCR product was amplified from all the tested F. oxysporum isolates，infected cucumber plants and soil under the optimized PCR conditions using primers Fc-1 and Fc-2. While no PCR product was obtained from other fungi, bacteria, healthy cucumber plants and non-infected soil. For the detection sensitivity, the minimal quantity of genomic DNA of purified F. oxysporum was 100fg and that of soil pathogens was 1000 spores per gram of soil. Furthermore, the PCR protocol enabled detection of F. oxysporum in symptomless cucumber root 6 days after inoculation with the pathogen. Therefore, this PCR-based method can be used to detect F. oxysporum rapidly, sensitively with reliability in infected cucumber plants and soil. Our detection protocol also allowed for early monitoring and diagnosis of F. oxysporum to facilitate disease management.
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