Establishment and Application of Real-time Quantitative PCR for Detecting of Streptomyces acidiscabies
CHEN Li-Da1, XIE Xue-Wen1, SHI Yan-Xia1, CHAI A-Li1, PAN Hao-Qin2, LI Lei1,*, LI Bao-Ju1,*
1 Insititute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, China; 2 Weifang University of Science and Technology, Shouguang 262700, China
Abstract：Potato common scab, a seed tuber-born disease, is caused by Streptomyces in temperate regions, which seriously threatens the potato (Solanum tuberosum) production and tuber quality. In this study, the specific primer pairs Aci-3/4 were designed according to the toxin gene sequence of Streptomyces acidiscabies, which is one of the pathogens causing potato common scab. A target fragment of 192 bp was specifically amplified using S. acidscabies genomic DNA as a template. The sensitivity of real-time quantitative PCR (qPCR) method obtained in this work showed 1 000 times higher than that of conventional PCR. The results of genomic DNA amplification of the tested samples showed that the cycle threshold (Ct) values in qPCR detection of diseased potato and diseased soil were in the range of 25~32, while conventional PCR did not detect DNA bands in diseased soil. These results suggested that the PCR detection method established in this work had the advantage of strong specificity and high sensitivity, which could quickly detect S. acidiscabies from diseased potato tissues and soil samples. This detection method for potato scab pathogens could provide an effective technical means for early monitoring and effective control of potato scab.
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