Development of a polyprobe for the simultaneous detection of four grapevine viroids in grapevine plants |
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| Authors: | Zhixiang Zhang Shan Peng Dongmei Jiang Song Pan Hongqing Wang Shifang Li |
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| Institution: | (1) State Key Laboratory of Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Yuanmingyuan West No. 2, 100193 Beijing, China;(2) Institute of Plant Virology, Fujian Agriculture and Forestry University, Fuzhou, 350002, Fujian Province, China;(3) Department of Fruit Science, College of Agronomy and Biotechnology, China Agricultural University, 100193 Beijing, China; |
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| Abstract: | This study aimed to develop a polyprobe for the simultaneous detection of four viroids that infect grapevine: Hop stunt viroid (HSVd), Australian grapevine viroid (AGVd), Grapevine yellow speckle viroid-1 and 2 (GYSVd-1, 2), using a non-isotopic dot blot hybridization technique. A polyprobe was constructed by cloning tandem
full-length sequences of HSVd, AGVd and GYSVd-1 into a single vector. The cRNA polyprobe detected all four viroids with similar
sensitivity to that obtained using individual probes. In addition, samples of 78 varieties from Beijing and Xinjiang were
analyzed using the polyprobe to survey the incidence of grapevine viroids in China. The result demonstrated that grapevine
viroids were detected in 56 (71.8%) varieties. In this study, a rapid, reliable and cost-effective approach to the simultaneous
detection of four grapevine viroids has been developed which has the potential for routine use in quarantine and certification
programs. |
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