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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Abbreviations
- AGVd:
-
Australian grapevine viroid
- CCR:
-
Central conserved region
- CEVd:
-
Citrus exotic viroid
- CTAB:
-
Cetyltrimethylammonium bromide
- EDTA:
-
Ethylenediaminetetraacetic acid
- EGME:
-
Ethylene glycol monomethyl ether
- GYSVd-1, 2:
-
Grapevine yellow speckle viroid 1 and 2
- HSVd:
-
Hop stunt viroid
- LAMP:
-
Loop-mediated isothermal PCR
- nt:
-
Nucleotide
- PAGE:
-
Polyacrylamide gel electrophoresis
- PVP-40:
-
Polyvinylpyrrolidone-40
- RT-PCR:
-
Reverse transcriptase polymerase chain reaction
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Acknowledgements
This study was supported by grants from the National Basic Research and Development Program of China (973 Program) (No. 2009CB119200), and National Natural Science Foundation OF China (31071765).
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Zhang, Z., Peng, S., Jiang, D. et al. Development of a polyprobe for the simultaneous detection of four grapevine viroids in grapevine plants. Eur J Plant Pathol 132, 9–16 (2012). https://doi.org/10.1007/s10658-011-9856-5
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DOI: https://doi.org/10.1007/s10658-011-9856-5