Abstract

GUERRERO GAMEZ, César Enrique et al. Detection of Three Citrus Viroids Species From Nuevo Leon and Tamaulipas, Mexico by Conventional and Real Time RT-PCR. Rev. mex. fitopatol [online]. 2013, vol.31, n.1, pp.20-28. ISSN 2007-8080.

Three protocols were developed based on realtime RT-PCR with SYBR Green system and were compared with conventional RT-PCR for detection of the Citrus exocortis viroid (CEVd), Hop stunt viroid (HSVd) and Citrus dwarf viroid (CDVd) using specific primers combinations; additionally, we conducted the analysis of citrus viroids from 90 citrus threes samples collected in orchards in the northeast of Mexico. Evaluated citrus samples comprised different species from the municipalities of General Teran, Montemorelos and Marin, Nuevo Leon, besides Rio Bravo, Victoria city and Hidalgo, Tamaulipas. Total RNA was extracted from the samples and analyzed by the techniques of multiple RT-PCR and real time RT-PCR with the SYBR Green system. We detected CEVd, HSVd and CDVd viroids individually and jointly in the samples. Using real-time RT-PCR with SYBR Green, we found a 41 % of CEVd positive samples, 42 % for HSVd and 49 % to CDVd. Comparing endpoint multiplex RT-PCR with realtime RT-PCR techniques, minimal differences were obtained in the results.

Keywords : RT-PCR; real-time RT-PCR; SYBR Green; CEVd; HSVd; CDVd.

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