Molecular characterization of new viral agents causing old citrus diseases by deep sequencing of small RNAs
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Cita bibliográficaVelázquez, K., Hervàs, J., Guerri, J., Vives, M.C. (2017). Molecular characterization of new viral agents causing old citrus diseases by deep sequencing of small RNAs. In V Simposio Internacional de Fruticultura Tropical y Subtropical.
There are some graft-transmissible citrus diseases such as incompatibility of varieties grafted on trifoliate rootstocks, impietratura and cristacortis that, although they have been described for many years in many citrus growing areas, their causal agent is still unknown and rapid detection methods could not be developed. Diagnosis can only be made by means of infectivity assays on indicator pl ants, resulting slow, expensive and non-specific and pose a limitation for the analysis of a large number of samples. These diseases are present in Spain and their diagnosis is mandatory in Sanitation and Certification Programs. To identify the causal agent asso ciated to such diseases and develop specific molecular detection methods, the small RNAs (sRNAs) fraction, extracted from Etrog citron plants infected with each of the above diseases, was analyzed by deep sequencing. The assembly of the sRNAs derived from each sample allowed the obtaining of several sequence fragments that were present in infected plants and not in healthy ones. In the case of incompatibility on trifoliate rootstocks disease, we determined the complete genome sequence of a new virus provisionally nam ed Trifoliate citrus bud union associated virus (TCBUaV), that showed homology with species of th e genus Cytorhabdovirus, whereas for impietratura and cristacortis diseases we obtained partial sequences of individual viral genomes that had no homology with an y virus available in the databases. Obtaining specific sequences of each pathogen has allowed us to develop specific and rapid molecular detection methods thereof by RT-PCR