Identification and characterization of viroids in 'Navelina ISA 315' sweet orange

To date, seven species of viroids have been described infecting Citrus. However, only Citrus exocortis viroid (CEVd), Hop stunt viroid (HSVd) and Citrus dwarfing viroid (CDVd) have been reported in this crop in Brazil. The aim of our work was to identify and characterize viroids infecting plants of 'Navelina ISA 315' sweet orange cultivar grafted on 'Rangpur' lime from field experiments located in Bebedouro, SP, that showed gumming in the bark and wood symptoms. Biological indexing was done on the indicator host 'Clemelin 1120' tangor plants that reacted with typical gumming symptoms. Viroid infection was assessed by graft-inoculation of buds from the field trees on 'Etrog' citron plants grafted on 'Rangpur' lime, followed by RNA extraction and sPAGE analysis. RNAs were subjected to RT-PCR with primers for citrus viroids. The full-length amplified products were sequenced and compared to those available in the GenBank. The trees were found to be infected with cachexia (Ca) variants of HSVd-Ca or HSVd-Ca plus CDVd. The results indicate that efforts have to be made to increase and stimulate the indexing programs, to maintain plants healthy and to develop sanitary programs focused on reducing the spread of viroids and other graft-transmissible agents.