Within a certification program of citrus, a survey covering 14 citrus fields not previously screened were randomly selected in seven areas of the Campania Region of southern Italy to evaluate the presence and distribution of citrus viroids in the region. During the last 3 years, 112 trees were visually inspected and sampled including 14 ‘Navelina’, 8 ‘Washington navel’, 13 ‘Biondo Commune’, 10 ‘Tarocco’, 6 ‘Valencia late’, 1 ‘Sanguinello’, and 1 ‘Vanilla’ sweet oranges (Citrus sinensis), four sour oranges (C. aurantium), 21 ‘Commune’ and 7 ‘Monreale’ clementines (C. clementina), 8 ‘Common’ and 2 ‘Avana’ mandarins (C. reticulata), 8 ‘Ovale di Sorrento’ and 6 ‘Zagara’ lemons (C. lemon), 2 Tangelos (C. reticulata × C. paradisi), and 1 grapefruit (C. paradisi). Nearly all (37 of 38) mandarin and clementine trees showed cachexia-like symptoms. Severe bark scaling that is characteristic of exocortis disease was observed on a single ‘Ovale di Sorrento’ lemon, the only tree incidentally found to be grafted onto the exocortis-sensitive trifoliate orange (Poncirus trifoliata) rootstock, since sour orange (C. aurantium) is the main rootstock used in Campania. Samples (104 of 112) that had been graft-inoculated into Arizona 861-S1 ‘Etrog citron’ (C. medica L.) on rough lemon (C. jambhiri Lush.) rootstock showed typical viroid symptoms. Viroid infection was confirmed by sequential polyacrylamide gel electrophoresis analysis (2) and molecular hybridization of imprinted membranes (1). Citrus exocortis viroid (CEVd), Hop stunt viroid (HSVd), and Citrus viroid III (CVd-III) were the most frequently detected, accounting respectively, for 67.9, 86.6, and 84.8% of the infected samples. Citrus viroid IV (CVd-IV) and Citrus bent leaf viroid (CBLVd) were found in 24.1 and 13.4% of the sources. CBLVd was detected in sweet orange (‘Tarocco’ and ‘Biondo Commune’) and lemon (‘Ovale di Sorrento’ and ‘Zagara’), while CVd-IV was detected in sweet orange (‘Tarocco’, ‘Biondo Commune’, and ‘Washington navel’), lemon (‘Ovale di Sorrento’ and ‘Zagara’), clementine (‘Commune’), and sour orange. Most trees (82.1%) were infected with more than one viroid species. The most frequent viroid combinations were CEVd + HSVd + CVd-III (40.2%), HSVd + CVd-III (16.1%), and CEVd + CBLVd + HSVd + CVd-III + CVd-IV (12.5%). To our knowledge, this is the first report of CEVd, CBLVd, HSVd, CVd-III, and CVd-IV in the Campania Region and the first report of CBLVd and CVd-IV in Italy. References: (1) A. Palacio et al. Eur. J. Plant Pathol.105:897, 2000. (2) R. F. Rivera-Bustamante et al. Anal. Biochem. 156:91, 1986.
A rapid isothermal assay for the detection of Hop stunt viroid in hop plants (Humulus lupulus), and its application in disease surveys