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Typing of Tomato Yellow Leaf Curl Viruses in Europe
Authors:Gian Paolo Accotto  Jesús Navas-Castillo  Emanuela Noris  Enrique Moriones  Diamantina Louro
Affiliation:(1) Istituto di Fitovirologia Applicata, CNR, Strada delle Cacce 73, 10135 Torino, Italy;;(2) Estación Experimental "ldquo"La Mayora"rdquo", CSIC, 29750 Algarrobo-Costa, Málaga, Spain;(3) Istituto di Fitovirologia Applicata, CNR, Strada delle Cacce 73, 10135 Torino, Italy;(4) Protecçâo das Culturas, Quinta do Marquês, 2780 Oeiras, Portugal
Abstract:Tomato yellow leaf curl disease is spreading in southern Europe, where it has quickly become a serious problem. In recent years, several virus isolates have been characterised. Although with some genetic variability, all isolates found in Europe belong to one of two species Tomato yellow leaf curl-Sardinia (TYLCV-Sar) or Tomato yellow leaf curl-Israel (TYLCV-Is). Several methods were tested to identify and type TYLCV isolates from field samples: (1) RFLP of a DNA fragment amplified from the coat protein gene; (2) PAGE of a fragment amplified from the C2 gene; (3) dot-blot hybridisation. All methods enabled the detection of the TYLCVs and provided good indications for attributing them to one species or the other. However, for typing purposes, the RFLP method was the most reliable, due to the easily recognisable pattern produced by the two virus species present in Europe. Dot-blot hybridisation is less expensive for identifying TYLCVs in large numbers of samples, particularly when a mixture of two probes is used. PAGE of the C2 fragment is the fastest of the methods tested.
Keywords:geminivirus  RFLP  PCR  PAGE  differentiation  dot-blot
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