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A rapid PCR method to discriminate between Tomato yellow leaf curl virus isolates
Author(s) -
MARTÍNEZCULEBRAS PEDRO VICENTE,
FONT ISABEL,
JORDÁ CONCEPCIÓN
Publication year - 2001
Publication title -
annals of applied biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.677
H-Index - 80
eISSN - 1744-7348
pISSN - 0003-4746
DOI - 10.1111/j.1744-7348.2001.tb00401.x
Subject(s) - tomato yellow leaf curl virus , biology , begomovirus , leaf curl , geminiviridae , virology , plant virus , virus
Summary Tomato yellow leaf curl virus (TYLCV) was recently divided into two different species: Tomato yellow leaf curl virus ‐Israel (TYLCV‐Is) and Tomato yellow leaf curl virus ‐Sardinia (TYLCV‐Sar). There are no rapid methods by which TYLCV viruses may be assigned to either TYLCV‐Is or TYLCV‐Sar species. In the present work, using an extensive alignment of begomovirus sequences, TYLCV‐specific primers were designed and tested which allow the specific amplification of DNA fragments from any isolate of TYLCV. Also, a primer was designed and tested which allows the specific amplification of TYLCV‐Sar. Furthermore, a combination of these primers was selected to develop a duplex PCR method, which has the potential to detect either TYLCV‐Is or TYLCV‐Sar. The PCR methods were also highly effective with minimal sample preparation and allowed direct amplification of TYLCV from infected leaf extracts. This approach may be used in the laboratory as a tool for rapid, large‐scale diagnostics of TYLCV‐infected samples.