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A Nested Polymerase Chain Reaction Protocol for in planta Detection of Fusicladium eriobotryae , Causal Agent of Loquat Scab
Author(s) -
GonzálezDomínguez Elisa,
León Maela,
Armengol Josep,
Berbegal Mónica
Publication year - 2015
Publication title -
journal of phytopathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.53
H-Index - 60
eISSN - 1439-0434
pISSN - 0931-1785
DOI - 10.1111/jph.12291
Subject(s) - biology , polymerase chain reaction , nested polymerase chain reaction , primer (cosmetics) , fungus , microbiology and biotechnology , real time polymerase chain reaction , virology , gene , genetics , botany , chemistry , organic chemistry
Scab caused by the fungus F usicladium eriobotryae is the most serious disease affecting loquat in S pain. Isolation of F . eriobotryae from infected tissue on culture media can be difficult due to its slow growth. A polymerase chain reaction ( PCR )‐based protocol was developed for F . eriobotryae ‐specific identification from pure culture or infected loquat tissues. The primer set was designed in the elongation factor 1‐ α gene ( EF 1‐ α ), and specificity and sensitivity for single and nested PCR were validated. The nested PCR assay resulted in 100% positive detection of F . eriobotryae in naturally and artificially infected tissues. This protocol can be useful for routine diagnosis, disease monitoring programmes and epidemiological research.