Abstract
Apple bull’s eye rot is caused by pathogenic Neofabraea species including N. malicorticis, N. perennans and N. alba. Fruits carrying this fungal quarantine disease are prohibited from entering China. The host plants for the pathogens include several Rosaceae fruits including apple and pear. Disease symptoms and pathogen morphology are often insufficient to determine the identity of the pathogen, particularly at the species level. In the current study, we analyzed the inter-species sequence variations in the β-tublin gene, and designed specific primers to allow PCR amplification of 554 bp fragments from pathogenic Neofabraea species. The PCR products were recovered and sequenced, and Blast search was conducted using the DNA sequences in the Genbank database. The results indicated precise PCR amplification of the target sequences from the host pathogen, which allowed unambiguous identification of the species.
Highlights
Apple anthracnose (Bull’s Eye Rot) has been a longstanding problem in the European and American apple industry for over a century [1], and contaminated fruits have been prohibited to enter China to prevent the fungal quarantine disease
This study used bacterial strains obtained from American Type Culture Collection (ATCC), Chinese Academy of Inspection and Quarantine (CAIQ), as well as isolates collected by Liaoning Entry-Exit Inspection and Quarantine Bureau (LNCIQ) from external sources (Table 1)
Faint bands were detected in a few strains, the sizes were clearly distinct from 554 bp. These results indicated that the β-tublin PCR primers were highly specific for the pathogenic Neofabraea species responsible for apple bull’s eye rot
Summary
Apple anthracnose (Bull’s Eye Rot) has been a longstanding problem in the European and American apple industry for over a century [1], and contaminated fruits have been prohibited to enter China to prevent the fungal quarantine disease. In recently year, it has occurred regularly overseas and caused significant economic losses. The pathogens causing apple bull’s eye rot are primarily three Neofabraea species, namely N. malicorticis, N. perennans and N. alba [3,4]. In 2009, we successfully applied this method and identified the pathogen isolated from US-imported apple as N. perennans
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