Development and application of a LAMP method for rapid detection of apple blotch caused by Marssonina coronaria

摘要

Marssonina coronaria is a devastating plant pathogen that causes apple blotch on apple. Early diagnosis of M. coronaria is the key to control apple blotch disease effectively. In this study, we developed a loop-mediated isothermal amplification (LAMP) assay for rapid detection of M. coronaria. The ribosomal DNA internal transcribed spacer (rDNA-ITS) sequence of M. coronaria was selected as the target for primer design. The LAMP reaction was optimal at 60 degrees C for 70 min based on the color change and gel electrophoresis. In specificity tests, the LAMP assay was able to distinguish M. coronaria from other apple pathogenic fungi. In sensitivity tests, the detection limit of the LAMP assay was 100 fg mu L-1 genomic DNA of M. coronaria. Furthermore, the LAMP assay was successfully applied to detect M. coronaria in diseased apple leaf samples from the field. Thus, our study provides a simple and efficient method for quick diagnosis of apple blotch caused by M. coronaria.