Identification of Monilinia fructigena, M. fructicola, M. laxa, and Monilia polystroma on Inoculated and Naturally...

Area of Science:

• Plant Pathology
• Molecular Biology
• Agricultural Science

Background:

• Quarantine regulations necessitate accurate identification of Monilinia fructigena, M. fructicola, M. laxa, and Monilia polystroma species in fruit trade.
• Existing identification methods may not be sufficiently rapid or specific for effective monitoring of imported and exported fruits.

Purpose of the Study:

• To develop and validate a timely and accurate molecular identification method for differentiating key Monilinia and Monilia species.
• To establish a multiplex polymerase chain reaction (PCR) assay for simultaneous detection of multiple quarantined fruit pathogens.

Main Methods:

• Random amplified polymorphic DNA (RAPD) analysis was employed to identify species-specific DNA markers.
• Sequencing of RAPD products was performed to design primers for PCR-based detection.
• A multiplex PCR assay was developed using a common reverse primer and species-specific forward primers.

Main Results:

• A multiplex PCR assay was successfully established, differentiating M. fructigena (402 bp), M. fructicola (535 bp), and M. laxa (351 bp).
• The method also identified Monilia polystroma (425 bp) and demonstrated specificity when tested on inoculated apples.
• The multiplex PCR accurately identified Monilinia species in naturally infected stone fruits, confirming its practical applicability.

Conclusions:

• The developed multiplex PCR method provides a rapid, specific, and reliable tool for identifying quarantined Monilinia and Monilia species.
• This assay is suitable for routine monitoring of imported/exported fruits and diagnosing infections in stone fruits.
• The method supports regulatory efforts to control the spread of economically important fruit pathogens.