Identification of Monilinia fructigena, M. fructicola, M. laxa, and Monilia polystroma on Inoculated and Naturally Infected Fruit Using Multiplex PCR

Plant Disease
Marie-José CôtéAllison J Meldrum

Abstract

Monilinia fructigena, M. fructicola, M. laxa, and Monilia polystroma each have a different regulatory status. To monitor imported and exported fruit for the presence of quarantined Monilinia or Monilia species, a timely identification method is required. Random amplified polymorphic DNA analysis was used to generate an M. fructigena-specific band that was characterized by sequencing. Using the sequence obtained, primers were designed to amplify bands in the same genomic region of M. fructicola and M. laxa. These bands were also characterized by sequencing. From all three sequences, a multiplex polymerase chain reaction (PCR) method based on a common reverse primer (MO368-5) and three species-specific forward primers (MO368-8R, MO368-10R, and Laxa-R2) was established for the differentiation of the three Monilinia species. The multiplex PCR was tested with additional isolates and consistently produced a 402-bp PCR product for M. fructigena, a 535-bp product for M. fructicola, and a 351-bp product for M. laxa. The method was also used with isolates of the recently characterized Monilia polystroma, and all isolates amplified a 425-bp PCR product. The identification method was shown to amplify a PCR product directly from inoculated ...Continue Reading

References

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Citations

Mar 25, 2020·Microbial Biotechnology·Youming ShenHaifei Li
Jan 30, 2014·World Journal of Microbiology & Biotechnology·Mónika FazekasImre J Holb
May 31, 2013·Phytopathology·Ruifang Gao, Guiming Zhang
Oct 8, 2011·PloS One·Meng-Jun HuChao-Xi Luo
May 1, 2015·Plant Disease·Jovana HrustićBrankica Tanović
May 3, 2016·Journal of Agricultural and Food Chemistry·Leandro Oliveira LinoBénédicte Quilot-Turion
Jul 1, 2017·Studies in Mycology·Y Marin-FelixP W Crous

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