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Development of species specific SCAR based molecular marker for the detection of Pseudocercospora eumusae, the causal agent of eumusae leaf spot disease in banana

  • Perumal Ganga Devi
  • Raman ThangaveluEmail author
Original Article
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Abstract

Eumusae leaf spot disease caused by Pseudocercopra eumusae is one of the destructive leaf spot diseases in banana. A diagnostic method based on conventional and quantitative real-time PCR (qPCR) assay was developed in the present study for the specific detection of P. eumusae from pure cultures and leaf spot tissues. The sequence characterized amplified region (SCAR) based molecular marker and TaqMan probe was designed and validated using the genomic DNA of different Pseudocercopra spp. and 11 other leaf spot pathogens of banana. The primers and TaqMan probe proved to be highly specific to the target pathogen P. eumusae and no cross reactivity was encountered with any of the other leaf spot pathogens tested. Both conventional and qPCR assay developed detected the target pathogen P. eumusae directly from single leaf spot tissue and thus simplifies the identification by eliminating the need for culturing of pathogen. However, the sensitivity of qPCR (1 pg/μL) is higher compared to conventional PCR (100 pg/μL). To the best of our knowledge, this is the first report on the use of SCAR based molecular marker for the specific and early detection of P. eumusae in banana.

Keywords

Pseudocercospora eumusae SCAR primers Conventional PCR qPCR assay TaqMan probe 

Notes

Acknowledgements

The authors are grateful to the Director, NRC for Banana and Tiruchirappalli for providing facilities to carry out this research work and also grateful to CIRAD, France for the supply of DNA of Mycosphaerella spp. for the study. The authors are also grateful to Dr. Jeff Daniells, Agri-Science Queensland, Australia, for carrying out English editing of this manuscript.

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Copyright information

© Società Italiana di Patologia Vegetale (S.I.Pa.V.) 2019

Authors and Affiliations

  1. 1.Division of plant pathologyICAR – National Research Centre for BananaTiruchirappalliIndia

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