Development of a RAPD protocol for typing strains of Phytophthora infestans in Mauritius

Abstract

In Mauritius, substantial decrease in potato yields due to late blight are reported annually and it has become necessary to track the genetic identity of local Phytophthora infestans strains. The Random Amplified Polymorphic DNA (RAPD) technique is a low-cost and simple genetic characterisation tool that has been widely used for molecular fingerprinting but which requires extensive optimisation. The aim of this study was to carry out a series of experiments to optimise a RAPD protocol for routine P. infestans typing. Amplifications performed with DNA template concentration of 30-50 ng/μl, primer concentration of 2.0 − 3.0 μM and MgCl₂ concentration of 3.0 mM gave the best RAPD profiles. These optimised conditions were used to carry out RAPD fingerprinting of 7 P. infestans isolates and codified data were used to construct a consensus dendogram, which grouped the 7 isolates into 3 distinct clusters, consistent with results obtained from other molecular characterisation techniques.

Keywords:

• Phytophthora infestans
• genetic fingerprinting
• rapid amplified polymorphic DNA
• RAPD optimisation
• molecular characterisation

Acknowledgements

The authors are thankful to all the lab staff who helped out during this project.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Additional information

Funding

This work was funded by and carried out at the Biosciences Laboratory, Department of Biosciences and Ocean Studies, Faculty of Science, University of Mauritius.