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Kasvitaudin sairastuttamia mansikanlehtiä

Application of recombinase polymerase amplification to diagnosis of Phytophthora diseases of strawberry

  • Environment and natural resources

Phytophthora is an increasing threat to agriculture and natural ecosystems. The Irish potato famine of the mid-1800s which lead to the death of a million humans in Europe and the recent decline of oak forest in California and kauri forests in New Zealand were all caused by Phytophthora species.

Similarly, strawberry crop also suffers heavy losses from Phytophthora fragariae and P. cactorum. Projects aiming at the eradication of Phytophthora have remained largely unsuccessful, however, projects aiming at the prevention of the Phytophthora spread through blocking initial introduction have the highest potential for success.

The aim of MSc Mustafa Munawar’s dissertation is to develop rapid detection means for strawberry Phytophthora pathogens which can assist in halting the spread of the pathogens. Mustafa utilized recombinase polymerase amplification (RPA) technology for the purpose. RPA is a rapid isothermal nucleic acid amplification technology that enables the amplification of target nucleic acid directly from crudely macerated plant tissues, excluding the need for laborious plant DNA extraction. Being isothermal, RPA can be done with a small portable device on-site and does not require expensive thermocyclers.

Mustafa Munawar’s dissertation describes devolvement of two rapid real-time RPA assays for P. cactorum and P. fragariae. Both assays can detect as low as 10 fg Phytophthora genomic DNA with reaction turnaround time less than 30 minutes. Moreover, the dissertation also presents a combination of RPA and PCR technologies (RPA-PCR couple) to expedite plant diagnostics and overcome amplification inhibitors. The technologies combination is expected to have vast applications, especially for amplifying targets from recalcitrant samples such as ancient samples, woody plants, or food.

The doctoral dissertation of MSc Mustafa Munawar, entitled  Application of recombinase polymerase amplification to diagnosis of Phytophthora diseases of strawberry will be examined at the Faculty of Science and Forestry on the 15th January 2021 in Kuopio and online. The opponent in the public examination will be Associate Professor Timo Hytönen, University of Helsinki, and the custos will be Professor Elina Oksanen, University of Eastern Finland. The public examination will be held in English.

Link to the event

Link to the dissertation