Development and Application of a Multiplex Marker Assay to Detect PVY Resistance Genes in Solanum tuberosum

Gregory L Elison,Jonathan L Whitworth,Darren G Hall,Richard G Novy

doi:10.1007/s12230-020-09777-1

Gregory L Elison, Jonathan L Whitworth + Show 2 more

Open Access

https://doi.org/10.1007/s12230-020-09777-1

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Abstract

Potato virus Y (PVY) is a major pathogen affecting potato production worldwide. Three independent genes (Ryadg,Rysto,and Rychc) conferring genetic resistance to all known strains of the virus are currently utilized by breeding programs to develop potato varieties containing extreme resistance to infection by PVY. These resistance genes are typically detected using primers specific for the generation of markers closely-linked to the resistance genes. In recent years there have been attempts to develop a multiplex PCR assay for all resistance genes, but all current published methods either omit at least one resistance gene or are unnecessarily complex and therefore difficult to incorporate successfully for marker-assisted selection. We have developed an assay which tests for the presence of all three sources of resistance in a single, easy to use PCR protocol. The multiplex PCR assay was applied to potato varieties, second-field year breeding germplasm, and a selection of test crosses having progeny segregating for multiple resistance genes to validate the methodology for use in marker-assisted selection in potato breeding programs.

Highlights

One of the most widespread and damaging potato viruses challenging growers today is Potato virus Y (PVY) which has a worldwide distribution and may cause yield losses of up to 70% (Karasev and Gray 2013)
Leaf samples were collected from 306 breeding clones in 2019, representing second year 12-hill plots of the Aberdeen potato breeding program which represent all material from that field with at least one parent known to be PVY resistant
A multiplex assay was developed with low cost (~ $0.30 / sample after extraction) and high rapidity which allows the simultaneous analysis of the presence of markers associated with three genes conferring extreme resistance (ER) to PVY in potato breeding clones and varieties

Summary

Introduction

One of the most widespread and damaging potato viruses challenging growers today is Potato virus Y (PVY) which has a worldwide distribution and may cause yield losses of up to 70% (Karasev and Gray 2013). Use of pesticides alone to prevent transmission of the virus have proved ineffectual (MacKenzie et al 2014), recent studies with crop oil and insecticides applied in seed crops on a routine schedule (e.g. every 4–5 days) have shown reduction in PVY infections (MacKenzie et al 2017). A more cost-efficient and environmentally friendly approach is to develop potato varieties with genetic resistance to the disease (Solomon-Blackburn and Barker 2001).

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