Detached-Leaf Method for Propagating Puccinia coronata and Assessing Crown Rust Resistance in Oat.

Abstract

The crown rust pathogen Puccinia coronata is an obligate biotroph with wind-disseminated propagules and numerous races. These characteristics make propagation of single-race cultures difficult. Genetic studies using single races in field and greenhouse environments are also problematic because pure cultures can easily become contaminated. In this study, we developed an isolated propagation system for P. coronata and tested its ability to assess host resistance. Oat (Avena sativa) leaf sections (10 cm each) were harvested, disinfested, and suspended in sterile plastic boxes by enclosing 3.5-cm linear sections of each leaf end between 4% agar blocks amended with various chemical constituents. The exposed sections (approximately 3 cm) were inoculated with P. coronata urediniospores suspended in water. Boxes were sealed and incubated in a lighted growth cabinet until the pathogen sporulated. Viable spores were produced on leaves in all treatments, whereas 6-benzylaminopurine (BAP) and kinetin treatments sustained the leaves longer and yielded the most viable spores. Based on these data, the BAP treatment was adopted and used for additional studies. Detached leaves of differential oat cultivars produced the same reactions as whole plants screened under standard conditions in a growth chamber. The proposed detached-leaf system should be useful for the propagation of numerous single-race cultures of P. coronata as well as evaluation of host resistance under highly controlled conditions.