Application of multi-phase experiments in plant pathology to identify genetic resistance to Diaporthe toxica in Lupinus albus

Phenotyping assays in plant pathology using detached plant parts are multi-phase experimental processes. Such assays involve growing plants in field or controlled-environment trials (Phase 1) and then subjecting a sample removed from each plant to disease assessment, usually under laboratory conditions (Phase 2). Each phase may be subject to non-genetic sources of variation. To be able to separate these sources of variation in both phases from genetic sources of variation requires a multi-phase experiment with an appropriate experimental design and statistical analysis. To achieve this, a separate randomization is required for each phase, with additional replication in Phase 2. In this article, Phomopsis leaf and pod blight (caused by Diaporthe toxica ) of Lupinus albus was used as a case study to apply a multi-phase experimental approach to identify genetic resistance to this pathogen, and demonstrate the principles of sound experimental design and analysis in detached plant part assays. In seven experiments, 250 breeding lines, cultivars, landraces, and recombinant in-bred lines from a mapping population of L. albus were screened using detached, inoculated leaves, and/or pods. The experimental, non-genetic variance in Phase 2 varied in magnitude compared to the Phase 1 experimental, non-genetic variance. The reliability of prediction for resistance to Phomopsis pod blight was high (mean of 0.70 in seven experiments), while reliability of prediction for leaf assays was lower (mean 0.35–0.51 depending on the scoring method used).