Suitable Methods of Inoculation and Quantification of Fusarium Root Rot in Lentil

Abstract

Lentil (Lens culinaris L. subsp. culinaris) is an important grain legume grown worldwide. As its popularity grows among consumers and more acres are produced, new root rot complexes have become more prevalent. This work sought to develop methods for studying root rot caused by Fusarium avenaceum in lentil using controlled environments. The objectives were to (i) find an effective and seed-safe sterilization technique, (ii) optimize the inoculation technique and lentil growing environment, and (iii) develop visual and automated disease scoring systems. Results showed the use of detergent and a low concentration (0.1%) of NaClO (the active ingredient in bleach) maintained germinability and effectively eliminated bacterial and fungal contamination on seeds. Other treatments, such as ethanol, reduced seed germination or failed to kill pathogenic fungi such as Fusarium spp. Placing inoculum at a moderate rate of 1 × 106 spores both directly on the seed and on top of the media covering the seed improved severity scores and reduced escapes compared with placement on top of the media only. Visual severity scoring systems and diagrammatic scales were developed for scoring the cotyledon region and roots. A computer vision algorithm was designed to improve the efficiency of scoring the cotyledon region and roots for disease severity using a simple RGB camera and lightbox. Visual and computer scores were best correlated when images were visually scored on a monitor, and multiple images were averaged. The scores generated from the computer vision algorithm had better correlations with visual scores for cotyledon rot (r = 0.92 and β1 = 0.96) than root rot (r = 0.62 and β1 = 0.67).