Evaluation of Ascochyta resistance in chickpea genotypes with quantitative polymerase chain reaction assay

Abstract

Ascochyta blight caused by Ascochyta rabiei is a globally important chickpea disease. Host resistance to Ascochyta blight is considered the most practical and effective means of control, but breeding has been hindered by a lack of effective resistance sources, and time-consuming, labour-intensive traditional methods to screen the resistance level of chickpea genotypes. This paper evaluated the progression of pathogen infection and the disease reaction of chickpea genotypes to Ascochyta blight by traditional and molecular methods. The resistance level of 84 chickpea genotypes was assessed by a quantitative polymerase chain reaction assay (qPCR) using a standard curve produced by various known amounts of pathogen DNA and compared with disease scores based on visual assessments 8 days after inoculation. Disease assessments revealed statistically significant differences between the resistance levels of chickpea genotypes, while the quantity of target DNA in the samples inoculated with the pathogen ranged from 0.004 to 83.37 ng. Our results showed a close relationship between the visual assessment of disease severity and the quantification of the target DNA in chickpea genotypes. The genotypes Tub-35, Tub-47, Tub-26, Tub-82, Tub-65 and Tub-69 were classified as highly resistant to Ascochyta blight based on the results of both assays used for screening chickpea genotypes. This qPCR analysis could be used to quantify disease progression in plant tissues and screen chickpea genotypes as a potential alternative to visual assessment of resistance levels in breeding programmes.