Project description:Root rot pathogens restrict pea and wheat production globally. In the EU, pea and pea-based cereal mixtures are being promoted; however, root rot pathogen dynamics in such mixtures are poorly understood. Winter pea and wheat were grown either in pure stands or in mixtures in the field in western France, and the severity of root rot in pea, wheat, and their mixtures, as well as the key pathogens associated with these crops, were assessed. Disease severity was moderate in pea and low in wheat, with no effect of sowing pattern. Didymella pinodella, a previously unreported pathogen in the pea-root rot complex in France, emerged as the most dominant pathogen in pea. It also occurred in low frequencies in wheat. Subsequent greenhouse aggressiveness tests showed that ten of the commonly grown pea cultivars in France lack resistance to D. pinodella. Among the Fusarium spp. isolated, F. avenaceum was the most frequent, occurring at similar frequencies in pea and wheat. In conclusion, D. pinodella may be an important pea root rot pathogen in France and there is a lack of resistance in the tested pea cultivars. In addition, F. avenaceum is a shared pathogen of wheat and pea.
Project description:Didymella pinodella is the major pathogen of the pea root rot complex in Europe. This wide host range pathogen often asymptomatically colonizes its hosts, making the control strategies challenging. We developed a real-time PCR assay for the detection and quantification of D. pinodella based on the TEF-1 alpha gene sequence alignments. The assay was tested for specificity on a 54-isolate panel representing 35 fungal species and further validated in symptomatic and asymptomatic pea and wheat roots from greenhouse tests. The assay was highly consistent across separate qPCR reactions and had a quantification/detection limit of 3.1 pg of target DNA per reaction in plant tissue. Cross-reactions were observed with DNA extracts of five Didymella species. The risk of cross contamination, however, is low as the non-targets have not been associated with pea previously and they were amplified with at least 1000-fold lower sensitivity. Greenhouse inoculation tests revealed a high correlation between the pathogen DNA quantities in pea roots and pea root rot severity and biomass reduction. The assay also detected D. pinodella in asymptomatic wheat roots, which, despite the absence of visible root rot symptoms, caused wheat biomass reduction. This study provides new insights into the complex life style of D. pinodella and can assist in better understanding the pathogen survival and spread in the environment.
Project description:Ascochyta blight is one of the most destructive diseases in field pea and is caused by either individual or combined infections by the necrotrophic pathogens Peyronellaea pinodes, Didymella pinodella, Ascochyta pisi and Ascochyta koolunga. Knowledge of disease epidemiology will help in understanding the resistance mechanisms, which, in turn, is beneficial in breeding for disease resistance. A pool of breeding lines and cultivars were inoculated with P. pinodes and D. pinodella to study the resistance responses and to characterize the underlying resistance reactions. In general, phenotypic analysis of controlled environment disease assays showed clear differential responses among genotypes against the two pathogens. The released variety PBA Wharton and the breeding line 11HP302-12HO-1 showed high levels of resistance against both pathogens whereas PBA Twilight and 10HP249-11HO-7 showed differential responses between the two pathogens, showing higher resistance against D. pinodella as compared to P. pinodes. OZP1604 had high infection levels against both pathogens. Histochemical analysis of leaves using diamino benzidine (DAB) showed the more resistant genotypes had lower accumulation of hydrogen peroxide compared to susceptible genotypes. The digital images of DAB staining were analyzed using ImageJ, an image analysis software. The image analysis results showed that quantification of leaf disease infection through image analysis is a useful tool in estimating the level of cell death in biotic stress studies. The qRT-PCR analysis of defense related genes showed that partially resistant genotypes had significantly higher expression of PsOXII and Pshmm6 in the P. pinodes treated plants, whereas expression of PsOXII, PsAPX1, PsCHS3 and PsOPR1 increased in partially resistant plants inoculated with D. pinodella. The differential timing and intensity of expression of a range of genes between resistant lines challenged with the same pathogen, or challenged with different pathogens, suggests that there are multiple pathways that restrict infection in this complex pathogen-host interaction. The combination of phenotypic, histochemical and molecular approaches provide a comprehensive picture of the infection process and resistance mechanism of pea plants against these pathogens.