Crop diseases are responsible for considerable yield losses worldwide and particularly in sub-Saharan Africa. To implement efficient disease control measures, detection of the pathogens and understanding pathogen spatio-temporal dynamics is crucial and requires the use of molecular detection tools, especially to distinguish different pathogens causing more or less similar symptoms. We report here the design a new molecular diagnostic tool able to simultaneously detect five bacterial taxa causing important diseases on rice in Africa: (1) Pseudomonas fuscovaginae, (2) Xanthomonas oryzae, (3) Burkholderia glumae and Burkholderia gladioli, (4) Sphingomonas and (5) Pantoea species. This new detection tool consists of a multiplex PCR, which is cost effective and easily applicable. Validation of the method is presented through its application on a global collection of bacterial strains. Moreover, sensitivity assessment for the detection of all five bacteria is reported to be at 0.5 ng DNA by μl. As a proof of concept, we applied the new molecular detection method to a set of 256 rice leaves collected from 16 fields in two irrigated areas in western Burkina Faso. Our results show high levels of Sphingomonas spp. (up to 100% of tested samples in one field), with significant variation in the incidence between the two sampled sites. Xanthomonas oryzae incidence levels were mostly congruent with bacterial leaf streak (BLS) and bacterial leaf blight (BLB) symptom observations in the field. Low levels of Pantoea spp. were found while none of the 256 analysed samples was positive for Burkholderia or Pseudomonas fuscovaginae. Finally, many samples (up to 37.5% in one studied field) were positive for more than one bacterium (co-infection). Documenting co-infection levels are important because of their drastic consequences on epidemiology, evolution of pathogen populations and yield losses. The newly designed multiplex PCR for multiple bacterial pathogens of rice is a significant improvement for disease monitoring in the field, thus contributing to efficient disease control and food safety.
Rice yellow mottle virus (RYMV) causes high losses to rice production in Africa. Several sources of varietal high resistance are available but the emergence of virulent pathotypes that are able to overcome one or two resistance alleles can sometimes occur. Both resistance spectra and viral adaptability have to be taken into account to develop sustainable rice breeding strategies against RYMV. In this study, we extended previous resistance spectrum analyses by testing the rymv1-4 and rymv1-5 alleles that are carried by the rice accessions Tog5438 and Tog5674, respectively, against isolates that are representative of RYMV genetic and pathogenic diversity. Our study revealed a hypervirulent pathotype, named thereafter pathotype T′, that is able to overcome all known sources of high resistance. This pathotype, which is spatially localized in West-Central Africa, appears to be more abundant than previously suspected. To better understand the adaptive processes of pathotype T′, molecular determinants of resistance breakdown were identified via Sanger sequencing and validated through directed mutagenesis of an infectious clone. These analyses confirmed the key role of convergent nonsynonymous substitutions in the central part of the viral genome-linked protein to overcome RYMV1-mediated resistance. In addition, deep-sequencing analyses revealed that resistance breakdown does not always coincide with fixed mutations. Actually, virulence mutations that are present in a small proportion of the virus population can be sufficient for resistance breakdown. Considering the spatial distribution of RYMV strains in Africa and their ability to overcome the RYMV resistance genes and alleles, we established a resistance-breaking risk map to optimize strategies for the deployment of sustainable and resistant rice lines in Africa.
The isolation, purification and accurate diagnosis of Pantoea and many other bacterial species that infect rice are essential for upstream studies. However, some isolates of Pantoea and other bacteria such as Sphingomonas have similar biochemical and morphological features on common culture media and are thus difficult to isolate selectively and accurately diagnose. We thus developed a semi-selective medium containing 65 g/l (65%) NaCl that allows growth of all Pantoea strains, but inhibits other microorganisms. It can be used to isolate and purify Pantoea spp. for preliminary diagnosis.
Information on comprehensive evaluation of agricultural innovations is often limited. This study provides an overview of multidisciplinary evaluation of a lowland rice variety, WITA 9 (released in Côte d’Ivoire in 1998), with respect to its agronomic performance, grain quality, resistance to diseases, adoption by farmers, impact on productivity and farmers’ income, and marketability. WITA 9 had the highest paddy yield among the tested varieties including an international check (IR 64) and recently developed varieties adapted to this country. WITA 9 had a higher amylose content (26–28%) than others tested. This study confirmed its resistance to bacterial leaf blight, Rice yellow mottle virus (RYMV), and rice blast. A household survey showed that the adoption rate was 24%, its paddy yield advantage was 0.7 t ha–1, and its adoption increased farmer’s income by US$ 91 ha–1 per season. A market study showed that consumers’ willingness to pay was higher for WITA 9 than any other locally produced rice variety and comparable to imported rice in one of two markets. We conclude that WITA 9 is an ideal innovation for enhancing productivity and rice import substitution in Côte d’Ivoire. An effective seed delivery system and enhancing farmers’ and consumers’ awareness of this variety are vital for accelerating impact. Graphical abstract