The rymv1-3 allele of the eIF(iso)4G-mediated resistance to Rice yellow mottle virus (RYMV) is found in a few Oryza glaberrima cultivars. The same resistance-breaking (RB) mutations emerged in the central domain of the VPg after inoculation of isolates of different strains. The RB mutations were fixed, often sequentially, at codons 41 and 52 which paralleled an increase in virus accumulation. RB mutations also emerged after inoculation of an avirulent infectious clone, indicating that they were generated de novo in resistant plants. Only virus isolates with a threonine at codon 49 of the VPg broke rymv1-3 resistance, those with a glutamic acid did not. A small subset of these isolates overcame rymv1-2 resistance, but following a specific pathway. Comparison with the RB process of rymv1-2, a resistance allele found in a few Oryza sativa cultivars, showed similarities in the mode of adaptation but revealed converse virulence specificity of the isolates.

Long-term cell cultures were used in coffee to study the cytological, genetic and epigenetic changes occurring during cell culture ageing. The objective was to identify the mechanisms associated with somaclonal variation (SV). Three embryogenic cell lines were established in Coffea arabica (2n = 4x = 44) and somatic seedlings were regenerated after 4, 11 and 27 months. Phenotyping and AFLP, MSAP, SSAP molecular markers were performed on 199 and 124 plants, respectively. SV were only observed from the 11 and 27-month-old cultures, affecting 30 and 94 % of regenerated plants, respectively. Chromosome counts performed on 15 plants showed that normal plants systematically displayed normal chromosome numbers and that, conversely, aneuploidy (monosomy) was systematically found in variants. The allopolyploid structure of C. arabica allowed aneuploid cells to survive and regenerate viable plants. No polymorphic fragments were observed between the AFLP and SSAP electrophoretic profiles of mother plants and those of the in vitro progeny. Methylation polymorphism was low and ranged between 0.087 and 0.149 % irrespective of the culture age. The number of methylation changes per plant-normal or variant-was limited and ranged from 0 (55-80 % of the plants) to 4. The three cell lines showed similar SV rate increases during cell culture ageing and produced plants with similar molecular patterns indicating a non random process. The results showed that cell culture ageing is highly mutagenic in coffee and chromosomal rearrangements are directly linked to SV. Conversely, the analysis of methylation and transposable elements changes did not reveal any relation between the epigenetic patterns and SV.

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 T9, 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 T9, 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.First reported 50 years ago, Rice yellow mottle virus (RYMV) is a major biotic constraint to rice cultivation in Africa (Séré et al. 2013). RYMV is a viral species of genus Sobemovirus that is responsible for high rice production losses in agroecosystems in most rice-growing countries of Africa (Kouassi et al. 2005;Traoré et al. 2015). Highly adapted to the two cultivated rice species, Asian rice Oryza sativa and African rice O. glaberrima, RYMV has a narrow host range that also includes other wild Poaceae species (Bakker 1974). RYMV possesses a single-stranded RNA genome that is organized into five open reading frames (ORF) (Fig. 1A). Highly diverse, RYMV is classified into six major strains with a strong geographical distribution. Strains S1 and S2/S3 are found in West and West-Central Africa whereas strains S4, S5, and S6 are present exclusively in East Africa (Pinel-Galzi et al. 2015). This spatial pattern of RYMV diversity is explained by the absence of seedborne transmission and of long-distance movement (Allarangaye et al. 2006;Fargette et al. 2006;Konaté et al. 2001). In addition to short-distance propagation that is mainly mediated by beetles, RYMV is transmitted by contact during agricultur...

Meloidogyne graminicola is a facultative meiotic parthenogenetic root-knot nematode (RKN) that seriously threatens agriculture worldwide. We have little understanding of its origin, genomic structure, and intraspecific diversity. Such information would offer better knowledge of how this nematode successfully damages rice in many different environments. Previous studies on nuclear ribosomal DNA (nrDNA) suggested a close phylogenetic relationship between M. graminicola and Meloidogyne oryzae, despite their different modes of reproduction and geographical distribution. In order to clarify the evolutionary history of these two species and explore their molecular intraspecific diversity, we sequenced the genome of 12 M. graminicola isolates, representing populations of worldwide origins, and two South American isolates of M. oryzae. k-mer analysis of their nuclear genome and the detection of divergent homologous genomic sequences indicate that both species show a high proportion of heterozygous sites (ca. 1–2%), which had never been previously reported in facultative meiotic parthenogenetic RKNs. These analyses also point to a distinct ploidy level in each species, compatible with a diploid M. graminicola and a triploid M. oryzae. Phylogenetic analyses of mitochondrial genomes and three nuclear genomic sequences confirm close relationships between these two species, with M. graminicola being a putative parent of M. oryzae. In addition, comparative mitogenomics of those 12 M. graminicola isolates with a Chinese published isolate reveal only 15 polymorphisms that are phylogenetically non-informative. Eight mitotypes are distinguished, the most common one being shared by distant populations from Asia and America. This low intraspecific diversity, coupled with a lack of phylogeographic signal, suggests a recent worldwide expansion of M. graminicola.

Nine accessions of wild Coffea arabica from Ethiopia were evaluated for resistance to Meloidogyne paranaensis. Two wellcharacterized susceptible and resistant cultivars were used as comparative controls. The experiments were conducted in a growth chamber using a clonal population of M. paranaensis (esterase phenotype P1) originating from Brazil. Resistance and susceptibility to the nematode were evaluated using the number of nematodes (eggs and J2) per plant, number of nematodes per gram of root and the reproduction factor (RF). All wild coffee accessions expressed a resistance response to M. paranaensis similar to that of the resistant control Nemaya (RF < 1.0). These results provide coffee breeders with material whose resistance can be transferred into commercial cultivars. Keywords: coffee, root-knot nematode, pathogenicity. RESUMO Avaliação da resistência de cafeeiros silvestres (Coffea arabica) a Meloidogyne paranaensisForam avaliados quanto à resistência a Meloidogyne paranaensis, nove acessos de cafeeiros silvestres incluindo dois cultivares bem caracterizados como testemunhas de suscetibilidade e resistência. Os experimentos foram realizados sob condições controladas em câmara de crescimento, utilizando uma população clonal de M. paranaensis (fenótipo de esterase P1), proveniente do Brasil. A resistência e a suscetibilidade ao nematóide foram avaliadas com base no número total de nematóides por planta (ovos + J2) e por grama de raiz e no fator de reprodução (FR). Todos os acessos mostraram resposta à infecção por M. paranaensis similar à da testemunha resistente (FR < 1,0). Com esses resultados, novos materiais, cuja resistência pode ser transferida aos cultivares comerciais, ficam disponíveis para os fitomelhoristas. Palavras-chave: café, nematóide das galhas, patogenicidade.

Root-knot nematodes (Meloidogyne spp.) threaten the livelihood of millions of farmers producing coffee worldwide. The use of resistant plants either as cultivars or rootstocks appears to be the single most effective method of control. A screening method was developed to evaluate large populations of plants for resistance to root-knot nematodes. Two coffee cultivars, one susceptible and the other resistant to Meloidogyne paranaensis, were grown under controlled conditions in two substrates: a commercial sieved potting compost and an inert substrate containing sand with a water-absorbent synthetic polymer. Plant growth and development and nematode multiplication were compared for two inoculation dates (2 and 8 weeks after planting) and two evaluation dates (eight and 13 weeks after inoculation). Root growth, but not nematode multiplication, was influenced by the choice of substrate. Evaluation of the differences in root weight and nematode numbers between the different cultivars, substrates and dates of inoculation suggested that an optimal condition could be defined. The best discrimination between susceptible and resistant plants was found in the experiment where inoculation occurred at 2 weeks after planting and evaluation occurred at 8 weeks after inoculation. Because the total duration of this experiment was only 3 months, highthroughput evaluation was possible, opening up new possibilities for screening large germplasm collections and studying the genetic control of root-knot nematode resistance in coffee.

Rice yellow mottle virus in Senegal is reported here for the first time. The near-complete genomic sequences of two isolates (Se1 and Se5) were obtained. A comparison with 18 sequences from West Africa revealed a new cluster with an isolate from Gambia, located at a basal position in the phylogenetic tree.

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