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Aiming to evaluate an indirect screening approach for Ceratocystis wilt resistance, the leaf disc (LD) method, two assays were conducted to assess its efficiency in discriminating resistant/susceptible genotypes. The first one used five and the second 16 cacao clones. For each assay, 40 discs of 2month-old leaves were cut, wounded at the midrib, laid down in wet sterilized foam and inoculated (20 μL-3 × 10 4 CFU/ mL suspension of Ceratocystis cacaofunesta spores). The number of perithecia formed per leaf disc was assessed 4 days after inoculation (DAI). To assess the correlation between the LD method and field resistance, 16 branches of the same 16 clones were randomly selected and inoculated (30 μL/ 3 × 10 4 CFU/mL). The lesion length, width and area were measured 60 DAI. In both assays it was possible to distinguish resistant from susceptible clones. The clonal heritabilities were high (h 2 clo ≥ 77.19) for the assays in the lab and in the field, the highest estimate being obtained for perithecia number (h 2 clo = 95.82). The correlation of perithecia number with lesion width was positive and highly significant (r = 0.65, pvalue = 0.009). Therefore, high genetic gains on field resistance can be achieved by indirectly selecting through the LD method.

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Phytopathogens are the main disease agents that promote attack of cocoa plantations in all tropical countries. The similarity of the symptoms caused by different phytopathogens makes the reliable identification of the diverse species a challenge. Correct identification is important in the monitoring and management of these pests. Here we show that matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS) in combination with multivariate data analysis is able to rapidly and reliably differentiate cocoa phytopathogens, namely Moniliophthora perniciosa, Phytophthora palmivora, P. capsici, P. citrophthora, P. heveae, Ceratocystis cacaofunesta, C. paradoxa, and C. fimbriata. MALDI-MS reveals unique peptide/protein and lipid profiles which differentiate these phytopathogens at the level of genus, species, and single strain coming from different hosts or cocoa tissues collected in several plantations/places. This fast methodology based on molecular biomarkers is also shown to be sufficiently reproducible and selective and therefore seems to offer a suitable tool to guide the correct application of sanitary defense approaches for infected cocoa plantations. International trading of cocoa plants and products could also be efficiently monitored by MALDI-MS. It could, for instance, prevent the entry of new phytopathogens into a country, e.g., as in the case of Moniliophthora roreri fungus that is present in all cocoa plantations of countries bordering Brazil, but that has not yet attacked Brazilian plantations. Graphical Abstract Secure identification of phytopathogens attacking cocoa plantations has been demonstrated via typical chemical profiles provided by mass spectrometric screening.

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RESUMO Diferentes espécies de Phytophthora são responsáveis por causar uma das doenças mais importantes do cacaueiro, a podridão parda, sendo P. palmivora a espécie predominante e bem distribuída mundialmente. Avaliou-se a morfologia e a agressividade de 100 isolados de P. palmivora, coletados em fazendas de 13 municípios da região cacaueira da Bahia, durante três anos consecutivos. Foram inoculados discos de folhas dos clones Scavina6 (SCA-6), padrão de resistência e SiC-23, padrão de suscetibilidade, com suspensões de cada um dos isolados, provenientes da coleção de Phytophthora Arnaldo Gomes Medeiros do Centro de Pesquisas do Cacau/CEPLAC, à concentração de 3 x 105 zoósporos/ mL. Foi utilizado o método patométrico escala de notas para avaliação da infecção sete dias após a inoculação. Avaliou- se o aspecto das colônias, a produção de esporângios e clamidósporos de cada isolado e foram mensuradas as estruturas de 61 isolados e a determinação do tipo de compatibilidade dos 100 isolados. Observou-se ampla variação na reação dos dois clones de cacaueiro aos isolados de P. palmivora, demonstrando diferentes graus de agressividade entre os mesmos. Esta agressividade não está relacionada ao local ou ano de coleta, pois isolados mais e menos agressivos foram obtidos nos mesmos municípios e anos de coleta. Houve também variação na morfologia das colônias. Os esporângios elipsóides a ovóides apresentaram dimensões médias de 28,3 – 50,5 x 18 – 36,7 ?m, com pedicelos curtos (1,5 – 4,4 µm) e relação comprimento largura variando de 1,1 a 2,2. Os clamidósporos, globosos terminais ou intercalares variaram de 18,4 – 43,5 ?m de diâmetro. Todos os 100 isolados são do tipo compatível A2. A variabilidade observada na agressividade dos isolados terá implicações nos estudos epidemiológicos e na seleção de genótipos resistentes a P. palmivora.

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