ABSTRACT The characterisation and evaluation of accessions are important steps in both the management of a germplasm collection and in a plant genetic improvement program. Carrying out characterisation and evaluation, whether morphological or molecular, requires observing the overall features of plants in the field, or collecting leaves for DNA extraction, which is time consuming and costly; whereas DNA extracted directly from the seeds allows access to the genome without the need for planting the seeds, which speeds up the selection of genotypes of interest and information on genetic variability. The aim was to develop a non-destructive method for extracting DNA from cashew seeds. The seeds were mechanically perforated using a mini-grinder, and a sample of the endosperm was removed. Various protocols and commercial kits for DNA extraction were tested, as well as materials to seal the opening made in the seed. It is possible to extract cashew tree DNA from the endosperm, preferably using commercial kits with cementitious adhesive as the sealing material, offering a germination percentage of 60% of the perforated seeds. program molecular field costly variability nondestructive non destructive minigrinder, minigrinder mini grinder, grinder mini-grinder removed tested seed material 60 6

RESUMO O meloeiro (Cucumis melo L.) é uma das hortaliças mais importantes para o Nordeste brasileiro. A caracterização de germoplasma é um processo eficiente no manejo dos Recursos Genéticos Vegetais, pois contribui para uma melhor conservação e utilização em programas de Melhoramento. Nesse estudo, avaliou-se a divergência genética entre subacessos de meloeiro procedentes da Agricultura Tradicional do Semiárido brasileiro, por meio de marcadores ISSR e RAPD, verificando o grau de associação com a classificação botânica. Foram analisados 26 subacessos conservados no Banco Ativo de Germoplasma de Cucurbitáceas para o Nordeste brasileiro, localizado na Embrapa Semiárido, em Petrolina-PE. A matriz binária dos dados moleculares foi utilizada para a obtenção das dissimilaridades genéticas, pelo complemento do índice de Jaccard. As dissimilaridades genéticas foram representadas em dendrogramas gerados pelo método UPGMA. Vinte oito primers ISSR e 26 primers RAPD foram polimórficos. Um total de 686 fragmentos foi obtido, sendo 451 polimórficos. O polimorfismo médio dos marcadores RAPD foi maior que o dos marcadores ISSR. O número de marcadores foi considerado adequado para determinar com precisão a diversidade genética. O dendrograma ISSR permitiu a formação de três grupos, o dendrograma RAPD, quatro grupos, e o dendrograma conjunto, três grupos. O estudo evidenciou a variabilidade genética presente entre os subacessos e a forte associação entre a caracterização feita pelos marcadores moleculares e a classificação taxonômica dos genótipos estudados.

ABSTRACT The melon (Cucumis melo L.) is one of the most important horticultural crops in the northeast of Brazil. The characterisation of germplasm is an efficient process for managing plant genetic resources, as it contributes to better conservation and improved use in breeding programs. In the present study, the genetic divergence between sub-accessions of the melon taken from traditional agriculture in the semi-arid region of Brazil was evaluated using ISSR and RAPD markers, verifying the level of agreement with the botanical classification. Twenty-six sub-accessions preserved in the Active Germplasm Bank of Cucurbitaceae for the northeast of Brazil, located at Embrapa Semiárido in Petrolina, Pernambuco, were analysed. The binary matrix of the molecular data was used to obtain the genetic dissimilarities using the complement of the Jaccard index. The genetic dissimilarities were represented in dendrograms generated using the UPGMA method. Twenty-eight ISSR primers and 26 RAPD primers were polymorphic. A total of 686 fragments were obtained, of which 451 were polymorphic. The mean polymorphism of the RAPD markers was greater than that of the ISSR markers. The number of markers was considered sufficient to accurately determine the genetic diversity. The ISSR dendrogram allowed the formation of three groups, the RAPD dendrogram, four groups, and the joint dendrogram, three groups. The study demonstrated the genetic variability between the sub-accessions, and the strong association between the characterisation made by the molecular markers and the taxonomic classification of the genotypes under study.

Abstract: Background and Aims: Cashew (Anacardium occidentale) is a native Brazilian tree, cultivated in various warm climates across the world. The cashew kernels production has been an important economic activity for many tropical countries, providing a variety of food with great nutrition value. The genetic diversity among the early dwarf cultivars used to production is poorly understood, as is the nutritional composition among nuts from those plants. This work aimed to evaluate the physicochemical characteristics of cashew kernels and to study genetic diversity among twelve cultivars to indicate superior genotypes for future breeding programs. Methods: Cashew kernels from twelve cultivars were evaluated for pH, titratable acidity, moisture, ash, lipids and proteins. The genetic variability was assessed using RAPD and ISSR molecular markers. Data from molecular and physicochemical analyses were used to estimate the genetic distances and Mantel test approach was applied to calculate Pearson’s correlation between the data. Key results: A high genetic variability was found among the cultivars. BRS 274 and BRS 275 were the closest genetically, while CCP 09 and BRS 189 were the most distant ones. On the other hand, EMBRAPA 51 and BRS 226 showed cashew kernels with more similar physicochemical characteristics, while BRS 189 and BRS 253 had the more different ones. Moreover, no correlation was found between the physicochemical and molecular results. Cashew kernels had pH close to neutrality, low acidity, considerable moisture content, and high lipid and protein contents. Conclusions: The cashew kernels from the cultivars studied here had low acidity and pH, considerable moisture and ash contents, and high protein and lipid contents. BRS 189, BRS 253, and CCP 1001 showed the best values for proteins, lipids, acidity, and pH. In addition, those cultivars have maximum distance genetic among them, which can be used as parents in future gene combinations in breeding programs.

Resumen: Antecedentes y Objetivos: El marañón (Anacardium occidentale) es un árbol nativo de Brasil y cultivado en varios climas cálidos en todo el mundo. La producción de nueces de marañón ha sido una actividad económica importante para muchos países tropicales, proporcionando una variedad de alimentos con un gran valor nutricional. La diversidad genética entre los primeros árboles de marañón enanos utilizados para la producción es poco conocida, así como la composición nutricional entre los frutos secos de estas plantas. Este trabajo tuvo como objetivo evaluar las características fisicoquímicas de los granos de marañón y estudiar la diversidad genética entre doce cultivares para indicar genotipos superiores para futuros programas de mejoramiento. Métodos: Se evaluó el pH, acidez titulable, humedad, ceniza, lípidos y proteínas de los granos de marañón de doce cultivares. La variabilidad genética se evaluó mediante marcadores moleculares RAPD e ISSR. Se utilizaron datos de análisis moleculares y fisicoquímicos para estimar las distancias genéticas y se aplicó el método de prueba de Mantel para calcular la correlación de Pearson entre los datos. Resultados clave: Se encontró una alta variabilidad genética entre los cultivares. BRS 274 y BRS 275 fueron los más cercanos genéticamente, mientras que CCP 09 y BRS 189 fueron los más distantes. Por otro lado, EMBRAPA 51 y BRS 226 mostraron granos con características fisicoquímicas más similares, mientras que BRS 189 y BRS 253 tuvieron los más diferentes. Además, no se encontró correlación entre los resultados fisicoquímicos y moleculares. Los granos de marañón tenían un pH cercano a la neutralidad, baja acidez, considerable contenido de humedad y alto contenido de lípidos y proteínas. Conclusiones: Los granos de marañón de los doce cultivares estudiados aquí tenían baja acidez y pH, contenido considerable de humedad y cenizas, y alto contenido de proteínas y lípidos. BRS 189, BRS 253 y CCP 1001 mostraron los mejores valores de proteínas, lípidos, acidez y pH. Además, esos cultivares tienen una distancia genética máxima entre ellos, que pueden usarse como progenitores en futuras combinaciones de genes en programas de mejoramiento genético.

RESUMO Anthurium affine é uma planta ornamental nativa do Brasil sendo muito apreciada pela sua longa duração. Para investigar a divergência genética disponível, 21 progênies de meio-irmãos A. affine foram avaliados utilizando descritores morfoagronômicos e moleculares. Os descritores morfoagronômicos utilizados foram: altura da planta, diâmetro da planta, hábito de crescimento, número de folhas, comprimento da folha, largura da folha, comprimento do pecíolo, diâmetro do pecíolo, ondulação da margem foliar, aparência geral, taxa de cobertura do solo, e marcadores do tipo Polimorfismo de DNA Amplificado ao Acaso (RAPD). Todos os descritores dentro da progênie apresentaram variabilidade e os descritores aparência geral, ondulação da margem foliar e taxa de cobertura do solo foram os que mais contribuíram para a divergência entre as plantas. Dos 45 primers testados, 19 apresentaram inequívocas bandas polimórficas (42.2%). Estas geraram 200 marcas moleculares, sendo 81 (40.5%) polimórficas. O número de bandas polimórficas por primers variou de um (OPB10) a oito (OPW11) e o polimorfismo variou de 16.6% (OPB10) a 70% (OPW05). Análises de agrupamento dos descritores morfoagronômicos e moleculares formaram cinco e quarto grupos, respectivamente. Os dendrogramas apresentaram diferenciação entre os acessos embora não haja associação entre os agrupamentos dos descritores morfoagronômicos e moleculares. Existe divergência genética dentro da progênie de meios-irmãos de A. affine, o que abre possibilidades para aumento da variabilidade.

ABSTRACT Anthurium affine is an ornamental Brazilian native plant very appreciated due to its long-lasting durability. To explore the genetic diversity available, 21 A. affine half-sibling progeny were evaluated using morphoagronomic and molecular descriptors. The morphoagronomic descriptors used were: plant height, plant diameter, growth habit, leaf number, leaf blade length, leaf blade width, petiole length, petiole diameter, leaf margin undulation, general appearance; soil cover rate and Random Amplified Polymorphic DNA (RAPD) markers the molecular one. All the descriptors within the progeny showed variability and general appearance, leaf margin undulation and soil cover rate are the ones that most contribute to the plants divergence. From 45 RAPD primers tested, 19 presented clear polymorphic bands (42.2%). Those generated 200 molecular markers, 81 (40.5%) of them being polymorphic. The number of polymorphic bands per RAPD primer ranged from one (OPB10) to eight (OPW11) and polymorphism ranged from 16.6% (OPB10) to 70% (OPW05). Grouping analysis of morphoagronomic and molecular descriptors formed five and four groups respectively. The dendrograms showed differentiation between accessions although there was no association between morphoagronomic and molecular groupings. There is genetic divergence within the half-sibling progeny of A. affine plants evaluated, which opens up possibilities for increase variability.

Resumo: O objetivo deste trabalho foi avaliar o uso de marcadores moleculares RAPD e ISSR para determinar a variabilidade genética entre genótipos de cajueiro (Anacardium spp.) e indicar possíveis cruzamentos promissores com base na variabilidade genética e no desempenho fenotípico do cajueiro. Foram avaliados, em nível molecular, dez híbridos - originados dos cruzamentos CCP 76 x BGC 589, CCP 76 x BRS 226, CCP 76 x HAC 276-1, CCP 76 x Embrapa 51, CCP 76 x BRS 253, CCP 76 x HAC222-4 e BRS226 x Embrapa 51 - e seus respectivos genitores. Os híbridos foram avaliados quanto à produção de castanha, à massa média da castanha, à incidência de castanhas furadas e à incidência de oídio na castanha (escala 0-4). Os marcadores RAPD e ISSR foram eficientes em determinar a variabilidade genética entre esses genótipos de cajueiro, tendo permitido o agrupamento de 21 grupos. Associados à caracterização fenotípica da castanha de caju quanto à produtividade, à massa e à sanidade da castanha, os marcadores utilizados são eficientes para identificar possíveis combinações capazes de proporcionar maior variabilidade genética e maior probabilidade de obtenção de genótipos transgressivos em populações segregantes.

Abstract: The objective of this work was to evaluate the use of RAPD and ISSR molecular markers to determine the genetic variability among cashew (Anacardium spp.) genotypes, and to indicate possible promising crosses based on cashew genetic variability and phenotypic performance. Ten hybrids - derived from the crosses CCP 76 x BGC 589, CCP 76 x BRS 226, CCP 76 x HAC 276-1, CCP 76 x Embrapa 51, CCP 76 x BRS 253, CCP 76 x HAC222-4, and BRS226 x Embrapa 51 - and their parents were assessed at the molecular level. The hybrids were evaluated for nut yield, mean nut weight, bored nuts, and powdery mildew on nuts (scale 0-4). The RAPD and ISSR markers were efficient in the determinaton of the genetic variability among cashew genotypes, allowing of the grouping of 21 clusters. Associated with the phenotypic characterization of cashew nut for yield, weight, and health, the used markers can efficiently identify possible combinations with higher genetic variability and higher probability of developing transgressive genotypes in segregating populations.