The site of self-incompatibility action in cupuassu (Theobroma grandiflorum)

Cupuassu (Theobroma grandiflorum (Willd. ex Spreng.) Schumann) is a fruit tree that is attracting attention in Brazil and also in other tropical countries. Its pulp is used to prepare ice-cream, juices, jellies and many other home-made sweets. This species has a very low fecundity, attributed to problems with pollination and self-incompatibility, noted as being restrictive to the agronomic productivity of the species. Controlled pollinations between compatible and incompatible parents were made and flowers were collected at three times: 24, 48 and 72 h after pollinations, during the floral seasons of 1995 and 1998. In flowers collected 24 h after pollination, in both compatible and incompatible crosses, around 70% of ovules showed egg apparatus without evidences of sperm cell delivery; nevertheless some pollen tubes had been observed. Ovaries collected at 48 h showed different behaviors according to the type of cross. In the compatibles, 35% of ovules did not show evidences of gamete fusion. In incompatible crosses this number rose to 50%. Collected ovaries with 72 h, from compatible and incompatible crosses, showed respectively 1.2% and 14.2% of the ovules with the egg apparatus and polar nuclei only. In these ovaries, for the two types of crosses, the presence of sperm nuclei was common and the zygote was still undivided. The incompatibility action is occurring after fertilization, in a late self-incompatibility action.

Localization of the cotyledon reserves of Theobroma grandiflorum (Willd. ex Spreng.) K. Schum., T. subincanum Mart., T. bicolor Bonpl. and their analogies with T. cacao L.

Cotyledon mesophyll cell morphology and lipid and protein synthesis of T. grandiflorum, T. subincanum and T. bicolor were analyzed and compared with T. cacao. These species possess foliar cotyledons folded around the hypocotyl radicle axis, typical of Sterculiaceae. Fruit size, morphology and weight are very distinct amongst the four species and so are the respective seeds. The main axis of the T. grandiflorum and T. bicolor seeds measured about 30 mm, while T. subincanum and T. cacao seeds measured 17 mm and 26 mm respectively. The seed weights of T. grandiflorum, T. bicolor, T. subincanum and T. cacao were 11.6 g, 9.4 g, 2.1 g and 3.0 g, respectively. The cotyledon mesophylls of the four species contained mainly polysaccharides and lipid-protein reserve cells. Theobroma cacao, T. grandiflorum and T. subincanum were composed of greater than 50% lipids. For the four species, lipid globules gradually accumulated adjacent to the cell wall, and these globules measured from 1 to 3 µm. TEM showed low-density proteins inside the central vacuole of the young mesophyll cells of T. cacao. The protein reserves of the mature cells were densely scattered amongst the lipid bodies, and a few starch granules occurred together with the cotyledon mesophyll of the four species. Polyphenolic cells were found throughout the mesophyll cells or aligned with the respective vascular bundles. Immature cells demonstrated the capacity to synthesize all these reserves, but gradually the pre-determined cells produced mainly lipid-protein reserves. Besides the unique characteristics of the T. cacao products, the lipid-protein synthesis capacities of T. grandiflorum, T. subincanum and T. bicolor suggest various possibilities for new industrialized food, pharmaceutical and cosmetic products.

Polyphenolic cells and their interrelation with cotyledon cells in seven species of Theobroma (Sterculiaceae)

The amount of cotyledon polyphenolic cells varies extensively within the Theobroma species. The polyphenolic compounds of these cells play a protective role and furthermore have an important function in the development of chocolate flavour. The morphology of the polyphenolic cells of the mesophyll is described and the development of these idioblasts in Theobroma cacao L., T. subincanum Mart., T. obovatum Klotzsch ex Bernoulli, T. grandiflorum (Willd. ex Spreng.) K. Schum., T. microcarpum Mart., T. bicolor Bonpl. and T. speciosum Willd. ex Spreng analysed. The total polyphenolic content in the seeds as determined by spectrophotometry showed a variation of about forty times. The alive, transparent polyphenolic cells are scattered throughout the cotiledonary mesophyll. However the polyphenolic cells of T. cacao and T. grandiflorum are also aligned perpendicularly with respect to the mesophyll borders and, in addition, both species display polyphenolic cells with a natural translucent purple colour. All the species analysed contained polyphenolic cells scattered throughout the parenchymal cells and also in a lengthwise association with vascular bundles. In T. bicolor and T. speciosum, the species with the lowest polyphenolic contents, these cells were mostly located around the vascular bundles. Using Scanning Electron Microscopy, the polyphenolic cells demonstrated a complex cytoarchitecture, and after fixing with glutaraldhyde, the polyphenolic secretion was shown to remain as a single unit or was organized into round droplets. Transmission Electron Microscopy displayed immature plastids from young mesophyll cells containing eletron-dense deposits similar to phenolic substances, suggesting that Theobroma plastids are involved in the synthesis of phenolics.

Performance polínica em cacaueiros (Theobroma cacao L.) autocompatíveis e autoincompatíveis

A importância econômica do cacaueiro se deve, principalmente, à produção de frutos, dos quais se extraem sementes que servem como matéria prima para a fabricação do chocolate. Vários fatores podem interferir na produção, porém as características reprodutivas do cacaueiro e sua influência na frutificação não têm sido intensamente estudadas. Além disso, alguns genótipos são autoincompatíveis, mas clones autocompatíveis são encontrados em populações comerciais. Com o objetivo de comparar a performance do pólen e o modo de reprodução entre clones autocompatíveis CCN51 e autoincompatíveis TSH1188, foram realizadas estudos de germinação in vitro (utilizando-se dois testes) e da razão pólen-óvulo (P:O). O Teste 1 (meio de cultura sem alguns elementos essenciais e com maior concentração de sacarose) proporcionou melhores resultados de germinação do pólen, com médias acima de 77%, enquanto que com o Teste 2 o máximo de pólen germinado foi, em média, de 39,95%. O percentual de pólen germinado in vitro foi maior em plantas autoincompatíveis. Com base na razão P:O, ambos os clones foram classificados como autógamos obrigatórios, o que foi considerado um desvio da classificação de Cruden.

Multivariate genetic divergence and hybrid performance of cacao (Theobroma cacao L.)

Genetic distances among cacao cultivars were calculated through multivariate analysis, using the D2 statistic, to examine racial group classification and to assess heterotic hybrids. A 5 x 5 complete diallel was evaluated. Over a five-year period (1986-1990), five cultivars of the S1 generation, pertaining to the Lower Amazon Forastero and Trinitario racial groups and 20 crosses between the corresponding S0 parents were analyzed, based upon five yield components - number of healthy and collected fruits per plant (NHFP and NCFP), wet seed weight per plant and per fruit (WSWP and WSWF), and percentage of diseased fruits per plant (PDFP). The diversity analysis suggested a close relationship between the Trinitario and Lower Amazon Forastero groups. A correlation coefficient (r) was calculated to determine the association between genetic diversity and heterosis. Genetic distance of parents by D2 was found to be linearly related to average performance of hybrids for WSWP and WSWF (r = 0.68, P < 0.05 and r = 0.76, P < 0.05, respectively). The heterotic performance for the same components was also correlated with D2, both with r = 0.66 (P < 0.05). A relationship between genetic divergence and combining ability effects was suggested because the most divergent cultivar exhibited a high general combining ability, generating the best performing hybrids. Results indicated that genetic diversity estimates can be useful in selecting parents for crosses and in assessing relationships among cacao racial groups.

Fragmentação mecânica de amêndoas de cupuaçu (Theobroma grandiflorum)

O cupuaçu (Theobroma grandiflorum) é uma fruta nativa da região amazônica brasileira cujas sementes convenientemente fermentadas, secas e torradas podem ser utilizadas para a obtenção de um produto similar ao chocolate. Este trabalho objetivou pesquisar uma forma alternativa de quebra da amêndoa seca sem torrar, através de um beneficiador de cilindros, seguida da utilização de um jogo de peneiras para a separação das frações fragmentadas. Os fatores estudados no rendimento da obtenção dos nibs (cotilédones fragmentados) e na verificação da pureza deste material (percentual de casca e gérmen) foram a umidade da amêndoa inteira e a velocidade diferencial dos cilindros de borracha do beneficiador. Pelos resultados obtidos, verificou-se que a umidade da amêndoa inteira foi a variável que apresentou maior influência na quebra. Os maiores percentuais de recuperação de nibs foram conseguidos na umidade de 5%. A variação dos níveis da velocidade diferencial não causou diferença significativa no rendimento de obtenção dos nibs. Em relação aos valores dos percentuais de casca e gérmen nos nibs, nenhuma variável estudada apresentou influência significativa nos valores desta resposta, mesmo assim todos os resultados foram bastante satisfatórios, apresentando valores abaixo de 1,75%. O rendimento deste processo alternativo está em torno de 90% de obtenção dos cotilédones encontrados em uma determinada massa de amêndoas. Como conclusão, o processo proposto constituiu-se em alternativa mecanizada tecnicamente válida, que apresentou bom rendimento de recuperação dos nibs, com percentuais de casca e gérmen abaixo do tolerado.

Perfil sensorial de pó de cacau (Theobroma cacao L.) alcalinizado

A Análise Descritiva Quantitativa(ADQ) foi empregada para caracterização das amostras de pós de cacau que representaram a amplitude do delineamento composto rotacional central 2 do processo de alcalinização dos "nibs" de cacau. As variáveis independentes foram faixas de temperatura de 60 a 120 ºC, de tempo de 30 a 150min e de concentração de K2CO3 de 1,22 a 4,78%. Foram avaliadas oito amostras de pó de cacau representativas das variações de cor e aceitabilidade do total das amostras obtidas experimentalmente e duas amostras de marcas comerciais. A análise do aroma foi feita diretamente nos pós de cacau alcalinizados e a dos demais atributos na forma de bebida achocolatada (2% do pó de cacau e 7% de açúcar em leite desnatado esterilizado). Doze provadores selecionados com base no seu poder de discriminação, reprodutibilidade e concordância com a equipe geraram em consenso, pelo método de rede (Kelly's Repertory Grid Method), três termos descritos para o aroma (alcalino, chocolate e queimado) e doze para os achocolatados (solubilidade, marrom, marrom avermelhado, chocolate, queimado, caramelo, doce, adstringente, alcalino, amargo, salgado e corpo). As avaliações das amostras foram feitas monadicamente com três repetições e em cabines individuais. Os dados obtidos foram submetidos a ANOVA, teste de Tukey e Análise de Componentes Principais. As avaliações dos aromas dos pós de cacau mostraram relação direta entre o aroma alcalino e os teores de álcali, temperatura e tempo do processo. De modo geral, os produtos com menores concentrações de K2CO3 (1,22 - 3,00% ) apresentaram aroma e sabor de chocolate mais fortes. Encontrou-se uma relação direta entre os teores de álcali no produto e o sabor alcalino, queimado e adstringente e uma relação inversa com a luminosidade da cor. Assim, as amostras com maior concentração de K2CO3 (4,78%), foram consideradas pela equipe sensorial, as de mais forte sabor e aroma alcalino, queimado e adstringente, assim como de cor marrom e marrom avermelhado mais escura. Todos os processos levaram à obtenção de amostras com alta solubilidade e fraco sabor amargo.

Datos experimentales de la cinética del secado y del modelo matemático para pulpa de cupuaçu (Theobroma grandiflorum) en rodajas

En este trabajo fueron obtenidos experimentalmente la cinética del secado de la pulpa de cupuaçu en forma de rodajas con espesor de 0,5 cm. El secado fue realizado utilizándose un secador laboratorial de bandejas, a las temperaturas de 50, 60 y 70 ºC y a la velocidad del aire de secado de 1,5 m/s. De acuerdo con los datos de la cinética del secado se puede observar que cuanto mayor es la temperatura del secado mayor es la velocidad del secado. El tiempo requerido para secar el producto hasta una humedad del 20%, fueron 9,2, 8,1 y 7,3 horas para las temperaturas de secado de 50, 60 y 70 ºC respectivamente. Las curvas experimentales del secado fueron ajustados al modelo difusional de Fick considerándose la muestra como una lámina infinita y al modelo de Page. Los dos modelos se ajustaron bien a los datos experimentales. Los coeficientes de difusión del modelo de Fick variaron de 1.171 a 1.561 m/s².

Nutritional value and antioxidant capacity of "cocoa honey" (Theobroma cacao L.)

Cocoa honey is considered as the liquid portion of cocoa pulp that is released from the fruit soon after it is cut open and can be used before fermentation by simple extraction due to its nutritional characteristics. The objective of the present study is to determine the biochemical characteristics of a cocoa by-product, "cocoa honey" (CH), produced in the State of Bahia-Brazil. The biochemical characterization was conducted to determine reducing sugars, total sugars, vitamin C, total dietary fiber, flavonoids, and total antioxidant activity using an EC50. It was observed that cocoa honey can be considered a source of bioactive compounds, can be consumed in natura or processed, and used as an ingredient in the chocolate industry and in other food products. However, it is necessary to use complementary methods, such as HPLC, to quantify the phenolic compounds of this by-product.

Bioactive compounds in different cocoa (Theobroma cacao, L) cultivars during fermentation

One component that contribute to the flavor and aroma of chocolate are the polyphenols, which have received much attention due to their beneficial implications to human health. Besides bioactive action, polyphenols and methylxantines are responsible for astringency and bitterness in cocoa beans. Another important point is its drastic reduction during cocoa processing for chocolate production and the difference between cultivars. Thus, the present study aimed to evaluate the modifications in monomeric phenolic compounds and methylxanthines during fermentation of three cocoa cultivars grown in southern Bahia. Cocoa beans from three cultivars were fermented and sun dried and monomeric phenolic compounds and methylxantines were determinated. The results showed that each cultivar have different amounts of phenolic compounds and the behaviour of them is different during fermentation. The amount of methylxantines varied but there was not a pattern for methylxantines behavior during process. In addition a huge reduction in phenolic compounds could be observed after drying. Differently of phenolic compounds, methylxantines did not have great modification after sun drying. So, the differences observed in this study between cultivars, take to the conclusion that the compounds studied in those cocoa cultivars have different behavior during fermentation and drying, which consequently, give to these cultivars differences in sensory characteristics.

Characterization of polyphenol oxidase in two cocoa (Theobroma cacao L.) cultivars produced in the south of Bahia, Brazil

Abstract The reactions leading to the formation of precursors of chocolate flavor are performed by endogenous enzymes present in the cocoa seed. Polyphenol oxidase (PPO) presence and activity during fermentation of cocoa beans is responsible for the development of flavor precursors and is also implicated in the reduction of bitterness and astringency. However, the reliability of cocoa enzyme activities is complicated due to variations in different genotypes, geographical origins and methods of fermentation. In addition, there is still a lack of systematic studies comparing different cocoa cultivars. So, the present study was designed to characterize the activity of PPO in the pulp and seeds of two cocoa cultivars, PH 16 and TSH 1188. The PPO activity was determined spectrophotometrically and characterized as the optimal substrate concentration, pH and temperature and the results were correlated with the conditions during the fermentation process. The results showed the specificity and differences between the two cocoa cultivars and between the pulp and seeds of each cultivar. It is suggested that specific criteria must be adopted for each cultivar, based on the optimal PPO parameters, to prolong the period of maximum PPO activity during fermentation, contributing to the improvement of the quality of cocoa beans.

Drying and germination of cupuassu (Theobroma grandiflorum (willd. Ex Spreng.) K. Schum.) seeds

The objective of this study was to verify the effect of drying on germination of cupuassu (Theobroma grandiflorum (Willd. ex Spreng) K. Schum.) seeds. Desiccation was in forced air oven, with temperature ranging from 23 to 33ºC. Sowing was carried out at 0.5cm of depth in plastic trays in sand and sawdust mixture (1:1), previously sterilized in hot water (100ºC), during 2h. Seeds were left to germinate in a laboratory with no temperature and relative humidity control, under natural light. It was quantified the seed moisture content, in four replications of 10 seeds; the germination percentage, performed during 30 days, with daily counts of the number of germinated seeds; the germination speed index; and number of days to the germination onset. The experimental design was completely randomized with four replications of 25 seeds. The reduction of moisture content from 58.6 to 37.8% did not affect seed germination and germination speed index; however, they were affected when moisture content was reduced to values below 30.7%. It was observed that only when moisture content was 16.1% seeds demanded more days to begin germination. Cupuassu seeds are classified as recalcitrant and they can be desiccated up to 37.8% with no reduction on germination.

Alterações fisiológicas após aplicação de silício em cacau e sua influência na preferência por pulgões

Os aspectos fisiológicos e bioquímicos influenciados pelo silício e que condicionam reações de resistência a pragas no cacaueiro ainda são pouco conhecidos. O objetivo deste experimento foi avaliar os efeitos da aplicação de silício sobre a fotossíntese, sobre o teor foliar de fenóis solúveis totais e a sua influência na preferência de Toxoptera aurantii em genótipos de cacau. O experimento foi conduzido em casa de vegetação, no delineamento de blocos casualizados, com quatro repetições, em arranjo fatorial 3 x 3, constituído por três genótipos (TSH 1188, CCN 51 e Catongo), duas doses de silicato de potássio (3 e 6 mL L-1), aplicadas por via foliar e um controle pulverizado somente com água. Foram realizadas avaliações das trocas gasosas, da fluorescência da clorofila a e do teor de compostos fenólicos foliar, além de um teste de preferência com chance de livre escolha com T. aurantii. Os genótipos TSH 1188 e CCN 51 apresentam maior eficiência fotoquímica e fotossintética, comparados com o Catongo. Independentemente do genótipo, a aplicação de silicato de potássio não alterou o índice de desempenho das plantas (PI ABS e PI TOTAL); no entanto, a dose 3 mL L-1 aumentou a fotossíntese líquida e o teor de fenóis solúveis totais. A dose 6 mL L-1 reduziu a preferência de T. aurantii pelas folhas do genótipo TSH 1188. A aplicação de silicato de potássio é promissora no aumento da resistência do cacaueiro a T. aurantii.

Agronomic performance of 27 cocoa progenies and plant selection based on productivity, self-compatibility and disease resistance

The objective of the present work was to evaluate 27 progenies of cocoa crosses considering the agronomic traits and select F1 plants within superior crosses. The experiment was installed in March 2005, in the Experimental Station Joaquim Bahiana (ESJOB), in Itajuipe, Bahia. The area of the experiment is of approximately 3 ha, with a total of 3240 plants. Thirteen evaluations of vegetative brooms, five of cushion brooms and 15 of number of pods per plant were accomplished. Thirty pollinations were made for each selected plant to test for self-compatibility. The production, based on the number of pods per plant, and resistance to witches´ broom indicated CEPEC 94 x CCN 10, RB 39 x CCN 51 and CCN 10 x VB 1151 as superior progenies. All selections tested were self-compatible. The analyses of progenies and individual tree data, associated to visual field observations, allowed the selection of 17 plants which were included in a network of regional tests to determine the phenotypic stability.

Notification of the first isolation of Cacipacore virus in a human in the State of Rondônia, Brazil

Flavivirus is a genus of arthropod-transmitted viruses of the family Flaviviridae, and in Brazil, up to eleven different Flavivirus have been isolated. We collected blood from farmers in the municipality of Theobroma, which is located 320km from the City of Porto Velho, the former capital of the Brazilian State of Rondônia. For viral isolation, we used newborn mouse brain, followed by RT-PCR with specific universal Flavivirus primers. We obtained fragments 958bp and 800bp in length. Based on BLAST, these sequences were 91% similar to a sequence of Cacipacore virus.