Finding the origin of domestication of cupuaçu requires more than genomics

Colli-Silva et al.1 recently proposed that cupuaçu (Theobroma grandiflorum) was domesticated in northwestern Amazonia as a selection from its sister species, cupuí (T. subincanum). This proposal ignores long-term research in taxonomy, botany, biogeography, and genetics about Theobroma, including cupuaçu. Our review of the research that was ignored and of Colli-Silva et al.'s results demonstrates that cupuaçu is a valid species, as they now agree2, but cupuí may be paraphyletic, the distribution of wild cupuaçu was not included in their samples so the origin of domestication continues unknown, precolonial archaeology lacks remains that can be attributed to either species, historical linguistics indicates that the term cupuaçu references the species' wild distribution and is a recent introduction in northwestern Amazonia, history suggests that cupuaçu started to be domesticated during the last 100-200 years, and the genomics results are not about the domestication of cupuaçu because it is a valid species and its wild distribution was not sampled.

Physicochemical, Rheological, and Nutritional Quality of Artisanal Fermented Milk Beverages with Cupuassu (Theobroma grandiflorum) Pulp and Flour.

The use of fruits and their by-products in food has dramatically impacted the food industry due to the nutritional benefits and the technological and sensory effects of food matrices. Therefore, this research aimed to evaluate the effects of adding cupuassu (Theobroma grandiflorum) pulp and flour on fermented milk beverages' physicochemical, microbial, and sensory properties during refrigerated storage (0, 7, 14, 21, and 28 days). Twelve formulations were realized with different percentages of cupuassu pulp (0, 5, 7.5, and 10% w/v) and flour (0, 1.5, and 3% w/v). The treatments with 3% cupuassu flour presented the highest percentages of protein, fat, fiber, and carbohydrates, compared with the samples containing pulp. On the other hand, the addition of pulp increased water retention capacity and color parameters (L*, a*, b*, and C*) and decreased pH and syneresis on day 0 of storage. During storage, the samples with pulp showed increases in pH values, consistency index, and apparent viscosity. In comparison, cupuassu flour addition decreased syneresis values and increased L* and b* during storage, as did pulp. In addition, sample HPHF (10% pulp and 3% cupuassu flour), based on just-about-right, penalty, and check-all-that-apply analyses, improved some sensory attributes of the fermented milk beverage, such as brown color, acid taste, bitter taste, cupuassu flavor, and firm texture. It can be concluded that cupuassu pulp and flour addition improves the physicochemical and sensory quality of fermented milk beverages and can provide nutritional value to the product.

Methane emission, intake, digestibility, performance and blood metabolites in sheep supplemented with cupuassu and tucuma cake in the eastern Amazon.

The use of co-products as a feed supplement for ruminants makes livestock sustainable and optimizes the use of available areas and animal performance. Furthermore, when cakes are used, the residual fat composition can influence ruminal metabolism and methane (CH4) production. This study aimed to assess the effects of a diet containing cupuassu (CUP; Theobroma grandiflorum) and tucuma (TUC; Astrocaryum vulgare Mart.) cakes on intake, digestibility, serum metabolites, performance, and CH4 emissions in confined sheep in the Amazon. Approximately 28 animals, Dorper-Santa Inês, castrated, with an average initial live weight (ILW) of 35 ± 2.3 kg, were distributed in metabolic cages, in a completely randomized design, with four treatments and seven replications: (1) Control (C40), without the addition of Amazonian cake and with 40 g of ether extract (EE)/kg of dietary dry matter (DM); (2) CUP, the inclusion of the CUP cake and 70 g of EE/kg; (3) TUC, the inclusion of the TUC cake and 70 g of EE/kg; and (4) Control (C80), without the addition of Amazonian cake and with 80 g of EE/kg of dietary DM, with roughage to concentrate ratio of 40:60. The use of the TUC cake as a feed supplement reduced the intake of DM, crude protein (CP), and EE compared to the inclusion of the CUP cake (p < 0.05); however, it increased the intake of neutral detergent fiber (NDF) by 32% (p < 0.01). The highest averages of DM (732 g/kg) and CP (743 g/kg) digestibility were presented in C40, while the highest digestibility of NDF was presented in TUC (590 g/kg). Albumin levels stayed above and protein levels were below the reference values, and the C40 diet also obtained below results for cholesterol, triglycerides and High Density Lipoprotein (HDL) (P < 0.05). Sheep fed CUP (91 g) and TUC (45 g) had lower daily weight gains (DWGs) than those fed with diets without the inclusion of cakes (C40 = 119 g; C80 = 148 g), and feed efficiency (FE) was also lower in CUP (84) and TUC (60) diets than in C40 (119) and C80 (137) diets. CH4 emissions were lower in animals fed TUC (26 L/day) and higher in C40 (35 L/day); however, TUC resulted in higher CH4 emissions in grams/body live weight (BW) gain/day (353 g/BW/day) vs. 183 g/BW/day (C40), 157 g/BW/day (C80), and 221 g/BW/day (CUP). The supplementation with cakes did not improve intake, digestibility and performance, did not compromise blood metabolites and did not reduce the enteric CH4 emission in confined sheep in the Amazon; however, the use of CUP cake showed similar results to the control treatments and did not increase CH4 emissions, as occurred with the inclusion of TUC cake.

Assessing the Effectiveness of Chemical Marker Extraction from Amazonian Plant Cupuassu (Theobroma grandiflorum) by PSI-HRMS/MS and LC-HRMS/MS.

Employing a combination of liquid chromatography electrospray ionization and paper spray ionization high-resolution tandem mass spectrometry, extracts from cupuassu (Theobroma grandiflorum) pulp prepared with either water, methanol, acetonitrile or combinations thereof were subjected to metabolite fingerprinting. Among the tested extractors, 100% methanol extracted preferentially phenols and cinnamic acids derivatives, whereas acetonitrile and acetonitrile/methanol were more effective in extracting terpenoids and flavonoids, respectively. And while liquid chromatography- mass spectrometry detected twice as many metabolites as paper spray ionization tandem mass spectrometry, the latter proved its potential as a screening technique. Comprehensive structural annotation showed a high production of terpenes, mainly oleanane triterpene derivatives. of the mass spectra Further, five major metabolites with known antioxidant activity, namely catechin, citric acid, epigallocatechin-3'-glucuronide, 5,7,8-trihydroxyflavanone, and asiatic acid, were subjected to molecular docking analysis using the antioxidative enzyme peroxiredoxin 5 (PRDX5) as a model receptor. Based on its excellent docking score, a pharmacophore model of 5,7,8-trihydroxyflavanone was generated, which may help the design of new antioxidants.

Forest-Fruticulture Conversion Alters Soil Traits and Soil Organic Matter Compartments.

Fruticulture in the Amazonian Rainforest is one of the main causes of deforestation, biodiversity loss, and soil erosion. Fruticulture plays a key role in the soil traits and soil organic matter (SOM) compartments by altering the soil ecosystem. Our aim was to assess the influence of Forest-Fruticulture conversion on soil traits, and SOM fractions in Brazil's Legal Amazon. The experiment was carried out in field conditions using four land uses as main treatments: Bixa orellana, Theobroma grandiflorum, Paullinia cupana, and the Amazon Rainforest. The soil physicochemical traits were analyzed using samples that were collected from 0-5, 5-10, and 10-20 cm soil depth by using grids (10 × 10 m) with 36 sampling points. Our results showed that the Fruticulture promoted an increase in bulk density, GMD, aggregate diameter, soil porosity, gravimetric moisture, sand, clay, carbon associated with humic acid, and, the sum of bases (K+, Ca2+, and Mg2+), while the Amazon Rainforest showed the highest values of silt, soil P content, SOC, p-SOC, m-SOC, carbon associated with fulvic acid, humine, and soil C stock. Overall, the fruticulture farming systems have negative effects on SOM compartments. The results of our study highlight the importance of considering fruticulture with endemic plant species by promoting soil fertility and soil aggregation.

Alternative Controlling Agent of Theobroma grandiflorum Pests: Nanoscale Surface and Fractal Analysis of Gelatin/PCL Loaded Particles Containing Lippia origanoides Essential Oil.

A new systematic structural study was performed using the Atomic Force Microscopy (AFM) reporting statistical parameters of polymeric particles based on gelatin and poly-ε-caprolactone (PCL) containing essential oil from Lippia origanoides. The developed biocides are efficient alternative controlling agents of Conotrachelus humeropictus and Moniliophtora perniciosa, the main pests of Theobroma grandiflorum. Our results showed that the particles morphology can be successfully controlled by advanced stereometric parameters, pointing to an appropriate concentration of encapsulated essential oil according to the particle surface characteristics. For this reason, the absolute concentration of 1000 µg·mL-1 (P1000 system) was encapsulated, resulting in the most suitable surface microtexture, allowing a faster and more efficient essential oil release. Loaded particles presented zeta potential around (-54.3 ± 2.3) mV at pH = 8, and particle size distribution ranging from 113 to 442 nm. The hydrodynamic diameter of 90% of the particle population was found to be up to (405 ± 31) nm in the P1000 system. The essential oil release was evaluated up to 80 h, with maximum release concentrations of 63% and 95% for P500 and P1000, respectively. The best fit for the release profiles was obtained using the Korsmeyer-Peppas mathematical model. Loaded particles resulted in 100% mortality of C. humeropictus up to 48 h. The antifungal tests against M. perniciosa resulted in a minimum inhibitory concentration of 250 µg·mL-1, and the P1000 system produced growth inhibition up to 7 days. The developed system has potential as alternative controlling agent, due to its physical stability, particle surface microtexture, as well as pronounced bioactivity of the encapsulated essential oil.

α-Amylase inhibition, cytotoxicity and influence of the in vitro gastrointestinal digestion on the bioaccessibility of phenolic compounds in the peel and seed of Theobroma grandiflorum.

The aim of this work was to evaluate the bioaccessibility, cytotoxicity, antioxidant and antidiabetic potential of peel and seeds of cupuassu (Theobroma grandiflorum). Thus, the extracts of cupuassu were evaluated for inhibition of α-amylase, cytotoxicity, and bioaccessibility after gastrointestinal digestion and probiotic fermentation (Lactobacillus delbrueckii, Lactobacillus jhonsoni, Lactobacillus rhamus and Bifidobacterium longum). Digestion increased concentrations of phenolics, showing bioaccessibility of up to 274.13% (total phenolics) and 1105.15% (ORAC). ß-carotene, quinic, tartaric, malic, citric, epicatechin, ethyl gallate, epigallocatechin gallate, gallic acid, pyrocatechol, vanillin, ramnetine were the main compounds while the epicatechin, ethyl gallate, gallic acid and pyrocatechol were the major effective phenolic compounds. The extracts did not show toxic effects and the cupuassu seeds showed 97% inhibition of α-amylase and 47.91% bioaccessibility of pyrocatechol. This study suggests that cupuassu extracts are sources of natural antioxidants with promising antidiabetic potential, and probiotics are able to increase phenolic compounds, responsible for health benefits.

Improvement of endoglucanase production by Aspergillus brasiliensis in solid-state fermentation using cupuaçu (Theobroma grandiflorum) residue as substrate.

AIMS: Optimize the production of Aspergillus brasiliensis endoglucanase in a solid-phase bioprocess using cupuaçu shell as substrate. METHODS AND RESULTS: The shells were supplemented with nitrogen and phosphorous and used as a substrate. The centesimal and inorganic composition of the residue was determined, and found to be rich in fibres, and possessed essential elements for fungal growth. In the initial cultivation of A. brasiliensis, endoglucanase activity of 7.35 U g-1 was obtained. A factorial experimental design was used to determine the most significant variables for the bioprocess. The interactions between moisture, temperature and nitrogen source were noteworthy (p < 0.05). From the rotational central composite design, the optimization of temperature and nitrogen supplementation was obtained, and this reached 40.50 U g-1 , which is an increase of more than five times the value obtained initially. The enzymatic extract was applied as the biocatalyst in the hydrolysis of cupuaçu shells and, after 48 h, it was possible to observe the production of reducing sugars. CONCLUSIONS: Cupuaçu shell can be used as a substrate for endoglucanase production by A. brasiliensis. The process was optimized for the cultivation temperature and the nitrogen source. The enzymatic extract can be applied in the hydrolysis of lignocellulosic biomass. SIGNIFICANCE AND IMPACT OF THE STUDY: Cupuaçu shells can be used to produce cellulases, a product of high added value that can generate economic and environmental benefits for communities and companies producing derivatives of the cupuaçu fruit.

Flavonoid/Polyphenol Ratio in Mauritia flexuosa and Theobroma grandiflorum as an Indicator of Effective Antioxidant Action.

Studies on polyphenols and flavonoids in natural products reveal benefits in the prevention of multiple diseases. Proper extraction, treatment of extracts, and quantification of polyphenols and flavonoids demand attention from the scientific community in order to report more specific biological action. Total polyphenol content (TPC) and total flavonoid content (TFC) (measured at three different times) of ethanol, methanol and acetone extracts of Mauritia flexuosa (aguaje) and Theobroma grandiflorum (copoazú) fresh pulp, from the Colombian Amazon region, were evaluated with the purpose of focusing in the polyphenol/flavonoid proportion and its effective antioxidant activity. This objective could help to explain specific flavonoid biological action based on higher flavonoid proportion rather than higher total polyphenol content. Differences in extracting solvents resulted in statistically significant different yields; the highest TPC was observed with acetone 70% in Mauritia flexuosa and ethanol 80% for T. grandiflorum. The best flavonoid/polyphenol ratio in M. flexuosa was about 1:2.4 and 1:12.8 in T. grandiflorum and the antioxidant efficacy was proportionally higher for flavonoids extracted from T. grandiflorum. HPLC analysis revealed 54 µg/g of the flavonoid kaempferol in M. Flexuosa and 29 µg/g in T. grandiflorum. Further studies evaluating this proportionality, in seeds or peel of fruits, as well as, other specific biological activities, could help to understand the detailed flavonoid action without focusing on the high total polyphenol content.

Enzymatic interesterification effect on the physicochemical and technological properties of cupuassu seed fat and inaja pulp oil blends.

The objective of this work was to evaluate the effect of enzymatic interesterification process in blends with different proportions (w:w) of cupuassu fat and inaja oil (80:20, 70:30, 60:40, 50:50 and 40:60). The interesterification reaction was carried out at 65 °C, agitation at 150 rpm, and enzyme concentration of 5% (w/w), for 6 h. Acidity index, melting point, consistency and solid fat content of the blends were characterized before and after the interesterification process. Fatty acid content was characterized in cupuassu fat and inaja oil and, nutritional quality indexes of atherogenicity (AI) and thrombogenicity (TI) were calculated. Enzymatic interesterification promoted a decrease in acidity (<0.6%) and changes in the blends' properties, making them suitable for food product preparation. All esterified blends (cupuassu seed fat:inaja pulp oil) presented suitable consistency properties, plasticity and spreadability to be used for the preparation of functional, table and soft table types of margarine and used in food preparation such as special fats.

Optimization of the spray drying process for developing cupuassu powder.

A central composite design was used to optimize the spray drying process for developing cupuassu (Theobroma grandiflorum) powder. The influence of inlet air temperature (Tinlet) (120-180 °C), feeding rate (Ffeed) (3-12 mL/min) and maltodextrin concentration (Cmalto) (10-40%) on the properties of moisture, water activity (aw), hygroscopicity, water solubility index (WSI), ascorbic acid (AA) and total phenolic compounds (TPC) were analyzed using response surface methodology and desirability function. In addition, powder morphology was assessed using scanning electron microscopy. The Tinlet and Ffeed parameters significantly influenced the moisture and aw, while the other responses (hygroscopicity, WSI, AA and TPC) were influenced by the Tinlet and Cmalto parameters. The highest Cmalto and intermediate Tinlet and Ffeed levels showed the best morphological characteristics (particles integrity, spheres shapes and smooth surfaces). The desirability function defined the optimal process conditions as Tinlet = 150 °C, Ffeed = 7.5 mL/min and Cmalto = 40%.

Monitoring and Optimization of Cupuaçu Seed Fermentation, Drying and Storage Processes.

Cupuaçu [Theobroma grandiflorum (Wild ex Spreng.) K. Schum] seeds have been employed for a long time in the Amazon region for food purposes. Similar to cocoa, processed cupuaçu pulp and seeds can be used to produce juices, ice creams, confectionary products and cupulate®, which is a similar product to chocolate. However, its market penetration requires the mastery of all processing stages to improve the food quality and safety and to make possible an efficient technology transfer to the local small farmers and communities. Based on the above, the current research work aimed at monitoring and optimizing the consecutive fermentation and drying processes of cupuaçu seeds over 7 days each, as well as storage for 90 days. A greenhouse structure incorporating the fermenter and solar drying terrace was designed to be inexpensive, versatile, easily scalable, and easy to maintain and operate by the local small farmers after a short period of training. This research effort also aimed at giving a vision for the future creation of an integrative and sustainable cupuaçu system covering the economic, social, cultural and environmental vectors. The experimental design comprised 5 batches of 100 kg of seeds each. Several microbiological and physicochemical parameters were performed and correlated with processing variables. Microbiological parameters encompassed viable counts of mesophilic microorganisms, coliforms, yeasts, and molds, whereas physicochemical measures included fermentation and drying temperature, pH, acidity, dry matter, ashes, water activity, color, total proteins, lipids and carbohydrates, and energy. The average seed fermentation temperature varied from ca. 28 to 44 °C, reaching the maximum on day 3 and a final value of ca. 31 °C. Regarding solar drying, the average seed temperatures ranged from ca. 24 °C (at the end) to 39 °C on day 3, and an initial value of ca. 29 °C. The average final seed pH value of drying was 5.34 and was kept during storage. During storage, results demonstrated the existence of significant correlations among several experimental parameters under scrutiny. Finally, bean viable counts obtained during storage unfolded acceptable values of total mesophilic bacteria well below the maximum limit. Viable counts of yeast and molds were generally found between 3 and 4 log(CFU/gsample), and total coliforms were also detected, although both were at acceptable levels and well beneath the established maximum limits for food safety.

Simultaneous selection of cupuassu tree and Brazilian mahogany genotypes in an agroforestry system in Pará state, Brazil

This study aimed to evaluate and select progenies and matrices of cupuassu-tree siblings, and to select Brazilian mahogany matrices with superior characteristics to be used in agroforestry systems (AFS). Twenty-five full cupuassu tree sibling progenies and one Brazilian mahogany half-sibling progeny were evaluated. The study was conducted for 14 years in a commercial property in Tomé Açu, Pará State, Brazil. The number of fruits and fruit yield per plant, as well as the rate of plants with symptoms of witches' broom disease, were used as response variables for cupuassu. For mahogany, the following response variables were used: total height, commercial height, crown height, diameter at breast height and commercial wood volume. There were five cupuassu tree progenies with interesting characteristics to be used in AFS. Based on the high values observed in the selection accuracy and heritabilities, a good potential for the selection of promising individuals in the cupuassu tree population is inferred. Based on the ranking of genotypic values and their agronomic performance, 10 matrices of cupuassu tree were selected. For Brazilian mahogany, three matrices showed excellent silvicultural performance. The matrices of these two species should be propagated vegetatively in order to evaluate clonal tests. (AU)

Combining Cupuassu (Theobroma grandiflorum) Puree, Pectin, and Chitosan Nanoparticles into Novel Edible Films for Food Packaging Applications.

Every year, the residues generated by the disposal of packaging materials produced from fossil fuels have been growing, denoting a major environmental problem that can be mitigated by the development of biodegradable materials from natural polymers, particularly edible films. This work aimed at the development of pectin films added by cupuassu puree and chitosan nanoparticles and to evaluate the improvement of the physical-mechanical performance of the composite films. The nanostructures displayed an average size of 110 nm and a zeta potential of approximately +40 mV. The films were produced by casting, and they exhibited manageability, homogeneity, and continuity. Based upon the mechanical analysis of maximum stress and elongation, it was concluded that the nanoparticles functioned as fillers, increasing the toughness of the pectin films. Water vapor permeability assays demonstrated that the nanostructured films containing cupuassu exhibited improved barrier properties. The glass transition temperature of the films was not strongly affected by the addition of nanoparticles. Conversely, the initial degradation temperature decreased with the addition of nanoparticles and cupuassu puree. The outcomes of this research pave a new route for the development of nonconventional food packaging materials.

Concentration of collagen fibers in the musculature of broiler chickens fed with cupuacu seed by-product / Concentração de fibras colágenas na musculatura de frangos alimentados com farinha de cupuaçu

In this essay, we quantify the concentration of collagen fibers in broiler chickens exposed to increasing concentrations of cupuacu seed by-product. Collection of material was carried out in five chickens per treatment at 70 days old in the groups: control, 5% and 10% inclusion of cupuacu seed by-product. Fragments of Thoracic Pectoralis (PT) and Iliotibial lateralis (ITL) muscles were prepared for light and electronic microscopy. The amount of collagen fibers in the muscle groups was 1.08±0.61% in the PTC group; 6.24±2.58% in PT5% and 7.30±2.75% in PT10%. In the Iliotibial Lateralis groups, the results were 6.96±3.14% in the ITLC; 7.43±4.22% in the ITL5% and 8.66±2.35% in ITL10%. The amount of collagen fibers in the ITL5% and ITL10% groups showed no significant statistical difference. However, when compared to the ILTC group, there was a significant statistical difference. The PT muscle responds to standard nutritional changes, unlike the ILT muscle, which requires a high-nutrient formulation. The use of 5% cupuacu seed by-product has proven to be a viable alternative source of animal feed, as it promotes an increase in the concentrations of collagen fibers in the musculature of broiler chickens and is possibly the determining factor in meat texture.(AU)

Neste estudo, foram quantificadas as concentrações de fibras colágenas de frangos expostos a crescentes concentrações de farinha de cupuaçu. A coleta de material foi realizada em cinco animais por tratamento, aos 70 dias de idade, nos grupos: controle, inclusão de 5% e de 10% de farinha de cupuaçu. Fragmentos dos músculos peitoral torácico (PT) e iliotibial lateral (ITL) foram preparados para microscopia de luz e eletrônica. A quantidade de fibras colágenas nos grupos foi: 1,08±0,61% no grupo PTC; 6,24±2,58% em PT5% e 7,30±2,75% em PT10%. Nos grupos iliotibial lateral, os resultados foram: 6,96±3,14% no ITLC; 7,43±4,22% no ITL5% e 8,66±2,35% em ITL10%. A quantidade de fibras colágenas nos grupos ITL5% e ITL10% não apresentou diferença estatística significativa. No entanto, quando comparados ao grupo ILTC, houve diferença estatística significativa. O músculo PT responde a mudanças nutricionais padrão, ao contrário do músculo ILT, que requer alta formulação nutricional. O uso de 5% de farinha de cupuaçu provou ser uma fonte alternativa viável de alimentação animal, pois promove um aumento nas concentrações de fibras de colágeno na musculatura de frangos de corte e é possivelmente um fator determinante na textura da carne.(AU)

The complete chloroplast genome of Theobroma grandiflorum, an important tropical crop.

Theobroma grandiflorum (Willd. ex Spreng.) K. Schum., an economically important tree in the family Malvaceae, is native to the Amazonian region of South America and it is widely cultivated as a tropical crop. Herein, we report the complete chloroplast genome of T. grandiflorum. The size of the complete chloroplast genome of T. grandiflorum is 160,606 bp with 36.88% GC content, which includes a pair of inverted repeat regions (IRs) of 25,496 bp separated by a large single-copy region (LSC) of 89,429 bp and a small single copy (SSC) region of 20,185 bp. A total of 126 genes were annotated, of which 82 were protein-coding genes, 36 were transfer RNA (tRNA) genes, and 8 were ribosomal RNA (rRNA) genes. A maximum-likelihood (ML) analysis supported a close relationship between T. grandiflorum and T. cacao. This study will provide useful information for further phylogenetic and evolutionary analysis of Malvaceae.

Substantial equivalence analysis in fruits from three Theobroma species through chemical composition and protein profiling.

Substantial equivalence studies were performed in three Theobroma spp., cacao, bicolor and grandiflorum through chemical composition analysis and protein profiling of fruit (pulp juice and seeds). Principal component analysis of sugar, organic acid, and phenol content in pulp juice revealed equivalence among the three species, with differences in some of the compounds that may result in different organoleptic properties. Proteins were extracted from seeds and pulp juice, resolved by two dimensional electrophoresis and major spots subjected to mass spectrometry analysis and identification. The protein profile, as revealed by principal component analysis, was variable among the three species in both seed and pulp, with qualitative and quantitative differences in some of protein species. The functional grouping of the identified proteins correlated with the biological role of each organ. Some of the identified proteins are of interest, being minimally discussed, including vicilin, a protease inhibitor, and a flavonol synthase/flavanone 3-hydroxylase. BIOLOGICAL SIGNIFICANCE: Theobroma grandiflorum and Theobroma bicolor are endemic Amazonian plants that are poorly traded at the local level. As close relatives of Theobroma cacao, they may provide a good alternative for human consumption and industrial purposes. In this regard, we performed equivalence studies by conducting a comparative biochemical and proteomics analysis of the fruit, pulp juice and seeds of these three species. The results indicated equivalent chemical compositions and variable protein profiles with some differences in the content of the specific compounds or protein species that may result in variable organoleptic properties between the species and can be exploited for traceability purposes.

Phenotypic and genotypic characterization and compatibility among genotypes to select elite clones of cupuassu / Caracterização fenotípica, genotípica e compatibilidade entre genótipos para seleção de clones elite de cupuaçuzeiro

ABSTRACT The expansion of the genetic base of cultivated materials is an ongoing activity of the cupuassu (Theobroma grandiflorum) breeding program. However, the parents involved need to be genotypically and phenotypically characterized to ensure compatibility of crossings, as well as to assist in the selection of more promising individuals for hybridization. This study aimed to identify and select T. grandiflorum clones that are compatible and genetically divergent using tools such as the estimates of genotypic, phenotypic, and combined distances, as well as the compatibility rates among clones. The genetic distance analysis of the clones was performed with 14 heterologous microsatellite primers of cocoa (Theobroma cacao) that amplify the DNA of cupuassu. Phenotypic characterization was based on 14 variables related to fruit production. The joint dissimilarity matrix was obtained by means of the sum of the phenotypic and molecular dissimilarity matrices. The intra- and inter-clonal compatibility was estimated through controlled crossings. A low correlation was noted between the dissimilarity matrices based on the molecular and agronomic data. As for compatibility, all clones were self-incompatible, with different compatibility rates when crossed. The compatibility index was strongly influenced by the degree of relationship of the clones. It was possible to identify and select the most promising sets of cupuassu clones to be used in breeding programs, despite their genetic relationship.

RESUMO A ampliação da base genética dos materiais de cultivo é trabalho contínuo do programa de melhoramento genético do cupuaçuzeiro (Theobroma grandiflorum). Há necessidade, entretanto, que os parentais envolvidos estejam caracterizados fenotípica e genotipicamente, para auxiliar na escolha dos indivíduos que serão hibridizados, bem como para garantir os mecanismos de rastreabilidade e proteção das cultivares. Este estudo teve como objetivo identificar e selecionar clones de cupuaçuzeiro inter-compatíveis e geneticamente divergentes, utilizando como ferramentas as distâncias genotípica, fenotípica e combinada mistas, assim como as taxas de compatibilidade entre os clones. Estimativas das distâncias genéticas entre os clones foram realizadas com base em 14 iniciadores microssatélites heterólogos de cacaueiro (Theobroma cacao) que amplificam o DNA do cupuaçuzeiro. Para a caracterização fenotípica foram empregadas 14 variáveis ​​relacionadas à produção de frutos. A matriz de dissimilaridade conjunta foi obtida por meio da soma das matrizes de dissimilaridade fenotípica e molecular. A compatibilidade intra e inter-clonal foi estimada através de cruzamentos controlados. Houve uma baixa correlação entre as matrizes de dissimilaridade com base nos dados moleculares e agronômicos. Quanto à compatibilidade, todos os clones foram auto-incompatíveis, contudo, compatíveis entre si, com diferentes taxas. O índice de compatibilidade foi fortemente influenciado pelo grau de relacionamento dos clones. Foi possível identificar e selecionar os conjuntos de clones de cupuaçuzeiro mais promissores para ser usados em melhoramento genético, apesar da ocorrência de relação genética entre eles.

First record of Phanuropsis laniger Johnson in Brazil and first record of Phanuropsis semiflaviventris Girault in Amazonas (Hymenoptera: Platygastridae) with notes of their hosts, stink-bugs of cupuaçu.

BACKGROUND: We report the first record and collection of Phanuropsis laniger Johnson, 1987, for Brazil since its original description. In addition we expand the distribution of Phanuropsis semiflaviventris Girault, 1916, for Amazonas, Brazil and present an updated list of records for both species with the occurrence of P. semiflaviventris in egg of stink-bugs on cupuaçu (Theobroma grandiflorum). NEW INFORMATION: First record of Phanuropsis laniger Johnson, 1987, for Brazil. We also expanded the distribution of Phanuropsis semiflaviventris Girault, 1916, for the state of Amazonas, Brazil.

Diversidade genética estimada com marcadores entre sequências simples repetidas em cultivos comerciais de Cupuaçuzeiro / Genetic diversity estimated using inter-simple sequence repeat markers in commercial crops of cupuassu tree

RESUMO:Quinze primers ISSR (entre sequências simples repetidas) foram utilizados para avaliar a diversidade genética entre e dentro de pomares comerciais de Theobroma grandiflorum (Willd. ex Spreng.) K. Schum. Para isso, foram analisados sessenta indivíduos, distribuídos nos três cultivos. Um total de 102 bandas foi amplificado, com uma porcentagem de 52,0% de polimorfismo em nível de espécie e média de 6,8 alelos por primer ISSR. A média do Índice de Conteúdo Polimórfico (PIC) foi de 0,55. Em relação aos índices de diversidade gênica de Nei (H) e de Shannon (I), os cultivos analisados apresentaram os valores: SAR H = 0,114 e I = 0,177; SSL H = 0,108 e I = 0,162 e SEC H = 0,104 e I = 0,156, considerados valores de moderados a baixos. A AMOVA revelou 34,91% da variância total entre os cultivos e 65,09% dentro deles. Os marcadores moleculares ISSR revelaram que há diversidade genética dentro de cada cultivo comercial estudado, portanto é possível selecionar genótipos superiores que poderão ser utilizados para originar cultivos mais uniformes. Esse resultado tem sido considerado de grande relevância, por fornecer ferramentas para a implementação de programas de melhoramento e delineamento de estratégias de conservação ex situ e in situ.

ABSTRACT:Fifteen ISSR (inter-simple sequence repeat) primers were used to evaluate the genetic diversity among and within commercial crops of T. grandiflorum (Willd. ex Spreng.) K. Schum. For this, 60 specimens were analyzed, distributed in three crops. A total of 102 bands were amplified, with a polymorphism percentage of 52.0% at species level and an average of 6.8 alleles per ISSR primer. The average for polymorphism information content (PIC) index was 0.55. In relation to the genetic diversity index of Nei (H), and Shannon (I), crops analyzed showed the following values: : SAR H = 0,114 e I = 0,177; SSL H = 0,108 e I = 0,162 e SEC H = 0,104 e I = 0,156, considered moderate to low values. AMOVA showed 34.91% of total variance among the crops, and 65.09% within them. The ISSR molecular markers revealed that there is genetic diversity within each commercial crops studied, thus is possible to select superior genotypes that can be used to give more uniform crops. This result has been considered of great relevance, to provide tools for breding implementation programs and design conservation strategies ex situ and in situ.